Article

Antioxidant, Antimutagenic, and Antitumor Effects of Pine Needles ( Pinus densiflora )

Authors:
To read the full-text of this research, you can request a copy directly from the authors.

Abstract

Pine needles (Pinus densiflora Siebold et Zuccarini) have long been used as a traditional health-promoting medicinal food in Korea. To investigate their potential anticancer effects, antioxidant, antimutagenic, and antitumor activities were assessed in vitro and/or in vivo. Pine needle ethanol extract (PNE) significantly inhibited Fe(2+)-induced lipid peroxidation and scavenged 1,1-diphenyl- 2-picrylhydrazyl radical in vitro. PNE markedly inhibited mutagenicity of 2-anthramine, 2-nitrofluorene, or sodium azide in Salmonella typhimurium TA98 or TA100 in Ames tests. PNE exposure effectively inhibited the growth of cancer cells (MCF-7, SNU-638, and HL-60) compared with normal cell (HDF) in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In in vivo antitumor studies, freeze-dried pine needle powder supplemented (5%, wt/wt) diet was fed to mice inoculated with Sarcoma-180 cells or rats treated with mammary carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA, 50 mg/kg body weight). Tumorigenesis was suppressed by pine needle supplementation in the two model systems. Moreover, blood urea nitrogen and aspartate aminotransferase levels were significantly lower in pine needle-supplemented rats in the DMBA-induced mammary tumor model. These results demonstrate that pine needles exhibit strong antioxidant, antimutagenic, and antiproliferative effects on cancer cells and also antitumor effects in vivo and point to their potential usefulness in cancer prevention.

No full-text available

Request Full-text Paper PDF

To read the full-text of this research,
you can request a copy directly from the authors.

... The total phenolic content can be determined by measuring the color intensity of the phenolic compound extract with the Folin-Ciocalteau reagent. As a result of many studies, it has been observed that the best solvent for the extraction of phenolics is the methanol: water mixture (Kwak et al., 2006). In this study, total phenolic substances were investigated with the Folin technique and it was found between 23.19 mgGAE g -1 and 379.44 mgGAE g -1 . ...
... Such a considerable variation is presumably due to the location and climatic conditions where it grows. Compared with other researchers, Kwak et al. (2006) found total phenolic contents as 57.71 mg g -1 dry weight basis in pine needles of Pinus densiflora extracted by 75% ethanol. This value was within the limits of our findings. ...
... For the antioxidant activity, they expressed that P. merkusii resin extracted in n-hexane had the lowest EC50 value (60.203 mg mL -1 ) compared with the other extracts and the EC50 value changed in relation to the solvent. Kwak et al. (2006) determined that the EC50 value was 95.12 mg mL -1 in the extracts with ethyl alcohol 75%. They concluded that pine needles would have potent antioxidative activity due to the high phenolic compounds. ...
Article
Full-text available
Pine resin has been used as a traditional health-promoting medicinal food in Turkey and some countries for centuries. In the present study, the antioxidant potential and the monoterpene profile of 13 pine resin samples purchased from herbalists in different provinces of Turkey were investigated. According to the analysis results, there were differences between the pine resin samples, especially in terms of color and antioxidant properties. The experimental results demonstrated that the pine resin had strong in-vitro antioxidant effects. Total phenolic, FRAP, and DPPH (EC50) values ranged between 23.19 and 379.44 mgGAE g-1 , 68.85 and 758.80 µmol Fe 2+ g-1 , 54.36 and 1006.97 µg g-1 , respectively. Total phenolic content values correlated well with the FRAP values. In contrast, it was found that there was a negative correlation between FRAP values and EC50 values (r=-0.719**) and between total phenolic substance and EC50 values (r=-0.688**). Also, the monoterpene compounds of the resin were determined by applying solid-phase microextraction (SPME) and gas chromatography-mass spectroscopy. Eight monoterpene compounds were identified in different pine resin samples, including α-Türkiye'den 13 Farklı Çam Reçine Örneğinin Antioksidan Özellikleri ve Monoterpen Bileşimi Öz: Çam reçinesi, yüzyıllardır Türkiye'de ve bazı ülkelerde geleneksel, sağlığı geliştirici tıbbi gıda olarak kullanılmaktadır. Bu çalışmada, Türkiye'nin farklı illerindeki aktarlardan satın alınan 13 adet çam reçinesi örneğinin antioksidan özellikleri ve monoterpen profili araştırılmıştır. Analiz sonuçlarına göre çam reçinesi örnekleri arasında özellikle renk ve antioksidan özellikler açısından farklılıklar tespit edilmiştir. Deneysel sonuçlar, çam reçinesinin güçlü in vitro antioksidan etkilere sahip olduğunu göstermiştir. Toplam fenolik, FRAP ve DPPH (EC50) değerleri sırasıyla 23.19 ve 379.44 mgGAE g-1 , 68.85 ve 758.80 µmol Fe 2+ g-1 , 54.36 ve 1006.97 µg g-1 arasında değişmiştir. Toplam
... Among traditional medicinal plants containing naturallyoccurring antioxidants, Pinus densiflora, known as Japanese red pine in western countries, is a species widely distributed in Korea (1)(2)(3), and the evergreen tree with needle leaves, belonging to the Pinaceae family (2), is indigenous to Eastern Asia. Pine needles have been widely used as food ingredients, food additives, and folk medicines in pine-based products due to their characteristic aroma and taste, and health-promoting properties (3,4). They have been shown to possess a wide spectrum of biological and pharmacological actions such as antioxidant, anti-mutagenic, antitumor, anti-inflammatory, antibacterial effects as well as protection against oxidative DNA damage, and apoptosis induced by hydroxyl radicals (4)(5)(6), which may induce preventive and therapeutic effects against degenerative diseases associated with aging. ...
... Pine needles have been widely used as food ingredients, food additives, and folk medicines in pine-based products due to their characteristic aroma and taste, and health-promoting properties (3,4). They have been shown to possess a wide spectrum of biological and pharmacological actions such as antioxidant, anti-mutagenic, antitumor, anti-inflammatory, antibacterial effects as well as protection against oxidative DNA damage, and apoptosis induced by hydroxyl radicals (4)(5)(6), which may induce preventive and therapeutic effects against degenerative diseases associated with aging. The effective components in pine needles are chlorophyll, carotenes, dietary fiber, terpenoids, phenolics, tannins, and alkaloids (6). ...
... Both reactions were centrifuged at 1,000 g for 10 min. The absorbance of the supernatants from A and B (A abs and B abs , respectively) at 540 nm was measured, and the percent hemolysis inhibition was calculated according to the following equation [4]: ...
Article
Full-text available
Antioxidant activities of water extracts obtained from dried pine needle (Pinus densiflora) were measured at 0, 4, 20, 100, 500, 1,000, and 1,200 ppm and compared with those of phenolic compounds of butylated hydroxyanisole, butylated hydroxytoluene, tert-butylhydroquinone, ferulic acid, and α-tocopherol. The activity was determined as the ability to scavenge 1,1-diphenyl-2-picrylhydrazyl radical and hydrogen peroxide, reductive power, and inhibition of lipid peroxidation in a linoleic acid system using the ferric thiocyanate method and thiobarbituric acid method, respectively. Pine needle water extract (PNWE) exhibited antioxidant activity in a concentration-dependent mode at the same parameters mentioned above, and a significant difference (P<0.05) was observed at 1,000 ppm. The protective activity of PNWE as a potent antioxidant in a non-cellular system was compared with that of phenolics at 150.67 μg/mL in the two assays using biological cellular systems, namely 2,2′-azobis(2-amidinopropane) dihydrochloride-initiated hemolysis and Fe²⁺-induced lipid peroxidation, using rat red blood cells and rat brain homogenate, respectively. The PNWE showed a strong power comparable to those of commercial phenolic compounds in biological systems. These results indicated that the protective activity of PNWE could be due to the presence of naturally-occurring phenolic compounds, which act as potent in vitro antioxidants in both non-cellular and cellular systems.
... In another study on determining pharmaceutical properties of little needles, it was found that pine needles could be effective in treatment of cardiovascular and skin diseases due to their affordability and the presence of bioactive substances [21]. It was hypothesis that pine needle extracts are more cytotoxic on to cancer cells than to normal cell in vitro, and the intake of pine needle powder exhibited significant antitumor effect in vivo [22]. This antitumor effect appears to be mediated through the antioxidative, antimutagenic, and antiproliferative properties of particular substances in pine needles [22]. ...
... It was hypothesis that pine needle extracts are more cytotoxic on to cancer cells than to normal cell in vitro, and the intake of pine needle powder exhibited significant antitumor effect in vivo [22]. This antitumor effect appears to be mediated through the antioxidative, antimutagenic, and antiproliferative properties of particular substances in pine needles [22]. In this context, the blood urea nitrogen and aspartate aminotransferase levels were reported to be significantly lower in pine needle supplemented rats. ...
... In this context, the blood urea nitrogen and aspartate aminotransferase levels were reported to be significantly lower in pine needle supplemented rats. These results clearly demonstrate that pine needles exhibit strong antioxidant, antimutagenic, and antiproliferative effects on cancer cells and also antitumor properties in vivo and point to their potential useful against cancer [22]. ...
... In addition, it can also be utilised to treat respiratory and congestion problems ("Pine Needles Information, Recipes and Facts," 2022). PNs powder is a rich source of vitamin A, essential oils and alkaloids, and its ethanol extract has antimutagenic solid, antitumor, antiproliferative and antioxidant properties (Ghosh and Ghosh, 2011;Kwak et al., 2006). A vastnumber of PNs species exist throughout the world, which can return nutrients to the forest, serve as mulching materials, or act as a source of nutrients when added to the soil. ...
... Chemical preservatives may undoubtedly efficiently inhibit the growth of microorganisms in food, but concerns about their safety and potential negative effects on health are mounting. Thus,researchers made efforts to utilise PNsextracts in the fields of pharmacology, microbiology, food preservation and medicine because of their alluring insecticidal, antibacterial, and antifungal activities (Feng et al., 2010;Ka et al., 2005;Kwak et al., 2006). Zeng et al. (2012) evaluated the antibacterial activities of water-soluble PNs-based plant extract on five food-borne bacteria, namely Staphylococcus aureus, Bacillus cereus, Proteus vulgaris, Escherichia coli and Bacillus subtilis. ...
Article
Full-text available
Pine needles (PNs) are one of the largest bio-polymer produced worldwide. Its waste, i.e., fallen PNs, is mostly responsible for forest fires and is a major challenge. In present article, we have reviewed differenteffortsmadeto tackle this situation.PNshave been used in various fields such asin composite, water purification industries,electronic devices, etc. Gasification is one of the appealing processes for turning PNs into bio-energy; pyrolysis technique has been employed to create various carbon-based water purification materials; saccharification combined with fermentation produced good yields of bio-ethanol; Pd or Ni/PNs biocatalyst showed good catalytic properties in variousreactionsand pyrolysis with or without catalyst is an alluring technique to prepare bio-fuel. Nano cellulose extracted from PNs showed appealing thermal and mechanical strength. The air quality of nearbyenvironment was examinedby studying the magnetic properties of PNs. Packing materials made of PNs showed exceptional ethylene scavenging abilities.
... It also helps to boost the immune system. Pine cone extract is thought to have various health benefits, including preventing the growth of HIV in human T cells, the growth of influenza, the growth of the herpes simplex virus, tumor activity, suppressing lymphoma malignancy that involves immune system cells, and boosting the immune system [44][45][46][47][48]. Terpenes, phenolic compounds, cinnamic acid, caffeic acid, chlorophyll components, inorganic and organic components, vitamins, and other chemical substances found in aqueous extract of pine cone from Pinus densiflora (Korean red pine) [44,49] can reduce Zn ions to Zn and operate as a topping and balancing specialist [50]. ...
... Pine cone extract is thought to have various health benefits, including preventing the growth of HIV in human T cells, the growth of influenza, the growth of the herpes simplex virus, tumor activity, suppressing lymphoma malignancy that involves immune system cells, and boosting the immune system [44][45][46][47][48]. Terpenes, phenolic compounds, cinnamic acid, caffeic acid, chlorophyll components, inorganic and organic components, vitamins, and other chemical substances found in aqueous extract of pine cone from Pinus densiflora (Korean red pine) [44,49] can reduce Zn ions to Zn and operate as a topping and balancing specialist [50]. In this study, ZnONPs were green produced using pine cone extract (PCE) and microwave irritation to shorten nanoparticle manufacture time. ...
Article
Full-text available
The biogenic synthesis of zinc oxide nanoparticles (ZnO NPs) with pinecone extract (PCE) as a reducing agent and antibacterial agent was explored. The current study aims to investigate the biosynthesis of ZnO NPs and their effect on photocatalytic dye degradation and antimicrobial properties. The physical, chemical, and morphological properties of biogenic ZnO NPs synthesized using PCE were investigated using advanced spectroscopy techniques such as Fourier transform infrared spectroscopy (FTIR), UV-visible spectroscopy, transmission electron microscopy (TEM) analysis, selected area electron diffraction (SAED), and X-ray diffraction (XRD) techniques. The photocatalytic degradation of methylene blue was measured spectrophotometrically using biogenic ZnO NPs as nanocatalysts, and decolonization of solution indicates dye degradation gradually as exposure duration increases. The antimicrobial properties of ZnONPs against the tested pathogenic strains were demonstrated using the disc diffusion method. The antimicrobial efficacy of ZnONPs was further explained using molecular docking analysis. Confirmation of the lowest binding energy was used to predict receptor binding site with NPs in order to understand the mechanistic approach. ZnONPs are likely to interact with pathogens via mechanical enfolding, which could be one of the major toxicity actions of ZnONPs against strains. Furthermore, the nontoxicity and biocompatibility of ZnO NPs were studied, revealing impressive hemocompatibility with red blood cells (RBCs) and no significant toxicity to Brine shrimps at lower ZnONP concentrations.
... Pine bark extracts are effective scavengers of free radicals and reactive oxygen species, can lower lipid levels in blood serum and may help to prevent disease and delay aging (Schafer et al., 2006;McGrath et al., 2015). PNs of form P. densiflora were used for folk medicine and for various disease prevention such as rheumatitis, hemorrhage, gastroenteric trouble, hypertension, and asthma (Kwak et al., 2006;Kim et al., 2020b). Recent scientific researches have shown that PNs form P. densiflora have antimicrobial, antiviral, antioxidant, anti-mutagenic, anti-thrombosis, antiasthmatic, and anti-inflammatory, and anti-cancer effects (Kwak et al., 2006;Park and Lee, 2011;Park et al., 2016;Ahn et al., 2018;Mostafa et al., 2018;Ha et al., 2020). ...
... PNs of form P. densiflora were used for folk medicine and for various disease prevention such as rheumatitis, hemorrhage, gastroenteric trouble, hypertension, and asthma (Kwak et al., 2006;Kim et al., 2020b). Recent scientific researches have shown that PNs form P. densiflora have antimicrobial, antiviral, antioxidant, anti-mutagenic, anti-thrombosis, antiasthmatic, and anti-inflammatory, and anti-cancer effects (Kwak et al., 2006;Park and Lee, 2011;Park et al., 2016;Ahn et al., 2018;Mostafa et al., 2018;Ha et al., 2020). However, the isolation and functional characterization of AMPs from PNs have been limited studies. ...
Article
Full-text available
Pine needles are used in several East Asian countries as food or traditional medicine. It contains functional components that exhibit a wide spectrum of pharmacological effects such as antioxidant, antimicrobial, anti-diabetic, and anti-inflammatory activities. We determined and characterized the novel antimicrobial peptides (AMPs) isolated from Pinus densiflora Sieb. et Zucc. The four active pine-needle (PN) peptides showed antimicrobial activity against foodborne bacteria with minimum inhibitory concentration (MIC) values within the range of 8–128 μg/ml. PN peptides showed no detectable hemolytic activity or cytotoxicity at the antimicrobial concentrations. The N -terminal amino acid sequence of the PN5 was identified using Edman degradation and Antimicrobial Peptide Database (APD) homology analysis showed that it was not identical to any other plant peptide. This suggests that PN5 can serve as an alternative therapeutic agent to be used in the food industry.
... To discuss the structure-activity relationships (SARs) of diterpenoids isolated from the P. densiflora cortex and leaves, 21 compounds (3-23) were divided into three groups based on abietane-type tricyclic phenolic (3-10), tricyclic (11)(12)(13)(14)(15)(16)(17), and labdane-type bicyclic diterpenoids (18)(19)(20)(21)(22)(23). Biological data of group 1 (3-10) ( Figure 3A and Figure S22) indicated that the methoxylation at C-7 (R 2 ) could improve the antiviral properties and reduce the cytotoxicity (when comparing compound 5 with 3 and 4). ...
... The antiviral profile of group 2 (11)(12)(13)(14)(15)(16)(17) indicated that the existence of one or two ketone groups at C-7 or C-13 significantly contributed to the anti-influenza activity of the abietane type (comparing compounds 13, 16, and 17 with 11, 12, 14, and 15). The final group (18)(19)(20)(21)(22)(23) demonstrated that substitution of the carboxyl group at C-4 clearly increased H1N1 inhibition. However, replacement of the carboxyl moiety with other groups significantly reduced the antiviral properties, while the cytotoxicity significantly increased (comparing compounds 20 and 23 with 18, 19 and 21). ...
Article
Full-text available
Pinus densiflora was screened in an ongoing project to discover anti-influenza candidates from natural products. An extensive phytochemical investigation provided 26 compounds, including two new megastigmane glycosides (1 and 2), 21 diterpenoids (3–23), and three flavonoids (24–26). The chemical structures were elucidated by a series of chemical reactions, including modified Mosher’s analysis and various spectroscopic measurements such as LC/MS and 1D- and 2D-NMR. The anti-influenza A activities of all isolates were screened by cytopathic effect (CPE) inhibition assays and neuraminidase (NA) inhibition assays. Ten candidates were selected, and detailed mechanistic studies were performed by various assays, such as Western blot, immunofluorescence, real-time PCR and flow cytometry. Compound 5 exerted its antiviral activity not by direct neutralizing virion surface proteins, such as HA, but by inhibiting the expression of viral mRNA. In contrast, compound 24 showed NA inhibitory activity in a noncompetitive manner with little effect on viral mRNA expression. Interestingly, both compounds 5 and 24 were shown to inhibit nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in a dose-dependent manner. Taken together, these results provide not only the chemical profiling of P. densiflora but also anti-influenza A candidates.
... 15,16 Various parts of conifers, such as needles, bark, cones, and pollen, are consumed and they have been shown to promote health and prevent some aging-related chronic diseases. 17 Essential oils obtained from conifer needles and bark have been widely used as bathing oils, ointments, or inhaling drugs for treating a wide range of disorders of neuralgic, infectious, and rheumatic origin. 18,19 Several studies have reported antioxidant, 9 antimicrobial, 20 antibacterial, 21 larvicidal, 22 antifungal, 19 herbicidal, 23 anti-inflammatory, 24 and free radical scavenging 25 activity of these oils. ...
... 27 Ethanol, methanol, and hexane extracts of conifer needles have been reported and they also possess considerable antioxidant, antimutagenic, antitumor, and anticancer activity. 16,17 Several methods, such as hydrodistillation, solvent extraction, simultaneous distillation−extraction, and solid-phase microextraction have been used for essential oil production. 28 Hydrodistillation is the most widely used method, although some water-soluble compounds are not obtained and thermolabile compounds can be degraded in the process. ...
Article
Full-text available
Extractives are an important class of compounds in plants because they contribute to many of their physicochemical properties such as color, odor, density, strength, permeability, and hygroscopicity. Moreover, they also possess significant biological activity and are thus an important part of the plants’ defense mechanisms against biotic and abiotic stresses. Tree needles are a rich source of extractives, counting for as much as 40% of their dry weight. In this study, chemical fingerprinting of essential oils and solvent extracts, obtained from the needles of four conifer tree species (i.e., pine, spruce, larch, and juniper), was performed by using ultrahigh-resolution Fourier transform ion cyclotron mass spectrometry. A wide variety of compounds were detected in the oil samples, including mono-, sesqui-, and diterpenes, terpenoids, fatty and resin acids, esters, and different phenolic compounds. Although the main compounds were present in all the four essential oil samples, large variations in their relative abundances were observed. In contrast, pine needle hexane and toluene extracts showed a high content of resin acids, including pinifolic acid, a rare labdane-type diterpene diacid, and its mono- and dimethyl esters. Thus, by selecting a suitable solvent, specific types of compounds may be isolated from tree needles for further biotechnological or medicinal applications.
... Exploration of natural products as anti-oxidant, antimicrobial or cytotoxic agents is a research field with great potential. Numerous groups are currently performing further activity-guided studies to find of new molecules of interest, including extraction prepared from divers parts of pine (Ustun et al., 2012;Cui et al., 2005;Kwak et al., 2006;Amalinei et al., 2014;Yesil-Celiktas et al ., 2009). ...
... Indeed, in agreement with the results of the present study, many of the phenolic compounds produced by the pine species have shown possess an inhibition of growth of human gastric and breast cells, cytotoxic and antiproliferative effects ( Kwak et al., 2006;Amalinei et al., 2014). ...
Article
Full-text available
p class="Abstract">New bioactive natural products, the phenolic composition and the biological activities of organic extracts from the needles of the Algerian Pinus coulteri were investigated. The analysis by HPLC-DAD of crude extract revealed the presence of 10 phenolic acids and nine flavonoids. In vitro anti-oxidant activities were performed using four different tests. The greatest antiradical activity was found in the ethyl acetate fraction (IC<sub>50 </sub>= 3.2 ± 0.3 µg/mL), whereas the diethyl ether fraction had the higher contents of total phenolics and flavonoids and exhibited a highest activity in reducing power and β-carotene–linoleic acid tests with EC<sub>50</sub>= 67.1 ± 0.4 μg/mL and 71.5 ± 0.2% of inhibition, respectively. Furthermore, a low to moderate antimicrobial activity according to all extracts was revealed against eight bacteria tested. The MIC value of chloroform fraction showed a strong degree of antibacterial activity (<0.09 mg/mL). The crude extract was found toxic with LC<sub>50</sub> value of 15.2 μg/mL by brine shrimp toxicity assay. The needle extract of P. coulteri is rich in valuable biologically active compounds and could represent a new resource of anti-oxidant agents for the treatment of diseases. Video Clip of Methodology: Disc diffusion test: 6 min 41 sec Full Screen Alternate </p
... Conifers have long been used in traditional medicine to treat various illnesses. Different parts of the conifer trees, such as needles, cones, and bark have been and still are consumed for improving health and preventing some age-related chronic diseases.[73][74][75] Resins obtained from conifers have been reported effective in treating skin ulcers.76,77 ...
Thesis
Full-text available
Extractives are a minor but essential compound class found in plants. They are predominant contributors to the physicochemical properties of plants such as taste, odour, colour, and durability. They also play a significant role in plants’ defence mechanisms against abiotic and biotic stresses. Coniferous trees, especially their needles, are a rich source of extractives, which have considerable utilization potential in many techno-chemical, pharmaceutical, and dietary applications. In this study, the needle extracts were obtained from four different conifer tree species (i.e., pine, spruce, juniper, and larch) by steam distillation, conventional Soxhlet extraction, or hydrothermal extraction, and their chemical composition was determined by using direct-infusion ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Two complementary ionization techniques, namely, electrospray ionization (ESI) and atmospheric pressure photoionization (APPI), were employed to target both polar and nonpolar compounds in the extracts. The use of FT-ICR MS allowed for the detection of an average of 3000 species, including compounds with diverse pharmaceutical and nutritional properties. Different extraction techniques resulted in the recovery of a wide variety of compounds, including terpene hydrocarbons (monoterpenes, sesquiterpenes, diterpenes, and triterpenes), terpenoids, fatty acids and other lipids, phenolic compounds, soluble carbohydrates as well as nitrogen alkaloids. Although the main compounds found in the steam distillation products (essential oils) were 8 terpenes and terpenoids, significant variation in their relative abundance was observed. The composition of the essential oil of spruce sprouts (i.e., new-born needles) differed considerably from that of the mature spruce needles. The lipophilic extracts showed high resin acid content, and some of the resin acids were exceptionally abundant in certain species, e.g., methyl pinifolate in pine needles. The hydrophilic extracts were concentrated with quinic acid, phenolic compounds, and monosaccharides. The obtained results show that conifer needles are a vast source of interesting bioactive compounds. The direct-infusion ESI/APPI FT-ICR MS allowed for the detection and further identification of hundreds of compounds solely based on accurate mass data. The use of ESI and APPI provided complementary information about the complex chemistry of the essential oil and solvent extract samples. Although the FT ICR MS technique provides molecular fingerprints for extremely complex mixtures, isomeric compounds cannot be separated unless the technique is coupled with liquid/gas chromatography or ion mobility, or by using tandem mass spectrometry.
... Pine needles, also known as bristle pine leaves and pine hairs, are the leaves of pine plants in the Pinaceae [25]. It is found that pine needles have remarkable pharmacological effects (such as lowering blood sugar, regulating blood lipids, inhibiting tumors, and anti-aging) [26][27][28][29]. It is also well-known that both fresh and fallen pine needles hardly rot. ...
Article
Full-text available
Polyolefin suffers from degradation during processing and application. To prolong the service life, antioxidants are needed in the packing formula of polyolefin products. The usage of natural antioxidants could avoid potential health hazards aroused by synthetic ones. Pine needles have long lives and hardly rot, suggesting their high resistance to degradation. To provide a new candidate of natural antioxidants and add more value to pine needles, pine needle extracts (PNE) were investigated as the antioxidant of linear low-density polyethylene (LLDPE). PNE-modified LLDPE (PE-PNE) exhibited much better short-term and long-term aging resistance than pure LLDPE (PE): Oxidation induction time (OIT) of PE-PNE was 52 times higher than that of PE, and the increments of carbonyl index (CI) of PE-PNE-1st samples placed under daylight and in the dark were approximately 75% and 63% of PE under the same conditions. It could be attributed to the attractive antioxidant capacity of PNE (IC50 of DPPH radical scavenging was 115 μg/mL). In addition, the PE-PNE sample showed high processing stability and maintenance of the mechanical property during multiple extrusions: only a 0.2 g/10 min decrease in melting flow rate was found after five extrusions; the tensile strength and elongation at break were almost unchanged. All results reveal that pine needle extracts could play a role in LLDPE stabilization. Moreover, as pine needles are mainly considered a kind of waste, the present study would benefit the budget-reducing polyolefin industry.
... D-limonene (17.8%), β-caryophllene (11.02%) and β-pinene (7.54%) as the principal components in pine cones (Yang et al., 2010). It has been reported that extracts obtained from pine cones have a reactive oxygen scavenging effect and lower serum lipids (Kartal & Öztürk, 2016;Kwak et al., 2006). It has been stated that reactive oxygen species, including superoxide anion, hydroxyl and ABTS radicals resulting from oxidative stress, can attack biological macromolecules in cells, tissues and organs, while high reactive oxygen concentrations in cells increase the probability of diseases such as diabetes, atherosclerosis, cancer, neurodegeneration and aging (Meng, 2010;Willcox et al., 2004). ...
Article
Full-text available
It was aimed to determine the phenolic compounds, mineral matters, antioxidant activities, ash, protein and color values of pine cone molasses. In conclusion, total sugar contents in pine cone molasses ranged from 69.86 to 73.14 %. In terms of the minerals, molasses was rich in K and Ca while the lowest contents of Se and Zn were found in pine cone molasses. The phenolic contents varied from 1294.1 to 1466.4 mg/L and DPPH (%) changed between 69.44 and 78.61 for P. nigra and P. brutia molasses respectively. The pH values varied from 3.91 to 4.24, and total acidity changed between 2.74 and 2.99 %. The viscosity varied from 270.4 to 368.8 mPa.s and L* values varied from 19.77 to 23.91 respectively. a* values were established between 6.77 and 8.41, while b* values are found between 5.99 and 6.74. The K, Ca, Mg, P and Mn contents of all the molasses samples has been detected at high levels. Oil yields were 0.178‐0.192 (w/w, %) for pine cone molasses. α‐terpineol was the main constituent in molasses and varied from 1863.4 to 1944.2 at a ppm level. The sizes not affected the physicochemical propeties of molasses. L* and a* values followed zero order,b* values followed first order kinetics. The pine cone molasses represent a potentially significant source of minerals, total sugars,essential oils and phenolic compunds. Nowadays, consumer interest and demand for potentially health‐promoting products have motivated the production of healty food products so these pine cone molasses (pekmez) are introduced as potentially healthy products of which the production stages and conditions that may advise the industrial production of cone molasses.
... Pines are conifers in the genus Pinus that are found in the Northern Hemisphere [1]. They have economic, ecological, and scientific importance, as they provide timber for construction, furniture, paneling, and flooring [2,3]; habitats and food for wildlife; and anticancer, antioxidant, and antimutagenic properties in their needles [4]. However, pine trees (such as P. tabuliformis Carr., P. thunbergii Parl., and P. koraiensis Siebold and Zucc.) are threatened by pine wilt disease (PWD), a devastating disease that kills trees within a few weeks to a few months from infection, with symptoms that are characterized by wilted and brown needles. ...
Article
Full-text available
Pine wilt disease (PWD) has caused extensive mortality in pine forests worldwide. The longicorn beetle Monochamus saltuarius, as the vector of the invasive species Bursaphelenchus xylophilus, plays an important role in the infection cycle. Although the gut microbiota and its contribution to health and disease have been extensively documented, it is unclear whether B. xylophilus affects the longicorn gut microbiota because of a lack of understanding of potential temporal changes in the microbial composition of the vector beetles. In this study, we collected beetles at the emergence and mating stages, and divided them into two groups according to whether they carried nematodes. Based on 16S rDNA sequence analysis, 174 bacterial species were identified that belonged to 112 genera, 53 families, and 8 phyla. Bursaphelenchus xylophilus increased the microflora abundance and diversity of the infected M. saltuarius. In addition, Firmicutes and Bacteroidetes were more abundant in infected M. saltuarius at the same developmental stage. Some of the bacteria in these two phyla were the key species in the co-occurrence network of intestinal flora and represented a unique module in the co-occurrence network of infected M. saltuarius. We found some high abundance colonies in the intestinal tract of infected M. saltuarius during the emergence period that were mostly related to metabolism. Compared with the emergence period, there were more similar microorganisms in the intestinal tract of M. saltuarius during the mating period. With the change in growth environment and continuous feeding, the intestinal microorganisms gradually stabilized and became single species.
... Needles, pollen, cones, and cortices derived from pine trees have been used as antibacterial, anti-inflammatory, and preservative agents in traditional medicine (29). Particularly, pine needles are crude drugs that are effective for preventing diseases such as rheumatism, gastroenteric trouble, and asthma (30,31). PN5, which consists of 11 amino acids, was isolated from pine needles and showed antibacterial activity against foodborne bacteria in a previous study (20). ...
Article
Full-text available
Antibiotic-resistant bacteria have become a public health problem. Thus, antimicrobial peptides (AMPs) have been evaluated as substitutes for antibiotics. Herein, we investigated PN5 derived from Pinus densiflora (pine needle). PN5 exhibited antimicrobial activity without causing cytotoxic effects. Based on these results, we examined the mode of action of PN5 against Gram-negative and -positive bacteria. PN5 exhibited membrane permeabilization ability, had antimicrobial stability in the presence of elastase, a proteolytic enzyme, and did not induce resistance in bacteria. Bacterial lipopolysaccharide (LPS) induces an inflammatory response in RAW 264.7 macrophages. PN5 suppressed proinflammatory cytokines mediated by NF-κB and mitogen-activated protein kinase signaling. In C57BL/6J mice treated with LPS and d-galactosamine, PN5 exhibited anti-inflammatory activity in inflamed mouse livers. Our results indicate that PN5 has antimicrobial and anti-inflammatory activities and thus may be useful as an antimicrobial agent to treat septic shock caused by multidrug-resistant (MDR) Escherichia coli without causing further resistance. IMPORTANCE Antibiotic-resistant bacteria are a global health concern. There is no effective treatment for antibiotic-resistant bacteria, and new alternatives are being suggested. The present study found antibacterial and anti-inflammatory activities of PN5 derived from Pinus densiflora (pine needle), and further investigated the therapeutic effect in a mouse septic model. As a mechanism of antibacterial activity, PN5 exhibited the membrane permeabilization ability of the toroidal model, and treated strains did not develop drug resistance during serial passages. PN5 showed immunomodulatory properties of neutralizing LPS in a mouse septic model. These results indicate that PN5 could be a new and promising therapeutic agent for bacterial infectious disease caused by antibiotic-resistant strains.
... 22 It has been well documented that the pine needle oil had antioxidant, antimicrobial and anticancer activities. 23,24 Pine needle oil treatment arrested the Hepg2 cycle cell and inhibited the proliferation of liver cancer cell line. 25 Other parts of the tree possess remarkable antioxidant, anti-inflammatory and antimicrobial activities. ...
Article
Introduction: Acetaminophen (APAP)-induced liver injury is one of the main causes of acute liver failure in the world. Pinus eldarica is specially distributed in the north of Iran and has been used for decades to treat wounds, pain, infection, fever, bronchitis and inflammation in many countries. This study was undertaken to evaluate the role of pine pollen extract (PE) on APAP-induced hepatotoxicity. Methods: This study was conducted in two separate parts: single- and multi- dose administration of PE. In a multiple dosing regimen, different doses of extract (10, 20, 30, 40 and 50 mg/kg) were orally administrated for 2 weeks and then on the 15th day, the animals received a single dose of APAP (600 mg/kg). In the second part, the highest dose of PE (50 mg/kg) was orally administrated half an hour after receiving the APAP at dose of 600 mg/kg. The negative and positive control groups were treated with normal saline and N-acetylcysteine (NAC), respectively. At the end of procedure, the biochemical parameters including alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) as well as pathological findings were evaluated. Results: A single and multiple oral dose of APAP increased the serum level of ALT, AST and ALP that were significantly attenuated by PE administration in both model. Indeed, exposure to APAP caused extensive necrosis and lymphocytic inflammation that were completely prevented by single- and multi-dose administration of PE, specially at a high dose. Conclusion: Finally, the present results indicate that PE could reverse the reduction of aminotransferases and improve histological changes of APAP-induced liver toxicity. Further research is needed to confirm this finding and reveal the exact mechanisms.
... Pinus eldarica, also named Tehran pine, is native to the South Caucasus between Europe and Asia and has been extensively grown in Iran. In traditional medicine, different parts of this plant (bark, needles, and nuts) have been used for the treatment of several conditions, including skin wounds, bronchial asthma, dermatitis, and skin irritation, and have reportedly shown antibacterial, antimutagenic, antitumor, anti-inflammatory, and glucose-lowering activities [8][9][10][11]. Essential oils and extracts from pine needles and bark are rich in terpenoids such as caryophyllene, limonene, α-pinene, and phenolic acids [12][13][14]. Pine extracts act as antioxidants by reacting with free radicals, thus protecting DNA against oxidative damage and apoptosis induced by hydroxyl radicals [15][16][17]. ...
Article
Full-text available
Lung cancer is one of the leading causes of cancer-related mortality worldwide. Although effective clinical drugs for treating advanced stages are available, interest in alternative herbal medicines has gained momentum. Herbal extracts are potent antioxidants that reportedly inhibit the growth of various cancer cell lines. In the present study, we investigated the effects of essential oils and hexane, methanolic, and aqueous extracts, obtained from various parts (bark, needles, and pollen) of Pinus eldarica against human lung cancer (A549) cells. First, the DPPH radical scavenging activities of P. eldarica extracts and essential oils were examined, which revealed that methanolic extracts presented higher antioxidant activity than the other extracts and essential oils. Next, A549 cells were exposed to various concentrations of the extracts and essential oils for 48 h. P. eldarica extracts/essential oil-treated lung cancer cells demonstrated a significant decrease in cell proliferation, along with an induction of apoptotic cell death, particularly, the pollen hexane extract, bark essential oil, and methanolic needle extract showed superior results, with IC50 values of 31.7, 17.9, and 0.3 μg/mL, respectively. In the cell cycle analysis, treatment of A549 cells with the methanolic needle and pollen hexane extracts led to apoptosis and accumulation of cells in the sub-G1 phase. Further, exposure to the bark essential oil and methanolic needle extract decreased the cell population in the G2/M phase. Notably, treatment with the pollen hexane extract, bark essential oil, and methanolic needle extract resulted in caspase-3 activation, poly (ADP-ribose) polymerase cleavage, Bcl-2 downregulation, and Bax and p53 regulation in A549 cells. Furthermore, these extracts and essential oils decreased the migration, and colony formation of A549 cells. These findings provide experimental evidence for a new therapeutic effect of P. eldarica against human lung cancer and suggest P. eldarica as a potential chemopreventive natural resource for developing novel cancer therapeutics.
... These results clearly demonstrated that phytoconstituents present in both green and fallen needles can act as promising immune-enhancers for the treatment of cancers of several origins. Earlier, P. densiflora needles have shown antitumor effects to rats incubated with mammary carcinogen, 7, 12-dimethylbenz[a]anthracene or to the mice injected with sarcoma-180 cells.25 ...
Article
Abstract Background: Pinus roxburghii has been used in the Himalayan region as folkloric remedy while nothing is yet known about its immunomodulatory potential. Methods: The crude extracts of green and fallen pine needles were subjected to sequential fractionation to get partially purified fractions. Human peripheral blood lymphocytes (PBL) were used to analyze immunoenhancing potential of these fractions. Microcytotoxicity assay was employed to investigate improved cancer cells killing capabilities of PBL against various cancer cell lines. GC-MS was carried out to identify the major compounds in the bioactive fractions. Results: The lymphocyte proliferation assay depicted the immunoenhancing potential of extracts and fractions of fallen and green needles of P. roxburghii. The ethyl acetate fractions of both fallen and green needles displayed highest mitogenic activity on human PBL. Both fractions heightened the expression of cell surface markers (CD3, CD8, and CD56) and significantly increased the production of cytokines (IL-2, IFN-γ and TNF-α). The enhanced intracellular granulysin (immunomarker for activated CTLs and NK cells) expression also confirmed immune stimulatory potential of these fractions on human lymphocytes. The ethyl acetate fractions of pine needles enhanced the cytotoxicity of PBL towards various cancer cells (HCT-116, HeLa, PC-3 and A549) as compared to untreated PBL. GC-MS analysis showed presence of major compounds like 3-α-mannobiose, octakis (trimethylsilyl) ether, methyloxime in ethyl acetate fraction of the green needles and cyclodecasiloxane, eicosamethyl in ethyl acetate fraction of the fallen needles. Conclusion: The bioactive fractions of the fallen and green needles of P. roxburghii stimulate cancer cells killing potential of human lymphocytes. Keywords: Anticancer, immunomodulation, granulysin, GC-MS, Pine needles.
... P. densiflora has the wide natural distribution area in Japan, Korea, North-Eastern China, and the extreme South-East of Russia. Pine needles contain several bioactive components such as α-pinene, caryophyllene, beta-pinene and bisbenzene, camphene, borneol, phellandrene, quercetin, kaempferol, and terpene, which have been reported to have antimicrobial, antimutagenic, and antioxidant effects [6][7][8]. Pine needles also contain calcium (28 mg/100 g) and various amino acids such as glutamic acid, phenylalanine, leucine, and lysine, as well as vitamins such as niacin, riboflavin, beta carotene, and thiamine [9,10]. Despite a rich source of bioactive compounds, some researchers indicated that high content of condensed tannins in pine needles might affect nutrient absorption by reducing protein digestibility in animals [11,12]. ...
Article
Full-text available
This study aimed to investigate the supplemental effects of fermented pine (Pinus densiflora) needle extract (FPNE) in laying hen diets on productive performance, egg quality, and serum lipid metabolites. A total of 108 40-week-old Hy-line brown laying hens were randomly assigned to one of the three dietary treatment groups: (1) basal diet + 0 mL FPNE/kg diet (CON), (2) basal diet + 2.5 mL FPNE/kg diet (T1), or (3) basal diet + 5 mL FPNE/kg diet (T2) for 6 weeks. Each group consisted of four replicates of nine hens each. Feed and water provided ad libitum. Results showed that dietary supplementation of FPNE increased egg production percentage (linear, p < 0.01 and quadratic, p < 0.05), egg mass (linear, p < 0.05), and feed intake (linear, p < 0.05) during the entire experimental period. In addition, dietary inclusion of FPNE significantly increased the eggshell color (linear, p < 0.01), egg yolk color (quadratic, p < 0.01), and eggshell breaking strength (linear, p < 0.05 and quadratic, p < 0.05) while the Haugh unit decreased (quadratic, p < 0.05). However, serum lipid profile did not differ among the dietary treatments (p > 0.05). Notably, antioxidant activity of egg yolk was improved by significantly decreasing the malondialdehyde content in egg yolks after 6 weeks of storage (linear, p < 0.001 and quadratic, p < 0.05). In summary, dietary inclusion of FPNE could improve laying performance and the antioxidant capacity of eggs.
... Geumgang to Chengsong and Yeongduk of Gyeongbuk province in Korea (Song et al., 2016). Pine needles have been found to have a lot of biological properties with anti-microbial, anti-mutagenic, anticarcinogenic, anti-inflammatory, anti-hyperlipidemic, anti-aging, anti-osteoclastic, and antioxidant activities (Choi et al., 1997;Kwak et al., 2006;Hwang et al., 2014;Jung et al., 2014;Venkatesan et al., 2017;Kim, 2018;Shim and Ma, 2018). Pine needles contain various kinds of valuable phytochemicals, such as phenolic acids, flavonoids, proanthocyanidins, lignans, phytosterols, resin, and essential oils (Zhang et al., 2011;Karapandzova et al., 2019;Metsämuuronen and Sirén, 2019). ...
Article
Full-text available
Five flavonol glycosides including quercetin 3-O-β-D-glucoside (QG), kaempferol 3-O-β-D-glucoside (KG), quercetin 3-O-(6″-O-acetyl)-β-D-glucoside (QAG), kaempferol 3-O-(6″-O-acetyl)-β-D-glucoside (KAG), and quercetin 3-O-(3″-O-p-coumaroyl)-β-D-glucoside (QCG) were isolated and purified from red pine (Pinus densiflora Sieb. et Zucc.) nee-dles, and identified by nuclear magnetic resonance and mass spectrometer spectral analyses. In addition, the quantification of the five flavonol glycosides in pine needles was performed by high-performance liquid chromatography analysis according to cultivar, growing district, harvest season, and thermal processing. The red pine needles had higher amounts of the five flavonol glycosides than the black pine needles except for QCG. There were no large differences in flavonoid composition and content among pine needles grown in three different areas. Levels of the five flavonol glycosides in red pine needles harvested during Spring ranged from 6.13 to 27.03 mg/100 g dry weight. Levels of two flavonol glycosides, QG and KG, gradually decreased with increasing harvest time, whereas the acylated flavonol glycoside, QCG, a predominant flavo-noid in pine needles, increased gradually with increasing harvest time. Two acetyl flavonol glycosides, QAG and KAG, increased steadily through Spring to Autumn, and then decreased gradually by Winter. Meanwhile heat treatments, such as roasting and steaming, increased the five flavonol glycosides during heating for 3 min, but then slowly decreased these when heating for 10 min. Microwave processing increased to some extent the five flavonol glycosides when heating for 3 min, and remained unchanged during the 10 min heating. These results suggest that the pretreated red pine needles with enhanced flavonoid content may be useful as potential sources for nutraceuticals and cosmeceuticals.
... Pinus in traditional medicine are used for respiratory system as antiseptic and expectorant, also for gastrointestinal disorders, urinary system diseases and for the treatment of skin diseases. Pine needles extracts showed effect against a range of bacteria as (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Bacillus cereus) (20) pine needles showed exhibit strong antioxidant, antimutagenic and also antitumor effects in vivo and point to their potential usefulness in cancer prevention (21). ...
Article
Full-text available
The detection qualitative of Phytochemical compounds of extracts was carried out using color reagents, total content of phenols and flavonoids was specified using folin– ciocalteu and aluminium chloride method, respectively, and antioxidant activity was determined through its ability to free radicals scavenging using DPPH radical. The efficacy of the plant extracts against pathogenic bacteria was studied by agar well diffusion method with different concentrations, and microdilution method was used to measure minimum inhibitory concentration (MIC) of all plant extracts. The results showed presence of tannins, phenols, and flavonoids in all extracts of both plants, while saponins were found in aqueous extracts only, cardiac glycosides and coumarins were absent in all plant extracts. Ethanolic extract of Pistacia atlantica recorded the highest content of phenols and flavonoids as 263.76 ± 0.53 (mg GAE/g Dw) and 46.83 ± 0.55 (mg RE/g Dw), respectively. While aqueous extract of Pinus canariensis recorded the lowest content of of phenols and flavonoids 30.11 ± 0.37 (mg GAE/g Dw) and 5.43 ± 0.38 (mg RE/g Dw), respectively. Both plants have been shown to have good antioxidant activity, as ethanolic extract of P. atlantica recorded the best ability to free radicals scavenging 90.27% ±1.51, ethanolic extracts of both plants were the most effective in inhibiting bacteria especially at high concentrations (500 mg/mL); the inhibition zone diameter of P. atlantica extract was reached against Shigella boydii 33.56 mm, while aqueous extract of P. canariensis was the most effective against Pseudomonas aeruginosa; the inhibition zone diameter was 21 mm. MIC ranged between 5.468 and 43.75 mg/mL depending on plant extract and bacterial species. This confirms the importance of plant extracts as a natural source of antibacterial to confront problems of increasing bacterial resistance to antibiotics that threaten public health.
... Pine needles have been traditionally used as supplements in various foods and folk medicine in Asia, specifically in Korea and China [15]. Pine needles exhibit several biological activities, including antimicrobial, antioxidant, antimutagenic, antitumor, and anticholesterol [1517]. ...
Article
Full-text available
Objective: This study was conducted to investigate the supplementary effect of a phytogenic blend (SPA: a mixture containing fermented Schisandra chinensis pomace, fermented Pinus densiflora needle extract, and Allium tuberosum powder in the ratio of 2 : 2 : 1) on egg production, egg quality, and blood and visceral organs properties in laying hens. Methods: A total of 135 Hy-line brown laying hens (48-wk-old) were randomly allocated to three dietary treatments with 5 replicates of 9 hens. The control group (CON) was fed a basal diet (no exogenous SPA addition) and the experimental groups were fed the basal diet containing SPA at the level of 0.1% and 0.3% for 6 weeks. Results: The feed intake was significantly improved in SPA supplemented groups as compared with the control (p < 0.05). However, egg production, daily egg mass, and feed conversion ratio were not different among the dietary treatments (p > 0.05). For egg quality traits, only Haugh unit (HU) was significantly improved in SPA (0.3%) (p < 0.05) as compared with other groups. However, HU was not affected during 4-wk of storage at 18℃ among the dietary treatments (p > 0.05). Furthermore, SPA supplementation did not affect the blood biochemical constituents except for the phosphate content, which was significantly higher in SPA groups than the CON group (p < 0.05). There were no significant differences in visceral organ characteristics and immune indicators (IgA, IgG, and IgM) in SPA or CON groups. Conclusion: This study suggested that the supplementation of SPA may have beneficial effects on feed intake and egg quality in laying hens.
... Such secondary metabolites possess several ecological and biological activities viz., anti-inflammatory, anti-oxidant and antimicrobial. In addition, they also exhibit a wide spectrum of actions such as anti-mutagenic, antitumor as well as protective against oxidative DNA damage, and apoptosis induced by hydroxyl radicals (Kwak et al., 2006;Jeong et al., 2009;Kim et al., 2010). These compounds also play an important role in imparting attractive colors to flowers that help in attracting pollinators. ...
Article
Full-text available
Environmental interventions and ecological adaptations harbor millions of valued substances and metabolites in plants which can be employed and commercialized for human benefits. Present study encompasses the untapped potential of pine needles of Indo-Himalayan region for the production of different metabolites and their pharmacological significance in terms of antioxidant and antimicrobial activity. Total phenolic and flavonoid content from the needles of ten pine species was quantified using three different solvent systems. Results revealed that out of 10 different selected Pinus species, Pinus taeda L. showed the highest concentration of total phenolics, Soluble-F phenolics and flavonoids content (approx. 147.02 mg/g, 141.08 mg/g and 21.91 mg/g, respectively) as compared to other species. On the other hand, Pinus greggii Engelm. ex Parl. showed the highest Bound-W phenolic content (approx. 3.62 mg/g). Among all the selected plant species, the needles of Pinus echinata Mill. exhibited the highest and Pinus thunbergii Parl. had the lowest ratio of total flavonoids to total phenolics. Most of these compounds were found to have effective antioxidant activities as well as antimicrobial activity, as estimated by oxygen radical absorbance capacity (ORAC) and disk diffusion test, respectively.
... P. densiflora is a needle-leaf tree distributed across eastern Asia. In recent years, pine needles have been reported to have antioxidant, antibacterial, anti-inflammatory, antimutagenic, and anti-tumor activities [5][6][7][8]. Several chemical compositions of pine needles have been identified to date [9]. ...
Article
Full-text available
This study aimed to investigate the optimal addition of terpene-based Pinus densiflora and Mentha canadensis extracts, with antioxidant and methane reduction effects, as feed supplements to ruminants. Two cannulated steers (450 ± 30 kg), consuming Timothy Hay and a commercial concentrate (60:40, w/w) twice daily (at 09:00 and 17:30) at 2% of body weight, with free access to water and a mineral block, were used as rumen fluid donors. In vitro fermentation experiments, with Timothy Hay as the substrate, were conducted with P. densiflora and M. canadensis extracts as supplements to achieve concentrations of 30, 50, and 70 mg/L on a Timothy Hay basis. Fibrobacter succinogenes decreased in proportion upon P. densiflora and M. canadensis extract supplementation at 50 mg/L, while the dry matter degradability of the feed was not significantly different (p < 0.05). Methane emission was significantly lower in the 50 and 70 mg/L treatment groups, for both extracts, at 12 h (p < 0.05). Based on methane production and antioxidant activity, our study suggests that 30 mg/L addition is the most appropriate level of supplementation.
... Pine trees from the genus Pinus are widely distributed in a variety of environments, ranging from areas at sea level to mountainous areas and from some of the coldest to the hottest environments on earth [1][2][3]. Pine forests are among the most valuable resources, with respect to timber, fuel, and various wood products [4][5][6]; native pine forests also provide important habitats or source of food for In this study, we examined the effects of two SA-derived SAR elicitors, acibenzolar-S-methyl (ASM) and methyl salicylic acid (MeSA), which effectively inhibited disease symptoms in PWN-infected pine trees. After treatment with these elicitors, the in vivo differentially expressed genes (DEGs) were characterised in B. xylophilus-infected pine trees. ...
Article
Full-text available
The pinewood nematode (PWN) Bursaphelenchus xylophilus causes pine wilt disease, which results in substantial economic and environmental losses across pine forests worldwide. Although systemic acquired resistance (SAR) is effective in controlling PWN, the detailed mechanisms underlying the resistance to PWN are unclear. Here, we treated pine samples with two SAR elicitors, acibenzolar-S-methyl (ASM) and methyl salicylic acid (MeSA) and constructed an in vivo transcriptome of PWN-infected pines under SAR conditions. A total of 252 million clean reads were obtained and mapped onto the reference genome. Compared with untreated pines, 1091 and 1139 genes were differentially upregulated following the ASM and MeSA treatments, respectively. Among these, 650 genes showed co-expression patterns in response to both SAR elicitors. Analysis of these patterns indicated a functional linkage among photorespiration, peroxisome, and glycine metabolism, which may play a protective role against PWN infection-induced oxidative stress. Further, the biosynthesis of flavonoids, known to directly control parasitic nematodes, was commonly upregulated under SAR conditions. The ASM- and MeSA-specific expression patterns revealed functional branches for myricetin and quercetin production in flavonol biosynthesis. This study will enhance the understanding of the dynamic interactions between pine hosts and PWN under SAR conditions.
... Pine needles: Pine needles (Pinus densiflora Siebold et Zuccarini) have long been used as a traditional health-promoting medicinal food. It has been shown that pine needle oil could induce DNA damage in a dose-dependent and have potential anticancer effects, antioxidant, antimutagenic, and antitumor activities (Kwak et al. 2006). ...
... Specifically, we investigated plant sap-derived EVs from four endemic plants collected from Bogildo and Hekseongsan in Korea: Dendropanax Morbifera (DM), Pinus densiflora (PD), Thuja occidentalis (TO), and Chamaecyparis obtusa (CO). It is known that DM and PD extracts have anticancer, antioxidant, antidiabetic, and anti-inflammatory activities, anti-melanogenic effects [25][26][27][28][29][30][31][32][33][34], whereas TO and CO extracts have antioxidant, anti-inflammatory, antimicrobial, and anti-asthmatic effects [35][36][37][38]. EVs isolated from these four kinds of plant saps were investigated for their anticancer effects on normal, low metastatic, and malignant cells. ...
Article
Full-text available
Edible plants have been widely used in traditional therapeutics because of the biological activities of their natural ingredients, including anticancer, antioxidant, and anti-inflammatory properties. Plant sap contains such medicinal substances and their secondary metabolites provide unique chemical structures that contribute to their therapeutic efficacy. Plant extracts are known to contain a variety of extracellular vesicles (EVs) but the effects of such EVs on various cancers have not been investigated. Here, we extracted EVs from four plants—Dendropanax morbifera, Pinus densiflora, Thuja occidentalis, and Chamaecyparis obtusa—that are known to have cytotoxic effects. We evaluated the cytotoxic effects of these EVs by assessing their ability to selectively reduce the viability of various tumor cell types compared with normal cells and low metastatic cells. EVs from D. morbifera and P. densiflora sap showed strong cytotoxic effects on tumor cells, whereas those from T. occidentalis and C. obtusa had no significant effect on any tumor cell types. We also identified synergistic effect of EVs from D. morbifera and P. densiflora saps on breast and skin tumor cells and established optimized treatment concentrations. Our findings suggest these EVs from plant sap as new candidates for cancer treatment.
... Consumption of different parts of Pinus species such as leaves (needles), shoots, cones, and their extracts has been reported to cure asthma, wound, bronchitis, common cold, and cough in humans (Kültür, 2007;Ugulu et al., 2009;Cakilcioglu et al., 2011). Various studies also exhibited the antioxidant and anticancer effects of different pine tree products (Kwak et al., 2006;Park et al., 2008;Zeng et al., 2014). Pine needles are reported to possess a wide variety of beneficial compounds typical of medicinal plants such as essential oils (α and β-pinene), phenolic substances, vitamin C, protein, fat, and phosphorus (Pfister et al., 1998;Xie et al., 2015). ...
Article
Full-text available
This study was proposed to examine the effects of pine needles powder (Pinus brutia) supplementation on growth performance, breast meat composition, and antioxidant status in broilers fed linseed oil-based diets. For this purpose, a total of 210, Ross-308 1-day-old male broiler chicks were allocated to 5 experimental groups each containing 42 birds. Broilers were fed a linseed oil-based basal diet supplemented with 0% (control), 0.25% (P1), 0.50% (P2), 0.75% (P3), and 1% (P4) pine needles powder. During the 42-D feeding period, no significant differences were observed between experimental groups for body weight gain, feed intake, and feed conversion ratio; however, carcass yield was increased linearly with pine needles powder supplementation. No marked changes in the breast meat chemical composition were observed among experimental groups. Supplemental pine needles powder linearly decreased the malondialdehyde concentration in breast meat and liver tissues; however, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of breast meat samples remained unaffected. No significant variation was observed among experimental groups for superoxide dismutase enzyme activity in blood erythrocyte lysates, but blood serum total oxidation status tended to decrease with pine needles powder supplementation. In conclusion, results suggested that pine needles powder supplementation to broiler diets could be a viable option to improve the animal antioxidant status and meat oxidative stability; however, supplementation of Pinus brutia needles powder up to 1% into broiler diets was not sufficient to efficiently curb the fat-induced oxidation in meat. Further investigation is needed to determine the full antioxidant potential of pine needles powder supplementation in poultry by comparing different pine species, evaluating the bioavailability of their active compounds and determining most effective dietary concentration for broiler meat production without any adverse effects.
... Pines are conifers in the genus Pinus that are found in the Northern Hemisphere 1 . They have economic, ecological, and scientific importance, as they provide timber for construction, furniture, paneling, and flooring 2,3 , habitats and food for wildlife, and (in their needles) agents with anticancer, antioxidant, and antimutagenic properties 4 . However, pine trees (such as Pinus densiflora, P. thunbergii, and P. koraiensis) are threatened by pine-wilt disease (PWD), a devastating disease that kills trees within a few weeks to a few months from infection, with symptoms that are characterized by wilted and brown-colored needles 5,6 . ...
Article
Full-text available
Pinus densiflora (Korean red pine) is a species of evergreen conifer that is distributed in Korea, Japan, and China, and of economic, scientific, and ecological importance. Korean red pines suffer from pine wilt disease (PWD) caused by Bursaphelenchus xylophilus, the pinewood nematode (PWN). To facilitate diagnosis and prevention of PWD, studies have been conducted on the PWN and its beetle vectors. However, transcriptional responses of P. densiflora to PWN have received less attention. Here, we inoculated Korean red pines with pathogenic B. xylophilus, or non-pathogenic B. thailandae, and collected cambium layers 4 weeks after inoculation for RNA sequencing analysis. We obtained 72,864 unigenes with an average length of 869 bp (N50 = 1,403) from a Trinity assembly, and identified 991 differentially expressed genes (DEGs). Biological processes related to phenylpropanoid biosynthesis, flavonoid biosynthesis, oxidation–reduction, and plant-type hypersensitive response were significantly enriched in DEGs found in trees inoculated with B. xylophilus. Several transcription factor families were found to be involved in the response to B. xylophilus inoculation. Our study provides the first evidence of transcriptomic differences in Korean red pines inoculated with B. xylophilus and B. thailandae, and might facilitate early diagnosis of PWD and selection of PWD-tolerant Korean red pines.
... and hypertension 13 . Recently, it was reported that pine needles have anti-oxidant, anti-mutagenic, anti-tumor, anti-bacterial, anti-inflammatory and memory enhancing activities [14][15][16][17][18] . Moreover, it also has been reported that bark and pollen of pine trees have anti-inflammatory and analgesic activities 19,20 . ...
Article
Full-text available
Purpose: To investigate the effects of pine needle extract (PNE) on the expression of proliferating cell nuclear antigen (PCNA) and Ki-67 during liver regeneration induced by 70% partial hepatectomy (PH) in rat. Methods: Forty-eight male rats (SD, 7 weeks) had surgery (70% PH). They were randomly divided into two groups. PH + PNE group was only provided PNE diluted in water (10%) for drinking and PH group was provided water from 5 days before surgery to the time of sacrifice. PNE was made by pressing and filtering. Animals were sacrificed at 12h, 24h, 36h, 60h, 84h, 168h after PH, respectively. The expressions of PCNA and Ki-67 were determined as proliferation indices. Results: Immunohistochemistry turned out to increase the expression of PCNA and Ki-67. PCNA expression of PH+PNE group increased up to twice of that of PH group. Western blot also seemed to increase the PCNA expression. These results indicated the promotion of cell proliferation in liver tissue and hepatic regeneration. Conclusions: Pine needle extract stimulates the expression of some mitotic proteins during liver regeneration induced by 70% PH in rats. It suggests that administration of pine needle extract could accelerate the liver regeneration after partial hepatectomy.
... Pine needles powder consists of antioxidant that could reduce metastasis in the body, assist in heavy metal detoxification, and have antiseptic and anti-inflammatory effects (Kwak et al. 2006;Guo et al. 2018). Moreover, phenol (C 6 H 5 OH) extract prepared from pine trees, which was used as a reliable food source of natural phenolic antioxidant, could make a better antioxidant category in the animals, poultry and the human body (Zheng et al. 2014;Kara et al. 2016). ...
Article
Full-text available
The aim of the study was to investigate the effect of pine needles and vitamin E powder on blood biochemistry and carcass characteristics of female Japanese quails. A total of 180 female Japanese quails were obtained from the open market and maintained at deep litter housing pens system for 42 days. After adaptation, all female quails were divided into the following groups, group 1, control QC; group 2, QE; supplemented with vitamin E at the rate of 150 mg L −1 ; group 3, QP; pine needles at the rate of 150 mg L −1 and group 4, QEP; the combination of both. The high-density lipoprotein cholesterol (HDLC) and total cholesterol (TC) were significantly (p < 0.05) affected by QE and QP groups as compared to QC and QEP groups. Triglyceride (TG), total protein (TP), glutathione peroxidase (GPx) and superoxide dismutase (SOD) significantly (p < 0.05) increased in all treatment groups except for the QC group. Aspartate transaminase was (AST) significantly (p < 0.05) increased and alanine transaminase (ALT) was decreased in treatment groups as compared to QC. The concentration of Zinc (Zn), female breast meat composition, and the content of crude fat were significantly (p < 0.05) higher in treatments groups than in QC. The addition of pine needles and vitamin E in female Japanese quail feed, improved female quail performance, and serum antioxidant system.
... Pine needles powder consists of antioxidant that could reduce metastasis in the body, assist in heavy metal detoxification, and have antiseptic and anti-inflammatory effects (Kwak et al. 2006;Guo et al. 2018). Moreover, phenol (C 6 H 5 OH) extract prepared from pine trees, which was used as a reliable food source of natural phenolic antioxidant, could make a better antioxidant category in the animals, poultry and the human body (Zheng et al. 2014;Kara et al. 2016). ...
Article
Full-text available
The aim of the study was to investigate the effect of pine needles and vitamin E powder on blood biochemistry and carcass characteristics of female Japanese quails. A total of 180 female Japanese quails were obtained from the open market and maintained at deep litter housing pens system for 42 days. After adaptation, all female quails were divided into the following groups, group 1, control QC; group 2, QE; supplemented with vitamin E at the rate of 150 mg L⁻¹; group 3, QP; pine needles at the rate of 150 mg L⁻¹and group 4, QEP; the combination of both. The high-density lipoprotein cholesterol (HDLC) and total cholesterol (TC) were significantly (p < 0.05) affected by QE and QP groups as compared to QC and QEP groups. Triglyceride (TG), total protein (TP), glutathione peroxidase (GPx) and superoxide dismutase (SOD) significantly (p < 0.05) increased in all treatment groups except for the QC group. Aspartate transaminase was (AST) significantly (p < 0.05) increased and alanine transaminase (ALT) was decreased in treatment groups as compared to QC. The concentration of Zinc (Zn), female breast meat composition, and the content of crude fat were significantly (p < 0.05) higher in treatments groups than in QC. The addition of pine needles and vitamin E in female Japanese quail feed, improved female quail performance, and serum antioxidant system.
... It damages cells and results in disequilibrium of the internal environment and diseases (Liu et al. 2011). Previous studies indicated that the major active ingredients, such as flavonoids and phenolic compounds in pine needles, have anti-microbial, anti-mutagenic, anti-oxidant and antitumoural functions (Choi et al. 1997;Choi et al. 2002;Kwak et al. 2006), which helps to explain the increased SOD and reduced MDA levels in this study when PNP was supplemented in the diet. ...
Article
Full-text available
A total of eighty 21-days old broilers with similar body weight were randomly allocated to four dietary treatments with five replicates of four chickens per replicate. Birds were fed diets containing 0% PNP (Control, CT), 1% PNP (PNP1), 3% PNP (PNP3) and 5% (PNP5) respectively. The results showed that the final body weight and body weight gain had no significant differences between groups. The PNP supplementation increased the weight of gizzard and rectums, but decreased small intestine weight (P < 0.05). Overall the weight indices of gizzards and rectums increased in the PNP treatments, while the weight indices of small intestine decreased compared with the CT. Broilers receiving 3% PNP had higher serum superoxide dismutase activity compared with CT. The serum malondialdehyde content was lower in PNP5 treatment compared with CT. The serum triglyceride contents were lower in the PNP3 and PNP5 treatments compared with CT. Total serum cholesterol of PNP1 and PNP3 treatments was lower than CT. In conclusion, this study showed that PNP supplementation reduced the serum cholesterol and triglycerides in broilers and it may improve the development of the digestive tract and antioxidant functions of broilers without altering growth performance.
... Essential oil content is higher in pine needle, has a special fragrance, mainly contains monoterpene, sesquiterpene and a small amount of terpene alcohols, esters, such as α-pinene, β-pinene, limonene, β-caryophyllene, etc ( Nikolic et al., 2007;Zeng et al., 2012;Satyal, Paudel et al. ,2013). The pine needle essential oil were reported to possess various biological activities, involving antimicrobial ( Lee et al., 2009, Amri et al., 2011Politeo et al., 2011), antifungall ( Satyal et al., 2013), antioxidant ( Kwak et al., 2006;Park and Lee, 2011;Emami et al., 2013;Chaudhary et al., 2014), anti-inflammatory ( Suntar et al., 2012), anti-tussive and expectorant, hypolipidemic ( Kim et al., 2012;Ko et al., 2013), anti-hypertension, anti-diabetic ( Joo et al., 2013), cardioprotective properties, anticancer ( Wei et al., 2008;Jo et al., 2012;Yousuf Dar., 2012) etc. Most pine trees which are evergreen, sustainable renewable are considered a source of bioactive compounds which have achieved important contributions to the discovery of pharmaceutical materials and other biomedical applications. ...
... Based on the biological function of frankincense, it possibly possesses anticancer characteristics. Pine needle (Pinus densiflora Siebold & Zucc.), is usually utilized as a herbal medicine, tea bag infusion and health supplement in East Asian countries, such as Korea and China (10). It is beneficial in the therapy of patients with coronary heart disease (CHD), neurodegenerative disorders and carcinoma. ...
Article
Full-text available
BC (BC), as the most common malignancy in women worldwide, is associated with high morbidity and mortality. However, chemoresistance and toxicity are the main causes that limit the success of treatment in aggressive BC cases. Thus, there is a vital need to identify and develop novel therapeutic agents. Frankincense, pine needle and geranium essential oils have been reported to play critical biological activities in cancer. However, to the best of our knowledge whether frankincense, pine needle and geranium essential oils have any effect on the progression of BC in MCF-7 cells remains unclear. In the present study, we assessed the possible effects of frankincense, pine needle and geranium essential oils on cell viability, proliferation, migration and invasion as well as the possible mechanisms. MCF-7 cells were treated with oils, and associations with BC were investigated. In the present study, we clearly revealed that frankincense, pine needle and geranium essential oils suppressed cell viability, proliferation, migration and invasion in human BC MCF-7 cells. Further data demonstrated that frankincense, pine needle and geranium essential oils induced apoptosis, but did not affect cell cycle progression. Consistent with the in vitro activities, frankincense essential oil was effective in inhibiting tumor growth and inducing tumor cell apoptosis in a human BC mouse model. In addition, these 3 essential oils modulated the activity of the AMPK/mTOR signaling pathway. In conclusion, the present study indicated that frankincense, pine needle and geranium essential oils were involved in the progression of BC cells possibly through the AMPK/mTOR pathway.
... b the translocation of NFκB p65 to the nucleus was assessed by immunofluorescence staining. Representative microscopic images are shown here antitumor and antibacterial effects [22][23][24]. The present study revealed that Pinus densiflora needle supercritical fluid extract inhibits the production of inflammatory molecules, NO, IL-6 and IL-1β in LPS-stimulated murine RAW 264.7 macrophage cells. ...
Article
Full-text available
Background Regulation of a persistently-activated inflammatory response in macrophages is an important target for treatment of various chronic diseases. Pine needle extracts are well known to have potent immunomodulatory effects. The current study was designed to evaluate the effects of Pinus densiflora needle supercritical fluid extract (PDN-SCFE) on bacterial lipopolysaccharide (LPS)-induced inflammatory response in RAW 264.7 murine macrophages. Methods Cytotoxic effect of PDN-SCFE was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The levels of nitric oxide (NO) and the corresponding enzyme, inducible nitric oxide synthase (iNOS), were quantified by Griess and immunoblotting methods, respectively. The levels of cytokines were quantified using commercial ELISA kits. Quantitative real-time PCR (qRT-PCR) analysis was performed to assess the mRNA expression of iNOS and cytokines. To elucidate the mechanism of action, the involvement of nuclear transcription factor-kappa B (NFκB), mitogen activated protein kinases (MAPKs) and Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathways were examined by an immunoblotting method. In addition, the cellular localization of NFκB was analyzed by immunofluorescence staining. Results MTT assay results indicated that PDN-SCFE is non-toxic to RAW 264.7 cells up to a maximum assayed concentration of 40 μg/mL. The PDN-SCFE exhibited a concentration-dependent inhibitory effect on LPS-induced NO production by down regulating the expression of iNOS. In addition, the extract suppressed the LPS-induced expression of interleukin-6 (IL-6) and interleukin-1β (IL-1β) but not tumour necrosis factor-α (TNFα). Mechanistic studies revealed that PDN-SCFE does not influence the NFκB and MAPK pathways. However, it showed a significant inhibitory effect on LPS-induced activation of STAT1 and STAT3 proteins in macrophages. Conclusion The present findings revealed that the anti-inflammatory activity of PDN-SCFE in LPS-challenged RAW 264.7 macrophages is probably caused by the suppression of the JAK-STAT signaling pathway. Graphical Abstract
Article
Full-text available
In this study, we investigated the anti-amnesic effect of Korean red pine (Pinus densiflora) bark extract (KRPBE) against amyloid beta1-42 (Aβ1-42)-induced neurotoxicity. We found that treatment with KRPBE improved the behavioral function in Aβ-induced mice, and also boosted the antioxidant system in mice by decreasing malondialdehyde (MDA) content, increasing superoxide dismutase (SOD) activities, and reducing glutathione (GSH) levels. In addition, KRPBE improved the cholinergic system by suppressing reduced acetylcholine (ACh) content while also activating acetylcholinesterase (AChE), regulating the expression of choline acetyltransferase (ChAT), postsynaptic density protein-95 (PSD-95), and synaptophysin. KRPBE also showed an ameliorating effect on cerebral mitochondrial deficit by regulating reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and ATP levels. Moreover, KRPBE modulated the expression levels of neurotoxicity indicators Aβ and phosphorylated tau (p-tau) and inflammatory cytokines TNF-α, p- IκB-α, and IL-1β. Furthermore, we found that KRPBE improved the expression levels of neuronal apoptosis-related markers BAX and BCl-2 and increased the expression levels of BDNF and p-CREB. Therefore, this study suggests that KRPBE treatment has an anti-amnestic effect by modulating cholinergic system dysfunction and neuroinflammation in Aβ1-42-induced cognitive impairment in mice.
Article
Forests cover one-third of the planet’s land surface. Forests are essential for maintaining a clean environment, food security and biodiversity. Forests are also a source of food for animals. In the processing of forest resources, the forest industry accumulates various forest biomass wastes containing valuable and nutritious biologically active substances. The article provides an overview of the current state of the art in applying forest industry waste to feed farm animals and poultry. The authors presented the characteristics of biological properties of plant forestry raw materials and described the mechanism of their action on the animal organism. The authors analysed the results of studies of domestic and foreign scientists on feeding animals and poultry with feed additives containing pine needles and pine nutshells. The authors also characterised the possible advantages and disadvantages of using feed additives. The analysis of Russian and foreign scientific literature in the area under study has shown that forest industry waste in feed production and animal feeding is relevant as the available fodder base does not always allow to satisfy the need of animals for nutrients and biologically active substances. It has been noted that pine needles are used worldwide as a feed additive, but in the countries with the most significant areas of forests and developed forest industry, their use is the most common and effective. The use of cedar nut shells in the feeding of farm animals and poultry is most produced in Russia, while this area is underdeveloped in the world. The literature review has confirmed that using forest industry wastes such as pine nut needles and shells in animal feed production is promising livestock production.
Article
Pinus roxburghii commonly known as “chir pine” belongs to the family Pinaceae and is one of the ornamental trees having tremendous medicinal values placed all over the temperate territory of the northern hemisphere including the Himalayas. There is a global interest in the scientific community to identify novel therapeutic agents that may fight against cancer. In the present study, ethyl acetate fractions (EAF) of green and fallen needles of Pinus roxburghii from the Himalayan region of Himachal Pradesh, India were investigated for immunostimulatory potential toward cancer reduction under both in vitro and in vivo conditions. The previous preliminary study is baselined for the novel idea of using ethyl acetate fractions of green and fallen needles of Pinus roxburghii to assess immunomodulation on Ehrlich ascites carcinoma (EAC) tumor model among six extracts and fractions. A 10 mg/kg body weight of EAF was injected intraperitoneally for 7 days for immunostimulation of mice prior to cancer induction and compared with standard immunostimulator levamisole. After 14 days of EAC inoculation, hematological, biochemical parameters and vital organs were analyzed. The in vitro data depicted that EAF of green and fallen needles of P. roxburghii possessed the highest immunoenhancing potential on human peripheral blood lymphocytes. Pre-treatment with EAF at 10 mg/kg of the body weight, enhanced the mean survival time to (% ILS=55) and (% ILS=58) respectively in percentage increased life span, whereas it was found to be (% ILS=78) for levamisole. Moreover, EAF of both needles indicated a reduction in tumor growth up to 70.3% while levamisole exhibited 85.4% compared to tumor control groups via immune boosting. Histological examinations revealed enhanced cell number and increased sizes of spleen and thymus indicating boosted immunological responses of EAFs. Hematological and biochemical parameters were recorded to be normal compared to tumor control. The study presented here proved that EAF of green and fallen needles of P. roxburghii have a novel, potential and rich source of immune-boosting phytochemicals for fighting tumor development. Therefore EAF of the needles may be used to develop novel alternative immune boosters which can fight against the development of various types of cancers.
Article
Full-text available
Background: Calcium hydroxide is a gold standard for root canal sealing material. However, there are many defects in calcium hydroxide that have prompted many researchers to look for alternative materials, including natural origin ingredients. Red pine (Pinus densiflora) is often used in dentistry for its antioxidant and antibacterial properties. The mixture of calcium hydroxide and red pine has never been achieved before. One of the criteria for material sealing is that the channel should be fully sealed with high flow ability and low viscosity. Therefore, this research is performed to analyze the viscosity value of the mixture of calcium hydroxide and red pine (Pinus densiflora). Purpose: Knowing the difference in viscosity of the combination of calcium hydroxide and red pine with a ratio of 1:1, 1:1.5, and 1:2. Methods: This study used four treatment groups, with each group consisting of 8 replications. Group 1 was a combination of calcium hydroxide and red pine with a 1: 1, group 2 the ratio was 1: 1.5, group 3 had the ratio 1: 2, and the positive control group used calcium hydroxide and a sterile aquadest. Calcium hydroxide in powder form and an extract of red pine in liquid form are mixed according to the ratio. The viscosity value is measured using a viscosity tester, namely the Brookfield Viscometer. Results: The control group had a lower viscosity than group 3, group 3, and group 2 than group 1. Conclusion: Combination of calcium hydroxide and red pine with a ratio of 1:2, the lowest viscosity was obtained compared to the ratio of 1: 1 and 1: 1.5
Article
Full-text available
Cancer is an uncontrolled division of cells that has been attributed to various factors due to the advancement in cell biology and widespread genomic studies. Traditional chemotherapy in which a single cause is targeted is not a viable option for cancer treatment as multiple factors like various genes or epigenetic factors are found to be responsible for cancer phenotype. Formation of tumor is due to the somatic mutations in genome, same phenotypic expression of various cancer types. Many times, chemotherapy drugs given in two different patients of similar types of cancer do not produce the same effect in both the patients due to different somatic mutations or different genes involved in the same cancer phenotype. Thus, mono-target therapy usually encounters more failures than successes. In the past decades, research has begun to understand the various genetic mutations involved in a variety of cancers and this knowledge could be used to guide a new generation of medicinal chemists to design and discover novel drug moieties which would be able to influence multiple targets involved in tumor expression. Combinatorial formulations (synthetic as well as natural) could be beneficial in such a scenario. In this review, mechanisms of cancer, conventional methods to treat cancer and various plant secondary metabolites for effective treatment of cancer have been discussed.
Article
Full-text available
Background: Calcium hydroxide is a gold standard for root canal sealing material. However, there are many defects in calcium hydroxide that have prompted many researchers to look for alternative materials, including natural origin ingredients. Red pine (Pinus densiflora) is often used in dentistry for its antioxidant and antibacterial properties. The mixture of calcium hydroxide and red pine has never been achieved before. One of the criteria for material sealing is that the channel should be fully sealed with high flow ability and low viscosity. Therefore, this research is performed to analyze the viscosity value of the mixture of calcium hydroxide and red pine (Pinus densiflora). Purpose: Knowing the difference in viscosity of the combination of calcium hydroxide and red pine with a ratio of 1:1, 1:1.5, and 1:2. Methods: This study used four treatment groups, with each group consisting of 8 replications. Group 1 was a combination of calcium hydroxide and red pine with a 1: 1, group 2 the ratio was 1: 1.5, group 3 had the ratio 1: 2, and the positive control group used calcium hydroxide and a sterile aquadest. Calcium hydroxide in powder form and an extract of red pine in liquid form are mixed according to the ratio. The viscosity value is measured using a viscosity tester, namely the Brookfield Viscometer. Results: The control group had a lower viscosity than group 3, group 3, and group 2 than group1. Conclusion: Combination of calcium hydroxide and red pine with a ratio of 1:2, the lowest viscosity was obtained compared to the ratio of 1: 1 and 1: 1.5.
Article
Full-text available
Diabetic and anemia-associated diabetic wounds increase the considerable morbidity and mortality in people, as reported by clinical studies. However, no anemia-associated diabetic wound dressing materials have been developed until now. Hence, this study aimed to develop a nanocomposite scaffold composed of chitosan (CS), poly (vinyl alcohol) (PVA), and phytogenic iron oxide nanoparticles (FeO NPs), for accelerated anemia-associated diabetic wound healing. The aqueous leaves extract of Pinus densiflora (PD) was utilized for the synthesis of iron oxide nanoparticles (FeO NPs). TEM and elemental analysis confirmed smaller size PD-FeO NPs (<50 nm) synthesis with the combination of iron and oxide. In addition, in vitro biological studies displayed the moderate antioxidant, antidiabetic activities, and considerable antibacterial activity of PD-FeO NPs. Further, the different concentrations of PD-FeO NPs (0.01, 0.03, and 0.05%) incorporated CS/PVA nanocomposites sponges were developed by the freeze-drying method. The porous structured morphology and the presence of PD-FeO NPs were observed under FE-SEM. Among nanocomposite sponges, PD-FeO NPs (0.01%) incorporated CS/PVA sponges were further chosen for the in vitro wound-healing assay, based on the porous and water sorption nature. Furthermore, the in vitro wound-healing assay revealed that PD-FeO NPs (0.01%) incorporated CS/PVA has significantly increased the cell proliferation in HEK293 cells. In conclusion, the CS/PVA-PD-FeO NPs (0.01%) sponge would be recommended for diabetic wound dressing after a detailed in vivo evaluation.
Article
Full-text available
The antioxidant activity of the essential oils, as well as of the organic and hydroethanolic extracts, of the fresh needles of 54 pine taxa was evaluated using the peroxy-oxalate and luminol chemiluminescence assays. Among all evaluated essential oils, P. canariensis and P. attenuata displayed the highest levels of activity. P. contorta var. murrayana, followed by P. nigra var. caramanica, exhibited the highest antioxidant capacity among the organic extracts, while the most active hydroethanolic extract was that of P. nigra subsp. nigra. Based on the overall levels of activity, the latter taxon was selected for phytochemical analysis targeting the isolation of the bioactive constituents. As such, the organic extract of P. nigra subsp. nigra was subjected to chromatographic separations to yield 11 secondary metabolites (1−11) that were evaluated for their antioxidant activity. Nonetheless, the isolated compounds were found to be less active than the crude extract, thus suggesting the potential role of synergism.
Chapter
Problems in transporting drug molecules to tumor sites in required dose or constitution lead to low efficacy and significant side effects. Shielding the drug molecules in micelles, liposomes, or nanoparticles is a major line of investigation to improve chemotherapeutic treatment. Though compatibility for proper envelopment of the drug and timely release at the tumor site are required of such a carrier, protecting its own physicochemical and morphological integrity during transport is another precondition.
Article
Pinus densiflora sieb. et zucc.(pine needle) is a traditional medicine used in several East Asian countries. However, the efficacy of pine needle has rarely been reported. In this study showed that the anti-proliferative effects and the mechanisms of hexane layer of pine needle MeOH extract (PNH) on gastric cancer cells. At first, PNH inhibited the proliferation of gastric cancer cells in a dose-dependent manner. Moreover, PNH treatment induced G1 phase cell cycle arrest through the increased p27KIP1 expression and decreased cyclin dependent kinase (CDKs) activity. Furthermore, PNH treatment induced premature senescence without oncogenic stress, through the expression of p27KIP1 and Skp2. Taken together, these results showed that PNH inhibited gastric cancer cell proliferation through the induction of G1-cell cycle arrest and premature senescence via induced p27KIP1 expression, as controlled by Skp2 reduction. Also, PNH could be a candidate for anti-gastric cancer treatment and may be useful in the development of anti-gastric cancer drugs.
Article
The significance of wood odors beyond flavoring effects for barbecues and wine aged in barrels has recently been under discussion. Wood has an immense influence on our physical and mental condition through our palate, such as positive health and mood-stimulating effects. As a result of increased public interest, the key odorants from various natural wood species were recently decoded. To gain profound insights into the contribution of single odorants to the overall scent of distinct wood types, this study compares several softwood species (Scots pine, incense cedar, European larch, Norway spruce, and white fir) by odor profile analysis. Nine odor-active constituents, which were previously detected in those woods, were quantified using stable isotope dilution analysis (SIDA). Odor activity values (OAVs) calculated on the basis of odor thresholds (OTs) determined in cellulose revealed hexanal, octanal, (E)-non-2-enal, p-cresol, vanillin, and thymoquinone as dominant odorants for wood odor. Recombination experiments by mixing the odorants in their naturally occurring concentrations in a cellulose matrix confirmed the successful characterization of the key odorants for Scots pine and incense cedar wood.
Article
Full-text available
Pinus densiflora needle extract (PDNE)is widely reported to have many pharmacological activities including antioxidant potential. However, the solvent system used for extraction greatly affects its antioxidant quality. Hence, in the present study, we investigated the effect of a different ratio (vol/vol)of ethanol to water (0–100%)in the extraction of PDNE with potent antioxidant capacity. The chemical assays, 2,2-diphenyl-1 picrylhydrazyl (DPPH)and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS), were conducted to assess the antioxidant potential of PDNE. Subsequently, the cytoprotective effect of PDNE was determined using tert-butyl hydroperoxide (TBHP)-challenged HepG2 cellular model. The needle extracts from 40% ethanol (PDNE-40)showed greater radical scavenging activity followed by 60%, 20%, 80%, 0% and 100% ethanol extracts. EC 50 value of the most active extract, PDNE-40, was 8.56 ± 0.51 μg/mL, relative to 1.34 ± 0.28 μg/mL of the standard trolox (for ABTS radical), and 75.96 ± 11.60 μg/mL, relative to 4.83 ± 0.26 μg/mL of the standard trolox (for DPPH radical). Either PDNE-20 or PDNE-40 pretreatment remarkably decreased the levels of reactive oxygen species (ROS), lipid peroxides and protein carbonyls in TBHP-challenged HepG2 cells. In addition, both PDNE-20 and PDNE-40 significantly reversed the decreased ratio of reduced (GSH)to oxidized (GSSG)glutathione. Moreover, these two extracts showed a significant inhibitory effect on TBHP-induced nuclear damage and loss of cell viability. In summary, the inclusion of 40% ethanol in water for extraction of Pinus densiflora needle greatly increases the antioxidant quality of the extract.
Article
We used GC and GC-MS systems to analyze the chemical composition of essential oils extracted from twigs of Pinus nigra arn. subsp. pallasiana (Lamb.) Holmboe (Black pine) naturally grown in Turkey. Twig samples were collected from four regions of Turkey (Adana, Adiyaman, Antalya and Elazig) and the twigs were hydrodistilled by clevenger apparatus. Forty three (Adana), 40 (Adiyaman), 29 (Antalya) and 44 (Elazig) components were identified representing 90.7%, 97.8%, 98.4% and 87.7%, respectively of all extracted oils. The main compounds of oil samples were germacrene D, α-pinene, δ-3-carene, β-pinene, limonene and β-caryophyllene. Here we discuss chemical differences with reference to forestry products and renewable resources, and we also assess intrapopulational means.
Article
Full-text available
Drug sensitivity assays were performed using a variation of a colorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] assay on V79, CHO-AuxB1, CHRC5, NCI-H460, and NCI-H249 cell lines following optimization of experimental conditions for each cell line. Results from this assay were compared with data assimilated simultaneously by clonogenic assay and by dye exclusion assay. Good correlation was observed using the CHO-AuxB1 cell line and the pleiotropic drug-resistant mutant CHRC5, with similar degrees of relative resistance observed with both the MTT and clonogenic assays. Good correlation was observed between the clonogenic and MTT assays for 1-h drug exposures, although the MTT assay was more sensitive to vinblastine. In general, the clonogenic assay was more sensitive when continuous drug exposures were utilized, although this was primarily related to the increased drug exposure time. While the use of the MTT assay in drug sensitivity testing of primary tumor samples is limited, since contaminating normal cells may also reduce the tetrazolium, the MTT assay can be semiautomated, and therefore it offers a valid, simple method of assessing chemosensitivity in established cell lines.
Article
Full-text available
The UV absorption method and the thiobarbituric acid (TBA) test for oxidation of an aqueous suspension of linoleate were compared. The absorption method depends on the formation of hydroperoxides having conjugated double bonds that absorb strongly at 233 nm. The absorption at 233 nm increased markedly during oxidation of linoleate catalyzed by either ascorbic acid or cupric ions. The concentration of ascorbic acid in the reaction mixture was also measured by UV absorption at 265 nm and pH 7.0. Color development in the TBA test also increased markedly with the extent of oxidation of linoleate. When ascorbic acid was the catalyst, UV absorption detected early stages of oxidation with greater sensitivity than the TBA test. The reverse was true when Cu⁺⁺ was the catalyst. In general, the relation between the two procedures will depend on whether copper is present when the TBA test is made.
Article
Full-text available
Increases in the intracellular levels of reactive oxygen species (ROS), frequently referred to as oxidative stress, represents a potentially toxic insult which if not counteracted will lead to membrane dysfunction, DNA damage and inactivation of proteins. Chronic oxidative stress has numerous pathological consequences including cancer, arthritis and neurodegenerative disease. Glutathione-associated metabolism is a major mechanism for cellular protection against agents which generate oxidative stress. It is becoming increasingly apparent that the glutathione tripeptide is central to a complex multifaceted detoxification system, where there is substantial inter-dependence between separate component members. Glutathione participates in detoxification at several different levels, and may scavenge free radicals, reduce peroxides or be conjugated with electrophilic compounds. Thus, glutathione provides the cell with multiple defences not only against ROS but also against their toxic products. This article discusses how glutathione biosynthesis, glutathione peroxidases, glutathione S-transferases and glutathione S-conjugate efflux pumps function in an integrated fashion to allow cellular adaption to oxidative stress. Co-ordination of this response is achieved, at least in part, through the antioxidant responsive element (ARE) which is found in the promoters of many of the genes that are inducible by oxidative and chemical stress. Transcriptional activation through this enhancer appears to be mediated by basic leucine zipper transcription factors such as Nrf and small Maf proteins. The nature of the intracellular sensor(s) for ROS and thiol-active chemicals which induce genes through the ARE is described. Gene activation through the ARE appears to account for the enhanced antioxidant and detoxification capacity of normal cells effected by many cancer chemopreventive agents. In certain instances it may also account for acquired resistance of tumours to cancer chemotherapeutic drugs. It is therefore clear that determining the mechanisms involved in regulation of ARE-driven gene expression has enormous medical implications.
Article
Full-text available
Human exposure to polycyclic aromatic hydrocarbons (PAH) occurs through complex mixtures such as coal tar. The effect of complex PAH mixtures on the activation of carcinogenic PAH to DNA-binding derivatives and carcinogenesis were investigated in mice treated topically with NIST (National Institute of Standards and Technology) Standard Reference Material 1597 (SRM), a complex mixture of PAH extracted from coal tar, and either additional benzo[a]pyrene (B[a]P) or dibenzo[a,l]pyrene (DB[a,l]P). In an initiation-promotion study using 12-O-tetradecanoylphorbol-13-acetate as the promoter for 25 weeks, the SRM and B[a]P co-treated mice had a similar incidence of papillomas per mouse compared with the group exposed to B[a]P alone as the initiator. PAH-DNA adduct analysis of epidermal DNA by 33P-post-labeling and reversed-phase high-performance liquid chromatography found the SRM co-treatment led to a significant decrease in the total level of DNA adducts and B[a]P-DNA adducts to less than that observed in mice treated with B[a]P alone at 6, 12 and 72 h exposure. After 24 and 48 h exposure, there was no significant difference in the levels of adducts between these groups. In the DB[a,l]P initiation-promotion study, the co-treated group had significantly fewer papillomas per mouse than mice treated with DB[a,l]P alone as initiator. Averaging over the times of exposure gave strong evidence that mice co-treated with SRM and DB[a,l]P had a significantly lower level of PAH-DNA adducts than mice treated with DB[a,l]P alone. Western immunoblots showed that both cytochrome P450 (CYP) 1A1 and 1B1 were induced by the SRM. These results are consistent with the hypothesis that two major factors determining the carcinogenic activity of PAH within a complex mixture are (i) the persistence of certain PAH-DNA adducts as well as total adduct levels, and (ii) the ability of the components present in the mixture to inhibit the activation of carcinogenic PAH by the induced CYP enzymes.
Article
This study was performed to examine the cytotoxic effects of the distilled pine-needle extracts against several cancer cell lines. First, cell lines including mice leukemic cancer cell line (L1210), sarcoma 180 and human monocyte-like cancer cells (U937) were tested using XTT methods in uitro. Pine-needle extracts were prepared by pressing the pine needles and distilling it at below 98^{\circ}C and then added to the growth medium in a final dilution of 10, 20, and 40 times. Growth of three kinds of cancer cells was significantly inhibited by more than 50% with the addition of the extracts. Fifty six to seventy six % of inhibition was shown with the 40 times dilution of the extracts. Greater inhibition was achieved with the 20 times dilution (81~90%) and the 10 times dilution (77~89%) of the extracts. Next, other human cancer cell lines including 3 kinds of breast cancer cell lines (T47D, MDA-MB-231 and MW7A) and one hepatoma cell line (SNU-354) were tested with the 20 times dilution of the extract. T47D and MDA-MB-231 cell lines showed lower inhibition (12%) with the addition of the extract. However, MH7A and SNU-354 cell lines showed 64% and 72% inhibition with the extract, respectively. These results suggest that the distilled pine-needle extracts have strong cytotoxic effect on certain cancer cell lines and the intensity of the effect may vary depending on the process of the pine needle.
Article
To develope natural food preservatives of pine needle (Pinus densiflora Seib et Zucc.) extract, pine needle sap, ethanol and ether extracts were prepared for investigation of antimicrobial activities against food-related bacteria and yeasts. All extracts exhibited growth inhibiting activities for most of microorganisms tested. However, in general, growth inhibiting activities were higher in ethanol extract than in sap or ether extract. Minimum inhibitory concentrations (MIC) of ethanol extract for Lactobacillus casei, Pseudomonas aeruginosa or Escherchia coli were as low as 0.1 mg/ml, whereas MIC of sap or ether extract for most bacteria and yeasts were 0.25-0.8 mg/ml, indicating that the ethanol extract showed the antimicrobial activity by 2.5 -8 times higher than the sap and ether extract. The antimicrobial activity of the ethanol extract was reduced by heating or alkali treatment. Moreover, growth of Pseudomonas aeruginosa was completely inhibited within 24 hours by the addition of at least 50 ppm of ethanol extract. These findings suggest that pine needle, specially the ethanol extrat may play a role for natural food preservatives.
Article
The preincubation of test compound, bacterial tester strain, and S-9 mix or buffer before pouring a minimal-glucose agar plate enhanced the sensitivity of mutation test and increased the spectrum of mutagens detected. Addition of NADH and ATP in S-9 mix enhanced the mutagenicity of some compounds. Addition of norharman in the preincubation mixture made it possible to detect a marginal or weak mutagenicity of certain types of mutagens. Addition of riboflavin revealed the mutagenicity of azo compounds. Glycosidase was required to detect the mutagenicity of glycosides or natural products.
Article
Antioxidative activities of 5 common edible seaweeds in Korea, three brown algae (seaweed fusiforme, sea mustard, sea tangle), one green algae (sea lettuce) and one red algae (laver), were examined. The antioxidative activities of ethanol extracts from these seaweeds were examined by measuring of inhibition rates against iron-induced linoleate peroxidation, DPPH (1,1-diphenyl -2-picrylhydrazyl) radical generation and MDA-BSA (malondialdehyde-bovine serum albumin) conjugation. Sea lettuce ethanol extract showed the strongest anti-oxidative activity among them, especially in inhibition against conjugation of lipid peroxide and protein. Second to sea lettuce, laver and sea tangle ethanol extracts showed high DPPH radical scavenging activity and inhibition against MDA-BSA conjugation. However, seaweed fusiforme and sea mustard ethanol extracts did not show antioxidative activities. Sea mustard contained the highest total flavonoids (11.33 mg/g dry wt) and sea lettuce contained the highest total polyphenol (8.97 mg/g dry wt) among these seaweeds. In addition, there was strong positive correlation between the antioxidative activity and total polyphenol content in these seaweeds, suggesting polyphenol compounds may contribute to antioxidative effect of seaweeds. From these data, it is suggested to consume much of seaweeds such as sea lettuce, laver and sea tangle to prevent age-related chronic diseases, and also develope neutraceutical products using polyphenol rich fraction from sea lettuce.
Article
This study was performed to observe cytotoxic effect of the pine needle extracts against cancer cell lines including human gastric carcinoma (KATOIII), human lung carcinoma (A549), human hepatocellular carcinoma (Hep3B) and human breast adenocarcinoma (MCF-7) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) and SRB (sulforhodamine B) method. The extracts were prepared by step-wise fractionation of ethanol extract of pine needles using diethylether, chloroform, ethylacetate, butanol and water. The growth of the cancer cells in medium containing pine needle extracts were significantly inhibited degree in proportion to the increase of the extract concentration. A significant shrinkage of Hep3B cells was observed when the cells were exposed into 0.5, 1 mg/mL of pinus rigida extract.
Article
METHODS for measuring antioxidants and appraising antioxidant activity appear to be of two general types. If the chemical nature of the antioxidant is known, one may strive for a test specific for the compound or group of interest; for example, the nitroprusside test for sulphydryl groups. Alternatively one may observe the inhibition of some natural oxidative process such as the β-oxidation of fats, as a function of the added antioxidant.
Article
Publisher Summary This chapter discusses the analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Analyses of the Folin-Ciocalteu (FC) type are convenient, simple, and require only common equipment and have produced a large body of comparable data. Under proper conditions, the assay is inclusive of monophenols and gives predictable reactions with the types of phenols found in nature. Because different phenols react to different degrees, expression of the results as a single number—such as milligrams per liter gallic acid equivalence—is necessarily arbitrary. Because the reaction is independent, quantitative, and predictable, analysis of a mixture of phenols can be recalculated in terms of any other standard. The assay measures all compounds readily oxidizable under the reaction conditions and its very inclusiveness allows certain substances to also react that are either not phenols or seldom thought of as phenols (e.g., proteins). Judicious use of the assay—with consideration of potential interferences in particular samples and prior study if necessary—can lead to very informative results. Aggregate analysis of this type is an important supplement to and often more informative than reems of data difficult to summarize from various techniques, such as high-performance liquid chromatography (HPLC) that separate a large number of individual compounds .The predictable reaction of components in a mixture makes it possible to determine a single reactant by other means and to calculate its contribution to the total FC phenol content. Relative insensitivity of the FC analysis to many adsorbents and precipitants makes differential assay—before and after several different treatments—informative.
Article
Antioxidative activity and total phenolic compounds of root, leaf and petiole of four accessions of Centella asiatica (L.) Urban, namely CA 01, CA 05, CA 08 and CA 11, were evaluated. Antioxidative activity of the extracts was measured using the ferric thiocyanate (FTC) method and thiobarbituric acid (TBA) test. The antioxidative activities were then compared with that of α-tocopherol (natural antioxidant) and butylated hydroxytoulene or BHT (synthetic antioxidant). The results showed that CA 01 and CA 05 had the highest antioxidative activities among the accessions tested. Results also showed that both leaf and root of C. asiatica had high antioxidative activity, which was as good as that of α-tocopherol. The total phenolic content, determined according to the Folin–Ciocalteu method, varied from 3.23 to 11.7 g/100 g dry sample, and showed strong association (r2=0.90) with antioxidative activity. The results suggest that phenolic compounds are the major contributors to the antioxidative activities of C. asiatica.
Article
Flavonoids, a group of phenolic compounds widely occurring in the plant kingdom, have been reported to possess strong antioxidant activity. In the present study, four flavonoids (quercetin, hesperetin, naringenin, rutin), chosen according to their structural characteristics, were tested in two different in vitro experimental models: (1) Fe2+-induced linoleate peroxidation (Fe2+-ILP), by detection of conjugated dienes; and (2) autooxidation of rat cerebral membranes (ARCM), by using thiobarbituric acid for assay of free malondialdehyde production. The results obtained were also interpreted in the light of flavonoid interactions, studied by differential scanning calorimetry, with dipalmitoylphosphatidylcholine (DPPC) vesicles as a biological membrane model. The antilipoperoxidant activity of the flavonoids tested can be classified as follows: rutin > hesperetin > quercetin ⪢ naringenin in the Fe2+-ILP test; quercetin > rutin ⪢ hesperetin > naringenin in the ARCM test. Quercetin, hesperetin, and naringenin interacted with DPPC liposomes causing different shifts, toward lower values, of the main transition peak temperature (Tm) typical for DPPC liposomes; however, no change in Tm of DPPC dispersion was observed in the presence of rutin. The hypothesis will be discussed that flavonoid capacity to modify membrane-dependent processes, such as free-radical-induced membrane lipoperoxidation, is related not only to their structural characteristics but also to their ability to interact with and penetrate the lipid bilayers.
Article
The DNA adducts of 7,12-dimethylbenz[a]anthracene (DMBA) previously identified in vitro and in vivo are stable adducts formed by reaction of the bay-region diol epoxides of DMBA with dG and dA. In this paper we report identification of several new DMBA-DNA adducts formed by one-electron oxidation, including two adducts lost from DNA by depurination, DMBA bound at the 12-methyl to the N-7 of adenine (Ade) or guanine (Gua) [7-methylbenz[a]anthracene (MBA-12-CH2-N7Ade or 7-MBA-12-CH2-N7Gua, respectively]. The in vitro systems used to study DNA adduct formation were DMBA activated by horseradish peroxidase or 3-methyl-cholanthrene-induced rat liver microsomes. The biologically-formed depurination adducts were identified by high-pressure liquid chromatography and by fluorescence line narrowing spectroscopy. Stable DMBA-DNA adducts were analyzed by the 32P-postlabeling method. Quantitation of DMBA-DNA adducts formed by microsomes showed about 99% as depurination adducts: 7-MBA-12-CH2-N7Ade (82%) and 7-MBA-12-CH2-N7Gua (17%). Stable adducts (1.4% of total) included one adduct spot that may contain adduct(s) formed from the diol epoxide (0.2%) and unidentified adducts (1.2%). Activation of DMBA by horseradish peroxidase afforded 56% of stable unidentified adducts and 44% of depurination adducts, with 36% of 7-MBA-12-CH2-N7Ade and 8% of 7-MBA-12-CH2-N7Gua. Adducts containing the bond to the DNA base at the 7-CH3 group of DMBA were not detected.(ABSTRACT TRUNCATED AT 250 WORDS)
Article
The term 'antimutagen' was originally used to describe an agent that reduces the apparent yield of spontaneous and/or induced mutations, regardless of the mechanisms involved. The 'antimutagens' include 'desmutagens' and 'bio-antimutagens'. In this article, our attention was focused on the bio-antimutagens affecting DNA repair in bacteria. Cobaltous chloride reduced the frequency of mutations in Escherichia coli induced by MNNG. The possibility that metal compound inhibits the growth of mutagen-treated cells was examined. The results clearly showed that the antimutagen surely reduces the mutation rate. The target of cobaltous chloride was found to be cellular factors including Rec A. Vanillin and cinnamaldehyde had strong antimutagenic activities against UV, 4NQO and AF-2. They stimulated Rec A-dependent recombination repair functions in the mutagen-treated cells. Among plant materials, tannins possess antimutagenic activity against UV-induced mutations in E. coli. It has been found that tannic acid stimulates the excision repair encoded by the uvrA gene thereby reducing the yield of mutants. Substances which are antimutagenic in bacterial systems also had antimutagenic activity in cultured mammalian cell systems. Vanillin reduced the frequency of mutagen-induced chromosomal aberrations.
Article
The methods for detecting carcinogens and mutagens with the Salmonella mutagenicity test were described previously (Ames et al., 1975b). The present paper is a revision of the methods. Two new tester strains, a frameshift strain (TA97) and a strain carrying an ochre mutation on a multicopy plasmid (TA102), are added to the standard tester set. TA97 replaces TA1537. TA1535 and TA1538 are removed from the recommended set but can be retained at the option of the investigator. TA98 and TA100 are retained. We discuss other special purpose strains and present some minor changes in procedure, principally in the growth, storage, and preservation of the tester strains. Two substitutions are made in diagnostic mutagens to eliminate MNNG and 9-aminoacridine. Some test modifications are discussed.
Article
Current guidelines and recommendations for genotoxicity testing of pharmaceuticals are disparate, both in terms of the most appropriate tests to use and the protocols to follow. Recent attempts have been made to standardise genotoxicity testing procedures, coinciding with the current review of the OECD guidelines and the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). However, as with other aspects of non-clinical safety assessment of pharmaceuticals, guidelines have been prepared by evaluation of general chemical data due to the lack of specific information on pharmaceuticals. To address this, a project was undertaken to collect and collate information specifically pertaining to the genotoxicity testing of pharmaceuticals in order to obtain a clear understanding of international strategy and procedures in the pharmaceutical industry. It is clear that the practices and regional variations are strongly influenced by national guidelines and do not necessarily follow companies' preferences. However, there is a surprising amount of variation in approach between companies on some issues. This is evident in how companies define a genotoxin. This ranges from a positive result in an in vivo assay as indicative of a genotoxin (43%) to any positive result in vitro or in vivo (30%). Indeed many companies (particularly in Japan) will terminate development on the strength of a clear positive result in an Ames test. There is much debate within the ICH process concerning tests to detect gene mutations in mammalian cells as part of a primary test battery. This survey shows that in general, the pharmaceuticals industry has severe doubts about these assays. Thirty-seven (78%) of the 47 participating pharmaceutical companies include an in vitro test to detect gene mutation in mammalian cells as part of their routine test battery. The HPRT test using Chinese hamster cells has the most widespread use, although there is only limited use of such tests in Japan. Compound development has been affected by the results of such tests, but usually only in terms of clarification of equivocal results in other genotoxicity tests in the test battery. The majority (63%) of companies do not support its use as a primary regulatory requirement, and 83% do not consider the mouse lymphoma assay (L5178Y) an acceptable replacement for in vitro mammalian cytogenetics. In conclusion, this survey has provided valuable information on the current modus operandi of the international pharmaceutical industry for consideration in current harmonisation initiatives.
Article
At the International Workshop on the Standardisation of Genotoxicity Test Procedures, in Melbourne (27-28 February 1993), the current international guidelines for the correct conduct of bacterial mutation assays were considered, and the major differences between them were examined. An attempt was made to construct a scientifically based, internationally harmonized protocol. The main points of agreement were as follows. The consensus opinion was that there are currently insufficient data to justify a preference for either the preincubation or plate-incorporation methodologies as the initial test. Whichever method is used there was consensus agreement that the bacterial test battery should consist of S. typhimurium TA1537, TA1535, TA98 and TA100. There was also consensus that the 3 strains TA97a, TA97 and TA1537 could be used interchangeably. Although it was not possible to achieve a consensus, the majority of the working group members agreed that strains for the detection of mutagens acting specifically on AT base pairs should be routinely included within the test battery. These strains may be S. typhimurium TA102 or E. coli WP2 strains (WP2 pKM101 and WP2 uvrA or WP2 uvrA pkM101). With regard to study design it was universally agreed that 5 doses of test compound should be used in each experiment, and a majority agreement was obtained for 3 plates per dose. The use of 2 plates per dose is acceptable ONLY if the experiment is repeated. It is recommended that the negative controls may consist of solvent control alone provided that historical data are available to demonstrate lack of effect of the solvent in question. Positive control compounds should be included in all experiments, although the nature of these control compounds need not be specified in the guidelines. There was consensus agreement that for non-toxic freely soluble test agents, an upper limit of 5 mg/plate should be tested (5 microliters per plate for liquids). For insoluble or toxic compounds, the recommendations were the same as those for other in vitro tests (see appropriate paper). A consensus agreement was reached on the need to carry out further tests if equivocal results are obtained in the initial test, although it was generally agreed that the design of the repeat study should be left flexible. As there are little or no data to support the use of an exact repeat assay, a majority of the group recommended that negative results in the first test should be further investigated by either conducting a modified repeat (e.g. S9 titration) or by conducting the alternative methodology.(ABSTRACT TRUNCATED AT 400 WORDS)
Article
The micronucleus test and sister-chromatic exchange (SCE) test were used to research the antimutagenic effect of pine needle extract. The results showed that the mutagenic effect of cyclophosphamide (CP) was inhibited by the pine needle extract. The micronucleus frequencies (MNF) of mouse bone marrow and human lymphocytes from peripheral blood were decreased with the effect of the extract (the dose was 2000 mg/kg or 5 mg/ml); the frequency of SCE in human lymphocytes was also reduced significantly, which indicated that the MNF and the SCE frequencies were negatively correlated with the dose of pine needle extract (r = -0.9782, -0.9587, -0.9765, respectively). This suggested that the pine needle extract was an effective antimutagen and it is important to choose the proper doses of pine needle extract for antitumor effect.
Article
The antitumor activity of the i.p. or p.o. administration of polysaccharide-protein complex, ATOM (antitumor organic substance Mie) prepared from cultured mycelia of Agaricus blazei (Iwade strain 101) "Himematsutake" examined against four kinds of established mouse tumors. ATOM was highly effective at the doses of 10 and 20 mg/kg/day x 10 on subcutaneously implanted Sarcoma 180 in mice, and was also active against Ehrlich ascites carcinoma, Shionogi carcinoma 42 and Meth A fibrosarcoma at doses of 50 and 100 mg/kg/day x 10. ATOM has no direct cytotoxic action on tumor cells in vitro. Thus the tumor growth-inhibitory effect of ATOM is apparently due to immunological host-mediated mechanisms. The number of peritoneal macrophages, the phagocytosis of polystyrene latex beads and the proportion of the third component of complement (C3)-positive fluorescent cells were increased in the mice treated with ATOM. These results suggest that the macrophage activa-tion and alterations of the C3 are necessary for the induction of an antitumor effect of ATOM.
Article
The radical modulation activity of lignins prepared from the cone of Pinus elliottii var. Elliottii was investigated, using ESR spectroscopy. These lignins produced radical(s) under alkaline conditions, and the radical intensity was increased with increasing pH. Lower concentrations of lignins slightly reduced the radical intensity of sodium ascorbate, whereas higher concentrations of lignins enhanced both the radical intensity and cytotoxic activity of sodium ascorbate. Lignins effectively scavenged superoxide anion, produced by hypoxanthine-xanthine oxidase reaction. Elliottii lignins significantly inhibited the human immunodeficiency virus (HIV)-induced cytopathic effect, in similar fashions to other natural, commercial and synthetic lignins. Pretreatment of mice with lignins significantly protected them from the lethal infection with E. coli. Crude alkaline extracts of Elliottii pine cone displayed similar magnitude of activity with lignins. These data further supports the medicinal efficacy of plant extracts.
Article
The antioxidative activity of a total of 92 phenolic extracts from edible and nonedible plant materials (berries, fruits, vegetables, herbs, cereals, tree materials, plant sprouts, and seeds) was examined by autoxidation of methyl linoleate. The content of total phenolics in the extracts was determined spectrometrically according to the Folin-Ciocalteu procedure and calculated as gallic acid equivalents (GAE). Among edible plant materials, remarkable high antioxidant activity and high total phenolic content (GAE > 20 mg/g) were found in berries, especially aronia and crowberry. Apple extracts (two varieties) showed also strong antioxidant activity even though the total phenolic contents were low (GAE < 12.1 mg/g). Among nonedible plant materials, high activities were found in tree materials, especially in willow bark, spruce needles, pine bark and cork, and birch phloem, and in some medicinal plants including heather, bog-rosemary, willow herb, and meadowsweet. In addition, potato peel and beetroot peel extracts showed strong antioxidant effects. To utilize these significant sources of natural antioxidants, further characterization of the phenolic composition is needed.
Article
Cancer chemoprevention involves intervention in the carcinogenic process by a natural or synthetic chemical that either blocks neoplasia development or arrests malignant phenotype progression. The chemopreventive test agent must experimentally be established as safe before a clinical trial. In our laboratory, inhibition of carcinogen-induced development of precancerous lesions in the mouse mammary gland organ culture model is used as a primary screen to select chemopreventive agents for in vivo efficacy evaluation. A nearly 75% correlation apparently exists between the efficacy observed in vitro and in vivo carcinogenesis. For in vivo experiments, MNU- and DMBA-induced mammary tumours in rats are the models of choice. Numerous agents have been identified and progressed to preclinical toxicity and clinical trials. More recently, combination chemoprevention has received considerable attention, since no known chemopreventive agent sufficiently reduces tumour incidence in rats. The sequence of events for establishing the experimental basis for chemoprevention of breast cancer is described.
Article
Radiation hazards in outer space present an enormous challenge for the biological safety of astronauts. A deleterious effect of radiation is the production of reactive oxygen species, which result in damage to biomolecules (e.g., lipid, protein, amino acids, and DNA). Understanding free radical biology is necessary for designing an optimal nutritional countermeasure against space radiation-induced cytotoxicity. Free radicals (e.g., superoxide, nitric oxide, and hydroxyl radicals) and other reactive species (e.g., hydrogen peroxide, peroxynitrite, and hypochlorous acid) are produced in the body, primarily as a result of aerobic metabolism. Antioxidants (e.g., glutathione, arginine, citrulline, taurine, creatine, selenium, zinc, vitamin E, vitamin C, vitamin A, and tea polyphenols) and antioxidant enzymes (e.g., superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidases) exert synergistic actions in scavenging free radicals. There has been growing evidence over the past three decades showing that malnutrition (e.g., dietary deficiencies of protein, selenium, and zinc) or excess of certain nutrients (e.g., iron and vitamin C) gives rise to the oxidation of biomolecules and cell injury. A large body of the literature supports the notion that dietary antioxidants are useful radioprotectors and play an important role in preventing many human diseases (e.g., cancer, atherosclerosis, stroke, rheumatoid arthritis, neurodegeneration, and diabetes). The knowledge of enzymatic and non-enzymatic oxidative defense mechanisms will serve as a guiding principle for establishing the most effective nutrition support to ensure the biological safety of manned space missions.
Article
Antioxidant activity of Pinus densiflora Sieb. et Zucc. (Pinaceae) was evaluated for potential to inhibit hydroxyl radicals, inhibit total reactive oxygen species generation in kidney homogenates using 2',7'-dichlorodihydro fluorescein diacetate (DCHF-DA) and scavenge authentic peroxynitrites. The methanolic extract of P. densiflora showed strong antioxidant activity in the tested model systems and thus fractionated with several solvents. The antioxidant activity potential of the individual fraction was in the order of ethyl acetate > n-butanol > water > dichloromethane fraction. The ethyl acetate soluble fraction exhibiting strong antioxidant activity was further puri fi ed by repeated silica gel and Sephadex LH-20 column chromatographies. An active lignan (+)-isolarisiresinol xylopyranoside, as well as two active flavonoids [kaempferol 3-O-beta-galactopyranoside and its 6"-acetyl derivative], were isolated.
Article
While a number of agents have been shown to induce mammary carcinogenesis in the rat, premalignant stages of the disease have been best characterized in chemically-induced models, specifically those initiated by either 7,12 dimethylbenz[alpha]anthracene (DMBA) or 1-methyl-1-nitrosourea (MNU). In general, it appears that epithelial cells in mammary terminal end buds or terminal ductules are the targets of carcinogenic initiation, and that a series of morphologically identifiable steps are involved in the development of mammary carcinoma. The premalignant steps include ductal hyperplasia of the usual type and carcinoma in situ of the cribriform or comedo type; atypical ductal hyperplasia has not been reported. Thus the histogenesis of lesions occurring in chemically induced mammary carcinogenesis in the rat is similar to that observed in the human; although, the spectrum of lesions observed in the rat is limited. Opportunities to investigate the biological and molecular characteristics of premalignant breast disease in the rat are presented.
Article
Thirty-six different extracts of six herbs and aromatic plants (fennel, common melilot, milfoil, lavandin cv. Super, spike lavender, and tarragon) were evaluated for their radical scavenging activity by the DPPH*, NBT/hypoxanthine superoxide, and *OH/luminol chemiluminescence methods, and for their antioxidant activity by the beta-carotene blenching test. The total phenolic content was also determined by the Folin-Ciocalteu method. The plant material included cultivated plants and their wastes after being distilled for essential oils. Both remarkably high phenolic content and radical scavenging activities were found for the ethyl acetate and dichloromethane fractions among the different plant extracts. In general, the distilled plant material was found to exhibit a higher phenolic content as well as antioxidant and radical scavenging activities than the nondistilled material. Ethyl acetate and dichloromethane extracts, and even some crude extract, of both distilled and nondistilled plants exhibited activities comparable to those of commercial extracts/compounds, thus making it possible to consider some of them as a potential source of antioxidants of natural origin.
Article
Oxidation of low-density lipoprotein (LDL) has been implicated in atherogenesis. Antioxidants that prevent LDL from oxidizing may reduce atherosclerosis. This study investigated LDL antioxidant activity in edible plant products for development of dietary supplementation to prevent atherosclerosis. Fifty-two kinds of edible plants were extracted using 70% aqueous ethanol solution, and the antioxidant activity of the extracts, which inhibit human LDL oxidation induced by copper ion, was determined on the basis of the oxidation lag time and represented as epigallocatechin 3-gallate equivalent. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenolic content were also measured for comparisons with antioxidant activity in LDL. Plant products showing the greatest activity in LDL oxidation assay were akamegashiwa (Mallotus japonicus) leaf, Japanese privet (Ligustrum japonicum) leaf, green tea [Camellia sinensis (L.) O. Kuntze], and astringent persimmon (Diospyros kaki). The present study revealed high levels of LDL antioxidant activity in plant products for which such activity levels are underestimated in the DPPH radical scavenging assay and Folin-Ciocalteu assay.
Development of the health foods containing extracts from Pinus strobus leaves
  • Lee
  • Yh
  • Ym Shin
  • Cha
  • Sh
  • Ys
  • Lee
  • Sy
Lee YH, Shin YM, Cha SH, Chol YS, and Lee SY: Development of the health foods containing extracts from Pinus strobus leaves. J Korean Soc Food Nutr 25, 379–383, 1996.
Standard Food Analysis
  • Chae Sk
  • Kang
  • Gs
  • Ma
  • Sj
  • Bang Kw
  • Oh
  • Mw
Chae SK, Kang GS, Ma SJ, Bang KW, Oh MW, et al.: Standard Food Analysis. Seoul, Korea: Jigu-moonwha Sa, 2003, pp 581–582.
Investigation of anti-aging effect and determination of chemical structure of pine nee-dle (Pinus densiflora) through animal experiment. Effects of ethyl ace-tate fraction on oxygen radicals and their scavenger enzymes in liver of SD rats
  • Choi Jh Di Lim
  • Park
  • Sh
  • Kim
  • Dw
  • Lee
  • Jh
Choi JH, Lim DI, Park SH, Kim DW, Lee JH, et al.: Investigation of anti-aging effect and determination of chemical structure of pine nee-dle (Pinus densiflora) through animal experiment. Effects of ethyl ace-tate fraction on oxygen radicals and their scavenger enzymes in liver of SD rats. Korean J Gerontol 11, 14–20, 2001.
Antimicrobial activity of pine needle extract and horseradish on the growth of Vibrio
  • Park
  • Kn
  • Lee
  • Sh
Park KN and Lee SH: Antimicrobial activity of pine needle extract and horseradish on the growth of Vibrio. J Korean Soc Food Sci Nutr 32, 185–190, 2003.
Introductory Oriental medicine at home. In Dongeubogam
  • Song
  • Hj
Song HJ: Introductory Oriental medicine at home. In Dongeubogam. Seoul, Korea: Kuk Il, 19913, pp 173–201.
Isolation of antioxidant, 2-hydroxy-5-methyl-3[2H]-furanone from pine needle
  • Boo Yc
  • Jeon
  • Co
  • Oh
  • Jy
Boo YC, Jeon CO, and Oh JY: Isolation of antioxidant, 2-hydroxy-5-methyl-3[2H]-furanone from pine needle. Korean Soc Appl Biol Chem 37, 310–314, 1994.
Experimental basis for the prevention of breast cancer
  • Metha
Metha RG: Experimental basis for the prevention of breast cancer. Eur J Cancer 36, 1275–1282, 2000.
Antimutagenic and antioxidative effects of ethanol extracts from garlic, ginger, green onion and red pepper
  • Lee
  • Ms
  • Kim
  • Sj
  • Kim
  • Sa
  • Woo Mk
  • Kwak
  • Cs
Lee MS, Kim SJ, Kim SA, Woo MK, Kwak CS, et al.: Antimutagenic and antioxidative effects of ethanol extracts from garlic, ginger, green onion and red pepper. Korean J Gerontol 13, 8–16, 2003.
Effect of hot water extracts of Saliva militiorrhiza Bge, Pinus persica Strokes, Angelica gigas Nakai and Pinus strobus on lipid oxidation
  • Kim Sm
  • Cho
  • Kim Ej Ys
  • Bae
  • Mj
  • Han
  • Jp
Kim SM, Cho YS, Kim EJ, Bae MJ, Han JP, et al.: Effect of hot water extracts of Saliva militiorrhiza Bge, Pinus persica Strokes, Angelica gigas Nakai and Pinus strobus on lipid oxidation. J Korean Soc Food Sci Nutr 27, 399–405, 1998.
Promotion Center through the Aging and Physical Culture Research Insti-tute, and SRC program (Aging and Apoptosis Research Center at Seoul Na-tional University) of the Address correspondence to
  • Acknowledgments
Acknowledgments and Notes This work was supported by the 2003 grants from the Agricultural R & D Promotion Center through the Aging and Physical Culture Research Insti-tute, and SRC program (Aging and Apoptosis Research