Immobilized lipase is a green and sustainable catalyst for hydrolysis of acidified oil. Glutaraldehyde is widely used for lipase immobilization while the appropriate strategy optimizes the catalytic performance of lipase. In this research, lipase from Candida rugosa (CRL) was immobilized on spherical silica (SiO2) by glutaraldehyde multipoint covalent treatments, including covalent binding method and adsorption-crosslinking method. The enzymatic stability properties and performance in hydrolysis of refined oil and acidified oil were studied. We confirmed that the residual activity decreased while the stability increased because of the influence on secondary structure of lipase after multipoint covalent treatments. In the comparison of different immobilization strategies in multipoint covalent treatment, SiO2-CRL (covalent binding method) showed lower loading capacity than SiO2-CRL (adsorption-crosslinking method), resulting in low activity. However, SiO2-CRL (covalent binding method) showed better reusability and stability. Immobilized lipase via covalent binding method was more potential in the application of catalytic hydrolysis of acidified oils.
Differently expressed genes (DEGs) across cervical (CC), endometrial (EC), and vulvar carcinoma (VC) may serve as potential biomarkers for these progressive tumor conditions. In this study, DEGs of cervical (CC), endometrial (EC), and vulvar carcinoma (VC) were identified by microarray analysis. The interaction network between the identified 124 DEGs was constructed and analyzed to identify the hub genes and genes with high stress centrality. DEGs, namely, CDK1 and MMP9, were found to show highest degree and highest stress centrality respectively from the gene interaction network of 124 nodes and 1171 edges. DEG CDK1 is found to be overlapping in both cervical and endometrial carcinomic conditions while DEG MMP9 is found in vulvar carcinomic condition. Further, as it is studied that many phytochemicals play an important role as medicinal drugs, we have identified phytochemicals from few widely available medicinal plants and performed comprehensive computational study to identify a multi-targeted phytochemical against the identified DEGs, which are crucially responsible for the progression of these carcinomic conditions. Virtual screening of the phytochemicals against the target DEG protein structures with PDB IDs 4Y72 and 1GKC resulted in identifying the multi-targeted phytochemical against both the proteins. The molecular docking and dynamics simulation studies reveal that luteolin can act as a multi-targeted agent. Thus, the interactional and structural insights of luteolin toward the DEG proteins signify that it can be further explored as a multi-targeted agent against the cervical, endometrial, and vulvar carcinoma.
We investigated the possible anticancer mechanisms of Pteris vittata [PV] n-hexane extract on MCF-7 [breast cancer cell line]. Cultured cell lines were treated with various concentrations of this extract ± Baf-A1 [autophagic inhibitor]. Cells’ viability, apoptotic markers [caspase-7, Bax, and Bcl-2], autophagic markers [light chain 3 [LC-3] and P62/SQSTM1]], and the tumor suppressor P53 and its mRNA were checked by their corresponding methods. Treated cell lines showed significant concentration and time-dependent reductions in cell viability in response to PV-n-hexane extract and also exhibited a concomitant induction of apoptosis [increased chromatin condensation, nuclear fragmentation, and pro-apoptotic Bax, and cleaved caspase-7 levels while decreased Bcl-2 levels] and autophagy [increased autophagosomes vacuoles, and LC3B II levels while decreased P62/SQSTM1 levels]. Moreover, PV-n-hexane extract-treated cells showed significant increases in the P53 and its mRNA levels. The addition of Baf-A1 reversed the PV-n-hexane extract autophagic effects and increased apoptotic cell percentage with a much increase in the cleaved caspase-7 and P53 protein and its mRNA levels. We concluded that the PV-n-hexane extract exhibits cytotoxic effects on the MCF-7 cell line with significant reductions in cell viability and concomitant autophagy and apoptosis induction. Inhibition of autophagy in the PV-treated MCF-7 cells enhances apoptosis via a p35-dependent pathway.
Cyclin-dependent kinase 12 (CDK12) has been found to regulate tumor progression. However, its function in gastric carcinoma (GC) remains controversial. This work aimed to explore the exact effect of CDK12 on GC progression. We detected the expression of CDK12 in GC cells and normal gastric mucosal epithelial cells. Then CDK12 function on GC cell proliferation, migration, and angiogenesis was researched by colony formation experiment, Transwell experiment, and angiogenesis assay. Moreover, CDK12 effect on the PI3K/AKT/mTOR pathway activity was explored by western blot. Further, we used LY294002 (10 μM) to treat GC cells to verify whether CDK12 regulates GC progression by activating the PI3K/AKT/mTOR pathway. Additionally, CDK12 effect on the expression of prognostic factors of GC was detected by western blot, including alkaline phosphatase (ALP) and Ki67. Quantitative real-time polymerase chain reaction and western blot were utilized to evaluate the expression of mRNAs and proteins. As a result, CDK12 was upregulated in GC cells. CDK12 overexpression facilitated the proliferation, migration, and angiogenesis of GC cells. However, CDK12 silencing showed an opposite result. CDK12 overexpression activated the PI3K/AKT/mTOR pathway, but CDK12 silencing inactivated it in GC cells. The blockage of the PI3K/AKT/mTOR pathway induced by LY294002 treatment counteracted the promotion of CDK12 on the proliferation, migration, and angiogenesis of GC. Further, CDK12 silencing suppressed the expression of ALP and Ki67 proteins in GC cells. Taken together, CDK12 promotes the proliferation, migration, and angiogenesis of GC by activating the PI3K/AKT/mTOR pathway. It may be a novel target for GC treatment.
Lipstatin, natural inhibitor of pancreatic lipase produced by Streptomyces toxytricini and used as an anti-obesity drug. Chemical mutagenesis was performed with different concentrations of N-methyl-N′-nitro-N-nitrosoguanidine (NTG) for strain improvement to obtain high yield of lipstatin. It was observed that the potential of the wild type strain to produce lipstatin (1.09 g/L) was very low. Selected mutants produced lipstatin in the range of 1.20–2.23 g/L at the flask level where maximum amount of lipstatin was produced by M5 mutant. For comparative study, both the parent and M5 mutant strain of S. toxytricini were grown at the lab scale bioreactor with suitable sources of carbon and nitrogen. Significant increase in the production of lipstatin was observed at the bioreactor level where the wild type strain produced 2.4 g/L of lipstatin, while through the NTG mutation, the production of lipstatin was 5.35 g/L. However, Dry Cell Weight (DCW) of the mutant strain was less in comparison with wild type strain and significant morphological differences were observed. Nearly 5 times increase in the production of lipstatin was achieved through NTG mutation and bioreactor-controlled conditions. It was determined that the NTG treatment might be beneficial for strain improvement to get a better candidate for lipstatin production on commercial scale.
TGFB1 cytokine is involved in normal mammary epithelial development as well as in breast tumorigenesis. It has role in both breast tumor suppression and progression. TGFB1 gene has several single nucleotide polymorphisms (SNPs) many of which modulate the activity of TGFB1. Our aim in this study was to analyze TGFB1 + 29 polymorphism in breast cancer individuals from North Indian population.
TGFB1 + 29 T/C polymorphism was analyzed using Sanger sequencing in 285 breast cancer patients and age matched 363 healthy controls from North Indian population. Next, transcript expression of 13 apoptotic genes, TRAIL, DR4, DR5, DcR1, DcR2, Bcl2, cytochrome c, Casp8L, Casp8, FlipS, FlipL, Casp3s and Casp3 were carried out in 77 breast tumor tissues obtained from 77 individuals.
TGFB1 + 29 CC genotype provided protection against the development of breast cancer (P = 0.012). This was mainly attributable to higher age group (> 45 years) women (P = 0.016). Individuals having CC protector genotype showed significantly higher expression of TGFB1 transcript compared to the TT and TC risk genotypes (P = 0.044). Furthermore, we observed that TGFB1 + 29 CC genotype showed increased TRAIL mediated apoptosis via the extrinsic pathway in breast tumor patients with age greater than 45 years (P = 0.027).
TGFB1 + 29 homozygous mutant CC genotype is related to protection against breast cancer in North Indian women population greater than 45 years of age.
Our previous findings demonstrated that Helichrysetin possessed promising anti-cancer activity. It was able to induce apoptosis in the A549 cell line. However, its mechanism of action is unknown. The present study aimed to unravel possible underlying molecular mechanisms of helichrysetin-induced apoptosis in A549 (human lung carcinoma) cells using comparative quantitative proteomics (iTRAQ labeled), followed by an exhaustive bioinformatics analysis. Our results suggested that DNA damage response (DDR) and cell cycle arrest were responsible for lung cancer cell death with helichrysetin treatment. Among proteins that changed in abundance were Nrf2 and HMOX1. They are oxidative stress-related proteins and were increased in abundance. BRAT1 was also increased in abundance, suggesting an increase in DNA damage repair, indicating the occurrence of DNA damage due to oxidative stress. However, several essential DDR downstream proteins such as p-ATM, BRCA1, FANCD2, and Rb1 that would further increase DNA damage were found to be dramatically decreased in relative abundance. Cell cycle-related proteins, p53, p21, and cyclin D1, were increased while cyclin A, cyclin E, and cdk2 were decreased. This is predicted to facilitate S-phase arrest. Furthermore, excessive DNA damage and prolonged arrest would in turn result in the induction of mitochondrial-mediated apoptosis. Based on these observations, we postulate that the effects of helichrysetin were in part via the suppression of DNA damage response which led to DNA damage and prolonged cell cycle arrest. Subsequently, this event initiated mitochondrial-mediated apoptosis in A549 lung cancer cells.
Biochemical and/or molecular mechanisms of arsenic or fluoride toxicity in experimental animals have been widely investigated in the recent past. However, their combined effects on target cells/organelle are poorly understood. The present study was executed to delineate their combined effects on mitochondrial function in the liver of rat. Female Wistar rats (140 ± 20 g) were force fed individually or in combination with sodium arsenate (4 mg/kg body weight) and sodium fluoride (4 mg/kg body weight) for 90 days. Thereafter, established markers of mitochondrial function viz. mitochondrial lipid peroxidation, oxidative phosphorylation, ATPase, succinic dehydrogenase, and caspase-3 activity were determined. Cytochrome C release and oxidative DNA damage were also estimated in the liver of respective groups of rats. The study showed significant differences in these results amongst the three groups. Observations on parameters viz. LPO, cytochrome-C, caspase-3, and 8-OHdG suggested an antagonistic relationship between these two elements. Results on ATPase, SDH, and ADP:O ratio indicated synergism. It is concluded that AsIII + F in combination may express differential effects on signalling pathways and proapoptotic/antiapoptotic proteins/genes that contribute to liver cell death.
Interaction of As and F with mitochondria
Multiple organs, including the testes, are damaged by iron overload. It has been shown that
N-acetyl cysteine (NAC) influences oxidative stress in iron overload. The present study
aimed to evaluate the roles of acetylated peptide (AOP) and NAC in the inhibition of ironoverload
induced-testicular damage. At the beginning of the experiment, NAC (150 mg /
kg) was given for a week to all 40 rats. Then, four groups were formed by dividing the
animals (10 rats/group). Group I included healthy control rats. Group II (iron overload)
was given intraperitoneal iron dextran (60 mg/kg/day) 5 days a week for 4 weeks. Group
III (NAC) was given NAC orally at a dose of 150 mg/kg/day for 4 weeks in addition to iron
dextran. Group IV (AOP) was given AOP orally at a dose of 150 mg/kg/day for 4 weeks
besides iron dextran. When the experiment time was over, testosterone serum level, testicular
B cell lymphoma-2 (BCL-2) and protein kinase B (PKB) protein levels, nuclear factor
kappa-B (NF-κB), and Beclin1 mRNA expression levels, and malondialdehyde (MDA),
and reduced glutathione (GSH) were determined by ELISA, quantitative reverse transcription-
PCR, and chemical methods. Finally, histopathological examinations and immunohistochemical
detection of claudin-1 and CD68 were performed. The iron overload group
exhibited decreased testosterone, BCL-2, PKB, claudin-1, and GSH and increased MDA,
NF-κB, Beclin1, and CD68, while both NAC and AOP treatments protected against the
biochemical and histopathological disturbances occurring in the iron overload model. We
concluded that NAC and AOP can protect against testes damage by iron overload via their
antioxidant, anti-inflammatory, antiapoptotic, and ant-autophagic properties. The NAC
and AOP may be used as preventative measures against iron overload–induced testicular
The aberrant expression of mRNAs participates in the pathogenesis of hepatic fibrosis. However, the precise mechanisms regulated by microRNAs (miRNAs) remain unclear. This study aims to investigate the functions about differentially expressed mRNAs (DEMs) in liver fibrosis and their regulatory mechanisms. The DEMs datasets about hepatic stellate cells (HSCs) obtained from hepatic fibrosis mice versus HSCs obtained from normal mice were downloaded from the GEO database (GSE120281). According to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of the GSE120281 datasets, ECM-receptor interaction was the most significant enrichment pathway that was correlated with hepatic fibrosis, and the fibronectin 1 (FN1) gene was upregulated most significantly in the signaling pathway. Downregulation of the expression of the FN1 gene by transfecting with FN1-siRNA alleviated the activity of HSCs. Four different bioinformatics web-based tools were used to predict that microRNA-96-5p (miR-96-5p) would directly target FN1, and a luciferase assay further confirmed this. Moreover, miR-96-5p was declined in activated HSCs and FN1, whereas laminin γ1 (LAMC1), collagen 1α1 (COL1A1) in the ECM-receptor interaction pathway, and the fibrosis marker α-smooth muscle actin (α-SMA) could be reduced by upregulation of the miRNA. Additionally, miR-96-5p expression was low in CCl4-induced liver fibrosis mice. Increased miR-96-5p expression alleviated liver fibrosis, improved liver function, and inhibited the expression of α-SMA, FN1, COL1A1, and LAMC1. In conclusion, this study indicated that upregulation of miR-96-5p could reduce HSC activation and relieve hepatic fibrosis by restraining the FN1/ECM-receptor interaction pathway.
Oral squamous cell carcinoma (OSCC), a global threatening disease, is reported mostly in the middle and elderly male population. Even though the exact cause of OSCC was not known, consumption of tobacco in any form has been reported in most of OSCC patients. OSCC is a massive invasive type of cancer which easily spreads to the distant organs. Hence treating it at appropriate time is necessary and the rate of OSCC incidence is also constantly increasing. At present, chemoradiation is the only therapy prescribed for OSCC patients which renders various side effects. Hence, the treatment with lesser side effect was of current research interest. Doxazosin (α1 adrenorecptor antagonist) had been proven to render anticancer effect in prostate, renal, hepatic, and ovarian cancers but its role in oral cancer cells was not been elucidated. Therefore, we have assessed the anticancer effect of doxazosin on oral squamous cancer cells via through the induction of apoptosis, and antioxidant property. The cytoprotective effect of doxazosin on normal Vero cells and anticancer effect on oral cancer KB cells were analyzed with MTT assay. Doxazosin antioxidant activity were analyzed by their reactivity with free radicals and metal ions by the method of FRAP, DPPH, chemilumiscence, and ORAC assay. The antioxidant levels were also assessed by TBARS, SOD, and glutathione levels, and later on apoptosis staining techniques like DCFH-DA, Rhodamine 123, and AO/EtBr stain were conducted. Apoptosis was confirmed by estimating the levels of apoptotic proteins in doxazosin-treated KB human oral cancer cells by ELISA method. The results from our study show that doxazosin is a potent antioxidant and it significantly induces apoptosis in human oral cancer by altering various cellular molecules at downstream signaling which has been depict in the results. Our study proves doxazosin as a potent anticancer drug which may be used in the treatment of oral carcinoma, if it is subjected to further research using human clinical trials.
The gut microbiota widely varies from individual to individual, but the variation shows stability over a period of time. The presence of abundant bacterial taxa is a common structure that determines the microbiota of human being. The presence of this microbiota greatly varies from geographic location, sex, food habits and age. Microbiota existing within the gut plays a significant role in nutrient absorption, development of immunity, curing of diseases and various developmental phases. With change in age, chronology diversification and variation of gut microbiota are observed within human being. But it has been observed that with the enhancement of age the richness of the microbial diversity has shown a sharp decline. The enhancement of age also results in the drift of the characteristic of the microbes associated with the microbiota from commensals to pathogenic. Various studies have shown that age associated gut-dysbiosis may result in decrease in tlongevity along with unhealthy aging. The host signalling pathways regulate the presence of the gut microbiota and their longevity. The presence of various nutrients regulates the presence of various microbial species. Innate immunity can be triggered due to the mechanism of gut dysbiosis resulting in the development of various age-related pathological syndromes and early aging. The gut microbiota possesses the ability to communicate with the host system with the help of various types of biomolecules, epigenetic mechanisms and various types of signalling-independent pathways. Drift in this mechanism of communication may affect the life span along with the health of the host. Thus, this review would focus on the use of gut-microbiota in anti-aging and healthy conditions of the host system.
Miliusa nilagirica, a rare tree species of Western Ghats, belongs to the Annonaceae family, a family with potential antioxidant and antidiabetic properties. This study is designed vividly to establish the relationship between the constituent phytochemicals and their hyperglycemic effects through the antioxidant traits of M. nilagirica in vitro. Phytochemical tests were conducted on dry powdered leaves and extracts of various methods to determine the existence of various constituents. The antidiabetic potential of leaf extracts was estimated by using the α-amylase inhibitory model and the antioxidant potential was estimated with various assays. The quantitative phytochemical screening of leaf parts shows the presence of carbohydrates (88.74 ± 0.65 mg GE/g sample), proteins (82.17 ± 2.52 mg BSAE/g sample), phenolics (40.44 ± 0.43 GAE/100 g), and flavonoids (66.05 ± 0.48 mg RE/g extract). Methanol extract of Soxhlet of M. nilagirica registered the strongest antioxidant activity in all assays, 75.66% inhibition (DPPH assay), 795.01 µmol/g (ABTS˙⁺ radical scavenging), 994.33 µmol/g (FRAP assay), 362.02 mg AAE/g extract (TAC assay), 47% inhibition (NO scavenging assay). In vitro α-amylase inhibition showed a highly noticeable reduction in ethyl acetate extract from Soxhlet (75.19%). HPLC and FTIR analyses on the extracts added strengths to the obtained results on the potentiality of M. nilagirica. From the results, it is evident that phytochemicals from M. nilagirica can be studied further, isolated, and incorporated as an alternative to synthetic supplements for hyperglycemia.
Because of the essential role of PLpro in the regulation of replication and dysregulates the host immune sensing, it is considered as a therapeutic target for novel drug development. To reduce the risk of immune evasion and vaccine effectiveness small molecular therapeutics are the best complementary approach. Hence, we used a structure-based drug designing approach to identify potential small molecular inhibitors for PLpro of SARS-CoV-2. Initial scoring and re-scoring of the best hits revealed that three compounds NPC320891 (2,2-Dihydroxyindene-1,3-Dione), NPC474594 (Isonarciclasine), and NPC474595 (7-Deoxyisonarciclasine) exhibit higher docking scores than the control GRL0617. Investigation of the binding modes revealed that alongside the essential contacts i.e. Asp164, Glu167, Tyr264, and Gln269 these molecules also target Lys157 and Tyr268 residues in the active site. Moreover, molecular simulation demonstrated that the reported top hits also possess stable dynamics and structural packing. Furthermore, the residues' flexibility revealed that all the complexes demonstrated higher flexibility in the regions 120-140, 160-180, and 205-215. The 120-140 and 160-180 lies in the fingers region of PLpro, which may open/close during the simulation to cover the active site and push the ligand inside. In addition, the total binding free energy was reported to be -32.65±0.17 kcal/mol for the GRL0617-PLpro, for the NPC320891-PLpro complex the TBE was -35.58±0.14 kcal/mol, for the NPC474594-PLpro the TBE was -43.72±0.22 kcal/mol while for NPC474595-PLpro complex the TBE was calculated to be -41.61±0.20 kcal/mol respectively. Clustering of the protein’s motion and FEL further revealed that In NPC474594 and NPC474595 complexes, the drug was seen to have moved inside the binding cavity along with the loop in the palm region harboring the catalytic triad thus justifying the higher binding of these two molecules particularly. In conclusion, the overall results reflect favorable binding of the identified strongly than the control drug thus demanding in vitro and in vivo validation for clinical purposes.
An alkaline serine protease gene from haloalkaliphilic actinobacteria, Nocardiopsis sp. Mit-7 (NCIM-5746) was cloned and overexpressed in E. coli BL21 under the control of the T7 promoter in the pET Blue1 vector. MW of the recombinant protease was determined as 34kDa by SDS-PAGE and the Mass Spectrometer (MALDI-TOF),.The role of aspartate, serine, and tryptophan was shown in the active site of the alkaline protease. The recombinant protease was optimally active at 65°C, higher than the native enzyme, and active over a pH range of 7.0-13.0, optimally at pH 9. The enzyme was further immobilized suggesting its reduced sensitivity against adverse conditions.
Keywords: Heterologous expression; MALDI; Recombinant alkaline Protease; Haloalkaliphilic actinobacteria; Protease structure
Probiotics are live microorganisms that can have beneficial effects on humans. Encapsulation offers them a better chance of survival. Therefore, nozzle-free electrospinning was introduced for their embedding in nanofibrous material. Probiotic Lactobacillus paragasseri K7 in lyophilized and fresh form, with and without inulin as prebiotic, was added to a polymer solution of sodium alginate (NaAlg) and polyethylene oxide (PEO). Conductivity, viscosity, pH, and surface tension were determined to define the optimal concentration and volume ratio for smooth electrospinning. The success of the formed nanoscale materials was examined by scanning electron microscope (SEM), while the entrapment of probiotics in the nanofibrous mats was detected by attenuated total reflection—Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). Spontaneous diffusion of bacteria from electrospun samples in PBS buffer pH 7.4 was studied by plate counting on MRS agar. By exposing polymer solutions containing L. paragasseri K7 and inulin to a high electric field, the nanofilm was formed on a polypropylene substrate, used as collecting material. When polymer solutions without inulin were used, the bead-like nanofibers may have become visible. The SEM results suggest that inulin, in addition to K7 strain, additionally lowers the conductivity of spinning macromolecular solution and hinders the nanofiber formation. The results of ATR-FTIR confirmed the presence of L. paragasseri K7 embedded in nanocomposites by the appearance of characteristic peaks. The samples containing the probiotic regardless of its form with inulin had similar surface composition, except that the sodium content was higher in the samples with fresh probiotic, probably due to greater and thus less easy embedding of the bacteria in NaAlg. Within 2 h, the largest amount of probiotic strain K7 was spontaneously released from the electrospun sample containing the inulin and probiotic in freeze-dried form (44%), while the amount released from the nanofibrous sample, which also contained the inulin and probiotic in fresh form, was significantly lower (21%). These preliminary results demonstrate the potential of nozzle-free electrospinning technology for the development of probiotic delivery systems for short-term use, such as feminine hygiene materials (tampons, pads, napkins).
Breast cancer is the second most common cancer after lung cancer in the world. Due to the anti-cancer properties of Berberine (Ber), in this study, the effect of combination therapy of Ber in the presence of blue LED irradiation and Valproic acid (Val) on the MDA-MB-231 breast cancer cell line was investigated. For this reason, after culturing the cells using different concentrations of Ber and Val, breast cancer cells were treated in both mono-treatment and combination therapy. In combination therapy, two modes were considered: (1) treatment with Val and then treatment with Ber in the dark or in presence of blue light irradiation (PDT)at a wavelength of 465 nm and energy of 30 J/cm² for 15 min, and (2) treatment with Ber in the dark or PDT and then treated with Val. In all cases, cell viability, morphological changes, and colonization were assessed. Evaluation of apoptosis was performed by fluorescence microscope and flow cytometry. According to the results, combination therapy has a higher mortality rate compared to mono-treatment, and in combination therapy, treatment of cells first with Ber (10 µg/mL)-PDT and then treatment with Val (250 µg/mL) caused a significant reduction (P < 0/05) in the survival rate of cancer cells. According to the findings, it can be said that the use of Ber-PDT in combination with Val, in addition to reducing the dose of the drug, has shown a synergistic effect which can suggest the potential of this strategy as a new treatment.
The pathogenesis of recurrent oral ulcers (ROU) is complex, with a long duration of illness and challenging to cure. According to traditional Chinese medicine (TCM),”heat accumulation in the heart-spleen” is one of the main causative factors. Jiaweidaochi powder (JWDCP) is based on the ancient Chinese medicine formula JWDCS, with the addition of Tongcao and gypsum and the removal of Mu Tong. It is generally used to treat “heat accumulation in the heart-spleen.” Previous studies have demonstrated that it effectively reduces recurrence rates and is anti-inflammatory in modulating immunity. The ROU rats’ model for JWDCP intervention treatment had been established, and histological tests revealed that JWDCP has a therapeutic effect on the pathological changes in the oral mucosa. In addition, the methylation levels of peripheral blood IFNG gene were detected by bisulfite sequencing PCR (BSP), and the methylation levels of the IFNG promoter region in the model group and each dose group were lower than those in the control group. However, no significant methylation differences were observed. Furthermore, the results of enzyme-linked immunosorbent assay (ELISA) and RNA quantitative polymerase chain reaction showed that JWDCP could reduce IFN-γ and IL-4 protein concentrations, with high GATA-3 mRNA production, T-bet mRNAproduction was upgraded, elevated IL-4 mRNA levels, and reduced IFN-γ mRNA levels after treatment (P < 0.001). The expression of transcription factor T-betmRNA and GATA-3 gene mRNA was accompanied by changes in IFN-γmRNA and IL-4mRNA, demonstrating that Th2 type differentiation in RAS suppresses the body’s immunity and that the imbalance of transcription factor expression further leads to Th1/Th2 drift. JWDCP is likely to reduce the protein concentration by regulating the imbalance of transcription factors and enhancing antioxidant capacity, thus achieving therapeutic effects. Treatment of recurrent oral ulcer models is not sufficient to reset IFNG methylation levels, correlating with the refractoriness of ROU, further confirming the complexity of epigenetic mechanisms and that epigenetic alterations in specific mediators may persist locally.
Enzymatic halogenation captures scientific interest considering its feasibility in modifying compounds for chemical diversity. Currently, majority of flavin-dependent halogenases (F-Hals) were reported from bacterial origin, and as far as we know, none from lichenized fungi. Fungi are well-known producers of halogenated compounds, so using available transcriptomic dataset of Dirinaria sp., we mined for putative gene encoding for F-Hal. Phylogenetic-based classification of the F-Hal family suggested a non-tryptophan F-Hals, similar to other fungal F-Hals, which mainly act on aromatic compounds. However, after the putative halogenase gene from Dirinaria sp., dnhal was codon-optimized, cloned, and expressed in Pichia pastoris, the ~63 kDa purified enzyme showed biocatalytic activity towards tryptophan and an aromatic compound methyl haematommate, which gave the tell-tale isotopic pattern of a chlorinated product at m/z 239.0565 and 241.0552; and m/z 243.0074 and 245.0025, respectively. This study is the start of understanding the complexities of lichenized fungal F-hals and its ability to halogenate tryptophan and other aromatic. compounds which can be used as green alternatives for biocatalysis of halogenated compounds.
Bordetella pertussis, the causative agent of whooping cough, is an opportunistic virulent bacterial pathogen that is resistant to a wide range of antibiotics due to a variety of resistance mechanisms. Looking at the increasing number of infections caused by B. pertussis and its resistance to diverse antibiotics, it is essential to develop alternative strategies to fight against B. pertussis. Diaminopimelate epimerase (DapF) is an important enzyme of the lysine biosynthesis pathway in B. pertussis that catalyzes the formation of meso-2, 6-diaminoheptanedioate (meso-DAP), which is an important step in lysine metabolism. Therefore, Bordetella pertussis diaminopimelate epimerase (DapF) becomes an ideal target for antimicrobial drug development. In the present study, computational modelling, functional characterization, binding studies, and docking studies of BpDapF with lead compounds were carried out using different in silico tools. In silico prediction results in the secondary structure, 3-D structure analysis, and protein-protein interaction analysis of BpDapF. Docking studies further showed the respective amino acid residues for ligands in the phosphate‑binding loop of BpDapF play a vital role in the formation of H‑bonds with these ligands. The site where the ligand was bound is a deep groove, which is regarded as the binding cavity of the protein. Biochemical studies indicated that Limonin (binding energy − 8.8 kcal/mol), Ajmalicine (binding energy − 8.7 kcal/mol), Clinafloxacin (binding energy − 8.3 kcal/mol), Dexamethasone (binding energy − 8.2 kcal/mol), and Tetracycline (binding energy − 8.1 kcal/mol) exhibited promising binding towards the drug target DapF of B. pertussis in comparison with the binding between other drugs and act as the potential inhibitors of BpDapF that eventually can reduce the catalytic activity of BpDapF.
Insulin resistance (IR) is one of the leading causes of Type 2 diabetes mellitus (T2DM). Inflammation, as a result of the disordered immune response, plays important roles in IR and T2DM. Interleukin-4-induced gene 1 (IL4I1) has been shown to regulate immune response and be involved in inflammation progress. However, there was little known about its roles in T2DM. Here, high glucose (HG)-treated HepG2 cells were used for T2DM investigation in vitro. Our results indicated that the expression of IL4I1 was up-regulated in peripheral blood samples of T2DM-patients and HG-induced HepG2 cells. The silencing of IL4I1 alleviated the HG-evoked IR through elevating the expressions of p-IRS1, p-AKT and GLUT4, and enhancing glucose consumption. Furthermore, IL4I1 knockdown inhibited inflammatory response by reducing the levels of inflammatory mediators, and suppressed the accumulation of lipid metabolites triglyceride (TG) and palmitate (PA) in HG-induced cells. Notably, IL4I1 expression was positively correlated with aryl hydrocarbon receptor (AHR) in peripheral blood samples of T2DM-patients. The silencing of IL4I1 inhibited the AHR signaling by reducing the HG-induced expressions of AHR and CYP1A1. Subsequent experiments confirmed that 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), an agonist of AHR, reversed the suppressive effects of IL4I1 knockdown on HG-caused inflammation, lipid metabolism and IR in cells. In conclusion, we found that the silencing of IL4I1 attenuated inflammation, lipid metabolism and IR in HG-induced cells via inhibiting AHR signaling, suggesting that IL4I1 might be a potential therapy target for T2DM.
Endophytes associated with medicinal plants are a potential source of valuable natural products. This study aimed to evaluate the antibacterial and antibiofilm activities of endophytic bacteria from Archidendron pauciflorum against multidrug-resistant (MDR) strains. A total of 24 endophytic bacteria were isolated from the leaf, root, and stem of A. pauciflorum. Seven isolates showed antibacterial activity with different spectra against four MDR strains. Extracts derived from four selected isolates (1 mg/mL) also displayed antibacterial activity. Among four selected isolates, DJ4 and DJ9 isolates exhibited the strongest antibacterial activity against P. aeruginosa strain M18, as indicated by the lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) (DJ4 and DJ9 MIC: 7.81 µg/mL; DJ4 and DJ9 MBC: 31.25 µg/mL). 2 × MIC of DJ4 and DJ9 extracts was found to be the most effective concentration to inhibit more than 52% of biofilm formation and eradicate more than 42% of established biofilm against all MDR strains. 16S rRNA-based identification revealed four selected isolates belong to the genus Bacillus. DJ9 isolate possessed nonribosomal peptide synthetase (NRPS) gene, and DJ4 isolate possessed NRPS and polyketide synthase type I (PKS I) gene. Both these genes are commonly responsible for secondary metabolites synthesis. Several antimicrobial compounds, including 1,4-dihydroxy-2-methyl-anthraquinone and paenilamicin A1, were detected in the bacterial extracts. This study highlights endophytic bacteria isolated from A. pauciflorum provide a great source of novel antibacterial compounds.
In this study, we aimed to explore long non-coding RNA (lncRNA) sustained low-efficiency dialysis (SLED1) correlated with Bcl-2 apoptosis pathway in acute myeloid leukemia (AML). This study further aimed to determine its role in the regulation of AML progression and its action as a potential biomarker for better prognosis. AML microarray profiles GSE97485 and probe annotation from the Gene Expression Omnibus (GEO) database from the National Center for Biotechnology Information (NCBI) were detected using the GEO2R tool (http://www.ncbi.nlm.nih.gov/geo/geo2r/). The expression of AML was downloaded from the TCGA database (http://cancergenome.nih.gov/). The statistical analysis of the database was processed with R software. Bioinformatic analysis found that lncRNA SLED1 is highly expressed in AML patients and is associated with poor prognosis. We found that the increased SLED1 expression levels in AML were significantly correlated with FAB classification, human race, and age. Our study has shown that upregulation of SLED1 promoted AML cell proliferation and inhibited cell apoptosis in vitro; RNA sequencing showed increased expression of BCL-2 and indicated that SLED1 might promote the development of AML by regulating BCL-2. Our results showed that SLED1 could promote the proliferation and inhibit the apoptosis of AML cells. SLED1 might promote the development of AML by regulating BCL-2, but the mechanism involved in the progression of AML is unclear. SLED1 plays an important role in AML progression, may be applied as a rapid and economical AML prognostic indicator to predict the survival of AML patients, and help guide experiments for potential clinical drag targets.
Orthopedic infections due to biofilm formation in biomaterial-based implants have become challenging in bone tissue engineering. In the present study, in vitro antibacterial analysis of amino-functionalized MCM-48 mesoporous silica nanoparticles (AF-MSNs) loaded with vancomycin is analyzed for its potential as a drug carrier for the sustained/controlled release of vancomycin against Staphylococcus aureus. The effective incorporation of vancomycin into the inner core of AF-MSNs was observed by alternation in the absorption frequencies obtained by Fourier transform infrared spectroscopy (FTIR). Dynamic light scattering (DLS) and high resolution-transmission electron microscopy (HR-TEM) results show that all the AF-MSNs had homogeneous spherical shapes with a mean diameter of 165.2 ± 1.25 nm, and there is a slight change in the hydrodynamic diameter after vancomycin loading. Furthermore, the zeta potential of all the AF-MSNs (+ 30.5 ± 0.54 mV) and AF-MSN/VA (+ 33.3 ± 0.56 mV) were positively charged due to effective functionalization with 3-aminopropyl triethoxysilane (APTES). Furthermore, cytotoxicity results show that the AF-MSNs have better biocompatibility than non-functionalized MSNs (p < 0.05), and results prove AF-MSNs loaded with vancomycin show better antibacterial effect against S. aureus than non-functionalized MSNs. Results confirm that bacterial membrane integrity was affected by treatment with AF-MSNs and AF-MSN/VA by staining the treated cells with FDA/PI. Field emission scanning electron microscopy (FESEM) analysis confirmed the shrinkage of bacterial cells and membrane disintegration. Furthermore, these results demonstrate that amino-functionalized MSNs loaded with vancomycin significantly increased the anti-biofilm and biofilm inhibitory effect and can be incorporated with biomaterial-based bone substitutes and bone cement to prevent orthopedic infections post-implantation.
Atherosclerosis (AS) is one of the most common and important vascular diseases. It is believed that the abnormal expression of circular RNAs (circRNAs) plays an important role in AS. Hence, we investigate the function and mechanism of circ-C16orf62 in AS development.
In this study, oxidized low-density lipoprotein (ox-LDL)-treated human macrophages (THP-1) were used as pathological conditions of AS in vitro. The expression of circ-C16orf62, miR-377 and Ras-related protein (RAB22A) mRNA was detected by real-time quantitative polymerase chain reaction (RT-qPCR) or western blot. Cell viability or cell apoptosis was assessed by cell counting kit-8 (CCK-8) assay or flow cytometry assay. The releases of proinflammatory factors were investigated using enzyme-linked immunosorbent assay (ELISA). The production of malondialdehyde (MDA) and superoxide dismutase (SOD) was examined to assess oxidative stress. Total cholesterol (T-CHO) level was detected, and cholesterol efflux level was tested using a liquid scintillation counter. The putative relationship between miR-377 and circ-C16orf62 or RAB22A was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.
circ-C16orf62 expression was elevated in AS serum samples and ox-LDL-treated THP-1 cells. Apoptosis, inflammation, oxidative stress and cholesterol accumulation induced by ox-LDL were suppressed by circ-C16orf62 knockdown. Circ-C16orf62 could bind to miR-377 and thus increased the expression level of RAB22A. Rescued experiments showed that circ-C16orf62 knockdown alleviated ox-LDL-induced THP-1 cell injuries by increasing miR-377 expression, and miR-377 overexpression lessened ox-LDL-induced THP-1 cell injuries by degrading RAB22A level.
In conclusion, circ-C16orf62 played a crucial role in the regulation of apoptosis, inflammation, oxidative stress and cholesterol accumulation in ox-LDL-treated human macrophages via mediating the miR-377/RAB22A axis, hinting that circ-C16orf62 might be involved in AS progression.
Feeds for aquaculture animals are designed to provide them with the greatest amount of nourishment they need to carry out their regular physiological activities, such as maintaining a potent natural immune system and boosting growth and reproduction. However, the problems that severely hamper this sector's ability to contribute to achieving global food security include disease prevalence, chemical pollution, environmental deterioration, and inadequate feed usage. The regulated release of active aquafeed components; limited water solubility, bioaccessibility, and bioavailability, as well as their potent odour and flavour, limit their utilisation. They are unstable under high temperatures, acidic pH, oxygen, or light. Recent advancements in nano-feed for aquaculture (fish/shrimp) have attract enormous attention due to its excellent nutritional value, defeating susceptibility and perishability. Encapsulation is a multifunctional smart system that could bring benefits of personalized medicine; minimize costs and resources in the preclinical and clinical study in pharmacology. It guarantees the coating of the active ingredient as well as its controlled release and targeted distribution to a particular area of the digestive tract. For instance, using nanotechnology to provide more effective fish/shrimps feed for aquaculture species. The review enables a perspective points on safety and awareness in aquafeeds that have been made by the advancements of nanosystem. Therefore, potential of nano-delivery system in aquafeed industry for aquaculture act as concluding remark on future directions.
The aim of the present study is to identify actinobacteria Streptomyces bacillaris ANS2 as the source of the potentially beneficial compound 2,4-di-tert-butylphenol, describe its chemical components, and assess its anti-tubercular (TB) and anti-cancer properties. Ethyl acetate was used in the agar surface fermentation of S. bacillaris ANS2 to produce the bioactive metabolites. Using various chromatographic and spectroscopy analyses, the potential bioactive metabolite separated and identified as 2,4-di-tert-butylphenol (2,4-DTBP). The lead compound 2,4-DTBP inhibited 78% and 74% of relative light unit (RLU) decrease against MDR Mycobacterium tuberculosis at 100ug/ml and 50ug/ml concentrations, respectively. The Wayne model was used to assess the latent/dormant potential in M. tuberculosis H37RV at various doses, and the MIC for the isolated molecule was found to be 100ug/ml. Furthermore, the molecular docking of 2,4-DTBP was docked using Autodock Vinasuite onto the substrate binding site of the target Mycobacterium lysine aminotransferase (LAT) and the grid box was configured for the docking run to cover the whole LAT dimer interface. At a dosage of 1 mg/ml, the anti-cancer activity of the compound 2,4-DTBP was 88% and 89% inhibited against the HT 29 (colon cancer) and HeLa (cervical cancer) cell lines. According to our literature survey, this present finding may be the first report on anti-TB activity of 2,4-DTBP and has the potential to become an effective natural source and the promising pharmaceutical drug in the future.
Several clinical studies have reported the analgesic effect of curcumin (Curc) in various situations such as rheumatoid arthritis, osteoarthritis, and postsurgical pain. Therefore, in this work, Curc-loaded electrospun nanofibers (NFs) are designed to evaluate their sustained release on analgesic effect duration in rats after epidural placement via repeated formalin and tail-flick tests. The Curc-loaded polycaprolactone/gelatin NFs (Curc-PCL/GEL NFs) are prepared through an electrospinning technique and introduced to the rat's epidural space after laminectomy. The physicochemical and morphology features of the prepared Curc-PCL/GEL NFs were characterized via FE-SEM, FTIR, and degradation assay. The in vitro and in vivo concentrations of Curc were measured to evaluate the analgesic efficacy of the drug-loaded NFs. Rat nociceptive responses are investigated through repeated formalin and tail-flick tests for 5 weeks after the placement of NFs. Curc had a sustained release from the NFs for 5 weeks, and its local pharmaceutical concentrations were much greater than plasma concentrations. Rat's pain scores in both early and late phases of the formalin test were remarkably decreased in the experimental period. Rat's tail-flick latency was remarkably enhanced and remained constant for up to 4 weeks. Our findings show that the Curc-PCL/GEL NFs can supply controlled release of Curc to induce extended analgesia after laminectomy.
The efficient and economical removal of fermentation inhibitors from the complex system of biomass hydrolysate was one of the basics and keys in bio-chemical transformation. In this work, post-cross-linked hydrophilic-hydrophobic interpenetrating polymer networks (PMA/PS_pc IPNs and PAM/PS_pc IPNs) were proposed to remove fermentation inhibitors from sugarcane bagasse hydrolysate for the first time. PMA/PS_pc and PAM/PS_pc IPNs can obviously enhance the adsorption performance towards fermentation inhibitors due to their higher surface area and hydrophilic-hydrophobic synergetic surface properties, especially PMA/PS_pc IPNs has higher selectivity coefficients of 4.57, 4.63, 4.85, 16.0, 49.43, and 22.69, and higher adsorption capacity of 24.7 mg/g, 39.2 mg/g, 52.4 mg/g, 9.1 mg/g, 13.2 mg/g, and 144.9 mg/g towards formic acid, acetic acid, levulinic acid (LA), 5-hydroxymethylfurfural (HMF), furfural, and acid-soluble lignin (ASL), respectively, in a lower total sugar loss of 2.03%. The adsorption kinetics and isotherm of PMA/PS_pc IPNs were studied to elucidate its adsorption behavior towards fermentation inhibitors. In addition, the cyclic utilization property of PMA/PS_pc IPNs was stable. Synthesizing PMA/PS_pc IPNs is a new strategy to provide an efficient adsorbent for the removal of fermentation inhibitors from lignocellulosic hydrolysate.
The sustainable development of human society in today’s high-tech world depends on some form of eco-friendly energy source because existing technologies cannot keep up with the rapid population expansion and the vast amounts of wastewater that result from human activity. A green technology called a microbial fuel cell (MFC) focuses on using biodegradable trash as a substrate to harness the power of bacteria to produce bioenergy. Production of bioenergy and wastewater treatment are the two main uses of MFC. MFCs have also been used in biosensors, water desalination, polluted soil remediation, and the manufacture of chemicals like methane and formate. MFC-based biosensors have gained a lot of attention in the last few decades due to their straightforward operating principle and long-term viability, with a wide range of applications including bioenergy production, treatment of industrial and domestic wastewater, biological oxygen demand, toxicity detection, microbial activity detection, and air quality monitoring, etc. This review focuses on several MFC types and their functions, including the detection of microbial activity.
Sustainable remediation of arsenic-fluoride from rice fields through efficient bio-extraction is the need of the hour, since these toxicants severely challenge safe cultivation of rice and food biosafety. In the present study, we screened an arsenic-fluoride tolerant strain AB-ARC of Acinetobacter indicus from the soil of a severely polluted region of West Bengal, India, which was capable of efficiently removing extremely high doses of arsenate and fluoride from the media. The strain also behaved as a plant growth-promoting rhizobacterium, since it could produce indole-3-acetic acid and solubilize phosphate, zinc, and starch. Due to these properties of the identified strain, it was used for bio-priming the seeds of the arsenic-fluoride susceptible rice cultivar, Khitish for testing the efficacy of the AB-ARC strain to promote combined arsenic-fluoride tolerance in the rice genotype. Bio-priming with AB-ARC led to accelerated uptake of crucial elements like iron, copper, and nickel which behave as co-factors of physiological and antioxidative enzymes. Thus, the activation of superoxide dismutase, catalase, guaiacol peroxidase, glutathione peroxidase, and glutathione-S-transferase enabled detoxification of reactive oxygen species (ROS) and reduction of the oxidative injuries like malondialdehyde and methylglyoxal generation. Overall, due to ameliorated molecular damages and low uptake of the toxic xenobiotics, the plants were able to maintain improved growth vigor and photosynthesis, as evident from the elevated levels of Hill activity and chlorophyll content. Hence, bio-priming with the A. indicus AB-ARC strain may be advocated for sustainable rice cultivation in arsenic-fluoride co-polluted fields.
Laccases are widespread multi-copper oxidases and generally classified into three-domain laccases and two-domain laccases. In this study, a novel laccase PthLac from Parageobacillus thermoglucosidasius harbored only one domain of Cu-oxidase_4 and showed no sequence relatedness or structure similarity to three-domain and two-domain laccases. PthLac was heterologously expressed in Escherichia coli, purified, and characterized. The optimum temperature and pH of PthLac on guaiacol were at 60 ℃ and pH 6, respectively. The effects of various metal ions on PthLac were analyzed. All the tested metal ions did not suppress the activity of PthLac, except for 10 mM Cu²⁺, which increased the activity of PthLac to 316%, indicating that PthLac was activated by Cu²⁺. Meanwhile, PthLac kept 121% and 69% activity when incubated at concentrations of 2.5 and 3 M NaCl for 9 h, suggesting the long-term halotolerancy of this enzyme. In addition, PthLac showed resistance to the organic solvents and surfactants, and displayed dye decolorization capacity. This study enriched our knowledge about one-domain laccase and its potential industrial applications.
Globally 80% type 2 diabetes mellitus (T2DM) patients suffer nonalcoholic fatty liver disease (NAFLD). The interplay of gut microbiota and endogenous metabolic networks has not yet been reported in the setting of T2DM with NAFLD. As such, this study utilized 16S rRNA gene sequencing to assess the changes in intestinal flora and nuclear magnetic resonance spectroscopy (¹H NMR) to identify potential metabolites in a T2DM with NAFLD rat model. Spearman correlation analysis was performed to explore the relationship between gut microbiota and metabolites. Results revealed that among T2DM with NAFLD rats, diversity indexes of intestinal microbiota were distinctly decreased while levels of 18 bacterial genera within the intestinal tract were significantly altered. In addition, levels of eight metabolites mainly involved in the synthesis and degradation of ketone bodies, the TCA cycle, and butanoate metabolism were altered. Correlation analysis revealed that gut bacteria such as Blautia, Ruminococcus torques group, Allobaculum, and Lachnoclostridium strongly associate with 3-hydroxybutyrate, acetone, acetoacetate, 2-oxoglutarate, citrate, creatinine, hippurate, and allantoin. Our findings can provide a basis for future development of targeted treatments.
Wound care management aims at stimulating and improving healing process without scar formation. Although various plants have been reported to possess wound healing properties in tribal and folklore medicines, there is a lack of scientific data to validate the claim. In this aspect, it becomes inevitable to prove the efficacy of naturally derived products at pharmacological levels. Couroupita guianensis as a whole plant has been reported to exhibit wound healing activity. The leaves and fruit of this plant have been utilized in folkloric medicine to cure skin diseases and infections for many years. However, to the best of our knowledge, no scientific studies have been conducted to verify the wound healing properties of C. guianensis fruit pulp. Therefore, the present study seeks to investigate the wound healing potential of C. guianensis fruit pulp using an excision wound model in Wistar albino male rats. This study indicated that the ointment prepared from crude ethanolic extract of C. guianensis fruit pulp facilitated wound contraction that were evidenced by a greater reduction in the wound area and epithelialization period and increased hydroxyproline content. The experimental groups treated with low and mid dose of C. guianensis ethanol extract (CGEE) ointments had shown a wound closure of 80.27% and 89.11% respectively within 15 days, which is comparable to the standard betadine ointment which showed 91.44% healing in the treated groups. Further, the extract influenced the expression of genes VEGF and TGF-β on post wounding days that clearly explained the strong correlation between these genes and wound healing in the experimental rats. The animals treated with 10% CGEE ointment showed a significant upregulation of both VEGF and TGF-β as compared with other test and standard groups. These findings provide credence to the conventional application of this plant in the healing of wounds and other dermatological conditions, and may represent a therapeutic strategy for the treatment of wounds.
To analyze the regulatory effects and key targets of the fat-soluble components of ginseng in lung cancer.
Gas chromatography-mass spectrometry and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform were used to analyze and identify the fat-soluble components of ginseng. Network pharmacology was used to analyze the therapeutic targets of the fat-soluble components of ginseng in lung cancer and screen key proteins. In vitro assays were conducted to verify the effects of the active fat-soluble components of ginseng on proliferation and apoptosis in lung cancer cells and to verify the regulation of key proteins.
Ten active fat-soluble components of ginseng were screened for follow-up. Network pharmacology showed 33 overlapping targets between the active fat-soluble components of ginseng and lung cancer, and functional enrichment of the targets showed involvement of response to nitrogen, hormone response, membrane raft, and positive regulation of external stimulus. Pathway enrichment analysis showed vascular endothelial growth factor (VEGF) signaling, adipocyte lipolysis regulation, chronic myelogenous leukemia, endocrine resistance, and NSCLC-related pathways. A protein-protein interaction network was constructed, and the top 10 targets were selected in accordance with their scores. Ultimately, five target genes (EGFR, KDR, MAPK3, PTPN11, and CTNNB1) were selected in combination with literature mining for subsequent experimental verification. Proliferation assays showed that the growth of lung cancer cells was significantly decreased in a concentration-dependent manner in the fat-soluble components of ginseng intervention group compared with controls. Flow cytometry showed that active fat-soluble components of ginseng promoted apoptosis in a concentration-dependent manner in lung cancer cells. Western blot and quantitative real-time PCR showed that levels of the five key proteins and mRNAs were significantly decreased in the intervention group; furthermore, histone protein and mRNA levels were significantly higher in the high-concentration intervention group compared with the low-concentration group.
The active fat-soluble components of ginseng inhibited the growth of lung cancer cells and promoted apoptosis. The underlying regulatory mechanisms may be related to signaling pathways involving EGFR, KDR, MAPK3, PTPN11, and CTNNB1.
The biosensor method has not yet been applied to study the fungus-acetone interaction. The first electrochemical (amperometric) study of Fusarium oxysporum f. sp. vasinfectum cells’ responses to acetone was performed to evaluate the initial stages of the metabolism of acetone in the cells of the micromycete. Using a laboratory model of a membrane microbial sensor based on the micromycete cells, it was found that the fungus had constitutive enzyme systems that were involved in acetone transport into fungal cells. The research showed that uninduced by acetone cells had degradative activity against acetone. A positive cooperativity of acetone binding with enzymes initiating acetone degradation was revealed. Oxygen concentration affected the activation of cell enzymes initiating acetone degradation, but the cells’ activity in the presence of acetone maintained even at low oxygen concentration. Kinetic parameters (the maximum rate of the cells’ response to substrate and the half-saturation constant) of the processes causing the fungal cells’ response to acetone were calculated. The results demonstrated the convenience of the biosensor method for assessing the potential of the micromycete as a substrate-degrading culture. In the future, the mechanism of response to acetone for microbial cells will be studied.
Dekkera bruxellensis has been studied for several aspects of its metabolism over the past years, which has expanded our comprehension on its importance to industrial fermentation processes and uncovered its industrial relevance. Acetate is a metabolite often found in D. bruxellensis aerobic cultivations, whereas its production is linked to decreased ethanol yields. In a previous work, we aimed to understand how acetate metabolism affected the fermentation capacity of D. bruxellensis. In the present work, we evaluated the role of acetate metabolism in respiring cells using ammonium or nitrate as nitrogen sources. Our results showed that galactose is a strictly respiratory sugar and that a relevant part of its carbon is lost, while the remaining is metabolised through the Pdh bypass pathway before being assimilated into biomass. When this pathway was blocked, yeast growth was reduced while more carbon was assimilated to the biomass. In nitrate, more acetate was produced as expected, which increased carbon assimilation, although less galactose was uptaken from the medium. This scenario was not affected by the Pdh bypass inhibition. The confirmation that acetate production was crucial for carbon assimilation was brought by cultivations in pyruvate. All physiological data were connected to the expression patterns of PFK1, PDC1, ADH1, ALD3, ALD5 and ATP1 genes. Other respiring carbon sources could only be properly used by the cells when some external acetate was supplied. Therefore, the results reported herein helped in providing valuable contributions to the understanding of the oxidative metabolism in this potential industrial yeast.
Public health is seriously jeopardized in developing countries due to poor sanitation and
the presence of persistent pollutants in natural water bodies. Open dumping, wastewater
discharge without proper treatment and atmospheric fallout of the organic and inorganic
pollutants are the main causes behind the poor condition. Some of the pollutants pose
a greater risk due to their toxicity and persistence. Such a class of pollutants are known
as chemical contaminants of emerging concern (CECC), including antibiotics and drug
residues, endocrine disruptors, pesticides and micro- and nano-plastics. Conventional
treatment methods cannot treat them properly and are often associated with several
disadvantages. However, the chronological development of techniques and materials
for their treatment has exhibited graphene as an efcient candidate for environmental
remediation. This current review considers the various graphene-based materials, their
properties, advancement in synthesis methods with time and their detailed application in
removing dyes, antibiotics and heavy metals. It has been discussed how graphene and its
derivatives exhibit unique electronic, mechanical, structural and thermal properties. In this
paper, the mechanism of adsorption and degradation using these graphene-based materials
has also been discussed vividly. In addition to this, a bibliographic analysis was performed
to identify the trend of research related to graphene and its derivatives in the adsorption
and degradation of pollutants round the globe refected by the publications. Therefore, this
review can be instrumental in understanding the fact that further development of graphenebased materials and their mass production can provide a very efective and economical
wastewater treatment method.
Lung cancer is the second most commonly diagnosed cancer and the leading cause of cancer-related death. Lung adenocarcinoma (LUAD) is the most common form of lung cancer and has a low 5-year survival rate. Therefore, much more research is needed to identify cancer biomarkers, promote biomarker-driven therapy and improve treatment outcomes. LncRNAs have been reported to participate in various physiological and pathological processes, especially in cancer, and thus have attracted much attention. In this study, lncRNAs were screened from the single-cell RNA-seq dataset CancerSEA. Among them, four lncRNAs (HCG18, NNT-AS1 and LINC00847 and CYTOR) were closely associated with the prognosis of LUAD patients according to Kaplan–Meier analysis. Further study explored the correlations between these four lncRNAs and immune cell infiltration in cancer. In LUAD, LINC00847 was positively correlated with the immune infiltration of B cells, CD8 T cells, and dendritic cells. LINC00847 decreased the expression of PD-L1, immune checkpoint blockade (ICB) immunotherapy-related gene, which suggests that LINC00847 is a potential new target for tumor immunotherapy.
In recent years, the uses of silver nanoparticles have increased, which lead to nanoparticles discharge into aquatic bodies which may, if not well controlled, have harmful effect on different organisms. This calls for the need to constantly evaluate the toxicity level of nanoparticles. In this study, green biosynthesized silver nanoparticles mediated by endophytic bacteria Cronobacter sakazakii (CS-AgNPs) were subjected to toxicity evaluation by brine shrimp lethality assay. The ability of CS-AgNPs to improve plant growth by nanopriming of Vigna radiata L seeds treated with different concentrations (1ppm, 2.5ppm, 5ppm and 10ppm) in order to enhance biochemical constituents was investigated, also its inhibitory effect to growth of phytopathogenic fungi Mucor racemose was examined. Results showed that Artemia salina treated with CS-AgNPs exhibited good hatching percentage and LC50 value of 688.41 µg/ml when Artemia salina eggs were exposed to CS-AgNPs during hatching. Plant growth was enhanced at 2.5ppm CS-AgNPs, with increased photosynthetic pigments, protein, and carbohydrate content. This study suggests that silver nanoparticles synthesized via endophytic bacteria Cronobacter sakazakii are safe to use and can be utilized as means of combating plant fungal pathogens.
MicroRNAs (MiRNAs) play pivotal roles in regulating gene expression, and serve as crucial biomarkers for diagnosis of a variety of disease. However, label-free and sensitive miRNA detection remains a huge challenge due to the low abundance. Herein, we developed an approach through integrating primer exchange reaction (PER) with DNA-templated silver nanoclusters (AgNCs) for label-free and sensitive miRNA detection. In this method, PER was used to amplify miRNA signals and produce single-strand DNA (ssDNA) sequences. The produced ssDNA sequences mediated DNA-templated AgNCs based signal generation by unfolding the designed hairpin probe (HP). The generated AgNCs signal was correlated with the dosage of target miRNA. Eventually, the established approach exhibited a low detection of limit of 47 fM with a great dynamic range of more than five orders of magnitude. In addition, the method was also utilized to detect the miRNA-31 expression in collected clinical samples from pancreatitis patients and demonstrated that miRNA-31 was upregulated in patients, showing a great promising of the method in clinical application.
It has been optimized thermal acid hydrolytic pretreatment and enzymatic saccharification (Es) in flask culture of Undaria pinnatifida seaweed, which is a prebiotic. The optimal hydrolytic conditions were a slurry content of 8% (w/v), 180 mM H2SO4, and 121°C for 30 min. Es using Celluclast 1.5 L at 8 U/mL produced 2.7 g/L glucose with an efficiency of 96.2%. The concentration of fucose (a prebiotic) was 0.48 g/L after pretreatment and saccharification. The fucose concentration decreased slightly during fermentation. Monosodium glutamate (MSG) (3%, w/v) and pyridoxal 5'-phosphate (PLP) (30 μM) were added to enhance gamma-aminobutyric acid (GABA) production. To further improve the consumption of mixed monosaccharides, adaptation of Lactobacillus brevis KCL010 to high concentrations of mannitol improved the synbiotic fermentation efficiency of U. pinnatifida hydrolysates.
A new zinc(II) complex formulated as [Zn(pipr-ac)2], where pipr-ac stands for piperidineacetate, was synthesized and structurally identified with the help of experimental and DFT methods. Frontier molecular orbital (FMO) analysis demonstrated that the new complex has higher biological activity compared to the free ligand. Molecular electrostatic potential (MEP) showed the nitrogen atoms and oxygen of carbonyl groups are the active sites of Zn(II) compound. Also, natural bond orbital (NBO) analysis confirmed the charge transfer from the ligating atoms to the metal ion and formation of four coordinated Zn(II) complex. MTT assay illustrated a noticeable cytotoxic activity of the new zinc(II) complex compared to cisplatin on K562 cell line. The CT-DNA and serum albumin (SA) binding of the Zn(II) complex were explored individually. In this regard, UV–Vis spectroscopy and florescence titration revealed the occurrences of fluorescence quenching of CT-DNA/SA by metal compound via static mechanism and creation of hydrogen bonds and van der Waals interactions between them. The binding was further confirmed by viscosity measurement and gel electrophoresis assay for CT-DNA and circular dichroism spectroscopy for SA. Moreover, molecular docking simulation demonstrated that the new compound binds mainly through hydrogen bonds to the groove of DNA and hydrogen bonds and van der Waals interactions to site I of SA.
Paracetamol is the most predominantly used antipyretic and analgesic drug. As paracetamol is metabolised mostly in the liver, both deliberate and unintentional overdoses of paracetamol are reported to provoke severe hepatotoxicity, including liver failure. Caesalpinia bonducella seed is well known for its medicinal and therapeutic properties. However, there is no report on its potential protective effects against paracetamol-instigated hepatotoxicity. Therefore, we studied the protective effects of aqueous seed extract of Caesalpinia bonducella (ASECB) on paracetamol-instigated hepatotoxicity in rats. Thirty female albino rats were divided into five groups: control, paracetamol-intoxicated, ASECB + paracetamol, silymarin + paracetamol, and ASECB alone. The rats were assessed for liver enzyme markers (alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transpeptidase), antioxidant activity (superoxide dismutase, catalase, reduced glutathione, glutathione peroxidase), lipid peroxidation (malondialdehyde), histopathological, cytokine levels (pro-inflammatory cytokines TNF-α and IL-6, and anti-inflammatory cytokine IL-10), and protein expression (pro-apoptotic markers caspase 3 and caspase 8 and anti-apoptotic marker Bcl-2) after the 8-day study period. Repercussions of paracetamol intoxication induced upregulation of liver enzyme markers, antioxidant depletion, malondialdehyde production, decreased expression of Bcl-2 and IL-10, and overexpression of apoptotic and pro-inflammatory mediators, which were attenuated by pre-treatment with ASECB. ASECB markedly mitigated paracetamol-instigated liver injury by suppressing caspase-8/3 signalling and inflammatory infiltration in liver tissue by significantly reducing TNF-α and IL-6. In conclusion, ASECB pre-treatment exerts potent liver protection against paracetamol-instigated hepatotoxicity evidenced by mitigation of oxidative stress, lipid peroxidation, inflammation, and apoptosis.
The major threat to mungbean (Vigna radiata L.) cultivation in the Indian subcontinent is yellow mosaic diseases (YMD), caused by Begomovirus containing bipartite genomes (DNA-A and DNA-B). In the current study, we address the epidemiology of begomoviruses infecting mungbean plants in three YMD hotspot regions of India. Full-length genomic components of the viruses from the symptomatic leaves were cloned by rolling circle amplification (RCA) and sequenced. Mungbean yellow mosaic virus (MYMV) was detected in Bihar and mungbean yellow mosaic India virus (MYMIV) in Assam and Orissa. Furthermore, we studied the population structure and genetic diversity of MYMV and MYMIV isolates of Vigna species reported to date from India. Interestingly, based on phylogenetics, we observed independent evolution of DNA-A and coevolution of DNA-B of MYMV and MYMIV. This finding is supported by the high mutation rate and recombination events in DNA-B, particularly in BV1 and BC1 genes over DNA-A, with high transition/transversion bias (R) for DNA-A over DNA-B. To investigate the effect of Begomovirus infection in plants, we constructed infectious clones (i.e. MYMV and MYMIV) and inoculated them to eight mungbean genotypes, cowpea (Vigna unguiculata L.) and tobacco (Nicotiana benthamiana) through agroinfiltration. The infected plants developed varying degrees of typical YMD symptoms. Based on the disease severity score and viral titre, mungbean genotypes were categorized as highly susceptible to MYMV (ML267) and MYMIV (K851) and immune to MYMV (PDM139, SML668) and MYMIV (Pusa Vishal). Conclusively, our findings may help prevent an epidemic of YMD in Vigna species and develop mungbean genotypes resistant to YMD via breeding programs.
Cancer is one of the fatal diseases and has high mortality worldwide, and the major drawback with the cure is the side effects from the chemotherapeutic agents. The increased multidrug resistance among microbial pathogens is a serious threat to plant and animal health. There is an urgent need for an alternative that can battle against pathogens and can be used for cancer treatment. Presently, actinomycetes were isolated from cave soil, and the crude extract obtained from the potent isolate was analyzed with gas chromatography-mass spectrometry (GC–MS) and high-performance thin layer chromatography (HPTLC) to identify bioactive metabolites. The crude extract was examined for in vitro antimicrobial activity on human pathogens and antifungal activity on plant pathogens. The isolate Streptomyces sp. strain YC69 exhibited antagonistic activity and antimicrobial activity in a dose-dependent manner, with the highest inhibition in Staphylococcus aureus. GC–MS revealed many bioactive compounds, and HPTLC depicted metabolite fingerprints. The antifungal activity exhibited a delayed lag phase in growth curve assay and distorted and collapsed cells of Fusarium oxysporum in scanning electron microscopy (SEM) images. In the MTT assay, the IC50 of 41.98 µg/ml against HeLa cells was obtained with clear evidence for deformed cells and blebbing of the cell membrane. The results from the current study suggest that the crude extract from Streptomyces sp. strain YC69 contains antimicrobial metabolites that can inhibit pathogenic microbes in plants and humans. The MTT assay results conclude that further studies on purification may lead to the use of Streptomyces sp. strain YC69 as a source for anti-oncogenic compounds.
Cell division is driven by nucleic acid metabolism, and thymidylate synthase (TYMS) catalyzes a rate-limiting step in nucleotide synthesis. As a result, thymidylate synthase has emerged as a critical target in chemotherapy. 5-Fluorouracil (5-FU) is currently being used to treat a wide range of cancers, including breast, pancreatic, head and neck, colorectal, ovarian, and gastric cancers The objective of this study was to establish a new methodology for the low-cost, one-pot synthesis of uracil derivatives (UD-1 to UD-5) and to evaluate their therapeutic potential in BC cells. One-pot organic synthesis processes using a single solvent were used for the synthesis of drug analogues of Uracil. Integrated bioinformatics using GEPIA2, UALCAN, and KM plotter were utilized to study the expression pattern and prognostic significance of TYMS, the key target gene of 5-fluorouracil in breast cancer patients. Cell viability, cell proliferation, and colony formation assays were used as in vitro methods to validate the in silico lead obtained. BC patients showed high levels of thymidylate synthase, and high expression of thymidylate synthase was found associated with poor prognosis. In silico studies indicated that synthesized uracil derivatives have a high affinity for thymidylate synthase. Notably, the uracil derivatives dramatically inhibited the proliferation and colonization potential of BC cells in vitro. In conclusion, our study identified novel uracil derivatives as promising therapeutic options for breast cancer patients expressing the augmented levels of thymidylate synthase
Early determination of infectious pathogens is vitally important to select appropriate antibiotics, and to manage nosocomial infection. Herein, we propose a target recognition triggered triple signal amplification–based approach for sensitive pathogenic bacteria detection. In the proposed approach, a double-strand DNA probe (capture probe) that is composed of an aptamer sequence and a primer sequence is designed for specific identification of target bacteria and initiation of following triple signal amplification. After recognition of target bacteria, primer sequence is released from capture probe to bind with the designed H1 probe, forming a blunt terminal in the H1 probe. Exonuclease-III (Exo-III enzyme) specifically recognizes the blunt terminal in H1 probe and degrades the sequence from 3′ terminal, resulting a single-strand DNA to induce the following signal amplification. Eventually, the approach exhibits a low detection limit of 36 cfu/mL with a broad dynamic range. The high selectivity endows the method a promising prospective for clinical sample analysis.
The aim of this research is to investigate the quantum geometric properties and chemical reactivity of atropine, a pharmaceutically active tropane alkaloid. Using density functional theory (DFT) computations with the B3LYP/SVP functional theory basis set, the most stable geometry of atropine was determined. Additionally, a variety of energetic molecular parameters were calculated, such as the optimized energy, atomic charges, dipole moment, frontier molecular orbital energies, HOMO–LUMO energy gap, molecular electrostatic potential, chemical reactivity descriptors, and molecular polarizability. To determine atropine’s inhibitory potential, molecular docking was used to analyze ligand interactions within the active pockets of aldo–keto reductase (AKR1B1 and AKR1B10). The results of these studies showed that atropine has greater inhibitory action against AKR1B1 than AKR1B10, which was further validated through molecular dynamic simulations by analyzing root mean square deviation (RMSD) and root mean square fluctuations (RMSF). The results of the molecular docking simulation were supplemented with simulation data, and the ADMET characteristics were also determined to predict the drug likeness of a potential compound. In conclusion, the research suggests that atropine has potential as an inhibitor of AKR1B1 and could be used as a parent compound for the synthesis of more potent leads for the treatment of colon cancer associated with the sudden expression of AKR1B1.
Silver nanoparticles (AgNPs) have gained great interest because of their specific and distinct properties. Chemically synthesized AgNPs (cAgNPs) are often unsuitable for medical applications due to requiring toxic and hazardous solvents. Thus, green synthesis of AgNPs (gAgNPs) using safe and nontoxic substances has attracted particular focus. The current study investigated the potential of Salvadora persica and Caccinia macranthera extracts in the synthesis of CmNPs and SpNPs, respectively. Aqueous extracts of Salvadora persica and Caccinia macranthera were prepared and taken as reducing and stabilizing agents through gAgNPs synthesis. The antimicrobial effects of gAgNPs against susceptible and antibiotic-resistant bacterial strains and their toxicity effects on L929 fibroblast normal cells were evaluated. TEM images and particle size distribution analysis showed that the CmNPs and SpNPs have average sizes of 14.8 nm and 39.4 nm, respectively. The XRD confirms the crystalline nature and purity of both CmNPs and SpNPs. FTIR results demonstrate the involvement of the biologically active substances of both plant extracts in the green synthesis of AgNPs. According to MIC and MBC results, higher antimicrobial effects were seen for CmNPs with a smaller size than SpNPs. In addition, CmNPs and SpNPs were much less cytotoxic when examined against a normal cell relative to cAgNPs. Based on high efficacy in controlling antibiotic-resistant pathogens without detrimental adverse effects, CmNPs may have the capacity to be used in medicine as imaging, drug carrier, and antibacterial and anticancer agents.