Article

Dietary Supplementation with Fresh Pineapple Juice Decreases Inflammation and Colonic Neoplasia in IL-10-deficient Mice with Colitis

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Abstract

Bromelain, a mixture of proteolytic enzymes typically derived from pineapple stem, decreases production of proinflammatory cytokines and leukocyte homing to sites of inflammation. We previously showed that short-term oral treatment with bromelain purified from pineapple stem decreased the severity of colonic inflammation in C57BL/6 Il10(-/-) mice with chronic colitis. Since fresh pineapple fruit contains similar bromelain enzymes but at different proportions, this study aimed to determine whether long-term dietary supplementation with pineapple (supplied as juice) could decrease colon inflammation and neoplasia in Il10(-/-) mice with chronic colitis as compared with bromelain derived from stem. Colitis was triggered in Il10(-/-) mice by exposure to the non-steroidal anti-inflammatory drug piroxicam. Mice with colitis were supplemented with fresh vs. boiled pineapple juice or bromelain purified from stem for up to 6 months. Experimental mice readily consumed fresh pineapple juice at a level that generated mean stool proteolytic activities equivalent to 14 mg bromelain purified from stem, while control mice received boiled juice with inactive enzymes. Survival was increased in the group supplemented with fresh rather than boiled juice (P = 0.01). Mice that received fresh juice also had decreased histologic colon inflammation scores and a lower incidence of inflammation-associated colonic neoplasia (35% versus 66%; P < 0.02), with fewer neoplastic lesions/colon (P = 0.05). Flow cytometric analysis of murine splenocytes exposed to fresh pineapple juice in vitro demonstrated proteolytic removal of cell surface molecules that can affect leukocyte trafficking and activation. These results demonstrate that long-term dietary supplementation with fresh or unpasteurized frozen pineapple juice with proteolytically active bromelain enzymes is safe and decreases inflammation severity and the incidence and multiplicity of inflammation-associated colonic neoplasia in this commonly used murine model of inflammatory bowel disease.

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... Due to the good anti-inflammatory properties of bromelain and promising in vitro results, high hopes are associated with its use in treating chronic inflammatory bowel disease (IBD). Numerous studies indicate that bromelain inhibits intestinal inflammation by modulating the expression/activity of compounds involved in inflammation [121][122][123][124][125]. Chronic inflammation of the intestines is a severe disease in patients with ulcerative colitis and Crohn's disease. ...
... Significantly, the effective anti-inflammatory activity of bromelain was associated with a lack of cytotoxicity in mice. The anti-inflammatory effect of bromelain in IBD was confirmed in subsequent in vitro studies by Hale et al. [125]. The authors again showed that long-term dietary supplementation with pineapple decreases colon inflammation and neoplasia in IL-10 −/− mice with chronic colitis. ...
... Proteolytic degradation of several cell surface molecules: CD44 ↓, CD45R ↓, CD62L ↓, CD8 ↓ Reduction the the clinical and histologic severity of IBD [123,125] Human cells from endoscopic colon biopsies from patients with ulcerative colitis, Crohn's disease ...
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Bromelain is a major sulfhydryl proteolytic enzyme found in pineapple plants, having multiple activities in many areas of medicine. Due to its low toxicity, high efficiency, high availability, and relative simplicity of acquisition, it is the object of inexhaustible interest of scientists. This review summarizes scientific reports concerning the possible application of bromelain in treating cardiovascular diseases, blood coagulation and fibrinolysis disorders, infectious diseases, inflammation-associated diseases, and many types of cancer. However, for the proper application of such multi-action activities of bromelain, further exploration of the mechanism of its action is needed. It is supposed that the anti-viral, anti-inflammatory, cardioprotective and anti-coagulatory activity of bromelain may become a complementary therapy for COVID-19 and post-COVID-19 patients. During the irrepressible spread of novel variants of the SARS-CoV-2 virus, such beneficial properties of this biomolecule might help prevent escalation and the progression of the COVID-19 disease.
... For instance, kiwifruit supports immune function and reduces the likelihood of developing coldor flu-like illnesses [20]. Papaya has anti-inflammatory and immunomodulatory properties and has been shown to have protective effects against breast cancer and prostate cancer [21][22][23][24][25]. Pineapple contains an enzyme called bromelain, which has been demonstrated to have anti-inflammatory, antithrombotic, fibrinolytic, anticancer, and immunomodulatory effects, in addition to being a wound healing and circulatory improvement agent [26,27]. Pineapple has also been found to help prevent colorectal cancer [28]. ...
... Numerous previous studies have demonstrated these fruits possess immunomodulatory properties. For example, kiwifruit may enhance immune function [20]; guava and papaya have anti-inflammatory and immunomodulatory properties [21,30]; pineapple has anti-inflammatory effect [27]; grape seed proanthocyanidins could improve functional activation of the immune system [33]. A large number of publications suggested that the immunomodulatory and anti-inflammatory properties of fruits can be attributed to the phytochemicals present in fruits [8][9][10]. ...
Article
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Fruits have been widely considered as the default “health foods” because they contain numerous vitamins and minerals needed to sustain human health. Fermentation strategies have been utilized to enhance the nutritive and flavor features of healthy and readily consumable fruit products while extending their shelf lives. A traditional fermented multi-fruit beverage was made from five fruits including kiwi, guava, papaya, pineapple, and grape fermented by Saccharomyces cerevisiae along with lactic acid bacteria and acetic acid bacteria. The immunomodulatory properties of the fermented multi-fruit beverage, in vivo nonspecific and ovalbumin (OVA)-specific immune response experiments using female BALB/c mice were performed. Administration of the fermented multi-fruit beverage reduced the calorie intake, thus resulting in a less weight gain in mice compared to the water (placebo)-fed mice. In the nonspecific immune study model, the fermented multi-fruit beverage enhanced phagocytosis and T cell proliferation but did not affect B cell proliferation and immunoglobulin G (IgG) production. Analysis of cytokine secretion profile also revealed that the fermented multi-fruit beverage enhanced proinflammatory cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and T helper (Th)1-related cytokine interferon (IFN)-γ production, thus creating an immunostimulatory effect. Nonetheless, in the specific immune study model, the results showed that the fermented multi-fruit beverage decreased the production of proinflammatory cytokines IL-6 and TNF-α production in OVA-immunized mice. Moreover, it also caused a decrease in the production of anti-OVA IgG1, which was accompanied by a decrease in Th2-related cytokines IL-4 and IL-5 production and an increase in Th1-related cytokine IFN-γ production, indicating that it may have the potential to shift the immune system from the allergen‐specific Th2 responses toward Th1-type responses. The results indicate that fermented multi-fruit beverage has the potential to modulate immune responses both in a nonspecific and specific manners.
... In the case of trypsin and chymotrypsin, their role is controversial as some evidence support the antitumoral effect, while others attribute to them a tumorigenic effect [12,13]. Preclinical and clinical studies showed that plant proteolytic enzymes act as antitumor and chemopreventive agents due to a variety of biological activities, encompassing pro-apoptotic [14-17] and anti-inflammatory effects [18]. In addition, these enzymes promote the inactivation of endogenous proteases and cytokines [19,20], modulate the expression of several adhesion molecules [21] and tumor suppressor proteins, and inhibit mitogen-activated protein kinases (MAPK)-regulated pathway [18,[22][23][24]. ...
... Preclinical and clinical studies showed that plant proteolytic enzymes act as antitumor and chemopreventive agents due to a variety of biological activities, encompassing pro-apoptotic [14-17] and anti-inflammatory effects [18]. In addition, these enzymes promote the inactivation of endogenous proteases and cytokines [19,20], modulate the expression of several adhesion molecules [21] and tumor suppressor proteins, and inhibit mitogen-activated protein kinases (MAPK)-regulated pathway [18,[22][23][24]. ...
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Previous studies showed that P1G10, a proteolytic fraction from Vasconcellea cundinamarcensis latex, reduced the tumor mass in animals bearing melanoma, increased in vitro DNA fragmentation and decreased cell adhesion. Here, we present some molecular and cellular events related to the antimetastatic effect induced by the CMS-2 fraction derived from P1G10 in metastatic melanoma B16-F10 and melanocyte Melan-a. Using difference gel electrophoresis and mass spectrometry, we identified four proteins overexpressed in tumor cells, all of them related to proliferation, survival, migration and cell invasion, that had their expression normalized upon treatment with CMS-2: nucleophosmin 1, heat shock protein 65, calcyclin binding protein and eukaryotic translation initiation factor 4H. In addition, some antioxidant and glycolytic enzymes show increased expression after exposure to CMS-2, along with an induction of melanogenesis (differentiation marker). The down regulation of cofilin 1, a protein involved in cell motility, may explain the inhibition of cell migration and dendritic-like outgrowth in B16-F10 and Melan-a, observed after CMS-2 treatment. Taken together, it is argued that CMS-2 modulates the expression of proteins related to metastatic development, driving the cell to a more differentiated-like state. These effects support the CMS-2 antimetastatic activity and place this fraction in the category of anticancer agent.
... Promising studies have examined potential therapies including resveratrol, curcumin, bromelain, green tea polyphenols, pomegranate, rutin, and Andrographis paniculata. [12][13][14][15][16][17][18][19] Food Allergy Herbal Formula-2 (FAHF-2) 20 is derived from Wu Mei Wan and consists of 9 Chinese herbal medicines. FAHF-2 has received Food and Drug Administration investigational new drug approval (FDA IND#77,468) under the botanical drug title for treating patients (including children) with multiple food allergies. ...
... Promising studies have examined potential therapies including resveratrol, curcumin, bromelain, tea polyphenols, pomegranate, and rutin. [12][13][14][15][16][17][18] Among these, one recently studied herbal therapy, HMPL-004, an aqueous ethanol herbal extract of Andrographis paniculata, has been shown to prevent colitis in animal models. 19,39 A pilot human clinical trial further confirmed the efficacy and safety of HMPL-004 in patients with ulcerative colitis. ...
Article
Crohn's disease (CD) is a chronic inflammatory disease with increasing incidence in children. Current medications have potentially serious side effects, hence increasing interest in alternative therapies. We previously developed an herbal formula, FAHF-2, based on a classical traditional Chinese herbal formula Wu Mei Wan that has long been used in China to treat colitis. We investigated FAHF-2's potential anti-inflammatory effects. FAHF-2 efficacy was tested in vivo in the CD45RbRAG1 transfer colitis model. Weight loss, colonic histology, and cytokine production from mesenteric lymph nodes were assessed. Human peripheral blood mononuclear cells (PBMCs) and colonic biopsies were obtained from children newly diagnosed with CD and controls and cultured with or without FAHF-2. Cytokine levels were measured by multiplex immunoassay. The effect of FAHF-2 on TNF-α-producing cells was determined by flow cytometry. NF-κB signaling was investigated in human lamina propria mononuclear cells upon FAHF-2 treatment by In-Cell Western. FAHF-2-treated mice had decreased weight loss, improved histology, and reduced TNF-α, IL-17, IL-6, and IFN-γ production. In vitro treated PBMCs produced less TNF-α, IFN-γ, and IL-12. FAHF-2 reduced the TNF-α-producing monocytes and T cells. Inflamed CD biopsies produced less TNF-α, IL-17, IL-6, and IL-1β. These effects are because of decreased NF-κB activation. FAHF-2 inhibited both adaptive and innate immune proinflammatory cytokine responses in PBMCs and inflamed CD mucosa due in part to blockage of NF-κB activation. FAHF-2 was effective in halting progression of colitis in a murine model. This study shows that FAHF-2 has potential as a novel treatment of CD.
... Preclinical studies indicate that bromelain exerts anti-inflammatory, antiedematous, antithrombotic/fibrinolytic, and immunomodulatory effects [15][16][17]. Relevant to the present investigation, bromelain has been shown to ameliorate experimental colitis in IL-10-deficient mouse and to decrease secretion of proinflammatory cytokines and chemokines in colon biopsies from patients with ulcerative colitis and Crohn's disease [18][19][20]. Considering the well-known association existing between intestinal inflammation and CRC, we investigated the possible antiproliferative/proapoptotic effects (and the mode of action) of this food component in a human colorectal carcinoma cell line and its potential chemopreventive effect in an animal model of colon cancer. In order to verify if the pharmacological effect of bromelain was due to its proteolytic activity, some in vitro experiments were performed by using proteolytically inactivated bromelain. ...
... Bromelain, a mixture of proteolytic enzymes derived from pineapple stem, has been reported to have beneficial effects in a variety of gastrointestinal diseases, including experimental inflammatory bowel disease [15][16][17][18][19][20]. In the present study, we have shown that bromelain exerts antiproliferative/proapoptotic actions in a colorectal carcinoma cell line and chemopreventive effects in a mouse model of experimental colon carcinogenesis in vivo. ...
Article
Colorectal cancer is an important health problem across the world. Here, we investigated the possible antiproliferative/proapoptotic effects of bromelain (from the pineapple stem Ananas comosus L., family Bromeliaceae) in a human colorectal carcinoma cell line and its potential chemopreventive effect in a murine model of colon cancer. Proliferation and apoptosis were evaluated in human colon adenocarcinoma (Caco-2) cells by the (3) H-thymidine incorporation assay and caspase 3/7 activity measurement, respectively. Extracellular signal-related kinase (ERK) and Akt expression were evaluated by Western blot analysis, reactive oxygen species production by a fluorimetric method. In vivo, bromelain was evaluated using the azoxymethane murine model of colon carcinogenesis. Bromelain reduced cell proliferation and promoted apoptosis in Caco-2 cells. The effect of bromelain was associated to downregulation of pERK1/2 /total, ERK, and pAkt/Akt expression as well as to reduction of reactive oxygen species production. In vivo, bromelain reduced the development of aberrant crypt foci, polyps, and tumors induced by azoxymethane. Bromelain exerts antiproliferative and proapoptotic effects in colorectal carcinoma cells and chemopreventive actions in colon carcinogenesis in vivo. Bromelain-containing foods and/or bromelain itself may represent good candidates for colorectal cancer chemoprevention.
... Bromelain can reduce the severity of colonic inflammation (Hale, 2004;Hale et al., 2005) and bromelain's anti-inflammatory function also results in proteolytic action by eradicating cell surface receptors that are implicated in leukocyte defects and activation (Hale et al., 2010). Bromelain also differs in the release of other chemokines, thus decreasing the incidence and frequency of persistent colitis. ...
Article
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Pineapple (Ananas comosus) is the third most important fruit in world production. The fruit is a source of balanced nutrients that provide the human body with valuable macro and micro-nutrients, including calcium, carbohydrates, vitamins, minerals, fiber, enzymes, and bioactive compounds that aid in the process of digestion and contribute to human health. Pineapple can be used as a supplementary nutritional fruit for good personal health. It possesses the biologically useful ingredient bromelain, which has demonstrated significant anti-inflammatory, antibiotic, anti-cancer, and anticoagulative properties. The multitude of potential uses of bromelain, combined with the effects of many other nutrients found in pineapple, allows us to appreciate not only its unquestionable taste but also the other benefits of this fruit. Antioxidants in pineapples help fight against free radicals, preventing cancers, heart diseases, and lowering cholesterol levels. Pineapple has abundant health benefits and also has the potential for breakthroughs in the food industry and agriculture sector. The present review will focus on the developments and future scopes of the medicinal and nutritional properties of pineapple.
... Но следует учитывать, что наиболее полезным для здоровья все же считается свежевыдавленный необработанный ананасовый фреш. [26] Именно в нем содержится наибольшее количество ценных ферментов, которые под воздействием температуры (пастеризации) исчезают из продукта. В то же время в замороженном напитке, как показывают исследования специалистов, большая часть полезных веществ сохраняется, что позволяет рекомендовать именно эту методику для долговременного хранения сока. ...
Article
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Ананасовый сок — уникальный продукт, который принесет неоценимую пользу всей семье. С его помощью можно утолить жажду и насытить организм уникальными микроэлементами, быстро избавиться от стресса и хандры и без лишних усилий вернуть стройность фигуры, избавиться от болей и воспалительных процессов. HTML версия статьи
... By BLAST analysis, these reductases likely originate from the Lachnospiraceae family of bacteria, and their examination could provide a glimpse into the microbial B-vitamin economy that importantly underpins host homeostasis, as humans are unable to de novo synthesize many essential B vitamins (77)(78)(79). Interestingly, fruit bromelain detected in 4 of 8 healthy patient fecal extracts is a pineapplederived cysteine protease we did not expect to encounter (80,81). This protease is commonly sold as an over-the-counter supplement or as a component of meat tenderizers, and its detection may be an artifact introduced through patients' diets. ...
Article
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The gut microbiota plays an important yet incompletely understood role in the induction and propagation of ulcerative colitis (UC). Organism-level efforts to identify microbes associated with UC have revealed the importance of community structure, but less is known about the molecular effectors of disease. We performed 16S rRNA gene sequencing in parallel with label-free data-dependent LC-MS/MS proteomics to characterize the stool microbiomes of healthy (n=8) and ulcerative colitis (n=10) patients. Comparisons of taxonomic composition between profiling techniques revealed major differences in community structure partially attributable to the additional detection of host, fungal, viral, and food peptides by metaproteomics. Differential expression analysis of metaproteomic data identified 176 significantly enriched protein groups between healthy and UC patients. Gene ontology analysis revealed several enriched functions with serine-type endopeptidase activity overrepresented in UC compared to healthy patients. Using a biotinylated fluorophosphonate probe and streptavidin-based enrichment, we show that serine endopeptidases are active in patient fecal samples and that additional putative serine hydrolases are detectable by this approach compared to unenriched profiling. Finally, as metaproteomic databases expand, they are expected to asymptotically approach completeness. Using ComPIL and de novo peptide sequencing, we estimate the size of the probable peptide space unidentified (“dark peptidome”) by our large database approach in order to establish a rough benchmark for database sufficiency. Despite high variability inherent in patient-derived samples, our analysis yielded a catalog of differentially enriched proteins between healthy and UC fecal proteomes. This catalog provides a clinically relevant jumping-off point for further molecular-level studies aimed at identifying the microbial underpinnings of UC.
... Hospitalization was a significant portion of the healthcare expenditures associated with this disease, accounting for 49% to 80% of the overall costs, whereas the expense of surgery represented 40-61% of all hospitalization costs [101]. Bromelain can reduce the severity of colonic inflammation [102,103] and bromelain's anti-inflammatory function also results in proteolytic action by eradicating cell surface receptors that are implicated in leukocyte defects and activation [104]. Bromelain also differs in the release of other chemokines, thus decreasing the incidence and frequency of persistent colitis. ...
Article
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Bromelain is a complex combination of multiple endopeptidases of thiol and other compounds derived from the pineapple fruit, stem and/or root. Fruit bromelain and stem bromelain are produced completely distinctly and comprise unique compounds of enzymes, and the descriptor “Bromelain” originally referred in actuality to stem bromelain. Due to the efficacy of oral administration in the body, as a safe phytotherapeutic medication, bromelain was commonly suited for patients due to lack of compromise in its peptidase efficacy and the absence of undesired side effects. Various in vivo and in vitro studies have shown that they are anti-edematous, anti-inflammatory, anti-cancerous, anti-thrombotic, fibrinolytic, and facilitate the death of apoptotic cells. The pharmacological properties of bromelain are, in part, related to its arachidonate cascade modulation, inhibition of platelet aggregation, such as interference with malignant cell growth; anti-inflammatory action; fibrinolytic activity; skin debridement properties, and reduction of the severe effects of SARS-Cov-2. In this paper, we concentrated primarily on the potential of bromelain’s important characteristics and meditative and therapeutic effects, along with the possible mechanism of action.
... Although new RCTs are urgently needed to better understand the effects of bromelain on the prevalence, severity and sternness of colitis, quality of life and markers of inflammation, this supplement decreased the harshness of colonic inflammation. This effect is probably due to its proteolytic action, which most likely eradicates the cell-surface receptors involved in leukocyte defection and activation [127]. ...
Article
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Bromelain is a mixture of proteolytic enzymes that is extracted primarily from pineapples. It is present in almost all the aerial parts of the plant—the peels, leaves, stems and fruit—although only the fruit and stems contain significant amounts. The market-demand for bromelain is quickly increasing, especially in the health sector as it can be used as a drug and/or nutraceutical. Although the complete molecular mechanism has not been fully identified, bromelain possesses several properties, including anti-inflammatory, anti-oedema, antithrombotic and fibrinolytic, mucolytic, anticancer and cicatrizing effects that have been evaluated in several clinical trials. However, one of the main limitations for the clinical use of this supplement is the varying composition of extracts, which leads to heterogeneity in results and, therefore, difficulty in making evidence-based prescriptions. In fact, different geographic locations, land and modes of cultivation, as well as extraction methods, can all give extracts with different activities, depending on the plant part used. Additionally, although novel extraction techniques have been developed to improve bromelain purification and extraction and give higher yields without loss in enzymatic activity, these methods are still expensive and challenging. This review will describe the state of the art in the main conventional and unconventional extraction and purification methods of bromelain and discuss the advantages and limitations of these strategies. Pharmacokinetic and pharmacodynamic profiles and the clinical applications that have arisen from randomized controlled clinical trials are also discussed. Finally, future perspectives for bromelain extracts will be presented.
... Diet might modulate inflammation by altering gut microbial ecology 21 . Gut bacteria are important not only in the absorption of certain vitamins and in the synthesis of bile acids (BA), but also have the potential to determine metabolic response to specific nutrients 22 , and to modify circulating pro-or anti-inflammatory mediators 23 . For example, trimethylamine, a precursor of trimethylamine-N-oxide, a pro-inflammatory metabolite that derives from choline and carnitine, is produced by bacteria 24,25 . ...
... Methylcobalamin, Acetyl-L-Carnitine, N-Acetyl Cysteine, Vitamin D, and Curcumin improve nerve regeneration in a number of conditions including nerve injuries (crush), nerve repair after transection and neuropathic conditions such as diabetes . Curcumin, Bromelain, and Serratopeptidase decrease the inflammatory response and have demonstrated improved recovery after surgery with less pain and earlier return to activity [61][62][63][64][65][66][67][68][69][70][71][72][73]. ...
... By BLAST analysis, these reductases likely originate from the Lachnospiraceae family of bacteria, and their examination could provide a glimpse into the microbial B-vitamin economy that importantly underpins host homeostasis, as humans are unable to de novo synthesize many essential B vitamins (55)(56)(57). Interestingly, fruit bromelain detected in 4 of 8 healthy patient fecal extracts is a pineapple-derived cysteine protease we did not expect to encounter (58,59). This protease is commonly sold as an over-the-counter supplement or as a component of meat tenderizers, and its detection may be an artifact introduced through patients' diets. ...
Preprint
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Mass spectrometry-based metaproteomics technologies enable the direct observation of proteins within complex multi-organism environments. A major hurdle in mapping metaproteomic fragmentation spectra to their corresponding peptides is the need for large peptide databases encompassing all anticipated species contained within a biological sample. As we cannot predict the taxonomic composition of microbiomes a priori , we developed the ComPIL database which contains a comprehensive collection of 4.8 billion unique peptides from public sequencing repositories to enable our proteomics analyses. We analyzed fecal samples from ulcerative colitis (UC) patients using a tandem mass spectrometry (LC-MS/MS) workflow coupled to ComPIL in search of aberrant UC-associated proteins. We found 176 host and microbial protein groups differentially enriched between the healthy (control) or UC volunteer groups. Notably, gene ontology (GO) enrichment analysis revealed that serine-type endopeptidases are overrepresented in UC compared to healthy volunteers. Additionally, we demonstrate the feasibility of serine hydrolase chemical enrichment from fecal samples using a biotinylated fluorophosphate (FP) probe. Our findings illustrate that probe-susceptible hydrolases from hosts and microbes are likely active in the distal gut. Finally, we applied de novo peptide sequencing methods to our metaproteomics data to estimate the size of the “dark peptidome,” the complement of peptides unidentified using ComPIL. We posit that our metaproteomics methods are generally applicable to future microbiota analyses and that our list of FP probe-enriched hydrolases may represent an important functionality to understanding the etiology of UC.
... Colitis in Il10 -/mice is characterized by colonic inflammation that is transmural but discontinuous, forming "skip lesions" similar to what is observed in humans with CD. Il10 -/mice with colitis also have an increased risk of dysplasia and invasive colon cancer relative to control mice [12,13,[16][17][18]. Aicda deficiency was previously reported to decrease inflammation-associated colon cancer in mice, based on the observation that adenocarcinoma had developed in 6 of 22 Il10 -/mice vs. 1 of 23 Il10 -/-Aicda -/mice by~1 year of age (p = 0.05; [19]). ...
Article
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Patients with inflammatory bowel disease (IBD) have an increased risk of colorectal cancer, particularly in ulcerative colitis (UC) when the majority of colon epithelial cells may be exposed to inflammation-associated mutagenesis. In addition to mutagenesis generated by oxidative stress, inflammation can induce activation-induced cytidine deaminase (Aicda), a mutator enzyme in the APOBEC family, within colon epithelial cells. This study tested the hypothesis that deletion of the Aicda gene could protect against the development of inflammation-associated colorectal cancers, using a model of UC-like colitis in “T/I” mice deficient in TNF and IL10. Results showed that T/I mice that were additionally Aicda-deficient (“TIA” mice) spontaneously developed moderate to severe UC-like colitis soon after weaning, with histologic features and colon inflammation severity scores similar those in T/I mice. Although the mean survival of TIA mice was decreased compared to T/I mice, multivariable analysis that adjusted for age when neoplasia was ascertained showed a decreased numbers of neoplastic colorectal lesions in TIA mice, with a trend toward decreased incidence of neoplasia. Aicda deficiency increased serum IL1α and slightly decreased IL12p40 and M-CSF, as compared with T/I mice, and led to undetectable levels of IgA, IgG1, IgG2a, IgG2b, and IgG3. Taken together, these studies show that Aicda deficiency can decrease the number of neoplastic lesions but is not sufficient to prevent the risk of inflammation-associated colorectal neoplasia in the setting of severe UC-like inflammation. The TIA model may also be useful for assessing the roles of antibody class-switch recombination deficiency and somatic hypermutation on regulation of microbiota and inflammation in the small intestine and colon, as well as the pathogenesis of colitis associated with hyper-IgM syndrome in humans. Further studies will be required to determine the mechanisms that drive early mortality in TIA mice.
... Some vegetables such as tomatoes, eggplants, and potatoes contain solanine, a glycoalkaloid, which was suggested to increase intestinal permeability and be detrimental for arthritogenic pathologies [21][22][23][24][25][26]. In contrast, other nutrients have been suggested to offer numerous health benefits [27], including long chain omega-3 polyunsaturated FA (PUFA) (chia seeds, flaxseeds, fatty fish) [28][29][30][31][32], monounsaturated FA (MUFA) (avocado, sesame) [33,34], antioxidants [35], phytochemicals [36], flavonoids [37,38], vitamin D [39], fruits with enzymatic proteins such as papain and bromelain (papaya, mango, pineapple) [40][41][42], ginger [43], turmeric [44,45], black pepper [46,47], green tea [48][49][50], and legumes [46,51]. However, only a few randomized double-blind placebo-controlled clinical trials have attempted to determine whether supplementation with these ingredients [31,34,35,37,38,43] or probiotics [52,53] are beneficial in RA patients. ...
Article
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Background Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease that affects synovial joints, leading to inflammation, joint destruction, loss of function, and disability. Although recent pharmaceutical advances have improved treatment of RA, patients with RA often inquire about dietary interventions to improve RA symptoms, as they perceive rapid changes in their symptoms after consumption of certain foods. There is evidence that some ingredients have pro- or anti-inflammatory effects. In addition, recent literature has shown a link between diet and microbiome changes. Both diet and the gut microbiome are linked to circulating metabolites that may modulate inflammation. However, evidence of the effects of an anti-inflammatory and probiotic-rich diet in patients with RA is scarce. There is also a need for biological data to support its anti-inflammatory effects. Methods The main goal of this study is to delineate the design process for a diet tailored to our RA population. To achieve this goal, we collected information on diet, supplements, cooking methods, and intake of different ingredients for each patient. Different groups were interviewed, and their feedback was assessed to design a diet that incorporates suggested anti-inflammatory ingredients in a manner that was easy for patients to adopt based on their lifestyles and backgrounds. Results We designed a diet that includes a high intake of potential anti-inflammatory ingredients. Feedback from highly motivated patients was critical in constructing an anti-inflammatory diet (ITIS diet) with elevated adherence. Conclusion In order to tailor our diet, we surveyed our patients on several different parameters. We obtained important feedback on how feasible our ITIS diet is for RA patients. Using this feedback, we made minor improvements and finalized the design of the ITIS diet. This diet is being used in an on-going pilot study to determine their anti-inflammatory effect in pain and joint swelling in RA patients.
... For a long time, proteolytic activity has been detected in fresh pineapple juice, whose main ingredient is bromelain, classified within the group of cysteine proteases (Hale et al., 2010). This enzyme group can modify the structure of the membrane of fat globules, releasing the oil and facilitating its extraction (Shah et al., 2005). ...
Article
Due high proportion of oil in Jatropha curcas seed (50–60%), it is interest to identify methods for their extraction. In addition, the residual paste is rich in proteins and could be used safely as raw matter for other processes. In the present work, we propose the use of a crude pineapple green extract (Ananas comosus L.) with a high content of enzymes with different proteolytic activities (1.18–5.90 IUT) to extract oil. A comparative extraction was made between the enzymatic treatment, a microwaves heat treatmet, the combination of both methods and the Soxhlet extraction. The oil yield with the crude extract, after 6 h of enzymatic hydrolysis was 46.9%, whereas after microwaving (3 min), the enzymatic hydrolysis treatment increased the yield to 54%. Oil was characterized physicochemically without showing difference between treatments. Fatty acids present in the J. curcas oil are the unsaturated fatty acids: oleic, linoleic, palmitoleic, as well as saturated ones like palmitic, stearic, and myristic. The use of enzymatic extracts combined with microwaves is a good alternative to improve the yields without altering the chemical and physical characteristics of the Jatropha oil.
... A recent meta-analysis has associated a high intake of fruit and vegetables with a reduction of UC in European population, suggesting that the intake of their bioactive compounds (fiber, antioxidant vitamins and phytochemicals such as polyphenols, carotenoids, isoflavones) could explain this inverse association [7]. In murine colitis models, which mimic human pathology, it has been demonstrated the preventive effect on pro-inflammatory intestinal process associated to the intake of mango [8] or pineapple [9] juices, polyphenols-rich extracts from orange [10] or its sub-products [11], apple [12] and pomegranate [13]. Polyphenols seems to be the main bioactive compounds involved in the anti-colitic action of fruits, due to its ability to inhibit some pivotal pro-inflammatory mediators as nuclear transcriptional Factor-κB (NF-κB) and specific cytokines (Tumor Necrosis Factor-α (TNF-α), and interleukin 1-β (IL-1β)) and the induction of antioxidant defence systems. ...
Article
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The potential anti-inflammatory effect of plant sterols (PS) enriched milk-based fruit beverages (PS, 1 g/100 mL) (MfB) with/without galactooligosaccharides (GOS, 2 g/100 mL) (MfB-G) in an experimental mice model of chronic ulcerative colitis was evaluated. Beverages were orally administered to mice every day by gavage to achieve PS and GOS doses of 35 and 90 mg/kg, respectively, and experimental colitis was induced by giving mice drinking water ad libitum containing 2% (w/v) dextran sulphate sodium (DSS) for 7 days, alternating with periods without DSS up to the end of the study (56 days). MfB beverage showed significant reduction of symptoms associated to ulcerative colitis and improved the colon shortening and mucosal colonic damage, but it was not able to reduce the increase of myeloperoxidase levels produced by DSS. MfB-G showed higher incidence of bloody feces and loss of stool consistency than MfB, as well as high levels of immune cells infiltration in colon tissue and myeloperoxidase. Therefore, PS-enriched milk-based fruit beverage could be an interesting healthy food to extend the remission periods of the diseases and the need to evaluate, in a pre-clinical model, the anti-inflammatory effect of the combination of bioactive compounds in the context of a whole food matrix.
... The efficacy of proteinases has been studied at least in three digestive disorders: ulcerative colitis, inflammatory bowel disease, and Crohn's disease. Oral bromelain was initially reported to reduce the severity of colon inflammation in a rodent model (Hale et al. 2005) and fresh pineapple juice decreases inflammation in IL-10deficient mice with colitis (Hale et al. 2010). Colon biopsies from patients with ulcerative colitis and Crohn's disease had decreased levels of inflammatory cytokines if treated with bromelain (Onken et al. 2008). ...
Book
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This book offers an overview of the diverse fields application of proteases (also termed proteolytic enzymes or proteinases), including food science and technology, pharmaceutical industries, and detergent manufacturing, reviewing the advances in the biotechnological application plant proteolytic enzymes over the last decade. In recent years, they have been the focus of renewed attention from the pharmaceutical and biotechnology industries, not only because of their activity on a wide variety of proteins but also because they are active over a range of temperatures and pHs. The main audience of this book are researchers working with plant proteases but also professionals from several industry segments such as food production and pharmaceutical companies.
... The efficacy of proteinases has been studied at least in three digestive disorders: ulcerative colitis, inflammatory bowel disease, and Crohn's disease. Oral bromelain was initially reported to reduce the severity of colon inflammation in a rodent model (Hale et al. 2005) and fresh pineapple juice decreases inflammation in IL-10deficient mice with colitis (Hale et al. 2010). Colon biopsies from patients with ulcerative colitis and Crohn's disease had decreased levels of inflammatory cytokines if treated with bromelain (Onken et al. 2008). ...
Chapter
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Proteases or proteinases are enzymes that catalyze cleavage of proteins at peptide bonds generating smaller peptides. Some of them are very specific in their choice of target site while others act rather nonspecifically and hydrolyze the protein substrate if conditions allow into short peptides. They must have appeared early in evolution along with proteins, to keep a balance between synthesis and protein degradation. Their early emergence is confirmed by their ubiquitous presence in most living forms including viruses, plants, and animals.
... The results suggested that the plant had antibacterial activity, antioxidant activity and anthelmintic activity. Hale et al., (2010) studied dietary supplementation with fresh pineapple juice decreases inflammation and colonic neoplasia in IL-10-deficient mice with colitis. The results demonstrated that long-term dietary supplementation with fresh or unpasteurized frozen pineapple juice with proteolytically active bromelain enzymes is safe and decreases inflammation severity. ...
Experiment Findings
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... 14 Pineapple juice is safe and decreases inflammation severity, incidence and multiplicity of inflammation-associated colonic neoplasia. 15 However, consumption of healthy diets such as fruits, fruit juices, acidic diet, and wine may as well result in surface damage, erosion, and decrease hardness of the restorative materials by their acidity. [16][17][18][19] Moreover, acidity can be an essential component in certain sour food. ...
Article
Objective: To evaluate surface roughness, hardness, and morphology changes of various bulk-fill resin composites eroded by different food-simulating liquids and beverages. Materials and methods: One hundred and thirteen specimens were fabricated in polytetrafluoroethylene cylindrical mold (10 mm in diameter and 4 mm in thickness). Before immersion, baseline data of roughness, Vicker's microhardness were recorded and surface characteristics were examined using scanning electron microscopy (SEM). Each product of specimens (SDR, Dentsply; SonicFill, Kerr; Tetric N-Ceram Bulk Fill, Ivoclar Vivadent AG; and Filtek Bulk Fill, 3M ESPE) were divided into 5 groups for spicy and sour soup, spicy soup (Tom Yum), pineapple juice, passionfruit juice, and deionized water (served as a control). Specimens were then alternately immersed in storage agents for 5 seconds and artificial saliva for 5 seconds over 10 cycles. Specimens were stored in artificial saliva for 22 hours. This process was repeated for 28 days. After immersion, surface hardness and roughness of specimens were evaluated at 7, 14, 21, and 28 days and data were analyzed by two-way repeated ANOVA and Tukey's HSD (α = 0.05). Surface morphology of specimens was also examined on day 28. Results: The SDR group had the most statistically significant decrement in hardness (25.65 ± 1.74 kg/mm(2) in mean difference) and increment in roughness (0.26 ± 0.10 μm in mean difference; P < .05). Passionfruit juice caused the most surface changes in bulk-fill resin composites. SEM photomicrographs showed surface changes of all resin composites in varying degrees. Conclusion: Acidic food-simulating liquids and beverages significantly increased the surface roughness and decreased surface microhardness of bulk-fill resin composites after evaluation at the end of the 28-day immersion period. Clinical significance: For restoration of the affected teeth in patients who consume acidic food and beverages, roughness and erosion of resin composites should be considered. All of bulk-fill resin-based composites except SDR may be suitable for restorations in these patients.
... This included interferon g, and TNF-a, but not IL1-b or IL6. In IL10-deficient mice, long term supplementation with pineapple juice or bromelain decreased histological markers of inflammation (Hale et al., 2010;Hale et al., 2005), which is dependent on its proteolytic activity. In addition, bromelain is also absorbed in the human gastrointestinal tract without loss of its proteolytic activity (Castell et al., 1997;Chobotova et al., 2010), and may cause its effects systemically. ...
Article
Kiwifruit are recognized as providing relief from constipation and symptoms of constipation-predominant irritable bowel syndrome (IBS-C). However, the underlying mechanisms, specifically in regards to gastrointestinal transit time and motility, are still not completely understood. This review provides an overview on the physiological and pathophysiological processes underlying constipation and IBS-C, the composition of kiwifruit, and recent advances in the research of kiwifruit and abdominal comfort. Additionally, gaps in the research are highlighted and scientific studies of other foods with known effects on the gastrointestinal tract are consulted to find likely mechanisms of action. While the effects of kiwifruit fiber are well documented, observed increases in gastrointestinal motility caused by kiwifruit are not fully characterized. There are a number of identified mechanism that may be activated by kiwifruit compounds, such as the induction of motility via protease-activated signaling, modulation of microflora, changes in colonic methane status, bile flux, or mediation of inflammatory processes.
... Bromelain notably reduce the harshness of colonic inflammation and when taken orally, it significantly reduces the severity of ulcerative colitis . The chief anti-inflammatory mechanism of bromelain comes out to be proteolytic in nature by which it eradicates cell surface receptors involved in leukocyte defection and activation (Hale et al. 2010). Bromelain also vary the emission of certain chemokines and thus reduces the prevalence and sternness of spontaneous colitis. ...
Article
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Bromelain is a concoction of sulfhydryl proteolytic enzymes. Depending upon the site of extraction it can be regarded as either stem bromelain (SBM) (EC 3.4.22.32) or fruit bromelain (FBM) (EC 3.4.22.33). Bromelain remain enzymatic active over a broad spectrumand endure a range of pH (5.5 to 8.0) and temperature (35.5 to 71 ºC). It is one of the extensively investigated proteolytic enzyme owing to its astonishing applications in various industries. This necessitated employing a strategy that result in highest purified bromelain in less steps and lowest cost. Use of modernistic approach such as membrane filtration, reverse micellar systems, aqueous two phase extraction and chromatographic techniques have shown promise in this regard. Besides its industrial applications, bromelain has been widely utilized as a potential phytomedical compound. Some of its reported actions include inhibition of platelet aggregation, anti-edematous, anti-thrombotic, anti-inflammatory, modulation of cytokines and immunity, skin debridement and fibrinolytic activity. It also assist digestion, enhance absorption of other drugs and is a potential postoperatively agent that promote wound healing and reduce postsurgical discomfort and swelling.
... To understand the pathogenesis of IBD, several animal models based on genetic modification have been developed, such as interleukin-10deficient (IL-10 (−/−)) mice [4]. These mice have defects in immune regulation and spontaneously develop chronic enterocolitis and adenocarcinoma through the dysplasia sequence, a process similar to clinical IBD-associated cancer [5]. ...
Article
Background: Interleukin-10-deficient (IL-10 (-/-)) mice spontaneously develop chronic colitis and adenocarcinoma through the dysplasia sequence. Autophagy malfunction is associated to inflammatory bowel disease (IBD) and colorectal cancer (CRC) pathogenesis. Autophagy is regulated by silent information regulator-1 (SIRT1), a NAD+-dependent histone deacetylase. Our aim was to investigate the expression changes of SIRT1-AMPK-autophagy pathway in the progression from chronic colitis to CRC. Methods: We studied C57BL/6-IL-10-deficient mice between 6 and 18weeks of age. Macroscopic and histological analysis, and characterization of inflammatory and tumor biomarkers were performed. Results: IL-10-deficient mice developed colitis from the age of 6weeks onward. The severity of inflammation and dysplasia, and the proliferative activity increased gradually with age. IL-10 (-/-) mice were characterized by improved levels of TNF-α and decreased expression of SIRT1. Moreover, our findings show an increase in p-AMPK expression and an activation of the autophagy in IL-10 (-/-) mice from all stages, evidenced by the accumulation of LC3-II protein, the increase in Beclin 1 expression and the reduction in Bcl-2 levels. Conclusions: SIRT1-AMPK-autophagy pathway may be involved in the maintenance of chronic inflammation and dysplasia development in the IL-10-deficient mice model. Modulation of this pathway could be a novel strategy for IBD and CRC treatment.
... Bromelain enzymes present in fresh pineapple juice have anti-inflammatory activities. Hale et al. [47] reported that long-term dietary supplementation with fresh or unpasteurized frozen pineapple juice with proteolytically active bromelain enzymes was safe and decreased inflammation severity and the incidence and multiplicity of inflammation-associated colonic neoplasia in murine model of inflammatory bowel disease. Fresh pineapple juice has healing power against acute tendon injuries. ...
... The vitamin contents of plain flavoured and spiced yoghurts and 0.06) and niacin (0.01, 0.01 and 0.24) contents above that the observed increased in the vitamin contents could be a justification of the report that carrot (Cohen et al., 2002) pepper fruit (Ihemeje et al., 2013) and pineapple (Hale et al;2010) are all rich in vitamin .similar trend of increase in vitamin content of flavoured and spiced yoghurts was respec-tively recorded by Amna et al. (2008) and Mbaeyi and Anyanwu (2010) The pH of the various yoghurt sample are presented in Table 3 result indicates that pineapple flavoured by the plain yoghurt (4.6) pepper fruit spiced yoghurt, ginger spiced yoghurt and carrot flavoured yoghurt had the same pH (4.7). ...
Article
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Production and quality evaluation of plain yoghurt, spiced yoghurt (pepper fruit, ginger) and flavoured yoghurt (carrot and pineapple) were carried out being proximate composition, mineral analysis, microbiological analysis, organoleptic evaluation and statistical analysis. Results show significant (p<0.05) nutritional enhancement of the plain yoghurt by the addition of spices and flavourings. The mineral content of the plain yoghurt were likewise increased. Organoleptically, the spiced and flavoured yoghurts were all acceptable by consumers but pepper fruit spiced yoghurt was the most preferred in terms of general acceptability. Key words: Yoghurt, pepper fruit, carrot and pineapple.
... Methylcobalamin, Acetyl-L-Carnitine, N-Acetyl Cysteine, Vitamin D, and Curcumin improve nerve regeneration in a number of conditions including nerve injuries (crush), nerve repair after transection and neuropathic conditions such as diabetes. Curcumin, Bromelain, and Serratopeptidase decrease the inflammatory response and have demonstrated improved recovery after surgery with less pain and earlier return to activity[55][56][57][58][59][60][61][62][63][64][65][66][67]. ...
... It is a good source of many nutrients such as vitamins B2 (riboflavin), C (ascorbic acid), manganese, and phytochemicals [4]. Consumption of pineapple and its components has been linked with immunity [5][6][7][8][9][10][11][12][13][14][15][16][17][18]. Some researchers also associated pineapple with cancer [19][20][21][22][23], wound healing [24][25][26], diabetes mellitus and liver disorders [27][28][29][30][31], diarrhea or other disorders of the gastrointestinal tract [32,33], and even tuberculosis [34]. ...
Article
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This randomized, controlled trial examined the effects of canned pineapple consumption on immunomodulation, nutritional status, and physical health of ninety-eight (98) school children with mean age of 8.44 ± 0.20. The study participants were divided into three groups: Group A (33) includes subjects who were not given canned pineapple, Group B (33) includes those who were given 140 g, and Group C (32) includes those given 280 g of canned pineapple for nine weeks. Each major group was further divided into two groups: normal (N) and underweight (U) based on 2007 WHO Growth Reference Standards. Sociodemographic, anthropometric, physical examination, dietary intake, hemoglobin level, and immunological data were analyzed. Results showed a decrease in incidence of viral and bacterial infections for both Group B and Group C (normal and underweight) after canned pineapple consumption. Granulocyte production increased by 0.77–26.61% for normal weight subjects and 14.95–34.55% for underweight. CD16+56 count augmented by 20.44–22.13% for normal weight and 3.57–15.89% for underweight subjects. Thus, intake of both one can (140 g) and two cans (280 g) of canned pineapple may shorten the duration and incidence of infection and may increase the production of granulocytes and CD16+56, but intake of two cans (280 g) demonstrated higher granulocyte and CD16+56 production. This trial is registered with Philippine Health Research Registry: PHRR140826-000225.
... Bromelain (pineapple enzyme, Ananas comasus) is an aqueous extract obtained from the stem and fruit of the pineapple plant that contains high levels of proteolytic enzymes and which composition varies depending on the source and purification method (1,2). Bromelain directly influences pain mediators such as bradykinin (3), although its analgesic properties are closely linked to its anti-inflammatory properties (4,5). ...
Article
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Objectives: To evaluate the anti-inflammatory and analgesic effect of Bromelain (pineapple extract) administered orally in the postoperative after extraction of impacted lower molars. Study Design: This is a prospective, placebo-controlled, unicentric, double-blind study; the sample size was 34 patients. The pre and postoperative outcomes, evaluated on the third (D3) and eighth day (D8), included inflamtion, pain and oral aperture, as well as the need for analgesics. One group received bromelain 150mg per day for three days and 100mg on days 4 to 7. The other group received placebo in the same dosage. All outcomes werrecorded quantitatively and analyzed with the Mann-Whitney U test for independent samples. Results: Although there were no statistically significant differences between the treatment groups, a trend towards less inflammation and improved oral aperture was observed in the group that received bromelain, compared to the group that received placebo. This trend can be attributed completely to random reasons, since there is no statistical difference in the results. Conclusions: Further studies are necessary to analyze different administration patterns and doses of bromelain for the use in the postoperative of impacted third molars. Key words:Tooth extraction, third molar, postoperative period, bromeline, clinical study.
... to the site of inflammation [123, 125]. In a recent study has shown that long-term dietary supplements with fresh pineapple juice decrease inflammation severity and multiplicity of inflammation-associated colonic neoplasia in the IL-10 −/− mice model of colitis [126] . These reports focus our attention on bromelain as a natural compound that can be developed as an effective therapy to IBD. ...
Article
Full-text available
Inflammatory bowel disease (IBD) can be divided into two major categories, ulcerative colitis (UC) and Crohn disease (CD). While the main cause(s) of IBD remain unknown, a number of interventional and preventive strategies have been proposed for use against CD and UC. Many reports have focused on the use of alternative natural medicines as potential therapeutic interventions in IBD patients with minimal side effects. While the use of alternative medicines may be effective in IBD patients that are refractory to corticosteroids or thiopurins, alternative treatment strategies are limited and require extensive clinical testing before being optimized for use in patients.
... bioactive phytochemicals to the body. Long-term dietary supplementation with pineapple juice decreased colon inflammation in mice [42], while dried plum was able to reduce atherosclerotic lesion in apolipoprotein E-deficient mice [43]. Bromelain, contained in pineapple [44] and anthocyanin [45] contained in blackcurrant have also been shown to inhibit TNF-and IL-6 secretion in vitro. ...
Article
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An emerging role of IL-17 in the inflammatory response associated with pathogenesis of neurodegeneration has been recently suggested. However, though diet represents a key factor in the modulation of inflammatory processes, evidence is not currently available on the nutritional regulation of IL-17 in humans. In a double blind, randomized, placebo controlled, crossover study, we investigated the effect of High Fat Meal (HFM) on IL-17 circulating levels in presence of a placebo (HFM-P) or with a Fruit Juice Drink (HFM-FJD) composed of pineapple, blackcurrant and plum in fourteen healthy overweight humans. Fasting in the morning subjects ingested a test meal providing 1344 Kcal. Ingestion of HFM-P induced an inflammatory response mediated by TNF-α (p < 0.001), IL-6 (p < 0.001) and IL-17 (p < 0.01). Plasma IL-17 concentration significantly increased at 1 h (+2.6 ± 1.1 pg/ml), remaining high at 4 h (+2.98 ± 1.2 pg/ml), 6 h (+2.38 ± 0.6 pg/ml) and 8 h (+2.8 ± 0.9 pg/ml) (ANOVA for time-course p=0.009). When the HFM was consumed in the presence of the FJD a marked inhibition of IL-17 response to the HFM was observed (ANOVA between treatment p=0.037). We provided, for the first time, evidence on the role of diet in modulating IL-17 production in healthy overweight subjects.
... The disease pathophysiology in these mice is similar to human CD, with the loss of intestinal mucosal tolerance to commensal bacteria, infiltration of activated macrophages, and increased differentiation of Th1 and Th17 cells [9,10]. The Il10 À/À mouse has been used for the investigation of in vivo anti-inflammatory effects of several food components [11][12][13][14][15]. However, the appropriate choice of food components for testing is necessary. ...
Preprint
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The use of dietary components as new antineoplastic agents has been highlighted due to their high biological activity against tumor cells, chemopreventive effects and relatively low toxicity. The present study evaluated the antitumor effects and chemoprotective potential of the bromelain enzyme complex (BL) alone and in combination with the chemotherapy drug doxorubicin (DOX) using the following in vitro tests: cell viability by Alamar blue in cells: AGP01, SKMEL-103 and CAL-27 lines; MTT assay and fluorescent labeling in murine sarcoma 180 (S-180); and the comet assay in human lymphocytes. The results showed cytotoxic effects of BL with IC50 (µg/mL) of 124.80 (AGP-01), 91.81 (SKMEL-103), 95.75 (CAL-27) and 25.27 (S-180). When incubated with the BL + DOX combination, the IC50s were (in µg/mL) 26.29 (AGP-01), 29.04 (SKMEL-103), 2.68 (CAL-27) and 6.11 (S -180), demonstrating combination indices ranging from synergistic to antagonistic. When evaluating the mechanism of cell death in S-180, an increase in the number of cells undergoing early apoptosis was observed after incubation with BL (100 µg/mL). In genotoxicity assays, isolated BL was not genotoxic in human lymphocytes, unlike DOX, which showed this activity. When combined (BL + DOX), BL modulated the DNA damage caused by the antineoplastic agent when compared to DOX alone, with ID values of 55.91% and FD of 33.65%, showing chemoprotective potential in this case. In conclusion, isolated BL exhibited antiproliferative effects on tumor cell lines and was not genotoxic to human blood cells, with positive prospects for its use in combination with DOX chemotherapy.
Article
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Bromelain is a key enzyme found in pineapple (Ananas comosus (L.) Merr.); a proteolytic substance with multiple beneficial effects for human health such as anti-inflammatory, immunomodulatory, antioxidant and anticarcinogenic, traditionally used in many countries for its potential therapeutic value. The aim of this updated and comprehensive review focuses on the potential anticancer benefits of bromelain, analyzing the cytotoxic, apoptotic, necrotic, autophagic, immunomodulating, and anti-inflammatory effects in cancer cells and animal models. Detailed information about Bromelain and its anticancer effects at the cellular, molecular and signaling levels were collected from online databases such as PubMed/MedLine, TRIP database, GeenMedical, Scopus, Web of Science and Google Scholar. The results of the analyzed studies showed that Bromelain possesses corroborated pharmacological activities, such as anticancer, anti-edema, anti-inflammatory, anti-microbial, anti-coagulant, anti-osteoarthritis, anti-trauma pain, anti-diarrhea, wound repair. Nonetheless, bromelain clinical studies are scarce and still more research is needed to validate the scientific value of this enzyme in human cancer diseases.
Article
Individuals with inflammatory bowel disease are at high risk of developing colitis-associated cancer; thus, strategies to inhibit disease progression should be investigated. The study aimed to explore the role of the synbiotic (probiotic VSL#3® and yacon-based concentrate) in a colitis-associated carcinogenesis model. IL-10−/− mice were induced to carcinogenesis with 1,2-dimethylhydrazine and divided into two experimental groups: control and synbiotic. Manifestations of colitis, colon histology, expression of antioxidant enzymes, production of organic acids and intestinal microbiota were evaluated. The use of the synbiotic showed benefits, such as the preservation of intestinal architecture, increased expression of antioxidant enzymes and the concentration of organic acids, especially butyrate. It was also observed different microbial community profiles between the groups during the study. Together, these factors contributed to mitigate the manifestations of colitis and improve intestinal integrity, suggesting the potential benefit of the synbiotic in intestinal diseases.
Article
Background: Chronically higher inflammation, which may partly result from diet and lifestyle, is implicated in risk for multiple chronic diseases. The dietary inflammatory index (DII) and empirical dietary inflammatory pattern (EDIP), developed to characterize dietary contributions to systemic inflammation, have several limitations. There are no scores to characterize contributions of lifestyle to inflammation. Objectives: To reflect dietary/lifestyle contributions to inflammation, we developed novel, inflammation biomarker panel-weighted, dietary (DIS) and lifestyle (LIS) inflammation scores in a subset (n = 639) of the Reasons for Geographic and Racial Differences in Stroke Study (REGARDS) cohort. Methods: We selected a priori 19 food groups and 4 lifestyle characteristics to comprise the DIS and LIS, respectively. We calculated the components' weights based on their strengths of association with an inflammation biomarker score [comprising high-sensitivity C-reactive protein (hsCRP), IL-6, IL-8, and IL-10] using multivariable linear regression. The sums of the weighted components constitute the scores, such that higher scores reflect, on balance, more proinflammatory exposures. We calculated the DIS, LIS, DII, and EDIP with cross-sectional data from the remaining REGARDS cohort ( n = 14,210 with hsCRP measurements) and 2 other study populations with hsCRP and/or an 8-component inflammation biomarker panel, and investigated their associations with circulating inflammation biomarker concentrations using multivariable logistic regression. Results: In REGARDS, those in the highest relative to the lowest DIS, LIS, DII, and EDIP quintiles had statistically significant 1.66-, 4.29-, 1.56-, and 1.32-fold higher odds of a high hsCRP concentration (>3 mg/dL), respectively (all P-trend < 0.001). Those in the highest relative to the lowest joint DIS/LIS quintile had a statistically significant 7.26-fold higher odds of a high hsCRP concentration. Similar findings were noted in the other 2 validation populations. Conclusion: Our results support that dietary and lifestyle exposures collectively contribute substantially to systemic inflammation, and support the use of our novel DIS and LIS.
Article
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The UV-protective activities of pineapple leaf extract in a zebrafish model have been evaluated by recording fin morphological changes. UV protective effects of pineapple leaf extract was evaluated using Kaplan–Meier analysis and Cox proportional hazards regression. The results showed that (1) the mean time of return to normal fin in the UV + pineapple leaf extract (100 and 400 μg/mL) groups are 3.90 and 2.50 days, respectively, more quickly than in the UV only group (4.12 days); (2) zebrafish fins in the pineapple leaf extract (100 and 400 μg/mL) groups are 1.44 and 4.29 times, respectively, more likely to return to recover than those in the UV only group; and (3) pineapple leaf extract has UV-absorbance abilities and significantly reduces p53 expressions and ROS generation in UV-exposed zebrafish embryos, which may attenuate UV-mediated apoptosis.
Article
Background of Study: Herbs as Indonesia’s cultural heritage have been used hereditary in many generations. One of them is temulawak (Curcuma xanthorrhiza Roxb). The immunomodulator (Immunostimulant) is a compound that can boost the immunity mechanism of the body, specifically as well as non-specifically. Rahman (2007) stated that temulawak is taken from rhizomes which consists of 64% starch, 1,6 – 22% curcumin and 1,48 – 1,63 % essential oil that believed to be able to improve the kidney’s work and to serve as anti-inflammatory. Because of these many benefits of temulawak and the phenomena of drinking herbs habits by postpartum women particularly in Javanese culture, then the researchers were encouraged to do a research on temulawak as a breast milk booster. Objective: To study the effect of giving temulawak on the breast milk production of postpartum mothers at independent midwife NINGSIH with the indicator of the babies’ weight gain, urination frequency, breastfeeding frequency and the length of babies’ sleep after being breastfed. Method: The research is a quasi-experiment and the research design is a Static Group Comparison in which observation method is applied. The population of this research is women in their postpartum time until the 14-day period after childbirth in Ningsih independent midwife during the research. The sample is all the qualified population who meets the requirement. The technique of the sampling is purposive sampling. The independent variable is the use of temulawak. The dependent variable is the milk production in breastfeeding in 14-day period after childbirth. The T-test is used for analyzing. The Result of the Study:The result of the study showed that the weight of the babies in the experimental group gained 535 grams in average and in the control group gained 270 in average. The urination frequency of the babies in the experimental group was 7.5 times in average and in the control group was 5.05 times. The breastfeeding frequency in the experimental group was 9.35 times in average and in the control group was 6.85 in average. After being breastfed, the babies in the experimental group spent 1.93 hours of sleeping in average, whereas the babies in the control group spent less than normal. In conclusion, there was only 1 person (5%) in the control group who had a good production of breast milk whereas in the experimental group, there were 14 persons (70%). Therefore, it can be concluded from the study that there was a significant effect on the use of temulawak to the production of breast milk of postpartum mothers with p value of 0,000 and alpha value of 0,05. Conclusion : There is a significant effect on the use of temulawak in increasing milk production in lactation in postpartum period
Article
This study was aimed to investigate the effects of bromelain on angiogenesis, nitric oxide, and matrix metalloproteinase-3 and -9 in rats exposed to 1200mV electrical burn injury. Thirty-five, male Wistar albino rats was divided into five groups (n=7 each), including control (untreated) group; electrical burn injury (EI) group; electrical burn injury+21.25mg/kg BW bromelain group (EIB1); electrical burn injury+42.50mg/kg BW bromelain group (EIB2); and electrical burn injury+85.00mg/kg BW bromelain group (EIB3). Rat models of electrical burns done by providing electricity in rats that had been anesthetized, a voltage of 1200mV and strong currents 15mA for 10s. The VEFG, NO, and MMP-9 levels were significantly greater in the EI group compared to the untreated group. Out of the 21.25mg/kg BW, 42.50mg/kg BW, and 85mg/kg BW doses of Bromelain extract, only the lowest doses prevented EI-induced increase in NO and MMP-9 level (P<0.05). Bromelain at the lowest dose (21.25mg/kg BW) act as anti-inflammatory and modulate the matrix metalloproteinase-9 in rats exposed to 1200mV electrical burn injury.
Article
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Bromelain is a cysteine protease enzyme present in the plants. In this study we isolated the enzyme from pineapple plant parts like leaves, green fruit, ripe fruits and immobilized it by using sodium alginate. We observed the effects of different activators and inhibitors, kinetic properties and different concentrations of substrate and estimated enzyme activity on free and immobilized bromelain from pineapple plant parts with hemoglobin as substrate. The protease enzyme showed an effective temperature of 40-60 o C and optimal temperature of 50-60 0 C, effective p H of 4.0-8.0 and optimal p H is 4.5-5.5. These all properties of enzymes will be useful in keeping enzyme in an active state in various industrial and medical preparations.
Article
In vitro and animals models have long been used to study human diseases and identify novel therapeutic approaches that can be applied to combat these conditions. Ulcerative colitis and Crohn's disease are the two main entities of inflammatory bowel disease (IBD). There is an intricate relationship between IBD features in human patients, in vitro and animal colitis models, mechanisms and possible therapeutic approaches in these models, and strategies that can be extrapolated and applied in humans. Malnutrition, particularly protein-energy malnutrition and vitamin and micronutrient deficiencies, as well as dysregulation of the intestinal microbiota, are common features of IBD. Based on these observations, dietary supplementation with essential nutrients known to be in short supply in the diet in IBD patients and with other molecules believed to provide beneficial anti-inflammatory effects, as well as with probiotic organisms that stimulate immune functions and resistance to infection has been tested in colitis models. Here we review current knowledge on nutritional and probiotic supplementation in in vitro and animal colitis models. While some of these strategies require further fine-tuning before they can be applied in human IBD patients, their intended purpose is to prevent, delay or treat disease symptoms in a non-pharmaceutical manner.
Article
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Infection with Helicobacter species is endemic in many animal facilities and may alter the penetrance of inflammatory bowel disease (IBD) phenotypes. However, little is known about the relative pathogenicity of H. typhlonius, H. rodentium, and combined infection in IBD models. We infected adult and neonatal IL10-/- mice with H. typhlonius, H. rodentium, or both bacteria. The severity of IBD and incidence of inflammation-associated colonic neoplasia were assessed in the presence and absence of antiHelicobacter therapy. Infected IL10-/- mice developed IBD with severity of noninfected (minimal to no inflammation) < H. rodentium < H. typhlonius <mixed H. rodentium + H. typhlonius (severe inflammation). Inflammation-associated colonic neoplasia was common in infected mice and its incidence correlated with IBD severity. Combined treatment with amoxicillin, clarithromycin, metronidazole, and omeprazole eradicated Helicobacter in infected mice and ameliorated established IBD in both infected and noninfected mice. Infection of IL10-/- mice with H. rodentium, H. typhlonius, or both organisms can trigger development of severe IBD that eventually leads to colonic neoplasia. The high incidence and multiplicity of neoplastic lesions in infected mice make this model well-suited for future research related to the development and chemoprevention of inflammation-associated colon cancer. The similar antiinflammatory effect of antibiotic therapy in Helicobacter-infected and -noninfected IL10-/- mice with colitis indicates that unidentified microbiota in addition to Helicobacter drive the inflammatory process in this model. This finding suggests a complex role for both Helicobacter and other intestinal microbiota in the onset and perpetuation of IBD in these susceptible hosts.
Article
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The pineapple plant (Ananas comosus) was shown to contain at least four distinct cysteine proteinases, which were purified by a procedure involving active-site-directed affinity chromatography. The major proteinase present in extracts of plant stem was stem bromelain, whilst fruit bromelain was the major proteinase in the fruit. Two additional cysteine proteinases were detected only in the stem: these were ananain and a previously undescribed enzyme that we have called comosain. Stem bromelain, fruit bromelain and ananain were shown to be immunologically distinct. Enzymic characterization revealed differences in both substrate-specificities and inhibition profiles. A study of the cysteine proteinase derived from the related bromeliad Bromelia pinguin (pinguinain) indicated that in many respects it was similar to fruit bromelain, although it was found to be immunologically distinct.
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Human peripheral blood mononuclear cells from healthy donors were treated ex vivo with the proteolytic enzyme bromelain and studied by flow cytometry. Bromelain-treated lymphocytes exhibited 60-90% reduced cell surface staining for CD44 and CD62-L molecules. While the staining for molecules CD16, CD56 and CD49d was unaffected, a moderate increase (10-40%) in expression of the beta(2)-integrins CD11a-c was seen. This selective modulation of cell adhesion molecules (CAM) was seen on T cells and NK cells, as well. The selective modulation of CAM may help explain some of the clinical effects observed after bromelain treatment in patients suffering from chronic inflammatory disease, HIV and cancer.
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Cell adhesion molecules are considered to be pivotal elements required for proper embryo development. The transmembrane glycoprotein CD44, which is expressed in numerous splice variants on the surface of many different cell types and tissues, has been suggested to be involved in several physiological processes such as cell-cell interactions, signal transduction, and lymphocyte homing and trafficking during embryogenesis and in the adult organism. Some splice variants are thought to play an important role in tumor progression. To investigate the physiological roles of CD44 in vivo, we abolished expression of all isoforms of CD44 in mice by targeted insertion of a lacZ/neo cassette into the reading frame of the leader peptide. CD44-deficient mice are viable without obvious developmental defects and show no overt abnormalities as adults. However, CD44-deficient lymphocytes exhibit impaired entry into the adult thymus, although lymphocyte development is apparently unaltered. Our data indicate that all splice variants of CD44 are dispensable for embryonic development and implicate a critical function for CD44 in lymphocyte recirculation.
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Bromelain, a mixture of cysteine proteases from pineapple stems, blocks signaling by the mitogen-activated protein (MAP) kinases extracellular regulated kinase 1 (ERK-1) and ERK-2, inhibits inflammation, and protects against enterotoxigenic Escherichia coli infection. In this study, we examined the effect of bromelain on Salmonella enterica serovar Typhimurium infection, since an important feature of its pathogenesis is its ability to induce activation of ERK-1 and ERK-2, which leads to internalization of bacteria and induction of inflammatory responses. Our results show that bromelain dose dependently blocks serovar Typhimurium-induced ERK-1, ERK-2, and c-Jun NH(2)-terminal kinase (JNK) activation in Caco-2 cells. Bromelain also blocked signaling induced by carbachol and anisomycin, pharmacological MAP kinase agonists. Despite bromelain inhibition of serovar Typhimurium-induced MAP kinase signaling, it did not prevent subsequent invasion of the Caco-2 cells by serovar Typhimurium or alter serovar Typhimurium -induced decreases in resistance across Caco-2 monolayers. Surprisingly, bromelain also did not block serovar Typhimurium-induced interleukin-8 (IL-8) secretion but synergized with serovar Typhimurium to enhance IL-8 production. We also found that serovar Typhimurium does not induce ERK phosphorylation in Caco-2 cells in the absence of serum but that serovar Typhimurium-induced invasion and decreases in monolayer resistance are unaffected. Collectively, these data indicate that serovar Typhimurium-induced invasion of Caco-2 cells, changes in the resistance of epithelial cell monolayers, and IL-8 production can occur independently of the ERK and JNK signaling pathways. Data also confirm that bromelain is a novel inhibitor of MAP kinase signaling pathways and suggest a novel role for proteases as inhibitors of signal transduction pathways in intestinal epithelial cells.
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Bromelain is a complex mixture of proteinases typically derived from pineapple stem. Similar proteinases are also present in pineapple fruit. Beneficial therapeutic effects of bromelain have been suggested or proven in several human inflammatory diseases and animal models of inflammation, including arthritis and inflammatory bowel disease. However, it is not clear how each of the proteinases within bromelain contributes to its anti-inflammatory effects in vivo. Previous in vivo studies using bromelain have been limited by the lack of assays to control for potential differences in the composition and proteolytic activity of this naturally derived proteinase mixture. In this study, we present model substrate assays and assays for cleavage of bromelain-sensitive cell surface molecules can be used to assess the activity of constituent proteinases within bromelain without the need for biochemical separation of individual components. Commercially available chemical and nutraceutical preparations of bromelain contain predominately stem bromelain. In contrast, the proteinase activity of pineapple fruit reflects its composition of fruit bromelain>ananain approximately stem bromelain. Concentrated bromelain solutions (>50 mg/ml) are more resistant to spontaneous inactivation of their proteolytic activity than are dilute solutions, with the proteinase stability in the order of stem bromelain>fruit bromelain approximately ananain. The proteolytic activity of concentrated bromelain solutions remains relatively stable for at least 1 week at room temperature, with minimal inactivation by multiple freeze-thaw cycles or exposure to the digestive enzyme trypsin. The relative stability of concentrated versus dilute bromelain solutions to inactivation under physiologically relevant conditions suggests that delivery of bromelain as a concentrated bolus would be the preferred method to maximize its proteolytic activity in vivo.
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Background & Aims: Interleukin (IL)-10 is an anti-inflammatory and immune regulatory cytokine. IL-10–deficient mice (IL-10−/−) develop chronic inflammatory bowel disease (IBD), indicating that endogenous IL-10 is a central regulator of the mucosal immune response. Prostaglandins are lipid mediators that may be important mediators of intestinal inflammation. In this study we assessed the role of prostaglandins in the regulation of mucosal inflammation in the IL-10−/− mouse model of IBD. Methods: Prostaglandin (PG) synthesis was inhibited with nonselective or cyclooxygenase (COX)-isoform selective inhibitors. Severity of inflammation was assessed histologically. Cytokine production was assessed by ribonuclease protection analysis and enzyme-linked immunosorbent assay. PGE2 levels were assessed by enzyme immunoassay. COX-1 and COX-2 expression was assessed by Western blot analysis. Results: Nonsteroidal anti-inflammatory drug (NSAID) treatment of wild-type mice had minimal effect on the colon. In contrast, NSAID treatment of 4-week-old IL-10−/− mice resulted in rapid development of colitis characterized by infiltration of the lamina propria with macrophages and interferon gamma–producing CD4+ T cells. Colitis persisted after withdrawal of the NSAID. NSAID treatment decreased colonic PGE2 levels by 75%. Treatment of IL-10−/− mice with sulindac sulfone (which does not inhibit PG production) did not induce colitis whereas the NSAID sulindac induced severe colitis. COX-1– or COX-2–selective inhibitors used alone did not induce IBD in IL-10−/− mice. However, the combination of COX-1– and COX-2–selective inhibitors did induce colitis. Conclusions: NSAID treatment of IL-10−/− mice results in the rapid development of severe, chronic IBD. Endogenous PGs are important inhibitors of the development of intestinal inflammation in IL-10−/− mice.
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latory effects. In this study, we show for the first time that extracellular proteases may also block signal transduction. We show that bromelain, a mixture of cysteine proteases from pineapple stems, blocks activation of ERK-2 in Th0 cells stimulated via the TCR with anti-CD3e mAb, or stimulated with combined PMA and calcium ionophore. The inhibitory activity of bromelain was dependent on its proteolytic activity, as ERK-2 inhibition was abrogated by E-64, a selective cysteine protease inhibitor. However, inhibitory effects were not caused by nonspecific proteolysis, as the protease trypsin had no effect on ERK activation. Bromelain also inhibited PMA-induced IL-2, IFN-g, and IL-4 mRNA accumulation, but had no effect on TCR-induced cytokine mRNA production. This data suggests a critical requirement for ERK-2 in PMA-induced cytokine production, but not TCR-induced cytokine production. Bromelain did not act on ERK-2 directly, as it also inhibited p21ras activation, an effector molecule upstream from ERK-2 in the Raf-1/MEK/ERK-2 kinase signaling cascade. The results indicate that bromelain is a novel inhibitor of T cell signal transduction and suggests a novel role for extracellular proteases as inhibitors of intracellular signal transduction pathways. The Journal of Immunology, 1999, 163: 2568 -2575.
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The thymus leukemia antigen (TL) is a nonclassical class I molecule, expressed abundantly on intestinal epithelial cells. We show that, in contrast to other major histocompatibility complex (MHC) class I molecules that bind CD8αβ, TL preferentially binds the homotypic form of CD8α (CD8αα). Thus, TL tetramers react specifically to CD8αα-expressing cells, including most intestinal intraepithelial lymphocytes. Compared with CD8αβ, which recognizes the same MHC as the T cell receptor (TCR) and thus acts as a TCR coreceptor, high-affinity binding of CD8αα to TL modifies responses mediated by TCR recognition of antigen presented by distinct MHC molecules. These findings define a novel mechanism of lymphocyte regulation through CD8αα and MHC class I.
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Fruit bromelain FA2, the main proteinase component of the juice of pineapple fruit, has been purified and characterized. 1.Efficient extraction of this enzyme from the crude material was possible using “ Cellulosin AP, ” a microbial polysaccharidase preparation containing cellulase, hemi-cellulase, and pectinase. The enzyme was purified mainly by successive applications of anion-exchange chromatography, yielding an apparently homogeneous protein as judged by several physical, chemical, and immunochemical criteria. Properties of FA2 include: molecular weight, 31, 000; isoelectric point, pH 4.6; absorbance at 280 nm of a 1% solution at pH 7.0 per cm, 19.2. 2. FA2 gave only alanine phenylthiohydantoin upon amino-terminal group analysis by the Edman procedure. Stepwise degradation yielded the amino-terminal sequence Ala-Val-Pro-Gln-Ser-Ile-Asp-Trp-Arg-Asp-Tyr-Gly-Ala. The amino acid composition of FA2 was not markedly different from that of stem bromelain, except for a much smaller lysine content and a smaller alanine content relative to glycine in FA2. FA2 contained neither amino sugars nor neutral carbohydrates as determined by several methods, so FA2 is not a glycoprotein. 3. By labeling the reactive cysteine residue (CYS) with [¹⁴C]iodoacetate, the following partial amino acid sequence has been determined. Asn-Glx-Asn-Pro-Cys-Gly-Ala-CYS 4. FA2 showed pH optima of 8.0 and 8.3 toward hemoglobin and casein, respectively. The Michaelis constant for α-N-benzoyl-L-arginine ethyl ester was 43 mM at pH 6.0 and 25°, FA2 cleaved bradykinin between Gly⁴-Phe⁵ and between Phe⁵-Ser⁶ at comparable rates; it hydrolyzed angiotensin II preferentially between Tyr⁴-lle⁵. 5. Similarities and differences found thus far between fruit bromelain FA2, stem bromelain SB1, and papain are tabulated.
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Treatment of T cells with the cysteine protease bromelain has been widely used to enhance the binding of human T cells to human E (autologous E rosettes) and has been shown to remove surface T cell CD44 molecules. Ligand binding to CD44 has been shown to markedly augment T cell activation. To study the activation potential of bromelain-treated CD44 T cells, we have compared the proliferation of sham- and bromelain-treated normal human PBMC to mitogenic CD2 mAb. We found that bromelain not only removed T cell CD44, but also removed the CD45RA isoform of CD45 as well as E2/MIC2, CD6, CD7, CD8, and Leu 8/LAM1 molecules. T cell proliferation in response to CD2 mAb was increased 325% in bromelain-treated PBMC compared to sham-treated PBMC (p < 0.005). Reciprocal treatment experiments using purified T cells and monocytes demonstrated that the enhancement of T cell CD2 activation by bromelain occurred only when T cells were treated with bromelain and was accompanied by increased adhesion of T cells to monocytes. These data demonstrate that expression of portions of the extracellular domains of the CD44, CD45RA, E2/MIC2, CD6, CD7, CD8, and Leu 8/LAM1 surface molecules are not required for CD2 activation of human T cells. Rather, the removal of these surface molecules by bromelain is associated with enhanced T cell-monocyte aggregation and enhanced CD2-mediated T cell activation. Taken together with data that CD44, E2/MIC2, CD6, and CD7 mAb inhibit CD2/lymphocyte function-associated Ag-3-mediated cellular interactions and also augment CD2-mediated triggering of T cells, these data suggest that members of the bromelain-sensitive group of surface molecules may comprise a set of CD2-associated adhesion ligands that acts in concert to modulate human T cell activation.
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The CD44 inhibitor Lutheran [In(Lu)]-related p80 molecule has recently been shown to be identical to the Hermes-1 lymphocyte homing receptor and to the human Pgp-1 molecule. We have determined the effect of addition of CD44 antibodies to in vitro activation assays of PBMC. CD44 antibodies did not induce PBMC proliferation alone, but markedly enhanced PBMC proliferation induced by a mitogenic CD2 antibody pair or by CD3 antibody. CD44 antibody addition had no effect upon PBMC activation induced by PHA or tetanus toxoid. CD44 antibody enhancement of CD2 antibody-induced T cell activation was specific for mature T cells as thymocytes could not be activated in the presence of combinations of CD2 and CD44 antibodies. CD44 antibody enhancement of CD2-mediated T cell triggering occurred if CD44 antibody was placed either on monocytes or on T cells. In experiments with purified monocyte and T cell suspensions, CD44 antibodies A3D8 and A1G3 augmented CD2-mediated T cell activation by three mechanisms. First, CD44 antibody binding to monocytes induced monocyte IL-1 release, second, CD44 antibodies enhanced the adhesion of T cells and monocytes in CD2 antibody-stimulated cultures, and third, CD44 antibodies augmented T cell IL-2 production in response to CD2 antibodies. Thus, ligand binding to CD44 molecules on T cells and monocytes may regulate numerous events on both cell types that are important for T cell activation. Given that recent data suggest that the CD44 molecule may bind to specific ligands on endothelial cells (vascular addressin) and within the extracellular matrix (collagen, fibronectin), these data raise the possibility that binding of T cells to endothelial cells or extracellular matrix proteins may induce or up-regulate T cell activation in inflammatory sites.
Article
The cluster of differentiation 44 (CD44) Ag system, originally described in brain and mature T cells, has been subsequently shown to be identical with the human lymphocyte homing-receptor defined by the Hermes-1 antibody and to be involved in T cell/endothelial cell interactions in synovium, mucosa, and lymph node. CD44 is also present on human E. On E, CD44 has been shown to be regulated by the In(Lu) dominant inhibitor gene and to express the Ina and Inb blood group Ag. Because human E have been shown to interact with human T cells via CD2 on T cells and LFA-3 on human E, we have studied the ability of human E and T lymphocyte CD44 Ag to participate in CD2/LFA-3 interactions between human E and T cells. In this study, we demonstrate that a mAb (A3D8) against the CD44, In(Lu)-related p80, lymphocyte homing-receptor molecule inhibited the binding of human E to human T cells. Whereas whole CD44 antibody molecules inhibited human E binding to T cells, saturating amounts of CD44 Fab fragments did not inhibit human E to T cell binding. Our data demonstrated that anti-CD44 antibody A3D8 acted at the level of the E to inhibit CD2/LFA-3 interactions between human E and T cells.
Article
The thiol protease bromelain has been shown to remove T-cell CD44 molecules from lymphocytes and to affect T-cell activation. We investigated the effect of a highly purified bromelain protease F9 (F9) on the adhesion of peripheral blood lymphocytes (PBL) to human umbilical vein endothelial cells (HUVEC). Preincubation of the lymphocytes with F9 reduced the adherence to about 20% of unstimulated and to about 30% of phorbol-dibutyrate (P(Bu)2) stimulated lymphocytes. Using flow cytometry, both crude bromelain and protease F9 reduced the expression of CD44, but not of LFA-1, on PBL. F9 was about 10 times more active than crude bromelain; at 2.5 micrograms/ml of F9 about 97% inhibition of CD44 expression was found. A mAb against CD44 was tested and found to block the F9-induced decrease in PBL-binding to HUVEC. The results indicate that F9 selectively decreases the CD44 mediated binding of PBL to HUVEC.
Article
The CD45 phosphotyrosine phosphatase is one of the most abundant glycoproteins expressed on immune cells. Previously, the serpentine twists and turns of the CD45 research field have tended to draw attention to CD45 either as a positive or negative regulator of immune cell function. This review draws heavily on CD45 knockout mouse data to emphasize that CD45 has both positive and negative actions in regulating receptor thresholds, and these roles vary according to cell lineage and developmental stage. Previously conflicting results are reconciled in a model suggesting how CD45 regulates the p56(lck)tyrosine kinase in T cell signalling and development.
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Leukocyte interactions with vascular endothelium are highly orchestrated processes that include the capture of free-flowing leukocytes from the blood with subsequent leukocyte rolling, arrest, firm adhesion, and ensuing diapedesis. These interactions occur under high shear stresses within venules and depend on multiple families of adhesion molecules. Many of the adhesion molecules involved are now identified. In addition, precise mechanisms underlying their regulation and our understanding of how different families of adhesion molecules work together is becoming clearer. Specifically, leukocyte/endothelial cell interactions such as capture, rolling, and firm adhesion can no longer be viewed as occurring in discrete steps mediated by individual families of adhesion molecules, but rather as a series of overlapping synergistic interactions among adhesion molecules resulting in an adhesion cascade. Although long thought to be mediated by distinct adhesion pathways, overlapping adhesion cascades mediate normal lymphocyte recirculation to peripheral lymphoid tissues and inflammation-induced leukocyte migration. These cascades thereby direct leukocyte migration, which is essential for the generation of effective inflammatory responses and the development of rapid immune responses.
Article
Genetic experiments indicate similarity between binding sites on MHC class I (MHCI) for CD8 and on MHCII for CD4, but the crystal structures of CD8/MHCI and CD4/MHCII complexes suggest critical differences between the interfaces in the two complexes. Biophysical analyses using ectodomains of co-receptors and MHC molecules demonstrate extremely fast kinetics and low-affinity interactions. Experiments with soluble multimeric MHC ligands suggest that CD4 and CD8 may differ in the mechanisms by which they promote the formation of ternary TCR/MHC/co-receptor complexes. Co-receptor-influenced duration of TCR signaling controls thymocyte selection. In naïve T cells, CD4/MHCII interactions may promote T-cell survival. Temporal and spatial analysis of TCR and CD4 co-clustering in the immunological synapse suggests that CD4 recruitment is regulated by the half-life of the initial TCR/MHCII complex. Diverse experimental systems have yielded conflicting data that have helped to formulate revised mechanistic models of co-receptor function.
Article
Bromelain is a natural proteinase preparation derived from pineapple stem that is marketed for oral use as a digestive aid and as an antiinflammatory agent. Bromelain treatment in vitro has been previously shown to selectively remove certain cell surface molecules that may affect lymphocyte migration and activation. This study reports the effects of bromelain on a broad range of cell surface molecules and on lymphocytes, monocytes, and granulocytes under physiologically relevant conditions. In vitro bromelain treatment of leukocytes in whole blood proteolytically altered 14 of 59 leukocyte markers studied. Constitutively expressed bromelain-sensitive molecules included CD7, CD8alpha, CD14, CD16, CD21, CD41, CD42a, CD44, CD45RA, CD48, CD57, CD62L, CD128a, and CD128b. The proteolytic effect of bromelain increased as the concentration of plasma decreased, with EC50 ranging from >1000 microg/ml for 100% plasma to approximately 1 microg/ml in the absence of plasma, indicating the presence of an inhibitor of bromelain in plasma. alpha2-macroglobulin purified from plasma mimicked the inhibitory effect of whole plasma on bromelain activity. If proteolysis is required for the antiinflammatory actions of oral bromelain, these data suggest that the required concentrations are more likely to be achieved locally in the gastrointestinal tract or in other tissue sites where the plasma concentration is low, rather than in the bloodstream. The cell surface molecules altered by bromelain are involved in leukocyte homing and cellular adhesion and activation. Thus bromelain could potentially exert an antiinflammatory effect by multiple mechanisms, including alterations in leukocyte migration and activation.
Article
Interleukin (IL)-10 is an anti-inflammatory and immune regulatory cytokine. IL-10-deficient mice (IL-10(-/-)) develop chronic inflammatory bowel disease (IBD), indicating that endogenous IL-10 is a central regulator of the mucosal immune response. Prostaglandins are lipid mediators that may be important mediators of intestinal inflammation. In this study we assessed the role of prostaglandins in the regulation of mucosal inflammation in the IL-10(-/-) mouse model of IBD. Prostaglandin (PG) synthesis was inhibited with nonselective or cyclooxygenase (COX)-isoform selective inhibitors. Severity of inflammation was assessed histologically. Cytokine production was assessed by ribonuclease protection analysis and enzyme-linked immunosorbent assay. PGE(2) levels were assessed by enzyme immunoassay. COX-1 and COX-2 expression was assessed by Western blot analysis. Nonsteroidal anti-inflammatory drug (NSAID) treatment of wild-type mice had minimal effect on the colon. In contrast, NSAID treatment of 4-week-old IL-10(-/-) mice resulted in rapid development of colitis characterized by infiltration of the lamina propria with macrophages and interferon gamma-producing CD4(+) T cells. Colitis persisted after withdrawal of the NSAID. NSAID treatment decreased colonic PGE(2) levels by 75%. Treatment of IL-10(-/-) mice with sulindac sulfone (which does not inhibit PG production) did not induce colitis whereas the NSAID sulindac induced severe colitis. COX-1- or COX-2-selective inhibitors used alone did not induce IBD in IL-10(-/-) mice. However, the combination of COX-1- and COX-2-selective inhibitors did induce colitis. NSAID treatment of IL-10(-/-) mice results in the rapid development of severe, chronic IBD. Endogenous PGs are important inhibitors of the development of intestinal inflammation in IL-10(-/-) mice.
Article
The marked diversity in the phenotype of intestinal neoplasia in murine models offers opportunities to model many characteristics of human CRC, including tumor progression, metastasis, gross morphology, and histology. However, the lack of a consistent model of metastasis is of particular concern in developing mouse models of human CRC. AOM-treated mice consistently develop metastasis, but the absence of control by known genetic mutations under carcinogen treatment makes this model system less desirable. The Smad3-/-, PI(3)Kγ-/-,90 and Apc1638N/+ mutants are the only mouse models reported to develop colonic adenocarcinomas that metastasize to the lymph nodes and liver, which are common metastatic sites of human CRCs. However, there has been difficulty reproducing this observation in similar strains from different laboratories. Neoplastic lesions in mice with specific genetic alterations often do not parallel the phenotype of human cancer. Lastly, the role of intestinal pathogens and their contribution to the inflammatory response and tumor initiation is generally underappreciated. Despite these limitations, mouse models are invaluable in approximating the pathogenesis of human intestinal cancer and thus provide an in vivo platform for identifying therapeutic targets and developing new strategies for prevention and treatment.
Article
Bromelain is a mixture of proteinases derived from pineapple stem that is marketed in health food stores as a "digestive aid". Orally administered bromelain was anecdotally reported to induce clinical and endoscopic remission of ulcerative colitis in two patients whose disease was refractory to multi-agent conventional medical therapy. However, the potential efficacy of bromelain in colitis has not yet been tested rigorously in either animals or humans. In this study, the clinical and histologic severity of inflammatory bowel disease (IBD) was determined in IL-10-/- mice treated orally with bromelain in vivo. Daily treatment with oral bromelain beginning at age 5 weeks decreased the incidence and severity of spontaneous colitis in C57BL/6 IL-10-/- mice. Bromelain also significantly decreased the clinical and histologic severity of colonic inflammation when administered to piroxicam-exposed IL-10-/- mice with established colitis. Proteolytically active bromelain was required for anti-inflammatory effects in vivo. Adverse effects of dermatitis, hair loss, and weight loss due to mucositis were rare, dose related, and were not seen in wild-type mice treated orally with up to 1000 mg bromelain/kg/day for 18 weeks. Although the exact mechanisms by which exogenous proteinases affect bowel inflammation have not yet been determined, the results justify additional studies of this complementary biologically based approach to treatment of IBD.
Article
Bromelain is a mixture of proteinases derived from pineapple stem that is marketed by health food stores as a "digestive aid". A number of studies suggest that bromelain may also have anti-inflammatory activity in vivo, including an anecdotal report describing potential efficacy in inflammatory bowel disease. We and others have previously shown that proteolytically active bromelain removes certain cell surface molecules and affects leukocyte migration, activation, and production of cytokines and inflammatory mediators in vitro. The purpose of this study was to determine whether ingested bromelain retains proteolytic activity within the murine gastrointestinal tract in vivo. The proteolytic activity of bromelain was determined in vitro using model substrates or immunofluorescence assays after administration of various doses and formulations orally to mice. Immune responses against bromelain were detected by enzyme immunoassays. When formulated in antacid, oral bromelain retained substantial proteolytic activity throughout the gastrointestinal tract. Bromelain concentrations within the colon were dependent on both dose and formulation and were sufficient to remove bromelain-sensitive molecules from both leukocytes and colon epithelial cells. Peak activity in the stool was observed 4 h after oral dosing. Although anti-bromelain IgG was detected in both serum and stool after long-term oral therapy, these antibodies did not prevent bromelain proteolytic activity within the gastrointestinal tract. These studies demonstrate that bromelain enzymes can remain intact and proteolytically active within the murine gastrointestinal tract. They provide further support for the hypothesis that oral bromelain may potentially modify inflammation within the gastrointestinal tract via local proteolytic activity within the colonic microenvironment.
Article
Bromelain is a natural mixture of proteolytic enzymes derived from pineapple stem that has been shown to have anti-inflammatory activity when administered orally. Although most proteins given orally without adjuvant (e.g., food) result in tolerance, we previously reported that long-term oral exposure to bromelain stimulated the development of high serum anti-bromelain antibody titers. The purpose of these studies was to further investigate the mechanisms responsible for the immunogenicity of oral bromelain. Results showed that repeated exposure was required for development of anti-bromelain antibodies, with strong antibody responses in all mice that received at least 12 doses of bromelain either orally or intragastrically over 3-6 weeks. Proteolytic activity was required for strong oral immunogenicity in the absence of conventional adjuvant, with strong serum antibody responses generated against proteolytically active bromelain and trypsin, but not against ovalbumin, lysozyme, or inactivated bromelain. Significantly higher anti-bromelain antibody titers were seen in IL-10-deficient versus wild-type mice, suggesting that simultaneous treatments that decrease IL-10 activity may further enhance systemic antibody responses following oral exposure. The antibodies generated did not affect the proteolytic activity of bromelain. The data demonstrate that proteolytically active antigens such as bromelain can stimulate both systemic and mucosal immune responses following repeated oral exposure. Further studies of the mechanisms involved in generation of immune responses following oral exposure to proteolytically active antigens can lead to a better understanding of mechanisms of oral tolerance and to the development of novel adjuvants for oral vaccines.
Article
Inflammatory bowel disease (IBD) is hypothesized to represent an aberrant immune response against enteric bacteria that occurs in a genetically susceptible host. Humans and mice with IBD are at markedly increased risk for colonic neoplasia. However, the long lead time required before development of inflammation-associated colon neoplasia in commonly used murine models of IBD slows the development of effective chemopreventative therapies. Neonatal coinfection with Helicobacter typhlonius and Helicobacter rodentium was used to trigger the onset of IBD in mice deficient in the immunoregulatory cytokine interleukin (IL)-10. The severity of colon inflammation and incidence of neoplasia was determined histologically. IL-10(-/-) mice demonstrated early onset, severe colon inflammation following neonatal infection with H. typhlonius and H. rodentium. The incidence of inflammation-associated colon neoplasia was approximately 95% at a mean age of 21 +/- 2 weeks. Mutation of endoglin, an accessory receptor for TGF-beta, did not affect the severity of IBD or the incidence of neoplasia in this model. The rapid onset of severe colon inflammation and multiple neoplastic lesions in the colons of IL-10(-/-) mice neonatally coinfected with H. typhlonius and H. rodentium makes this model well-suited for investigating the mechanisms involved in inflammation-associated colon cancer as well as its chemoprevention.
Article
Oral bromelain has been anecdotally reported to decrease inflammation in ulcerative colitis (UC). Proteolytically active bromelain is known to decrease expression of mRNAs encoding pro-inflammatory cytokines by human leukocytes in vitro. To assess the effect of bromelain on mucosal secretion of cytokines in inflammatory bowel disease (IBD), endoscopic colon biopsies from patients with UC, Crohn's disease (CD), and non-IBD controls were treated in vitro with bromelain or media, then cultured. Secretion of pro-inflammatory cytokines and chemokines was measured. Significant increases in granulocyte colony-stimulating factor (G-CSF), interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF) were detected in the media from actively inflamed areas in UC and CD as compared with non-inflamed IBD tissue and non-IBD controls. In vitro bromelain treatment decreased secretion of G-CSF, granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-gamma, CCL4/macrophage inhibitory protein (MIP)-1beta, and TNF by inflamed tissue in IBD. Bromelain may be a novel therapy for IBD.
Article
Bromelain, a mixture of proteases derived from pineapple stem, has been reported to have therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease. The purpose of this work was to understand potential mechanisms for this anti-inflammatory activity. Exposure to bromelain in vitro has been shown to remove a number of cell surface molecules that are vital to leukocyte trafficking, including CD128a/CXCR1 and CD128b/CXCR2 that serve as receptors for the neutrophil chemoattractant IL-8 and its murine homologues. We hypothesized that specific proteolytic removal of CD128 molecules by bromelain would inhibit neutrophil migration to IL-8 and thus decrease acute responses to inflammatory stimuli. Using an in vitro chemotaxis assay, we demonstrated a 40% reduction in migration of bromelain- vs. sham-treated human neutrophils in response to rhIL-8. Migration to the bacterial peptide analog fMLP was unaffected, indicating that bromelain does not induce a global defect in leukocyte migration. In vivo bromelain treatment generated a 50-85% reduction in neutrophil migration in 3 different murine models of leukocyte migration into the inflamed peritoneal cavity. Intravital microscopy demonstrated that although in vivo bromelain treatment transiently decreased leukocyte rolling, its primary long-term effect was abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion were correlated with rapid re-expression of the bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo bromelain treatment and minimal re-expression of CD128 over the time period studied. Taken together, these studies demonstrate that bromelain can effectively decrease neutrophil migration to sites of acute inflammation and support the specific removal of the CD128 chemokine receptor as a potential mechanism of action.
Handbook of Proteolytic Enzymes Hale et al. Page 10 Inflamm Bowel Dis Author manuscript; available in PMC 2011 December 1. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript 10 Purification and characterization of a proteinase from pineapple fruit, fruit bromelain FA2
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Barrett, AJ.; Rawlings, ND.; Woessner, JF., editors. Handbook of Proteolytic Enzymes. San Diego: Academic Press; 1998. p. 566-567. Hale et al. Page 10 Inflamm Bowel Dis. Author manuscript; available in PMC 2011 December 1. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript 10. Yamada F, Takahashi N, Murachi T. Purification and characterization of a proteinase from pineapple fruit, fruit bromelain FA2. J Biochem. 1976; 79:1223–1234. [PubMed: 956152]
AVMA guidelines on euthanasia: Jun 2007 update Available at: http://www.avma.org/issues/animal_welfare/euthanasia
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Handbook of proteo-lytic enzymes
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Barrett AJ, Rawlings ND, Woessner JF (eds.). Handbook of proteo-lytic enzymes. San Diego: Academic Press; 1998:566–567.