Bioactive Peptides

The recent increase in extensively drug resistant bacterial pathogens and the associated increase of morbidity and mortality demonstrate the immediate need for new antibiotic backbones with novel mechanisms of action. Here, we report the development of the PepSAVI-MS pipeline for bioactive peptide discovery. This highly versatile platform employs mass spectrometry and statistics to identify bioactive peptide targets from complex biological samples. We validate the use of this platform through the successful identification of known bioactive peptides from a botanical species, Viola odorata. Using this pipeline, we have widened the known antimicrobial spectrum for V. odorata cyclotides, including antibacterial activity of cycloviolacin O2 against A. baumannii. We further demonstrate the broad applicability of the platform through the identification of novel anticancer activities for cycloviolacins by their cytotoxicity against ovarian, breast and prostate cancer cell lines.

Diabetic nephropathy (DN) poses a significant health challenge, necessitating novel therapeutic approaches. In this study, we isolated proteins from cell-free supernatant (CFS) from the culture of the lactic acid bacteria Lactobacillus brevis RAMULAB49 strain. The proteins were subjected to simulated in vitro gastrointestinal digestion using gut enzymes-pepsin, pancreatin, and trypsin. The hydrolysates were filtered using 3kDa threshold ultra-centrifugal filters and were desalted using C18 disks. This was followed by nLC-ESI MS/MS tandem mass spectrometry-based identification of peptides, leading in the identification of a of 258 unique peptides across three enzyme combinations. The resultant sequences were made into peptide library construction based on their, bioactivity scores, allergenicity, toxicity, and antidiabetic potential, a total of 10 peptides was constructed and modeled in 3D. On the other hand, 266 DN associated genes were identified using a network pharmacology approach. The resultant protein-protein (PPI) network was analysed using the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment approaches, resulting in identification of critical pathways, ERK1, PI3K-Akt, EGRF and TNF signaling as significantly involved in DN, where, ERK1 emerging the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Low-molecular-weight peptides derived by digestion of body wall proteins from some sea cucumber species have wound-healing and health-promoting properties. Sea cucumber body composition varies widely by species, growth environment, age and season, so it is unknown if all species contain the requisite bioactive peptides. For the first time, small peptide (1-3 kDa) fractions have been isolated from the whole-body wall of the sea cucumber Isostichopus badionotus and its constituent collagen, and tested for wound healing capacity in vitro. The body wall comprised around 50% collagen, whose composition and structure were like, but not identical to, other sea cucumber collagens. Ultrafiltered digests (1-3 kDa) of the pure collagen and a further purified fraction of it, as well as a 1-3 kDa digest of whole-body wall, had potent antioxidant activities and promoted rapid wound healing in a keratinocyte scratch wound assay. Gene expression studies suggested that the wound-healing actions of the individual 1-3 kDa fractions differed significantly. Low-molecular-weight peptides derived

Brewers' spent grain (BSG) is the major by-product of the brewing industry which remain largely unutilized despite its nutritional quality. In this study, the effects of fermentation on BSG antioxidant potential were analyzed. A biotechnological protocol including the use of xylanase followed by fermentation with Lactiplantibacillus plantarum (Lactobacillus plantarum) PU1, PRO17, and H46 was used. Bioprocessed BSG exhibited enhanced antioxidant potential, characterized by high radical scavenging activity, long-term inhibition of linoleic acid oxidation and protective effect toward oxidative stress on human keratinocytes NCTC 2544. Immunolabelling and confocal laser microscopy showed that xylanase caused an extensive cell wall arabinoxylan disruption, contributing to the release of bound phenols molecules, thus available to further conversion through lactic acid bacteria metabolism. To clarify the role of fermentation on the antioxidant BSG potential, phenols were selectively extracted and characterized through HPLC-MS techniques. Novel antioxidant peptides were purified and identified in the most active bioprocessed BSG.

Hydrolysed Collagen Tripeptides (CTPs) are minimal bioactive signal units - exemplified by Gly-Pro-Hyp that outperform generic collagen hydrolysates as they preserve sequence integrity, cell recognition, and downstream signaling.

Following oral intake, CTPs are absorbed intact via PEPT1, appear in plasma within 30-60 min, and reach dermal tissue where they engage fibroblast receptors (e.g., CD44, α2β1, DDR1/2) and activate TGF-β/Smad and MAPK/ERK pathways.

This drives a consistent signal → synthesis → remodeling cascade, upregulating type I collagen, elastin, and hyaluronic acid, with clinically relevant outcomes - skin hydration / elasticity gains and wrinkle-depth reduction - typically observed within 8-12 weeks at ~1,000-2,500 mg/day.

Performance is spec-dependent: high Gly-Pro-Hyp content (≥50%) and Pro + Hyp 25-30% correlate with superior pharmacokinetics and effect sizes, while undefined peptides or free amino acids lack targeted signaling and reproducibility.

In multi-ingredient systems, CTPs synergize with elastin peptides and hyaluronic acid rebuild the dermal tension-and-recoil network. Collectively, Gly-Pro-Hyp - enriched CTPs constitute a precision nutricosmetic modality coupling rapid systemic delivery with receptor-mediated ECM regeneration for visible anti-aging benefits.

Bioactive peptides are short polymers from proteins that have a positive impact functionally for human health. This paper provides a review of bioactive peptides derived from various sources of meat and the generation of bioactive peptides. Bioactive peptides can be produced from a variety of animal species, such as bovine, poultry, and porcine. They can be obtained by hydrolysis from their parent protein sequence to release bioactive peptides. Some of the methods are enzymatic hydrolysis using enzymes from animals, commercial proteinase, or enzymes from plants. In addition, microorganism fermentation with starter lactic acid bacteria may also be used. Some of the functions of bioactive peptides that are beneficial for humans health include antimicrobials, antithrombotic drugs such as the prevention of blood clots and strokes, antioxidants that coun teract free radicals, and antihypertensive drugs that can lower blood pressure.

Cervical cancer is among the leading causes of death in women. Chemotherapy options available for cervical cancer include highly cytotoxic drugs such as taxol, cisplatin, 5-florouracil, and doxorubicin, which are not specific. In the current study, we have identified a new peptide conjugate (Fur4-2-Nal3-Ala2-Phe1-CONH2) (conjugate 4), from screening of a small library of tripeptide-conjugates of furan, as highly potent anticancer compound against human cervical cancer cells (HeLa cells) (IC50 = 0.15 ± 0.05 µg/mL or 0.28 +/− 0.09 µM). Peptides were constructed on Rink amide resin from C- to N-terminus followed by capping by α-furoic acid moiety. The synthesized peptides were purified by recycling RP-HPLC, and structures of all the peptides were confirmed by using FABMS/ESIMS, 1H- NMR, 13C-NMR, and HR-FABMS. Conjugate 4 was furthermore found to be specifically active against human cervical cancer cells since it did not inhibit the proliferation of other human normal cells (HUVEC (huma...

Background: Antimicrobial peptides (AMPs) constitute an armory of innate regulators of crucial importance in the gastrointestinal (GI) tract. It is becoming evident that AMPs, especially human β-defensins (hBDs) produced by colonocytes, shape the composition of the microbiota and help maintain gut homeostasis. Defective bacterial clearance and subsequent modulation of gut microbiota, due to abnormal defensin expression, is associated with pathogenesis of various GI diseases including Inflammatory Bowel Disease (IBD). Serotonin or 5-hydroxytrytophan (5-HT) is a neurotransmitter and hormone that contributes to the regulation of various physiological functions in GI tract. Changes in gut 5-HT signaling are observed in IBD as well as in experimental colitis. Enterochromaffin (EC) cells of the human GI tract are the largest producers of 5-HT, and 5-HT biosynthesis depends on the rate-limiting tryptophan hydroxylase 1 (Tph1). We have previously demonstrated Tph1-deficient (Tph1 -/-) mice, which have significantly lower gut 5-HT levels, exhibited reduced severity of colitis compared to their wild-type (Tph1 +/+ ) littermates; while replenishing 5-HT led to increased severity of colitis. As EC cells are situated in the epithelial layer and epithelial cells express 5-HT receptors, we hypothesize that 5-HT can influence β-defensin production directly by acting on the adjoining epithelial cells in relation to innate defense in the GI tract. Aims: To elucidate the role of 5-HT in β-defensin production from intestinal epithelial cells in the context of regulating innate immune response. Methods: We utilized human colonic HT-29 cells to assess the level of hBD expression upon stimulation with 5-HT (10 -3 , 10 -7 , 10 -11 M). Quantitative PCR (qPCR) and conventional ELISA were performed to measure gene expression and protein levels in culture supernatant, respectively. Additionally, we measured mouse β-defensin levels in colonic tissues of naïve Tph1 +/+ and Tph1 -/-. Results: 5-HT, in a dose dependent manner, regulates hBD production by HT-29 cells, where higher 5-HT concentrations down-regulated both mRNA and protein levels. Moreover, we observed significantly higher levels of total mBD in the colonic tissues of Tph1 -/-compared to Tph1 +/+ , suggesting an inhibitory role of 5-HT in defensin production. Conclusions: Our results illustrate that 5-HT released from EC cells can modulate hBD production from intestinal epithelial cells. These results exemplify novel information on the interaction between 5-HT and hBD in relation to intestinal innate immune response and

The aims of the present study were to characterize physicochemical characteristics and chemical structures by Fourier Transform Infrared (FTIR), and mark dissolved protein content, microstructure, and moisture content of potassium caseinate prepared by drying methods and acid strength. The experiment was arranged according to factorial complete randomized design with triplicates, while data from FTIR and microstructure analysis was presented descriptively. The results demonstrated that acids and drying methods for preparing potassium caseinate could increase antioxidant activity, a* score (reddish) and b* score (yellowish). Specifically, freeze-drying method coupled with acid treatments accounted for reducing moisture content but improved viscosity and microstructural properties. Briefly, we could argue that drying techniques and acids established noticeable effects on the quality of potassium caseinate.

Three synthetic peptides, ILAP, LLAP and MAGVDHI, derived from a Palmaria palmata protein hydrolysate were assessed for their antidiabetic potential in vitro and in vivo. In addition to inhibiting dipeptidyl peptidase-IV in a cell-based in situ assay all three peptides significantly increased the half-life of the incretin hormone glucagon-like peptide-1 (GLP-1). ILAP and LLAP mediated a significant increase (p < 0.001) in insulin secretion from BRIN-BD11 cells compared to the glucose control, while MAGVDHI had no insulinotropic activity at an eqimolar concentration (10–6 M). A significant increase in the concentration of cyclic adenosine monophosphate production in BRIN-BD11 cells mediated by ILAP (p < 0.001) and LLAP (p < 0.01) compared to the basal control, would indicate that insulin secretion may be mediated by membrane based activation. Furthermore, ILAP and LLAP acted as glucose-dependent insulinotropic polypeptide (GIP) secretagogues, stimulating a significant increa...

Prevalence of type 2 diabetes and overweight/obesity are increasing globally. Food supplementation as a preventative option has become an attractive option in comparison to increased pharmacotherapy dependency. Hydrolysates of fish processing waste and by‐products have become particularly interesting in a climate of increased food wastage awareness and are rapidly gaining traction in food research. This review summarizes the available research so far on the potential effect of these hydrolysates on diabetes and appetite suppression. Scopus and Web of Science are searched using eight keywords (fish, hydrolysate, peptides, satiating, insulinotropic, incretin, anti‐obesity, DPP‐4 [dipeptidylpeptidase‐4/IV]) returning a total of 2549 results. Following exclusion criteria (repeated appearances, non‐fish marine sources [e.g., macroalgae], and irrelevant bioactivities [e.g., immunomodulatory, anti‐thrombotic]), 44 relevant publications are included in this review. Stimulation of hormone se...

MicroRNAs (miRNAs) are a family of small noncoding RNAs that regulate gene expression. Due to their important activity in the fine-tuning of protein translation, abnormal expression of miRNAs has been linked to many human diseases, making the targeting of miRNAs attractive as a novel therapeutic strategy. Accordingly, researchers have been heavily engaged in the discovery of small molecule modulators of miRNAs. With an interest in the identification of new chemical space for targeting miRNAs, we developed a highthroughput screening (HTS) technology, catalytic enzyme-linked click chemistry assay (cat-ELCCA), aimed at the discovery of small molecule ligands for pre-miR-21, a miRNA that is frequently overexpressed in human cancers. From our HTS campaign, we found that natural products, a source of many impactful human medicines, may be a promising source of potential pre-miR-21-selective maturation inhibitors. Herein we describe our first efforts in natural product inhibitor discovery leading to the identification of a depsipeptide class of natural products as RNA-binding inhibitors of Dicer-mediated miRNA processing.

Eight molecules, four peptides (SPs) and four lipopeptides (LPs) derived by rational design from surfactin, a well-known secreted biosurfactant from Bacillus subtilis, were produced employing Fmoc-based solid-phase synthesis. These new peptides were tested to evaluate their potential biosurfactant and biological activities, aiming at possible applications in industrial, biological, pharmaceutical, and medical use. Five molecules (SP1, SP2, SP4, LP5, and LP8) presented potential for medical uses, mainly due to their drug delivery properties as suggested by their synergistic activity with the antibiotic vancomycin against Staphylococcus aureus. All synthetic peptides showed low toxicity against Vero cell cultures, in assays of hemolysis, and in different cytotoxicity assays. In addition, we found that three peptides (SP1, LP6, and LP7) had potential technological and industrial use because of their emulsifying capacity, low toxicity, and ability to physically stabilize solutions. Thes...

Casein is considered to be the main source of protein in milk; therefore, many studies have been conducted to identify casein-derived bioactive peptides and their physiological effects. Casein is inactive within the parent protein but can be liberated by various proteases and enzymatic hydrolysis during microbial fermentation and gastrointestinal digestion. Once absorbed, casein exhibits different bioavailabilities in the body. Specifically, casein-derived peptides function as angiotensin converting enzyme (ACE) inhibitor in the cardiovascular system; thus, they are expected to reduce and prevent hypertension. Additionally, casein-derived peptides behave as opioid-like peptides in the nervous system, which impacts relaxation. These peptides are also expected to modulate various aspects of immune functions. Finally, caseinophosphopeptide (CPP) and glycomacropeptide (GMP) may exhibit a number of nutritional effects such as the absorption of calcium, iron or zinc. Many studies have been conducted to evaluate casein-derived peptides due to their multifunctional properties and the results of these studies have contributed to the development of a wide variety of functional dairy products. The purpose of this paper was to review the generation of bioactive peptides, their absorption and metabolism, and their specific bioactive effects.

Angiotensin I converting enzyme (ACE) inhibitory peptides can induce antihypertensive effects after oral administration. By means of an ACE inhibitory peptide database, containing about 500 reported sequences and their IC 50 values, the different proteins in pea and whey were quantitatively evaluated as precursors for ACE inhibitory peptides. This analysis was combined with experimental data from the evolution in ACE inhibitory activity and protein degradation during in vitro gastrointestinal digestion. Pea proteins produced similar in silico scores and were degraded early in the in vitro digestion. High ACE inhibitory activity was observed after the simulated stomach phase and augmented slightly in the simulated small intestine phase. The major whey protein b-lactoglobulin obtained the highest in silico scores, which corresponded with the fact that degradation of this protein in vitro only occurred from the simulated small intestine phase on and resulted in a 10-fold increase in the ACE inhibitory activity. Whey protein obtained total in silico scores of about 124 ml/mg, compared to 46 ml/mg for pea protein, indicating that whey protein would be a richer source of ACE inhibitory peptides than pea protein. Although b-lactoglobulin is only partially digested, a higher ACE inhibitory activity was indeed found in the whey (IC 50 = 0.048 mg/ml) compared to the pea digest (IC 50 = 0.076 mg/ml). In silico gastrointestinal digestion of the highest scoring proteins in pea and whey, vicilin and albumin PA2, and b-lactoglobulin, respectively, directly released a number of potent ACE inhibitory peptides. Several other ACE inhibitory sequences resisted in silico digestion by gastrointestinal proteases. Briefly, the quantitative in silico analysis will facilitate the study of precursor proteins on a large scale and the specific release of bioactive peptides.

Bioactive peptides are the functional concept that refer to fragments of proteins that provide both functional and nutritional benefits as a functional component. Milk has been widely exploited as a source of diverse bioactive peptides. Bioactivity of peptides depends on amino acid composition and sequence specific to bioactivity. The production of bioactive peptides can be carried out through classical and bioinformatics approaches. In the classical approach, enzymatic hydrolysis, simulated digestion and hydrolysis by microbial enzymes are included. In the bioinformatics approach, databases, QSAR modeling, molecular docking and design of experiments (DoE) with response surface methodology are employed for identification and characterization of bioactive peptides from milk proteins. Milk-derived peptides have been identified to be antihypertensive, anti-diabetic, cytomodulatory, antioxidative, antimicrobial, and mineral binding peptides with wide applications as nutraceuticals in commercial products and food preservatives in food industry. Hence, this review has attempted to provide insight into the approaches and applications of milk-derived bioactive peptides with nutraceuticals properties in the development of functional foods.

Fluoride Toxicity due to the presence of higher levels of fluoride in drinking water (>1.5ppm) may be serious problems in health of the children and adult in general. In the state of Rajasthan, almost all districts have high Fluoride (up to 18.0 ppm) in their drinking / ground water sources and about 11 million of the populations are at risk. In the present study, 53 children were selected from the rural area of the eastern regions (Dausa district) in the Rajasthan India, where fluoride content in water is 5.5 ± 1.2 ppm. Moreover, age matched controls were selected from the Jaipur district where fluoride content in water was less than 1.5 ppm. 3.0 ml of blood sample were taken to investigate oxidative stress parameters namely, lipid peroxide level (LPO), superoxide dismutase, catalase, glutathione peroxidase and reduced glutathione content. Moreover, serum Fe, Cu, Zn, Se and fluoride were investigated. Increased LPO and depleted antioxidant levels were observed in subjects along with the alteration in trace elements. The concentration of fluoride in serum was significantly correlates with their water concentration. On the basis of the results it may conclude that fluoride exposure promote oxidative stress and alteration in trace elements. These alterations may induce pathological conditions in fluoride exposed children. These biochemical markers may be used as the detection of early fluorosis. However, further in depth studies is required for the understanding of pathophysiological role of fluoride.

The consumption of amaranth grain, a primary crop from Andes, has become popular among celiac patients, since it does not promote allergic reactions to their intestinal mucosa. However, it has been observed a high prevalence of Diabetes Mellitus among Celiac patients and vice-versa. A ready-to-eat amaranth snack was developed and presented high nutritive value and good functional characteristics. Considering that the treatment of Diabetes Mellitus is based on the control of glucose levels, and the digestion and absorption rate of starch from foods directly influence the rise of glucose levels, the aim of this study was to investigate the glucose profile of the amaranth snack examining its diges tion in vitro. In order to achieve this purpose an enzymatic hydrolysis was performed, where samples were collected during the digestion period to determinate the amount of glucose released in the system. White bread was used as a reference product. The results demonstrated that amaranth snack has high digestibility, with a Hydrolysis Index of 103%. Probably the digestion profile observed in the amaranth snack was due to the process undergone by the amaranth grain: milling and later gelatinization promoted by extrusion cooking. These treatments might have promoted a drastic rupture and exposure of amaranth grain starch content to the enzymatic attack. For its fast digestion the amaranth snack is recommended to athletes to help restore carbohydrates after exercises; when consumed by diabetic celiac patients, should be assessed on a dietetic program, where the diet is direct towards control of glucose levels.

Context: SARS-CoV-2, a member of family Coronaviridae and the causative agent of COVID-19, is a virus which is transmitted to human and other mammals. Aims: To analyze the B-cell epitope conserved region and viroinformatics-based study of the SARS-CoV-2 lineage from Indonesian B.1.1.7 isolates to invent a vaccine nominee for overcoming COVID-19. Methods: The sequences of seven Indonesian B.1.1.7 isolates, Wuhan-Hu-1 isolate, and WIV04 isolate were extracted from the GISAID EpiCoV and GenBank, NCBI. MEGA X was employed to understand the transformations of amino acid in the S protein and to develop a molecular phylogenetic tree. The IEDB was implemented to reveal the linear B-cell epitopes. In addition, PEP-FOLD3 web server was utilized to perform peptide modeling, while docking was performed using PatchDock, FireDock, and the PyMOL software. Moreover, in silico cloning was developed by using SnapGene v.3.2.1 software. Results: In this study, the changes of amino acid in all seven Ind...

 Functional foods and nutraceuticals have garnered attention for their potential role in promoting health and preventing disease. With increasing global interest in diet-based interventions, there is a need to evaluate their efficacy in managing chronic diseases such as cardiovascular diseases, diabetes, and obesity. Functional foods provide health benefits beyond essential nutrition, whereas nutraceuticals include bioactive compounds in concentrated medicinal forms. This systematic review explores their classification, mechanism of action, health benefits, and regulatory aspects. The studies included were pulled up from PubMed, Scopus, and Web of Science. The findings suggest that functional foods and nutraceuticals play a role in the prevention of cardiovascular disease, cancer, metabolic disorders, and neurodegenerative conditions. However, challenges such as bioavailability, genetic variability, and regional regulatory differences persist. Standardized regulations and personalized nutrition strategies are essential for optimizing their efficacy.

Food waste and by-products are protein-rich sources and represent an important alternative in the search for new strategies to produce compounds with bioactivity from protein hydrolysates. Several studies have shown that by-products and wastes from food processing industries can be utilised as potential sources of bioactive compounds that have an important application in the treatment of various disorders. Bioactive peptides, defined as small fractions of a certain number of amino acids encrypted in proteins, have high potential as a safe, natural, and cost-effective alternative to synthetic drugs to prevent or treat these diseases. By-products and wastes represent a relatively inexpensive source, so their utilisation for the production of bioactive peptides not only leads to a reduction in production costs but is also very important for the development of high value-added nutritional byproducts. This practice also contributes to reduce the problem of waste disposal. In this context, the aim of this study was to review the current studies on various food processing by-products and wastes that can be used in the production of bioactive peptides, the processes of obtaining protein hydrolysate from these products and the health benefits of these peptides.

Context: SARS-CoV-2, a member of family Coronaviridae and the causative agent of COVID-19, is a virus which is transmitted to human and other mammals. Aims: To analyze the B-cell epitope conserved region and viroinformatics-based study of the SARS-CoV-2 lineage from Indonesian B.1.1.7 isolates to invent a vaccine nominee for overcoming COVID-19. Methods: The sequences of seven Indonesian B.1.1.7 isolates, Wuhan-Hu-1 isolate, and WIV04 isolate were extracted from the GISAID EpiCoV and GenBank, NCBI. MEGA X was employed to understand the transformations of amino acid in the S protein and to develop a molecular phylogenetic tree. The IEDB was implemented to reveal the linear B-cell epitopes. In addition, PEP-FOLD3 web server was utilized to perform peptide modeling, while docking was performed using PatchDock, FireDock, and the PyMOL software. Moreover, in silico cloning was developed by using SnapGene v.3.2.1 software. Results: In this study, the changes of amino acid in all seven Ind...

Background: The escalating global threat of antimicrobial resistance (AMR) necessitates the development of novel antimicrobial agents, innovative strategies, and representative infection models to combat AMR bacterial infections. Host defence peptides (HDPs) and their derivatives have been proposed as complements to conventional antibiotics due to their antibacterial activity and modulation of the immune response.
Objectives: This study investigated the novel use of the HDP-derived synthetic antibacterial and anti-biofilm peptide (SAAP)-148 as a pretreatment in epithelial tissue models to prevent colonization by AMR bacteria. The combined activities of SAAP-148 pretreatment with post-infection halicin to treat infections were also explored.
Methods: Employing cultured human skin equivalents (HSEs) and primary bronchial epithelial cells (PBECs) as models of tissue infection, we examined the prophylactic and therapeutic effects of SAAP-148, both singularly and in combination with the repurposed antibiotic halicin, against AMR bacteria. We additionally interrogated the response of HSE and PBEC cultures to SAAP-148 treatment via confocal microscopy and quantitative PCR of native HDPs and inflammatory cytokine genes.
Results: Our findings demonstrated that pretreatment with SAAP-148 significantly reduces colonization of HSEs and PBECs by AMR Staphylococcus aureus and Pseudomonas aeruginosa. Confocal microscopy revealed differential uptake and localization of SAAP-148 in these tissues, correlating with its distinct activity in these tissues. SAAP-148 exposure temporarily increased expression of the HDPs cathelicidin (CAMP) and β-defensin 1 (DEFB1), and the cytokine IL-8 (CXCL8), which did not correlate with the transient antibacterial activity observed. Sequential treatment with SAAP-148 prior to infection with AMR S. aureus and post-infection halicin treatment demonstrated synergistic activity in HSEs, whereas this combined activity was indifferent in PBEC cultures.
Conclusions: These results support SAAP-148 as a candidate for pre-infection prophylaxis and synergistic antibiotic therapy with halicin in skin, broadening the potential of both agents to address AMR bacterial infection.

Amaç: Bu çalışmanın amacı; protein parçalandığında aktif hale geçen ve insansağlığı üzerinde birçok yararlı etkileri olduğu bilinen biyoaktif peptitlerin MCF-7meme kanseri hücre dizileri üzerine olan etkilerini incelemektir.Gereç ve Yöntem: Bu çalışmada keçi sütünün yapısal özellikleri pH, karbohidrat,yağ, protein, nem, kuru madde ve kül açısından incelenmiştir. Keçi sütü kazein veserum fraksiyonlarına ayrıldıktan sonra pepsin, papain ve tripsin enzimleri ile ayrıayrı muamele ederek enzimatik olarak parçalanarak süt peptitleri elde edildi. Bupeptit molekülleri MCF-7 hücre dizileri ile 24 saat ve 48 saat farklıkonsantrasyonlarda inkübe edildi. İnkübasyon sonrası MCF-7 hücrelerinin canlılıkoranları 3-[4,5-dimetiltiazol-2-il]-2,5-difenil tetrazoliyum bromür canlılık testiyapılarak tayin edildi. Ölen MCF-7 hücrelerinden proteinler izole edildikten sonrakütle spektroskopisi sisteminde kütlelerine göre ayrılan proteinler ve peptitler tek tekkarakterize edildi.Bulgular: Keçi sütü serum pro...

Fluoride Toxicity due to the presence of higher levels of fluoride in drinking water (>1.5ppm) may be serious problems in health of the children and adult in general. In the state of Rajasthan, almost all districts have high Fluoride (up to 18.0 ppm) in their drinking / ground water sources and about 11 million of the populations are at risk. In the present study, 53 children were selected from the rural area of the eastern regions (Dausa district) in the Rajasthan India, where fluoride content in water is 5.5 ± 1.2 ppm. Moreover, age matched controls were selected from the Jaipur district where fluoride content in water was less than 1.5 ppm. 3.0 ml of blood sample were taken to investigate oxidative stress parameters namely, lipid peroxide level (LPO), superoxide dismutase, catalase, glutathione peroxidase and reduced glutathione content. Moreover, serum Fe, Cu, Zn, Se and fluoride were investigated. Increased LPO and depleted antioxidant levels were observed in subjects along with the alteration in trace elements. The concentration of fluoride in serum was significantly correlates with their water concentration. On the basis of the results it may conclude that fluoride exposure promote oxidative stress and alteration in trace elements. These alterations may induce pathological conditions in fluoride exposed children. These biochemical markers may be used as the detection of early fluorosis. However, further in depth studies is required for the understanding of pathophysiological role of fluoride.

A search in a database of potential bioactive short sequences in food proteins reveals that bioactive peptides with a variety of beneficial effects for cardiovascular health are present in the sequence of common bean proteins, including bioactive sequences with antioxidant properties. A protein isolate, the storage protein phaseolin and a lectin extract from Phaseolus vulgaris L. var. Jamapa, were hydrolyzed by treatment with pepsin and pancreatin in order to investigate the possible release of peptides with antioxidant and metal chelating properties. Antioxidant activity was determined in Caco-2 cells exposed to a free radical generator, and iron and copper chelating activities were determined using colorimetric methods. The highest antioxidant activity, 71% inhibition, was found in the hydrolyzed protein isolate. Copper and iron chelating activities were highest in the lectin and phaseolin hydrolysates, 53% and 81%, respectively. Thus, experimental data indicates, as suggested by the database search, that antioxidant peptides are abundant in pepsin-pancreatin hydrolysates, which may represent a valuable health-promoting property in common bean.

Because consumption of whey protein hydrolysates is on the increase, the possibility that prolonged ingestion of whey protein hydrolysates affect the digestive system of mammals has prompted us to evaluate the enzymatic activities of pepsin, leucine-aminopeptidase, chymotrypsin, trypsin, and glutaminase in male Wistar rats fed diets containing either a commercial whey isolate or a whey protein hydrolysate with medium degree of hydrolysis and to compare the results with those produced by physical training (sedentary, sedentary-exhausted, trained, and trained-exhausted) in the treadmill for 4 weeks. The enzymatic activities were determined by classical procedures in all groups. No effect due to the form of the whey protein in the diet was seen in the activities of pepsin, trypsin, chymotrypsin, and leucine-aminopeptidase. Training tended to increase the activity of glutaminase, but exhaustion promoted a decrease in the trained animals, and consumption of the hydrolysate decreased it even further. The results are consistent with the conclusion that chronic consumption of a whey protein hydrolysate brings little or no modification of the proteolytic digestive system and that the lowering of glutaminase activity may be associated with an antistress effect, counteracting the effect induced by training in the rat.

In view of the recent findings of stimulatory effects of GHRH analogs, JI-34, JI-36 and JI-38, on cardiomyocytes, pancreatic islets and wound healing, three series of new analogs of GHRH(1-29) have been synthesized and evaluated biologically in an endeavor to produce more potent compounds. "Agmatine analogs", MR-356 (N-Me-Tyr 1 -JI-38), MR-361(N-Me-Tyr 1 , D-Ala 2 -JI-38) and MR-367(N-Me-Tyr 1 , D-Ala 2 , Asn 8 -JI-38), in which Dat in JI-38 is replaced by N-Me-Tyr 1 , showed improved relative potencies on GH release upon subcutaneous administration in vivo and binding in vitro. Modification with N-Me-Tyr 1 and Arg 29 -NHCH 3 as in MR-403 (N-Me-Tyr 1 , D-Ala 2 , Arg 29 -NHCH 3 -JI-38), MR-406 (N-Me-Tyr 1 , Arg 29 -NHCH 3 -JI-38) and MR-409 (N-Me-Tyr 1 , D-Ala 2 , Asn 8 , Arg 29 -NHCH 3 -JI-38), and MR-410 (N-Me-Tyr 1 , D-Ala 2 , Thr 8 , Arg 29 -NHCH 3 -JI-38) resulted in dramatically increased endocrine activities. These appear to be the most potent GHRH agonistic analogs so far developed. Analogs with Apa 30 -NH 2 such as MR-326 (N-Me-Tyr 1 , D-Ala 2 , Arg 29 , Apa 30 -NH 2 -JI-38), and with Gab 30 -NH 2 , as MR-502 (D-Ala 2 , 5F-Phe 6 , Ser 28 , Arg 29 , Gab 30 -NH 2 -JI-38) also exhibited much higher potency than JI-38 upon i.v. administration. The relationship between the GH-releasing potency and the analog structure is discussed. Fourteen GHRH agonists with the highest endocrine potencies were subjected to cardiologic tests. MR-409 and MR-356 exhibited higher potency than JI-38 in activating myocardial repair in rats with induced myocardial infarction. As the previous class of analogs, exemplified by JI-38, had shown promising results in multiple fields including cardiology, diabetes and wound healing, our new, more potent, GHRH agonists should manifest additional efficacy for possible medical applications.

The main objective of our rational drug design is to predict the best novel tripeptide leads for antitubercular effect from the designed set of templates with various descriptors such as Boman index, lipinski rules and molecular dynamics simulations. We built 63 candidate molecules from chemical templates and subjected to Boman index through online antimicrobial peptide database. The best resulting 13 templates derived from this database were passed them through empirical lipinski filters to assess drug like properties using various molecular descriptors namely molecular mass, partition coefficient, Hydrogen bond donors and Hydrogen bond acceptors respectively. Later, we conducted molecular dynamics on the 13 templates with 20 nano second simulations of total atoms at an average temperature 298.15K in 100 steps with applying constraints to all bonds in pure water. Finally, the conformational stability of the candidates were analyzed with measurement of total potential energy, total ...

Microalgae are a relatively underutilised source of valuable nutritional compounds and biochemicals. Their high protein contents make them an interesting target for the generation of bioactive peptides for the functional food industry. Here, the nitrogenous components of the red microalga Porphyridium purpureum and the diatom- Phaeodactylum tricornutum were quantified, and their SDS-PAGE profiles analysed. Proteinswere isolated from these species by solubilisation at alkaline pH followed by precipitation at acidic pH. The protein extracts were hydrolysed with the food-grade proteolytic preparations Alcalase 2.4 L and Flavourzyme 500 L. The P. purpureum and P. tricornutum hydrolysates were examined for their in vitroanti-diabetic and antioxidant properties. The microalgal-derived protein hydrolysates had dipeptidyl peptidase (DPP) IV inhibitory activity, with IC50 values of 2.28 ± 0.36 and 2.68 ± 0.19 mg/mL for P. purpureum and P. tricornutum, respectively. The oxygen radical absorba...

Dipeptides with a C terminal Pro inhibit dipeptidyl peptidase IV (DPP-IV), a key enzyme in incretin hormone processing. It was hypothesised that tri-and tetrapeptides with a proline at the C-terminus may also be DPP-IV inhibitors. Therefore, an in silico hydrolysis approach was used to release short (4 ≤ amino acids) C terminal Pro peptides from the individual caseins which constitute Pro rich substrates. This was achieved using theoretical digestion of caseins with a prolyl oligopeptidase activity. Fifteen peptides were subsequently selected for in vitro DPP-IV inhibitory analysis. Stability of these peptides to gastrointestinal enzymes was also evaluated in silico and the predicted breakdown peptides were assessed for their DPP-IV inhibitory and antioxidant potential. New DPP-IV inhibitors were identified, the most potent being Phe-Leu-Gln-Pro (IC 50 65.3 ± 3.5 µM). A low in vitro antioxidant (2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging) activity was also associated with the peptides studied. The strategy presented highlights the utility of employing an in silico approach for the prediction of food-derived peptides with a potential role in glycaemic management for subsequent development of functional foods.

The potential of quinoa to act as a source of dipeptidyl peptidase IV (DPP-IV) inhibitory and antioxidant peptides was studied. A quinoa protein isolate (QPI) with a purity of 40.73 ± 0.90% was prepared. The QPI was hydrolysed at 50C for 3 h with two enzyme preparations: papain (P) and a microbial papain-like enzyme (PL) to yield quinoa protein hydrolysates (QPHs). The hydrolysates were evaluated for their DPP-IV inhibitory and oxygen radical absorbance capacity (ORAC) activity. Protein hydrolysis was observed in the QPI control, possibly due to the activity of quinoa endogenous proteinases. The QPI control had significantly higher DPP-IV half maximal inhibitory concentrations (IC 50 ) and lower ORAC values than QPH-P and QPH-PL (P < 0.05). Both QPH-P and QPH-PL had similar DPP-IV IC 50 and ORAC values. QPH-P had a DPP-IV IC 50 value of 0.88  0.05 mg mL -1 and an ORAC activity of 501.60  77.34 µmol Trolox equivalent (T.E.) g -1 . To our understanding, this is the first study demonstrating the in vitro DPP-IV inhibitory properties of quinoa protein hydrolysates. QPHs may have potential as functional ingredients with serum glucose lowering properties.

A significant amount of information is available linking consumption of milk proteins with the beneficial modulation of key outcomes associated with diet related conditions such as cardiovascular disease, obesity and type two diabetes. While intact milk proteins per se may have some direct health promoting effects, the functional food potential of dairy proteins is mainly attributed to the peptides formed following hydrolysis. Evidence/emerging evidence for the potential role of dairy peptides as hypotensive, mineral binding, immunomodulatory, anticancer, opioid agonist and antagonist, antimicrobial, gut health enhancing, hypo-cholesterolemic, insulinotrophic and neuromodulatory agents now exists. Some information outlining the biofunctional potential of caseinophosphopeptides (CPPs) is presented herein.

Milk protein derived peptides have numerous well-documented bioactive properties. The conventional approach for the generation, identification and validation of bioactive peptides (BAPs) has involved (i) protein hydrolysis, (ii) bioactivity screening and (iii) validation in vivo. The low potency (in comparison to conventional drugs), susceptibility to breakdown during gastrointestinal transit and low intestinal permeability are key challenges in the development of highly bioactive food protein hydrolysates/peptides. However, the generation of potent and effective health enhancing hydrolysates/peptides can benefit from a range of in silico techniques including the application of structure bioactivity relationship modelling (e.g., quantitative structure activity relationship (QSAR) modelling), molecular docking and design of experiments (DOE) approaches to optimise BAP production and identification. Some examples of how these approaches have been employed in BAP discovery and generation will be outlined.

Background: The workflow used for the discovery of novel milk protein-derived bioactive peptides (BAPs) has significantly evolved over the past number of years. Many of the approaches currently described in the literature for the study of milk protein-derived BAPs have taken similar routes as those conventionally used in drug discovery. However, because milk protein-derived BAPs are prepared from dietary sources and in a food-grade manner, they differ from drugs in that they are generally present in a complex mixture as opposed to drug active compounds which are generally synthetically produced, having a relatively high level of purity. Scope and approach: This review assesses the utilisation of drug discovery methods for the identification of novel milk protein-derived BAPs. Different methods relevant to drug discovery have already proven effective for the discovery of new BAPs from milk proteins. Key findings and conclusions: However, reversing the workflow, starting from peptide identification in vivo to in vitro screening followed by conventional drug discovery approaches may have several merits. These include (a) the design of more targeted studies, (b) the development of novel approaches for the generation of milk protein hydrolysates and (c) the discovery of milk BAPs which may be bioavailable in humans. These BAPs may find applications in the commercial generation of dietary ingredients with health promoting effects in humans.

Hen egg lysozyme was hydrolyzed with pepsin in situ on a cation-exchange column to isolate antioxidant peptides. The most cationic fraction was eluted with 1 M NaCl. Five positively charged peptides f(109-119) VAWRNRCKGTD, f(111-119) WRNRCKGTD, and) IRGCRL were identified using tandem mass spectrometry. Using ORAC-FL, all five peptides presented antioxidant activity with values of (1,970; 3,123; 2,743; 2,393 and 0.313 µmol Trolox/ µmol peptide) respectively. Using method TBARS in zebrafish larvae, all five synthetic peptides were found to efficiently inhibit lipid peroxidation (36.8; 51.6;55.56; 63.2; 61.0 % inhibition of lipid peroxidation) respectively. None of the five peptides were toxic in zebrafish eggs and larvae at concentrations lower than 50 µg/ml. Concentrations higher than 50µg/ml were toxic for both zebrafish eggs and larvae.

Hydrolysates, which are used in hypoallergenic infant formulas, have been found to possess immune modulating effects. For an optimal utilization of hydrolysates, the working mechanisms and responsible proteins underlying the effects should be elucidated. In this study, the immunomodulating activity of whey and soy hydrolysates was studied by quantifying TLR activation and assessing cytokine production in hydrolysate stimulated dendritic cells. The responsible protein fraction was identified and characterized by gel electrophoresis. The immune effects under gastrointestinal conditions were studied by digesting the hydrolysates in an in vitro infant digestion model, after which the digests were analyzed. In both soy and whey hydrolysates, TLR activation and cytokine production in dendritic cells were induced by a fraction containing protein aggregates larger than 1000 kDa, which were formed by electrostatic interactions and disulfide bonds. Only soy aggregates remained intact during duodenal digestion, and maintained the TLR activating capacity. Soy and whey protein aggregates larger than 1000 kDa possess immunomodulatory properties, but only soy aggregates remain under intestinal digestion conditions. This knowledge is important for a better understanding of the effects of hydrolysates.

Wheat hydrolysates are used for medical nutrition to provide undernourished patients a readily digestible protein source for instance to recover from chemotherapy induced damage to the intestine. Another potential route for hydrolysates to interfere with chemotherapy induced intestinal damage and inflammation is via the immune system since there is evidence that Toll-like receptors (TLRs) are involved in this process. Here we determined the capacity of three wheat hydrolysates to modulate immunity by interfering with TLR signaling. We studied possible inhibiting effects on TLR2, TLR4, and TLR9. All wheat hydrolysates had TLR modulating effects but only one had strong TLR2 inhibiting effects, both attenuating TLR2/1 and TLR2/6 signaling in a reporter cell system. Furthermore, this TLR2 blocking hydrolysate reduced IL-6 production in human dendritic cells. Application of Reversed Phase-Ultra High Performance Liquid Chromatography combined with mass-spectrometry revealed that the presence of peptide WQIPEQSR is associated with the TLR2 inhibiting capacity of wheat hydrolysates. Our study demonstrates TLR2 inhibiting capacities of a wheat hydrolysate. As not all hydrolysates have TLR2 inhibiting capacity, our technology platform with TLR expressing cells can be instrumental in identification of these bioactive hydrolysates. As chemotherapy induced intestinal inflammation is often TLR2 dependent, our findings provide a good start for further research to investigate whether these hydrolysates might contribute to the management of mucositis in cancer patients receiving chemotherapy. peptide Size (Dalton)

CNPq), pela concessão da bolsa de estudos (processo no. 142219/2004-5). À Fundação de Amparo à Pesquisa de São Paulo (FAPESP), pelo auxílio à pesquisa (processo no. 2006/01735-0). À professora Primavera Borelli, e ao grupo do Laboratório de Hematologia do Departamento de Análises Clínicas e Toxicológicas (FCF/USP), em especial às alunas de pós-graduação Amanda Rabello Crisma e Karina Nakajima, pela colaboração nas análises hematológicas. À doutora Vanda Jorgetti, e ao grupo do Laboratório de Investigação Médica 16 (LIM-16, FM/USP), pela colaboração nas análises histomorfométricas dos fêmures. À professora Eliana Parisi Alvares, e ao grupo do Laboratório de Morfofisiologia do Sistema Digestivo do Departamento de Biologia Celular e do Desenvolvimento (ICB/USP), em especial ao técnico de laboratório Cruz Alberto Rigonati, pela colaboração nas análises histológicas do intestino. À professora Elizabeth Aparecida Ferraz da Silva Torres, e ao grupo do Laboratório de Bromatologia do Departamento de Nutrição (FSP/USP), em especial à técnica de laboratório e aluna de pós-graduação Geni Rodrigues Sampaio, pela colaboração nas análises dos ácidos graxos de cadeia curta. À professora Lilian Rose Marques de Sá, do Laboratório de Gastroenteropatia Comparada e Experimental do Departamento de Patologia (FMVZ/USP), pela colaboração nas análises histológicas do fígado e baço. À professora Susana Marta Isay Saad, e a aluna de pós-graduação Flávia Carolina