-
International Dental Journal May 2024The aim of this study was to investigate the effect of bovine trypsin on the adhesion and pH of dental plaque biofilms.
OBJECTIVE
The aim of this study was to investigate the effect of bovine trypsin on the adhesion and pH of dental plaque biofilms.
METHODS
A multispecies dental plaque biofilm model and a single-species dental plaque biofilm model were established in vitro. Three groups were tested: (1) blank control group (aseptic ultrapure water); (2) negative control group (1M Tris-HCl buffer, pH = 7.4); and (3) experimental group (bovine trypsin). Adhesion ability was measured using an automatic microplate reader and visualised by confocal laser scanning microscopy (CLSM). The pH was measured using a pH meter. The expression of gtfB, gtfC, and gtfD was analysed using quantitative real-time polymerase chain reaction.
RESULTS
Adhesion ability in the experimental group was significantly lower than that in the blank group and the negative control group (P < .05); readhesion ability in the experimental group was inhibited for a certain period of time (24-hour multispecies biofilms were inhibited from 4 to 8 hours, and the 48- and 72-hour multispecies biofilms were inhibited from 2 to 6 hours; P < .05). The decrease in pH was inhibited for a certain period of time (24-hour multispecies biofilms were inhibited from 2 to 8 hours, and the 48- and 72-hour multispecies biofilms were inhibited from 1 to 8 hours; P < .05). Expression levels of gtfB, gtfC, gtfD, and ldh in the experimental group were significantly lower than those in the blank group (P < .05).
CONCLUSIONS
Bacterial adhesion, and readhesion, decreasd pH, and expression of adhesion- and acid-related genes by Streptococcus mutans in biofilms could be reduced by bovine trypsin for a certain period of time.
PubMed: 38760192
DOI: 10.1016/j.identj.2024.04.025 -
Scientific Reports May 2024Biofloc (BF) stands out as a promising system for sustainable shrimp farming. Optimizing various culture conditions, such as stocking density, carbohydrate source, and...
Influence of stocking density on the growth, immune and physiological responses, and cultivation environment of white-leg shrimp (Litopenaeus vannamei) in biofloc systems.
Biofloc (BF) stands out as a promising system for sustainable shrimp farming. Optimizing various culture conditions, such as stocking density, carbohydrate source, and feeding management, is crucial for the widespread adoption of the BF system. This study compares the growth performance of white-leg shrimp (Litopenaeus vannamei) in culture ponds at low density (LD) with 50 organisms/m and high density (HD) with 200 organisms/m. Post-larvae of white-leg shrimp were stocked for 16 weeks in both LD and HD groups. The LD group exhibited a superior survival rate, growth rate, and feed consumption compared to the HD group. The BF from the LD system recorded a significantly higher protein content (16.63 ± 0.21%) than the HD group (15.21 ± 0.34%). Heterotrophic bacterial counts in water did not significantly differ with stocking density. However, Vibrio count in water samples was higher in the HD group (3.59 ± 0.35 log CFU/mL) compared to the LD group (2.45 ± 0.43 log CFU/mL). The whole shrimp body analysis revealed significantly higher protein and lipid content in the LD group. In contrast, the total aerobic bacterial count in shrimp from the HD group was high, with the identification of Salmonella enterica ssp. arizonae. Additionally, Vibrio counts in shrimp samples were significantly higher in the HD group (4.63 ± 0.32 log CFU/g) compared to the LD group (3.57 ± 0.22 log CFU/g). The expression levels of immune-associated genes, including prophenoloxidase, transglutaminase, penaiedin 3, superoxide dismutase, lysozyme, serine proteinase, and the growth-related gene ras-related protein (rap-2a), were significantly enhanced in the LD group. Conversely, stress-related gene expression increased significantly in the HD group. Hepatopancreases amylase, lipase, and protease were higher in the LD group, while trypsin activity did not differ significantly. Antioxidant enzyme activity (catalase, glutathione, glutathione peroxidase, and superoxide dismutase) significantly increased in the LD group. The histological structure of hepatopancreas, musculature, and female gonads remained similar in both densities. However, negative effects were observed in the gills' histology of the HD group. These results suggest that increasing stocking density is associated with significantly negative biological, microbial, and physiological effects on white-leg shrimp under the BF system.
Topics: Animals; Penaeidae; Aquaculture; Vibrio; White
PubMed: 38750082
DOI: 10.1038/s41598-024-61328-4 -
Clinical and Translational Science May 2024Inflammatory bowel disease (IBD) is characterized by a chronically dysregulated immune response in the gastrointestinal tract. Bone marrow multipotent mesenchymal...
Inflammatory bowel disease (IBD) is characterized by a chronically dysregulated immune response in the gastrointestinal tract. Bone marrow multipotent mesenchymal stromal cells have an important immunomodulatory function and support regeneration of inflamed tissue by secretion of soluble factors as well as through direct local differentiation. CXCR4 is the receptor for CXCL12 (SDF-1, stromal-derived factor-1) and has been shown to be the main chemokine receptor, required for homing of MSCs. Increased expression of CXCL12 by inflamed intestinal tissue causes constitutive inflammation by attracting lymphocytes but can also be used to direct MSCs to sites of injury/inflammation. Trypsin is typically used to dissociate MSCs into single-cell suspensions but has also been shown to digest surface CXCR4. Here, we assessed the regenerative effects of CXCR4 and CXCR4 MSCs in an immune-deficient mouse model of DSS-induced colitis. We found that transplantation of MSCs resulted in clinical improvement and histological recovery of intestinal epithelium. In contrary to our expectations, the levels of CXCR4 on transplanted MSCs did not affect their regenerative supporting potential, indicating that paracrine effects of MSCs may be largely responsible for their regenerative/protective effects.
Topics: Animals; Receptors, CXCR4; Mesenchymal Stem Cells; Colitis; Mesenchymal Stem Cell Transplantation; Disease Models, Animal; Regeneration; Mice, Inbred C57BL; Mice; Dextran Sulfate; Intestinal Mucosa; Chemokine CXCL12; Bone Marrow Cells
PubMed: 38742709
DOI: 10.1111/cts.13821 -
Journal of Translational Medicine May 2024Glycosylation is an enzyme-catalyzed post-translational modification that is distinct from glycation and is present on a majority of plasma proteins. N-glycosylation...
BACKGROUND
Glycosylation is an enzyme-catalyzed post-translational modification that is distinct from glycation and is present on a majority of plasma proteins. N-glycosylation occurs on asparagine residues predominantly within canonical N-glycosylation motifs (Asn-X-Ser/Thr) although non-canonical N-glycosylation motifs Asn-X-Cys/Val have also been reported. Albumin is the most abundant protein in plasma whose glycation is well-studied in diabetes mellitus. However, albumin has long been considered a non-glycosylated protein due to absence of canonical motifs. Albumin contains two non-canonical N-glycosylation motifs, of which one was recently reported to be glycosylated.
METHODS
We enriched abundant serum proteins to investigate their N-linked glycosylation followed by trypsin digestion and glycopeptide enrichment by size-exclusion or mixed-mode anion-exchange chromatography. Glycosylation at canonical as well as non-canonical sites was evaluated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) of enriched glycopeptides. Deglycosylation analysis was performed to confirm N-linked glycosylation at non-canonical sites. Albumin-derived glycopeptides were fragmented by MS3 to confirm attached glycans. Parallel reaction monitoring was carried out on twenty additional samples to validate these findings. Bovine and rabbit albumin-derived glycopeptides were similarly analyzed by LC-MS/MS.
RESULTS
Human albumin is N-glycosylated at two non-canonical sites, Asn and Asn. N-glycopeptides were detected at both sites bearing four complex sialylated glycans and validated by MS3-based fragmentation and deglycosylation studies. Targeted mass spectrometry confirmed glycosylation in twenty additional donor samples. Finally, the highly conserved Asn in bovine and rabbit serum albumin was also found to be glycosylated.
CONCLUSIONS
Albumin is a glycoprotein with conserved N-linked glycosylation sites that could have potential clinical applications.
Topics: Glycosylation; Glycoproteins; Humans; Glycopeptides; Amino Acid Sequence; Tandem Mass Spectrometry; Animals; Molecular Sequence Data; Albumins; Cattle; Chromatography, Liquid
PubMed: 38741158
DOI: 10.1186/s12967-024-05000-5 -
Journal of Affective Disorders May 2024The therapeutic response to lithium in patients with bipolar disorder is highly variable and has a polygenic basis. Genome-wide association studies investigating lithium...
BACKGROUND
The therapeutic response to lithium in patients with bipolar disorder is highly variable and has a polygenic basis. Genome-wide association studies investigating lithium response have identified several relevant loci, though the precise mechanisms driving these associations are poorly understood. We aimed to prioritise the most likely effector gene and determine the mechanisms underlying an intergenic lithium response locus on chromosome 21 identified by the International Consortium on Lithium Genetics (ConLiGen).
METHODS
We conducted in-silico functional analyses by integrating and synthesising information from several publicly available functional genetic datasets and databases including the Genotype-Tissue Expression (GTEx) project and HaploReg.
RESULTS
The findings from this study highlighted TMPRSS15 as the most likely effector gene at the ConLiGen lithium response locus. TMPRSS15 encodes enterokinase, a gastrointestinal enzyme responsible for converting trypsinogen into trypsin and thus aiding digestion. Convergent findings from gene-based lookups in human and mouse databases as well as co-expression network analyses of small intestinal RNA-seq data (GTEx) implicated TMPRSS15 in the regulation of intestinal nutrient absorption, including ions like sodium and potassium, which may extend to lithium.
LIMITATIONS
Although the findings from this study indicated that TMPRSS15 was the most likely effector gene at the ConLiGen lithium response locus, the evidence was circumstantial. Thus, the conclusions from this study need to be validated in appropriately designed wet-lab studies.
CONCLUSIONS
The findings from this study are consistent with a model whereby TMPRSS15 impacts the efficacy of lithium treatment in patients with bipolar disorder by modulating intestinal lithium absorption.
PubMed: 38735581
DOI: 10.1016/j.jad.2024.05.050 -
Nutrients Apr 2024Vulvovaginal candidiasis (VVC) is the most common cause of vaginal discharge among women. The present study aimed to investigate the synergistic anticandidal effect of...
Vulvovaginal candidiasis (VVC) is the most common cause of vaginal discharge among women. The present study aimed to investigate the synergistic anticandidal effect of lactobacillus cultures supplemented with plant extracts. Among 600 isolates of lactic acid bacteria, 41 isolates exhibited inhibitory activity against ATCC10231. Six out of 41 cell-free supernatants demonstrated the most potent antibacterial and anticandidal activities. They also inhibited the clinical isolates of causing VVC and non-. The synergistic effect between 84/7 and 89/4 was demonstrated by the lowest fractional inhibitory concentration index (FICI = 0.5). The synbiotic culture of bacterial combination, cultured with Jerusalem artichoke () extract, also exhibited the strongest inhibition against the tested . Biofilm formation decreased after 12 h of incubation in the selected cell-free supernatants of this synbiotic culture. The anticandidal activity of crude extracts was lost after treatment with proteinase K and trypsin but not with heating conditions, suggesting that it may be a heat-stable substance. In conclusion, the combination of 84/7 and 89/4 with may be a promising candidate for inhibiting infection and biofilm formation, with the potential use as ingredients in vaginal biotherapeutic products.
Topics: Candida albicans; Plant Extracts; Female; Humans; Candidiasis, Vulvovaginal; Synbiotics; Vaginal Discharge; Biofilms; Lactobacillus; Limosilactobacillus reuteri; Lactobacillus crispatus; Antifungal Agents
PubMed: 38732618
DOI: 10.3390/nu16091372 -
Foods (Basel, Switzerland) May 2024Lima beans () and adzuki beans () are some of the most nutritious underutilized pulses that are significant in being used as basic ingredients for the preparation of...
Lima beans () and adzuki beans () are some of the most nutritious underutilized pulses that are significant in being used as basic ingredients for the preparation of various food products. The present study aimed to determine the impact of soaking and germination on nutritional and bioactive components, in vitro protein digestibility, reducing power, metal chelating capacity, antioxidant activity, and anti-nutritional components of lima and adzuki beans. The findings showed that during the germination treatment, the in vitro protein digestibility of lima and adzuki beans increased by 14.75 and 10.98%, respectively. There was an increase in the antioxidant activity of lima beans by 33.48% and adzuki beans by 71.14% after 72 h of germination, respectively. The reducing power assay of lima and adzuki beans indicated an increase of 49.52 and 36.42%, respectively, during germination. Similarly, the flavonoid and metal chelating activity increased in lima and adzuki beans after 72 h of germination. In contrast, the anti-nutrients, such as phytic acid, tannin content, and trypsin inhibitor activity, decreased significantly < 0.05 after 72 h of germination. These results are encouraging and allow for utilizing the flour obtained from the germinated beans in functional bakery products, which can contribute to eradicating protein deficiency among some population groups. At the same time, promoting soaking and germination of the beans as a way to enhance the nutritional quality and reduce anti-nutrients can contribute to the interest in these underutilized pulses. They could be seen as an additional tool to improve food security.
PubMed: 38731793
DOI: 10.3390/foods13091422 -
Foods (Basel, Switzerland) Apr 2024A novel fibrinolytic enzyme was produced by the liquid fermentation of . The enzyme was purified from the culture supernatant by hydrophobic interactions, gel...
A novel fibrinolytic enzyme was produced by the liquid fermentation of . The enzyme was purified from the culture supernatant by hydrophobic interactions, gel filtration, and ion exchange chromatographies. It was purified by 241.02-fold, with a specific activity of 3619 U/mg and a final yield of 10.02%. SDS-PAGE analysis confirmed the purity of the enzyme, showing a single band with a molecular weight of 19.5 kDa. The first nine amino acids of the N-terminal of the purified enzyme were A-T-Y-T-G-G-S-Q-T. The enzyme exhibited optimal activity at a temperature of 42 °C and pH 7.6. Its activity was significantly improved by Zn, K, Ca, Mn, and Mg while being inhibited by Fe, Fe, Al, and Ba. The activity of the enzyme was completely inhibited by ethylenediamine tetraacetic acid (EDTA), and it was also dose-dependently inhibited by phenylmethylsulfonyl fluoride (PMSF) and soy trypsin inhibitor (SBTI). However, inhibitors such as N-α-tosyl-L-phenylalanine chloromethyl ketone (TPCK), aprotinin, and pepstatin did not significantly affect its activity, suggesting that the enzyme was a serine-like metalloproteinase. The enzyme acted as both a plasmin-like fibrinolytic enzyme and a plasminogen activator, and it also exhibited the capability to hydrolyze fibrinogen and fibrin. In vitro, it demonstrated the ability to dissolve blood clots and exhibit anticoagulant properties. Furthermore, it was found that the enzyme prolonged activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT), and reduced the levels of fibrinogen (FIB) and prothrombin activity (PA). Based on these studies, the enzyme has great potential to be developed as a natural agent for the prevention and treatment of thrombotic diseases.
PubMed: 38731663
DOI: 10.3390/foods13091292 -
Animals : An Open Access Journal From... Apr 2024An 8-week feeding trial was performed to investigate the effects of dietary bile acids on growth, glucose metabolism, and intestinal health in spotted seabass () reared...
An 8-week feeding trial was performed to investigate the effects of dietary bile acids on growth, glucose metabolism, and intestinal health in spotted seabass () reared at high temperatures (33 °C). The fish (20.09 ± 1.12 g) were fed diets supplemented with bile acids: 0 (Con), 400 (BA400), 800 (BA800), and 1200 (BA1200) mg/kg, respectively. The results showed that the growth was promoted in fish at the BA800 treatment compared with the control ( < 0.05). Increased enzyme activities and transcripts of gluconeogenesis in the liver were observed, whereas decreased enzyme activities and transcripts of glycolysis, as well as glycogen content, were shown in the BA800 treatment ( < 0.05). The transcripts of bile acid receptors in the liver were up-regulated in the BA800 treatment ( < 0.05). A bile acid supplementation of 800 mg/kg improved the morphological structure in the intestine. Meanwhile, intestinal antioxidant physiology and activities of lipase and trypsin were enhanced in the BA800 treatment. The transcripts of genes and immunofluorescence intensity related to pro-inflammation cytokines (, , and ) were inhibited, while those of genes related to anti-inflammation ( and ) were induced in the BA800 treatment. Furthermore, transcripts of genes related to the NF-κB pathway in the intestine (, , , and ) were down-regulated in the BA800 treatment. This study demonstrates that a dietary bile acid supplementation of 800 mg/kg could promote growth, improve glucose metabolism in the liver, and enhance intestinal health by increasing digestive enzyme activity and antioxidant capacity and inhibiting inflammatory response in .
PubMed: 38731303
DOI: 10.3390/ani14091299 -
BMC Research Notes May 2024Bovine seminal plasma proteins perform several functions related to sperm function. Changes in the expression pattern or abundance of seminal proteins are related to...
OBJECTIVES
Bovine seminal plasma proteins perform several functions related to sperm function. Changes in the expression pattern or abundance of seminal proteins are related to changes in the fertilizing capacity of bulls. Considering the role of seminal plasma proteins in sperm function and animal reproduction, we investigated changes in the protein abundance profile in response to sperm morphological changes using a proteomic approach.
DATADESCRIPTION
In our present investigation, we employed liquid chromatography coupled with mass spectrometry to elucidate the proteomic composition of seminal plasma obtained from Nellore bulls exhibiting varying percentages of sperm abnormalities. Following semen collection, seminal plasma was promptly isolated from sperm, and proteins were subsequently precipitated, enzymatically digested using porcine trypsin, and subjected to analysis utilizing the Acquity nano UHPLC System in conjunction with a mass spectrometer. This dataset encompasses a total of 297 proteins, marking the inaugural instance in which a comparative profile of seminal plasma proteins in young Nellore bulls, categorized by their sperm abnormality percentages, has been delineated using LC-MS/MS. The comprehensive nature of this dataset contributes pivotal proteomic insights, representing a noteworthy advancement in our understanding of the reproductive biology of the Nellore breed.
Topics: Animals; Male; Cattle; Semen; Proteome; Spermatozoa; Tandem Mass Spectrometry; Proteomics; Seminal Plasma Proteins; Chromatography, Liquid
PubMed: 38730318
DOI: 10.1186/s13104-024-06796-7