Enzymatic Digestion Research Articles

Optimal human ovarian follicle isolation: A review focused on enzymatic digestion.

The damage or depletion of ovarian reserves due to aging or cancer treatment can increase the need for fertility preservation techniques. One of the most common ways of supporting fertility in prepubertal girls and women who require immediate cancer treatment is through ovarian tissue cryopreservation and re-transplantation following cancer treatment. However, a more appropriate method should be employed in diseases such as leukemia, where malignant cells may be present in cryopreserved tissue, instead of ovarian tissue transplantation. Human ovarian follicle isolation for in vitro culture or the use of artificial ovaries for their growth can decrease the risk of reintroducing cancer cells into these individuals. Here we review the methods for the isolation of human ovarian follicles.

Impact of porcine brush border membrane enzymes on INFOGEST in vitro digestion model: A step forward to mimic the small intestinal phase

Brush border membrane (BBM) enzymes greatly affect the bioaccessibility and bioavailability of food nutrients. Despite their physiological importance, a step simulating the final stage of intestinal digestion has not yet been included in the harmonized protocols for in vitro digestion, primarily due to the challenges of replicating the dynamics of intestinal degradation. Herein, we propose an advancement toward a more physiologically relevant method, complementing the harmonized static gastric-duodenal digestion INFOGEST model with the missing small intestinal phase. BBM hydrolase activity, incubation time, at pH 7.2 were established to reproduce the small intestinal conditions. Skim milk powder, as a model of protein food, was subjected to the in vitro static digestion. Immediately after the duodenal phase, digesta were supplemented with BBM vesicles purified from pig jejunum. To comply with the dynamic nature of intestinal digestion and balance the spontaneous inactivation of hydrolases, BBM supplements were added every two hours throughout 6 h incubation time. Peptide degradation was monitored at each stage of digestion by amino acid analysis, free α-amino group assay, HPLC, LC-MS/MS. Hydrolysis by BBM peptidases led to a significant increase of free amino acids, reflecting the known level of amino acid adsorption (> 90 %) in humans after eating milk proteins. LC-MS/MS analysis demonstrated that BBM hydrolases erode progressively the peptides released by gastro-duodenal processing up to stable sequence motifs. The approach described is particularly relevant when the endpoint is identifying the peptide sequences that cannot be further hydrolysed by digestive enzymes or to determine the amino acid bio-accessibility.

A Review on Pharmacological Potential of Medicago sativa Linn

Background: Medicago sativa Linn. in Hindi, is called lasunghas, and in English, it is called alfalfa. It is a well-known crop of the Fabaceae family. We can eat its sprouts as a salad vegetable. This plant has a wide range of uses as a feed crop and is tolerant to climate change. Objective: In this review, the authors discussed the active chemical constituents and pharmacological uses of Medicago sativa Linn. in various diseases. Methods: This review article gathered all the information after going through the research papers from different databases like Taylor & Francis, ResearchGate, Elsevier, PubMed, Wiley Online Library, and PMC. Results: Alfalfa has been shown to be an important origin of phytochemicals like amino acids, coumarins, alkaloids, carotenes, flavonoids, organic acids, digestive enzymes, phytosterols, saponins, phytoestrogens, phenolic compounds, polyamines, and a few other volatile organic compounds like ketones, terpenes, alcohols and furanoids. This plant is useful in CNS, anxiety, oxidative stress, hepatotoxicity, inflammation, cancer, atherosclerosis, diabetes, burn wounds, vitiligo, reproductive system, myocardial infraction, bacteria, xanthine oxidase and metabolic syndrome. Conclusion: This review may be helpful in knowing of Medicago sativa Linn. with greater therapeutic potential for treatment of various diseases.

Modulating the assembly of starch-lecithin inclusion complexes and its underlying mechanism for controlling starch digestibility under the dense system

Starch-lipid inclusion complexes were used to regulate starch digestibility by changing the binding affinity of digestive enzymes such as α-amylase and α-glucosidase. However, when above a critical aggregation concentration (CAC) of starch and lipid molecules, they tended to self-assemble into double helices or normal micelles respectively, rather than interacting with each other to form starch-lipid complexes. Thus, how to enhance the complexation efficiency under a dense system is the key approach for the improvement of starch bioaccessibility and nutritional function. Herein, based on the dimethyl sulfoxide (DMSO) reverse solvent method, the effect of water/DMSO ratios and starch chain structures on the interaction and assembly behaviors between starch and lecithin molecules in high concentrations were systematically investigated. The results revealed that water/DMSO in a ratio of 1.5:1 promoted the interactions between cavities of starch helices and long alkyl chains of lecithin molecules, with higher level of complexed lecithin content, relative crystallinity (RC), enthalpy (ΔH) (Tp＞100°C), ordered-aggregation structures and resistant starch (RS) content. In this kind of solvent, starch samples with higher content of amylose and longer chains (degree of polymerization (DP) of B2 chains and B3+ chains) is advantageous to the formation of VII-type inclusion starch-lecithin complexes, with a rise in RS from 14.17% to 43.64%. It provided an effective method to modulate starch digestibility under the dense system of starch and lecithin via modulating the water/DMSO ratios and starch fine structures and lay the foundation for the large-scale preparation and application of starch-lecithin complexes.

Seasonal variations of allocation strategy in carbon, nitrogen and phosphorus among tissues of male Mytilus coruscus are regulated by the gametogenic cycle: a case study in a mariculture system

This study examines the seasonal allocation strategy of carbon, nitrogen, and phosphorus in tissues of male Mytilus coruscus within a mariculture system at Gouqi Island, providing insights into the mussels' reproductive and nutritional adaptations. The reproductive cycle of mussels is bifurcated, with a resting phase in summer and active gametogenesis from autumn to spring. Digestive enzyme activity in mussels correlates negatively with food conditionality, especially when food is scarce in winter, enzyme activity is heightened to enhance nutrient extraction, whereas in summer, with an abundance of food, enzymatic activity is conserved for other physiological processes. This suggests an adaptive modulation to optimize energy allocation between growth, maintenance, and reproduction. Allometric growth analysis reveals that smaller mussels exhibit uniform nutrient allocation across tissues, focusing on somatic growth. In contrast, larger, mature mussels prioritize phosphorus utilization in gonads, indicative of reproductive investment. This shift aligns with the predictive adaptive response model, anticipating future reproductive demands by adjusting resource allocation. The study also identifies seasonal fluctuations in nutrient demand and allocation among tissues, with gonadal tissues showing preferential accumulation of nitrogen and phosphorus during mature phases. Although mussels preferentially allocate nutrients to the shell only in autumn, a consistent preferential accumulation of nitrogen and phosphorus to this critical structural component, highlighting the importance of these elements in biomineralization and shell maintenance. This research elucidates the complex interplay between environmental factors, physiological processes, and nutrient dynamics in male M. coruscus, and underscore the importance of considering seasonal variations in nutrient allocation for sustainable mariculture practices.

Effects of different feeding rates on growth performance, intestinal digestive enzyme activity, and intestinal microbial composition of red swamp crayfish (Procambarus clarkii)

The study aimed to investigate the effects of different feeding rates on the growth, muscle composition, intestinal digestive enzyme activities, and intestinal microbiota composition of juvenile crayfish (Procambarus clarkii). A total of 240 crayfish with an initial body weight of 8.33±0.50 g were randomly distributed into five groups, and each group had three replicates. The five groups corresponded to five different feeding rates of 2%, 4%, 6%, 8%, and 10% body weight per day (BW/day). The feeding trial lasted for six weeks. The results showed that the final body weight (FBW), weight gain rate (WGR), and specific growth rate (SGR) first increased when the feed rate increased from 2% to 6 % (P<0.05) and then showed a decreased trend with the further increase of the feeding rate, but no significant differences were observed. The feed conversion ratio (FCR) increased with a rise in feeding rate, which reached a peak at the feeding rate of 10% (P<0.05). The survival rate (SR), hepatosomatic index (HSI), and condition factor (CF) were unaffected by the feeding rates (P>0.05). The moisture, crude protein, crude fat, and ash contents of crayfish muscle remained unchanged among all feeding rate groups. Intestinal activities of lipase and amylase increase with the feeding rate (up to 6%) increasing (P<0.05). The ACE and Chao indices in the groups fed at 2% of feeding rate were significantly lower than those at a feeding rate of 10% (P<0.05). The relative abundance of Firmicutes in the groups fed at feeding rates of 4%, 6%, and 8% was higher than that in other groups (P<0.05), Citrobacter in the group fed at feeding rate of 6% was lower than that in different groups (P<0.05). The intestinal microbiota primarily functions in connection to amino acid, carbohydrate, and vitamin metabolism. Therefore, the optimum feeding rate could be 6.0% of the feeding rate of juvenile P. clarkii.

Characterization of poly(A) and poly(T) tail lengths in plasmid DNA by liquid chromatography high-resolution mass spectrometry.

Rapid advances in messenger RNA (mRNA) technology necessitate effective analytical methods. This study describes the development of a novel in vitro method using ion-pair reversed-phase liquid chromatography coupled to high-resolution mass spectrometry (IP-RP-LC-HRMS) for assessing the poly(A) and complementary poly(T) tail lengths directly into the DNA template used to manufacture mRNA. Briefly, after the validation of poly(A) tail length in the plasmid by Sanger sequencing, double-stranded DNA fragments containing these tails in the plasmid of interest were amplified by thepolymerase chain reaction (PCR), purified on silica column, and digested with restriction enzymes ClaI and HindIII. Gel or capillary electrophoresis confirmed sample quality and enzymatic digestion efficiency. Subsequently, poly(A) and complementary poly(T) tails were extracted and analyzed by LC-MS to determine their length and heterogeneity at a single-nucleotide resolution. Three DNA templates containing poly(A) tail lengths of 60A-G, 95A, or 108A were studied. LC-MS results correlated well with Sanger sequencing, identifying major populations of 60A-G, 95A, or 108A. Surprisingly, unlike Sanger sequencing, LC-MS analysis revealed minor poly(A) populations with lengths longer or shorter than the theoretically encoded poly(A) tail length. This finding could be explained by (i) the slippage of bacterial DNA polymerase I during plasmid replication in bacterial culture, which occurs on repeat mononucleotide sequences, or (ii) the slippage of Q5® High-Fidelity DNA Polymerase during PCR amplification. In conclusion, the method is easy, rapid, and accurate and could replace Sanger sequencing to assess the poly(A) and complementary poly(T) tail lengths in plasmid DNA.

Effect of fasting and re-feeding cycles on growth, glucose level, glycogen level, and digestive enzyme activity of Nile tilapia juveniles (Oreochromis niloticus) for cost-effective aquaculture

Effect of fasting and re-feeding cycles on growth, glucose level, glycogen level, and digestive enzyme activity of Nile tilapia juveniles (Oreochromis niloticus) for cost-effective aquaculture

Study on anti-adhesion effect and mechanism of dynamic and static stress stimulation during early healing process of rat Achilles tendon injury

To investigate the anti-adhesive effect and underlying mechanism of dynamic and static stress stimulation on the early healing process of rat Achilles tendon injury. Achilles tendon tissues of 15 male Sprague Dawley (SD) rats aged 4-6 weeks were isolated and cultured by enzyme digestion method. Rat Achilles tendon cells were treated with tumor necrosis factor α to construct the Achilles tendon injury cell model, and dynamic stress stimulation (dynamic group) and static stress stimulation (static group) were applied respectively, while the control group was not treated. Live/dead cell double staining was used to detect cell activity, ELISA assay was used to detect the expression of α smooth muscle actin (α-SMA), and real-time fluorescence quantitative PCR was used to detect the mRNA expression of collagen type Ⅰ (COL1A1), collagen type Ⅲ (COL3A1), and Scleraxis (SCX). Thirty male SD rats aged 4-6 weeks underwent Achilles tendon suture and were randomly divided into dynamic group (treated by dynamic stress stimulation), static group (treated by static stress stimulation), and control group (untreated), with 10 rats in each group. HE staining and scoring were performed to evaluate the healing of Achilles tendon at 8 days after operation. COL1A1 and COL3A1 protein expressions were detected by immunohistochemical staining, α-SMA and SCX protein expressions were detected by Western blot, and maximum tendon breaking force and tendon stiffness were detected by biomechanical stretching test. In vitro cell experiment, when compared to the static group, the number of living cells in the dynamic group was higher, the expression of α-SMA protein was decreased, the relative expression of COL3A1 mRNA was decreased, and the relative expression of SCX mRNA was increased, and the differences were all significant ( P<0.05). In the in vivo animal experiment, when compared to the static group, the tendon healing in the dynamic group was better, the HE staining score was lower, the expression of COL1A1 protein was increased, the expression of COL3A1 protein was decreased, the relative expression of SCX protein was increased, the relative expression of α-SMA protein was decreased, and the tendon stiffness was increased, the differences were all significant ( P<0.05). Compared with static stress stimulation, the dynamic stress stimulation improves the fibrosis of the scar tissue of the rat Achilles tendon, promote the recovery of the biomechanical property of the Achilles tendon, and has obvious anti-adhesion effect.

Prospective and challenges of locally applied repurposed pharmaceuticals for periodontal tissue regeneration

Periodontitis is a persistent inflammatory condition that causes periodontal ligament degradation, periodontal pocket development, and alveolar bone destruction, all of which lead to the breakdown of the teeth’s supporting system. Periodontitis is triggered by the accumulation of various microflora (especially anaerobes) in the pockets, which release toxic substances and digestive enzymes and stimulate the immune system. Periodontitis can be efficiently treated using a variety of techniques, both regional and systemic. Effective therapy is dependent on lowering microbial biofilm, minimizing or eradicating pockets. Nowadays, using local drug delivery systems (LDDSs) as an adjuvant therapy to phase I periodontal therapy is an attractive option since it controls drug release, resulting in improved efficacy and lesser adverse reactions. Choosing the right bioactive agent and mode of delivery is the foundation of an efficient periodontal disease management approach. The objective of this paper is to shed light on the issue of successful periodontal regeneration, the drawbacks of currently implemented interventions, and describe the potential of locally delivered repurposed drugs in periodontal tissue regeneration. Because of the multiple etiology of periodontitis, patients must get customized treatment with the primary goal of infection control. Yet, it is not always successful to replace the lost tissues, and it becomes more challenging as the defect gets worse. Pharmaceutical repurposing offers a viable, economical, and safe alternative for non-invasive, and predictable periodontal regeneration. This article clears the way in front of researchers, decision-makers, and pharmaceutical companies to explore the potential, effectiveness, and efficiency of the repurposed pharmaceuticals to generate more economical, effective, and safe topical pharmaceutical preparations for periodontal tissue regeneration.

Force-triggered density gradient sedimentation and cocktail enzyme digestion treatment for isolation of single dermal papilla cells from follicular unit extraction harvesting human hair follicles.

Hair follicles (HFs) are dynamic structures which are readily accessible within the skin that contain various pools of stem cells with broad regenerative potential, such as dermal papilla cells (DPCs), dermal sheath cells, and epithelial HF stem cells. DPCs act as signalling centres for HF regeneration. The current method for isolating human DPCs are inefficient. These methods struggle to obtain freshly isolated original DPCs and do not maintain the characteristics of DPCs effectively. In this study, two simple but more efficient methods were explored. Force-triggered density gradient sedimentation (FDGS) and cocktail enzyme digestion treatment (CEDT) were used to isolate purified DP spheres from human HFs, obtaining purified freshly isolated original DPCs from DP spheres. The expression profiles of isolated DPCs were tested, and gene expression of DPC-specific markers were analyzed using immunofluorescence staining, RT-qPCR and western blot. The 10% Ficoll PM400 was determined as the optimal concentration for FDGS method. Primary DPCs, DSCs and HFSCs were isolated simultaneously using the FDGS and CEDT method. The expression profiles of fresh DPCs isolated using the FDGS and CEDT methods were similar to those of traditionally isolated DPCs. DP-specific markers were expressed at significantly higher levels in freshly isolated DPCs than in traditionally isolated DPCs. Compared to traditional methods, the presented laboratory protocols were able to isolate fresh DPCs with high efficiency, thereby improving their research potential.

Abstract 4118084: Targeting Monoamine Oxidase B in Dystrophic mdx Hearts Dampens Inflammation and Fibrosis

Background: Cardiomyopathy is a major cause of death for Duchenne muscular dystrophy (DMD) patients. Current treatments cannot prevent cardiac tissue remodeling. Oxidative stress, inflammation and fibrosis are early events in DMD, preceding heart dysfunction. Monoamine Oxidase B (MAOB), which forms H 2 O 2 by oxidizing amines, is overactivated in inflammatory conditions and overcomes cell antioxidant defenses, thus altering redox homeostasis and eliciting harmful effects. We showed that targeting MAOB with inhibitors (iMAOB) improves skeletal muscle function in dystrophic mice by lowering oxidative stress, and reduces inflammation in murine models of sepsis and arthritis by dampening NF-kB, a redox-sensitive transcription factor. Aim: We explore the therapeutic potential of iMAOB to alleviate cardiomyopathy using dystrophin-deficient mdx mice. We hypothesize that iMAOB treatment could dampen oxidative stress, inflammation and fibrosis in dystrophic mdx hearts by modulating the phenotype of cells that are crucial in cardiac remodeling. Methods: Three-month-old mdx mice, that already show signs of fibrosis but no cardiac dysfunction, were orally treated with iMAOB or vehicle for one month (n≥ 6). Heart ventricular mononucleated cells were obtained by enzymatic digestion. Myeloid, endothelial and fibroblast cells were isolated by cell sorting by FACS and analysed by RT-PCR. Oxidative stress, inflammation and fibrosis were measured in whole tissue by immunohistochemistry. Results: The expression of proinflammatory and profibrotic genes was markedly increased in myeloid [Interleukin (Il)-1b, Il-6, Tgf-b and Spp1, the gene coding for osteopontin], endothelial [Il-1b, Il-6, mmp2 and nos3] and cardiac fibroblast cells [Tgf-b, Spp1, Timp1 and Col1] isolated from mdx hearts, as compared to wild type mice. All these genes were significantly dampened by iMAOB treatment. In parallel, once again iMAOB treatment blunted the increase in oxidative stress, inflammation and fibrosis observed in mdx cardiac sections. Of notice, the differences were not linked to changes in the percentage of the various cell types, as they were unmodified amongst wild type, mdx and iMAOB-treated mdx hearts. Conclusions: We show that iMAOB can positively affect the phenotype of cells that are important in cardiac tissue remodeling. Our data suggest that iMAOB can be a viable therapeutic tool in DMD cardiomyopathy. As iMAOB are already in clinical use, such approach could be easily translated to patients.

Ultrastructure Expansion Microscopy (U-ExM) of the Fission Yeast Schizosaccharomyces pombe.

Among widely used super-resolution microscopy techniques, including stimulated emission depletion (STED), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM), expansion microscopy (ExM) is unique in achieving increased resolution through a physical manipulation of the actual sample rather than optics or postprocessing. Originally developed for applications in neuroscience, ExM now has a solid foothold across many fields and model systems, and has been adapted to work for organisms with cell walls, including budding and fission yeasts, through the inclusion of a pre-expansion enzymatic digestion step. A variant of the ExM technique optimized for preserving the architecture of protein complexes, ultrastructure expansion microscopy (U-ExM), enables super-resolution imaging of full 3D volumes at increased throughput using conventional microscopes and can be readily combined with commonly used antibodies, dyes, and stains. Here, we present its application to the fission yeast Schizosaccharomyces pombe.

Pancreatic Heterotopia in the Biliary Cyst Wall of a Pregnant Patient and Literature Review.

Pancreatic heterotopia is the presence of pancreatic tissue in a different anatomical location with no connection to the main pancreas. The most common locations in the gastrointestinal system are duodenum, stomach, and Meckel diverticulum. However, it is quite rare in the hepatobiliary system. Here we present a patient with pancreatic heterotopia who we treated with left hemihepatectomy and extrahepatic bile duct resection for Todani type 4a bile duct cyst. Heterotopic pancreatic tissue was present in dilated cystic bile ducts on pathological examination. A 22-year-old 26-week pregnant patient presented to the emergency department with right upper quadrant pain. There was massive dilatation of the left lobe bile ducts and common bile duct, and multiple stones were observed distal to the common bile duct. The patient was hospitalized for definitive surgical treatment 4 months after giving birth. The patient underwent left hemihepatectomy, extrahepatic bile duct resection, and hepaticojejunostomy. The postoperative course was uneventful. Pathological examination of the specimen showed heterotopic pancreatic tissues in the wall of the dilated cystic intrahepatic and extrahepatic bile ducts. One of the most accepted hypotheses in the etiology of biliary tract cysts involves the abnormal union of the pancreatic duct and common bile duct. This lesion is unique in the absence of pancreaticobiliary junction anomaly. We think that cystic dilation of intrahepatic and extrahepatic bile ducts is likely secondary to recurrent inflammation due to the release of digestive enzymes by heterotopic pancreatic tissue in the biliary tract. It is also the first example in the literature diagnosed during pregnancy.

Development of an Automated, Ultra-Rapid Bottom-Up Proteomics Workflow Utilizing Alginate-Based Hydrogels.

A new approach to sample preparation and enzymatic digestion in bottom-up proteomics has been developed using alginate-based hydrogel entrapment of enzymes. This hydrogel facilitates rapid and room-temperature digestions with multienzyme capabilities. Three methodologies were tested: within microcentrifuge tubes, in situ pipette tips, and automated robotic liquid handling. Factorial experimental design identified a 1 h, room temperature, pepsin-trypsin dual-enzyme digestion as optimal for sequence coverage and protein group identification, comparable to a gold-standard overnight proteomic protocol. This method promises significant advancements in proteomic analysis by enhancing reusability, speed, throughput, convenience, and cost-effectiveness, without hindering digestion efficiency.

Comparison between PCR-RFLP and sequencing techniques in the analysis of Paracoccidioides spp. biodiversity: limitations and insights into species and variant differentiation.

The study of Paracoccidioides spp. faces significant challenges due to limitations inherent in the molecular biology techniques employed. Recently, new species were described whose geographical and genetic distributions were investigated. The phylogenetic studies have revealed that genotypes originally thought to be exclusive in specific regions from South American countries are now being found in other areas of the continent. This finding indicates a broader geographic distribution of these genotypes than previously recognized. To evaluate two molecular biology techniques employed to identify genotypes of Paracoccidioides spp. strains from a Brazilian culture collection previously identified only by mycological methods. DNA samples from 35 Paracoccidioides spp. strains maintained in a Brazilian culture collection were subjected to amplification and enzymatic digestion with PCR-RFLP of tub1 gene, followed by sequencing of gp43 Exon 2 loci. Strains with species identification discrepancies had their tub1 sequences determined to verify possible nucleotide mutations. The genotypic characterization of Paracoccidioides spp. using PCR-RFLP of the tub1 gene identified 22 isolates as P. brasiliensis sensu stricto, two as P. americana, four as P. restrepiensis, and eight as P. lutzii. Sequencing of the gp43 Exon 2 loci revealed discrepancies in the identification of four P. venezuelensis isolates, previously characterized as P. brasiliensis sensu stricto by PCR-RFLP of tub1. The sequencing of tub1 from P. brasiliensis sensu stricto and P. venezuelensis isolates revealed nucleotide differences in the pyrimidine class (C and T) in their sequences, specifically at the position 176bp. These molecular tools were able to establish the genetic diversity within the Paracoccidioides genus, crucial for taxonomy and epidemiology studies. The finding of presence of P. venezuelensis in Brazil, previously thought to be exclusive to Venezuela, highlights genetic connections and evolutionary divergences within the genus. While the PCR-RFLP of tub1technique showed limitations in identifying P. venezuelensis, sequencing of the gp43 Exon 2 loci was able to accurately identify this genotype. Thus, our findings contribute to the understanding of the molecular epidemiology of PCM and emphasize the need for precise species characterization in mycological research.

Genotyping of human isolates from human toxoplasmosis outbreak: Restriction Fragment Length Polymorphism and Sanger Sequencing.

Toxoplasmosis is a parasitic disease caused by infection with Toxoplasma gondii, capable of infecting a wide range of hosts. The parasite exhibits a broad genetic diversity, necessitating genotypic characterization for genotype identification and associations with epidemiological information. Therefore, the Restriction Fragment Length Polymorphism (RFLP) technique is used for characterization. This study aimed to perform genotypic characterization of isolates from pregnant women infected during a human toxoplasmosis outbreak, comparing the RFLP and Sanger Sequencing methodologies. For this purpose, six human isolates were subjected to conventional PCR, Multiplex PCR, Nested PCR, Enzymatic Digestion, and Sanger Sequencing. Additionally, the standard strains GTI (Type I), PTG (Type II), and CTG (Type III) were also subjected to the same techniques described above. Subsequently, the amplified DNA products were compared with the standard strains. As a result, it was observed that Sanger Sequencing provides the same information as RFLP PCR, as well as the possibility of cost reduction for genotypic characterization, and providing greater agility in issuing results. Additionally, Sanger Sequencing of T. gondii isolates allows for detailed evaluation of nucleotide sequences, including the assessment of SNPs and enzymatic restriction sites, which the RFLP technique does not.

High-Order Multimodal Interaction Network for Efficient Prediction of Drug-Drug Interaction

The pharmaceutical industry places a lot of focus on preventing drug-drug interactions (DDIs). Medication interaction detection has been the primary emphasis of machine learning-based DDI prediction methods. Since studies have shown that DDIs can cause different future occurrences, it is more beneficial to predict DDI connected events while examining the mechanism behind combination medicine consumption or adverse reactions. The areas of medication development and illness diagnostics are seeing increased usage of a developing approach that predicts DDIs-associated occurrences. We examine potential interactions between the two medications as well as the kinds of interactions that can occur in this research. Additionally, we provide a strategy that relies on learning and use High-Order Multimodal Interaction Network (HOMIN) to forecast DDIs by learning feature representations. The purpose of this work is to provide HOMIN-DDI, a new method for drug-drug interaction predictions that we have developed utilizing a HOMIN architecture. First, we develop feature vectors from medication categories, targets, paths, and digestive enzymes, and we use the Jaccard similarity to evaluate how similar drugs appear. After that, we build a new HOMIN using the feature representation to predict DDIs. The results of the experiments show that the drug categories feature type, when used to the HOMIN -DDI approach, is effective. Furthermore, compared to employing only one feature, combining numerous features yields more useful and informative results. Compared to other algorithms, HOMIN-DDI performs better when it comes to predicting DDIs.

Artemisinin supplementation improves growth, lipid metabolism, and intestinal microbiota of golden pompano (Trachinotus ovatus) fed high cottonseed meal diets

This study aimed to investigate the effect of artemisinin supplementation to high cottonseed meal (CSM) diet on the growth, lipid metabolism and intestinal microbiota of golden pompano (Trachinotus ovatus). Five diets were formulated, including the control (CM, with CSM replacing 16 % of fishmeal) and experimental groups (CA0, CA2, CA4, and CA8, with CSM replacing 36 % of fishmeal and supplemented with 0 %, 0.20 %, 0.40 %, and 0.80 % of artemisinin, respectively). 375 fishes (initial weight 22.45 ± 0.07 g) were randomized into five groups of triplicates each and fed in tanks for 8 weeks. Results indicated that 0.8 % artemisinin improved final weight, weight gain rate and specific growth rate, reduced the serum lipid levels under the condition of 36 % fishmeal replaced with CSM. In the liver, artemisinin supplementation alleviated hepatocyte vacuolization and lipid deposition induced by the high level CSM replacement fishmeal diet, reduced the activity of lipogenesis-related enzymes and increased lipolysis-related enzyme activity. In the intestine, supplementation with 0.8 % artemisinin significantly increased the activities of intestinal digestive enzymes. Supplementation with 0.8 % artemisinin significantly increased the diversity and stability of the intestinal microbiota of the golden pompano, while also reducing the abundance of potentially harmful Vibrio associated with high CSM levels. In conclusion, 0.8 % artemisinin supplementation mitigated the negative effects of high CSM diets on growth, lipid metabolism, and liver health in golden pompano by enhancing lipid metabolism, intestinal digestive enzyme activity, and modulating intestinal microbiota.

Moringa oleifera Leaf Meal: A Sustainable Approach for Poultry Production: A Review

Poultry production is among the fastest ways to address the shortage of animal protein supply and intake in many nations. With the prohibition of antibiotics as growth promoters in the poultry industry due to rising bacterial resistance and growing public concern over health and food safety, the sector has been prompted to seek safe alternatives. This shift has led to a focus on finding natural antibiotic substitutes and optimizing feed management to enhance chicken health and growth. Consequently, phytogenics have emerged as promising natural alternatives to antibiotics in the poultry sector. Moringa oleifera has attracted significant research interest recently as a natural supplement with numerous health advantages for poultry. Known for its antimicrobial, antioxidant, anti-inflammatory, and cholesterol-lowering properties, Moringa also aids in activating digestive enzymes in the stomach due to its wealth of bioactive compounds. This review emphasizes the potential impact of Moringa oleifera as a natural feed additive on growth performance, nutrient digestibility, blood biochemical profile, antioxidant effects, immune response, and egg production in poultry.