High Amino Acid Research Articles

Biochemical and Proteome Analysis Reveal Different Nutritional Compound Compositions and Chloroplast Development Situations between Purple-Red and White-Yellow Tea Plant Cultivars

Across the world, most of the main tea plant cultivars are green-leaf cultivars, but colored-leaf cultivars have become increasingly popular in recent years. In this research, the differences between purple-red and white-yellow tea plant cultivars were compared on biochemical and proteome levels. White-yellow cultivars had significantly high amino acid (AA) content and low polyphenols/amino acid (PP/AA), while purple-red cultivars showed high PP/AA. Comprehensive analysis of all nutritional compounds revealed that most of them showed significant positive correlations, except AA, and that there were significant negative correlations between AA and other compounds. The nutritional compounds of some individual cultivars differed from the average trend of the classification, using color as a criterion. Twenty-one differential proteins were detected in the purple-red and yellow-white cultivars. Among these proteins, there were 16 upregulated proteins and 5 downregulated proteins in purple-red cultivars. Most of these proteins act in the photosynthetic system. This indicated that tea plants with purple-red leaves performed better in photosynthesis than yellow-white tea plant cultivars. The chloroplast development of white-yellow tea plant cultivars was obstructed and may introduce AA accumulation.

Metabolomics and flavoromics analysis of chemical constituent changes during roasting of germinated Sacha inchi (Plukenetia volubilis L.).

This study examined the changes in metabolites together with the flavor profiles of germinated Sacha inchi seeds during roasting by using gas chromatography. The results indicated that roasting partially increased the browning index, amino acid levels, total phenolic content, and antioxidant capacity, but slightly decreased the levels of reducing sugars. Oxidized and rancid compounds were significantly decreased at a 180°C roasting temperature. Pyrazine, furan, and pyrrole were Maillard reaction products that were increased at 180°C of roasting. Roasting at 145°C for 45min after germination for 4days was determined to be the optimal conditions for roasting germinated Sacha inchi seeds, which reduced the off-flavor and burned taste. The roasted germinated Sacha inchi seed contains higher amino acids than raw seed, which could be used as an alternative source for food products and supplements. In addition, the roasted germinated seeds at 4days were recommended for food applications.

Principal Component and Path Analysis for Trait Selection Based on the Assessment of Diverse Lentil Populations Developed by Gamma-Irradiated Physical Mutation.

Lentil is a notable legume crop valued for its high protein, vitamin, mineral, and amino acid (lysine and tryptophan) content. This crop has a narrow genetic base due to the formation of gene pool barriers during interspecific hybridization within and across species. Mutagenesis may be seen as a novel and alternative breeding technique for the production of new diversity. For the identification of new alleles, the creation of mutants followed by selection in subsequent generations would be necessary. Induction of mutation in lentil cv. Moitree by gamma rays therefore produced high variation for the majority of quantitative measures examined. Henceforth, principal component analysis (PCA) and path coefficient analysis were conducted to identify and exclude redundant mutant genotypes with similar traits as the success of breeding is dependent on understanding the relationship between morpho-agronomic traits and seed yield. As shown by the findings of this research, the total quantity of pods per mutant plant should be given considerable priority. The identified mutant genotypes, such as lines 24, 43, 28, 33, and 10, may be used as parents in future breeding or released directly following trials.

Novel Bat Lyssaviruses Identified by Nationwide Passive Surveillance in Taiwan, 2018-2021.

Bat lyssaviruses were identified in Taiwan’s bat population during 2016–2017. The lyssavirus surveillance system was continuously conducted to understand the epidemiology. Through this system, the found dead bats were collected for lyssavirus detection by direct fluorescent antibody test and reverse transcription polymerase chain reaction. Three bats were identified as positive during 2018–2021. A novel lyssavirus, designated as Taiwan bat lyssavirus 2, was detected in a Nyctalus plancyi velutinus. This lyssavirus had less than 80% nucleotide identity in the nucleoprotein (N) gene with other lyssavirus species, forming a separate branch in the phylogenetic analysis. The other two cases were identified in Pipistrellus abramus (Japanese pipistrelles); they were identified to be similar to the former lyssavirus identified in 2016–2017, which was renominated as Taiwan bat lyssavirus 1 (TWBLV-1) in this study. Even though one of the TWBLV-1 isolates showed high genetic diversity in the N gene compared with other TWBLV-1 isolates, it may be a TWBLV-1 variant but not a new species based on its high amino acid identities in the nucleoprotein, same host species, and same geographic location as the other TWBLV-1.

Potential of Nile Tilapia (Oreochromis niloticus) as an Alternative Complementary Food Ingredient for Stunting Children

BACKGROUNDS:The Result of the Basic Health Research 2018 stunting prevalence in Indonesia is 30.8%. One of the potential local foodstuffs in Indonesia is nile Tilapia fish (Oreochromis niloticus). Nile Tilapia fish has a high nutrient content, especially protein, calcium and monounsaturated fatty acids.
 OBJECTIVES: Analyzed of nutrient content and heavy metal content of lead in Nile Tilapia.
 METHODS:The study was a laboratory study using raw materials of Nile Tilapia fish . Then analyzed the content of proximate, fatty acids, amino acids and lead heavy metals. Statistical tests use descriptive tests.
 RESULTS:The total protein content of 18.46 grams, and the calcium content of 74.38 grams in 100 grams of material. Total fatty acids of Nile Tilapia 30.39%, Palmitic acid 7.87%, stearic acid 4.30%, oleic acid 8.13%, Linoleic Acid 3.67% of total fatty acids. Total amino acids is 21.56%, Arginine 1.88%, Leucine 1.69%, Lysine 1.84%, Aspartic Acid 2.16%, Glutamic Acid 3.45% of the total amino acids.
 CONCLUSION:High nutrient content in Nile Tilapia are proteins, calcium, oleic fatty acids, palmitic, linoleic and stearic acids. The highest amino acids are aspartic acid, glutamic acid, lysine arginine and leucine.

Initiation of muscle protein synthesis was unrelated to simultaneously upregulated local production of IGF-1 by amino acids in non-proliferating L6 muscle cells.

BackgroundIGF-1 is considered an important regulator of muscle protein synthesis. However, its role in stimulation of muscle protein synthesis by amino acids (AA) is not clear, despite pronounced alterations in IGF-1 mRNA expression and signaling in muscle tissues by feeding. This study evaluates the role of locally produced IGF-1 and IGF-1 signaling when skeletal muscle protein synthesis is activated by increased amino acid availability in confluent, non-proliferating cells.MethodsL6 skeletal muscle cells were subjected to amino acid starvation (24 h, 0.14 mM) followed by 18 h amino acid refeeding in Low AA (0.28 mM) or High AA concentrations (9 mM). Protein synthesis rates were estimated by L-[U-14C]-phenylalanine incorporation into cellular proteins. IGF-1 and IGF-1 receptor mRNA expression were quantified by real time PCR. SiRNA knockdown, antibodies and chemical inhibitors were used to attenuate muscle IGF-1 production and signaling.ResultsHigh AA concentrations (9mM) increased IGF-1 mRNA expression (+ 30%, p<0.05) and increased L-[U-14C]-phenylalanine incorporation compared to Low AA in confluent, non-proliferating muscle cells. Blocking IGF-1 signaling by chemical inhibitors reduced IGF-1 mRNA upregulation (~50%, p< 0.01), without decrease of protein synthesis. SiRNA knockdown of IGF-1 reduced protein synthesis, mainly explained by reduced cell proliferation. High AA or IGF-1 inhibitors did not change IGF-1 receptor mRNA expressions.ConclusionAmino acids increased IGF-1 mRNA expression and stimulated muscle protein synthesis. However, simultaneous upregulation of IGF-1 mRNA did not relate to increased protein synthesis by amino acids. The results indicate that increased IGF-1 mRNA expression is rather a covariate to amino acid initiation of protein synthesis in non-proliferating muscle cells; effects that may be related to unrecognized metabolic activities, such as transport of amino acids.

Blood and faecal biomarkers to assess dietary energy, protein and amino acid efficiency of utilization by growing and finishing pigs

BackgroundDiet evaluation and optimization is a slow and expensive process and it is not possible to do it at a farm level. This study aimed to use the blood serum metabolite (BSM) and faecal volatile fatty acid (VFA) profiles as potential biomarkers to identify changes in protein, amino acid and energy dietary content in growing and finishing pig diets at farm level.ResultsTwo studies were conducted. The first study (S1) included 20 pens of 11 pigs (87.0 ± 4.10 kg; 18 weeks old) assigned to 5 diets: control (C1), high or low crude protein (HP1 and LP1, respectively), and high or low net energy (HE1 and LE1, respectively). The second study (S2) included 28 pens of 11 pigs (41.3 ± 2.60 kg; 12 weeks old) assigned to 7 diets: control (C2), high or low crude protein (HP2 and LP2, respectively), high or low amino acid (HA2 and LA2, respectively), and high or low net energy (HE2 and LE2, respectively). Pigs were followed for 10 (S1) and 20 (S2) days, and blood and faecal samples were collected at 20 (S1) and 14 (S2) weeks of age. Data were analysed using general linear models and receiver operating characteristic curve analysis. Urea nitrogen showed the best results as a biomarker. Urea nitrogen was higher in pigs fed high protein diets, HP1 (13.6 ± 0.95 mg/dL) and HP2 (11.6 ± 0.61), compared to those fed low protein diets, LP1 (6.0 ± 0.95) and LP2 (5.2 ± 0.61; P < 0.001), showing good discrimination ability (Area under the curve (AUC) = 98.4 and 100%, respectively). These differences were not observed between diets LA2 (6.5 ± 0.61) and HA2 (8.7 ± 0.61; P > 0.05; AUC = 71.9%), which were formulated based on the ideal protein profile but with no excess of protein. Creatinine, triglycerides, branched-chain fatty acids, albumin, propionic acid, and cholesterol showed differences between at least 2 diets but only in one of the studies.ConclusionsUrea nitrogen showed high accuracy to detect excess of crude protein in growing and finishing pig diets. Other biomarkers like BCFA showed promising results and need to be further studied.

Amino acid catabolism regulates hematopoietic stem cell proteostasis via a GCN2-eIF2α axis

Hematopoietic stem cells (HSCs) adapt their metabolism to maintenance and proliferation; however, the mechanism remains incompletely understood. Here, we demonstrated that homeostatic HSCs exhibited high amino acid (AA) catabolism to reduce cellular AA levels, which activated the GCN2-eIF2α axis, a protein synthesis inhibitory checkpoint to restrain protein synthesis for maintenance. Furthermore, upon proliferation conditions, HSCs enhanced mitochondrial oxidative phosphorylation (OXPHOS) for higher energy production but decreased AA catabolism to accumulate cellular AAs, which inactivated the GCN2-eIF2α axis to increase protein synthesis and coupled with proteotoxic stress. Importantly, GCN2 deletion impaired HSC function in repopulation and regeneration. Mechanistically, GCN2 maintained proteostasis and inhibited Src-mediated AKT activation to repress mitochondrial OXPHOS in HSCs. Moreover, the glycolytic metabolite, NAD+ precursor nicotinamide riboside (NR), accelerated AA catabolism to activate GCN2 and sustain the long-term function of HSCs. Overall, our study uncovered direct links between metabolic alterations and translation control in HSCs during homeostasis and proliferation.

Early high dose versus low dose parenteral amino acids administration in the preterm infants: A meta-analysis

Objective: Preterm infants may face difficulty on getting the nutrition required for growth due to incompetence of sucking and immaturity of the gastrointestinal. Preterm infants require higher amounts of amino acids and protein to support the growth of lean body mass and tissue. Administrating amino acid supplementation during the first hour of life is the key to preventing early neonatal malnutrition and can be beneficial in the growth of the neonates. Objectives: Aim of this study is to determine the effects of early high dose parenteral amino acids administration versus low dose in preterm infants from all collected studies. Methods: This is a meta-analysis study that studies were collected MedLine, PubMed, and Cochrane Central Register of Controlled Trials comparing early high dose parenteral amino acids versus low dose. High-quality studies, as assessed by Jadad Criteria, were used to evaluate outcomes such as anthropometric data, length of hospital stays, and morbidities. Results: A total of 9 Randomized controlled trials (RCTs) with total of 960 participants were included in the analysis. Analysis of these studies showed a statistically significant reduction in the time to regain birth weight in the group of early high-dose amino acids administration in the preterm infants. Reductions were 0.79 day (MD 0.79 day, 95% CI 0.06 to 1.52 day; participants = 655; studies = 6; I2 = 0%) (P= 0.03). It also showed a statistically significant reduction in the length of stay with a reduction of 2.09 days in the early high-dose parenteral amino acids groups (MD 2.09 days, 95% CI 1.01 to 3.17 day; participants = 500; studies = 5; I2 = 0%) (P= 0.0002). No significant difference in morbidity of each group was found. Conclusion: Administration of early high-dose amino acids reduced the time to regain birth weight and the length of hospitalization in preterm infants. The analysis did not show any significant increase in the risk of morbidity.

Expression of fatty acid synthase genes and their role in development and arboviral infection of Aedes aegypti

BackgroundFatty acids are the building blocks of complex lipids essential for living organisms. In mosquitoes, fatty acids are involved in cell membrane production, energy conservation and expenditure, innate immunity, development and reproduction. Fatty acids are synthesized by a multifunctional enzyme complex called fatty acid synthase (FAS). Several paralogues of FAS were found in the Aedes aegypti mosquito. However, the molecular characteristics and expression of some of these paralogues have not been investigated.MethodsGenome assemblies of Ae. aegypti were analyzed, and orthologues of human FAS was identified. Phylogenetic analysis and in silico molecular characterization were performed to identify the functional domains of the Ae. aegypti FAS (AaFAS). Quantitative analysis and loss-of-function experiments were performed to determine the significance of different AaFAS transcripts in various stages of development, expression following different diets and the impact of AaFAS on dengue virus, serotype 2 (DENV2) infection and transmission.ResultsWe identified seven putative FAS genes in the Ae. aegypti genome assembly, based on nucleotide similarity to the FAS proteins (tBLASTn) of humans, other mosquitoes and invertebrates. Bioinformatics and molecular analyses suggested that only five of the AaFAS genes produce mRNA and therefore represent complete gene models. Expression levels of AaFAS varied among developmental stages and between male and female Ae. aegypti. Quantitative analyses revealed that expression of AaFAS1, the putative orthologue of the human FAS, was highest in adult females. Transient knockdown (KD) of AaFAS1 did not induce a complete compensation by other AaFAS genes but limited DENV2 infection of Aag2 cells in culture and the midgut of the mosquito.ConclusionAaFAS1 is the predominant AaFAS in adult mosquitoes. It has the highest amino acid similarity to human FAS and contains all enzymatic domains typical of human FAS. AaFAS1 also facilitated DENV2 replication in both cell culture and in mosquito midguts. Our data suggest that AaFAS1 may play a role in transmission of dengue viruses and could represent a target for intervention strategies.Graphical

Antigenicity Alternations of Variant PEDV S Protein Disclosed by Linear B Cell Epitope Mapping.

The spike protein (S) plays a crucial role in porcine epidemic diarrhea virus (PEDV) infection and induces neutralizing antibodies. Mutations of the S protein are supposed to provide the main antigenic shift leading to the antigenic escape of PEDVs. It is therefore a significant question how much accumulation of antigenic shift could lead to the antigenic escape of the variant PEDV. To provide an answer in the study, B cell epitopes (BCEs) on the S protein of the PEDV vaccine strain CV777 (SCV777) and variant strain SD2014 (SSD2014) were mapped using biosynthetic peptides and rabbit anti-PEDV S serum. Seventy-nine and 68 linear BCEs were identified from SCV777 and SSD2014, respectively. While 66.2% of the BCEs of SSD2014 could be recognized by anti-SCV777 serum and 67.1% of SCV777 BCEs could be recognized by anti-SSD2014 serum, more than 40% of the BCEs identified using anti-SCV777 serum on SCV777 could not be recognized by anti-SSD2014 serum and vice versa. The completely shared BCEs took low percentages of 29.4% and 25.3% for SSD2014 and SCV777, respectively. These results indicate a low conservation of antigenicity of the S protein compared to a relatively high amino acid sequence similarity of 92.2% between the two strains. The study provided a BCE shift reference of PEDV antigenic escape and surveillance control.

Effects of fattening diets on the nutritional quality and flavor of the adult female Chinese mitten crab (Eriocheir sinensis)

The effects of different fattening diet combinations on the growth, nutritional value, and flavor of Eriocheir sinensis were compared. Four different fattening diet combinations were evaluated: A1, consisting of 80% iced trash fish (ITF) + 20% formulated diet (FD); A2, consisting of 80% ITF + 20% soybean and corn (SC); B1, consisting of 80% Cipangopaludina cathayensis (CC) + 20% FD; and B2, consisting of 80% CC + 20% SC. After 28 days of fattening, the relevant indices were analyzed to identify the best fattening diet combination. There was no significant difference in the growth indices of E. sinensis among the four groups. Interestingly, crabs fed the ITF diets had higher crude protein contents in edible tissues than those fed the CC diets. Crabs fattened with the A2 diet combination had the best yellowness index of the hepatopancreas, while those in the B1 group had greater fatty acid (FA) compositions and higher contents of polyunsaturated FAs. Crabs fattened with the B2 diet combination had the highest amino acid (AA) scores and chemical scores. Meanwhile, the total content of free AAs, sweet and umami AAs, and equivalent umami concentration were highest in the B1 group, indicating better flavor. In conclusion, ITF and SC improved the color of the hepatopancreas of E. sinensis, while CC with supplementary FD or SC improved the nutritional value and flavor of edible parts of E. sinensis. These results provide strategies for fattening of E. sinensis to meet consumer demands.

Characterization of Chicken α2A-Adrenoceptor: Molecular Cloning, Functional Analysis, and Its Involvement in Ovarian Follicular Development.

Adrenoceptors are suggested to mediate the functions of norepinephrine (NE) and epinephrine (EPI) in the central nervous system (CNS) and peripheral tissues in vertebrates. Compared to mammals, the functionality and expression of adrenoceptors have not been well characterized in birds. Here, we reported the structure, expression, and functionality of chicken functional α2A-adrenoceptor, named ADRA2A. The cloned chicken ADRA2A cDNA is 1335 bp in length, encoding the receptor with 444 amino acids (a.a.), which shows high amino acid sequence identity (63.4%) with its corresponding ortholog in humans. Using cell-based luciferase reporter assays and Western blot, we demonstrated that the ADRA2A could be activated by both NE and EPI through multiple signaling pathways, including MAPK/ERK signaling cascade. In addition, the mRNA expression of ADRA2A is found to be expressed abundantly in adult chicken tissues including thyroid, lung, ovary and adipose from the reported RNA-Seq data sets. Moreover, the mRNA expression of ADRA2A is also found to be highly expressed in the granulosa cells of 6–8 mm and F5 chicken ovarian follicles, which thus supports that ADRA2A signaling may play a role in ovarian follicular growth and differentiation. Taken together, our data provide the first proof that the α2A-adrenoceptor is functional in birds involving avian ovarian follicular development.

Histopathological liver steatosis linked with high parenteral glucose and amino acid supply in infants with short bowel syndrome.

Steatosis is a common feature of intestinal failure-associated liver disease (IFALD) in adult and older pediatric patients receiving long-term parenteral nutrition (PN). There are limited clinical data concerning steatosis in infants with short bowel syndrome (SBS). We investigated early histopathological steatosis and its association to PN. In this retrospective study, 31 patients with SBS had a diagnostic liver biopsy taken at the median age of 5 (IQR 3-8) months. Follow-up biopsy was available for 24 patients at the median age of 29 (IQR 14-52) months. We evaluated the biopsies for steatosis and other histopathological signs of IFALD and compared results with patient characteristics, PN composition, and liver biochemistry. Diagnostic biopsies revealed steatosis in 8 (26%) patients. At the age of 3 months, patients with steatosis had received higher amounts of parenteral glucose: median 15.1 (IQR 12.4-17.2) vs 12.3 (8.7-14.4) g/kg/d (P = 0.04), amino acids: 2.9 (2.5-3.4) vs 2.2 (1.6-2.7) g/kg/d (P = 0.03), and energy: 87 (80-98) vs 73 (54-79) kcal/kg/d (P = 0.01) than those without steatosis. We detected no significant differences in parenteral lipid intake between the groups. Steatosis also associated with increased serum bile acid (P = 0.02), alanine aminotransferase (P = 0.0002), and aspartate aminotransferase (P = 0.001) levels. In this cohort, high parenteral glucose, amino acid, and energy provision associated with liver steatosis in infants with SBS. We recommend monitoring of bile acid and transaminase levels while aiming for PN with balanced macronutrient supply according to current recommendations to protect the liver from steatosis.

Acanthaster plancias a Potential Source of Protein and Amino Acids for Fish Feed Ingredients

Acanthaster planci becomes a pest for coral reef ecosystems if its presence is excessive. Knowing the nutrient and amino acid content of Acanthaster planci as fish feed ingredients is the goal of this study. The test sample was taken in the waters of Tolo Bay, Process that was dried for 5 days and then ground until smooth. Samples were taken and analyzed for their nutrient and amino acid content. A duplo test was performed for each sample. The results showed that the nutrient content of Acanthaster planci flour included crude protein, crude fat extract without nitrogen and ash, respectively, at 28.58, 8.79, 2.94, and 45.56%. In addition, Acanthaster planci contains high essential amino acids including leucine, valine, isoleucine, methionine, and tryptophan respectively at 29888.85, 16142.82, 14805.09, 3272.04 and 2267.67 mg/kg dry weight. High protein and essential amino acids indicate Acanthaster planci flour has potential as fish feed ingredients.

Salvianolic Acid Modulates Physiological Responses and Stress-Related Genes That Affect Osmotic Stress Tolerance in Glycine max and Zea mays

Drought is a serious threat worldwide to soybean and maize production. This study was conducted to discern the impact of salvianolic acid treatment on osmotic-stressed soybean (Glycine max L.) and maize (Zea mays L.) seedlings from the perspective of physiochemical and molecular reactions. Examination of varied salvianolic acid concentrations (0, 0.1, 1, 5, 10, and 25 μM) on soybean and maize seedling growth confirmed that the 0.1 and 1 μM concentrations, respectively, showed an improvement in agronomic traits. Likewise, the investigation ascertained how salvianolic acid application could retrieve osmotic-stressed plants. Soybean and maize seedlings were irrigated with water or 25% PEG for 8 days. The results indicated that salvianolic acid application promoted the survival of the 39-day-old osmotic-stressed soybean and maize plants. The salvianolic acid-treated plants retained high photosynthetic pigments, protein, amino acid, fatty acid, sugar, and antioxidant contents, and demonstrated low hydrogen peroxide and lipid contents under osmotic stress conditions. Gene transcription pattern certified that salvianolic acid application led to an increased expression of GmGOGAT, GmUBC2, ZmpsbA, ZmNAGK, ZmVPP1, and ZmSCE1d genes, and a diminished expression of GmMIPS2, GmSOG1, GmACS, GmCKX, ZmPIS, and ZmNAC48 genes. Together, our results indicate the utility of salvianolic acid to enhance the osmotic endurance of soybean and maize plants.

Investigation of active site amino acid influence on carbon and chlorine isotope fractionation during reductive dechlorination.

Reductive dehalogenases (RDases) are corrinoid-dependent enzymes that reductively dehalogenate organohalides in respiratory processes. By comparing isotope effects in biotically catalyzed reactions to reference experiments with abiotic corrinoid catalysts, compound-specific isotope analysis (CSIA) has been shown to yield valuable insights into enzyme mechanisms and kinetics, including RDases. Here, we report isotopic fractionation (ε) during biotransformation of chloroform (CF) for carbon (εC = -1.52± 0.34‰) and chlorine (εCl = -1.84± 0.19‰), corresponding to a ΛC/Cl value of 1.13± 0.35. These results are highly suppressed compared to isotope effects observed both during CF biotransformation by another organism with a highly similar RDase (>95% sequence identity) at the amino acid level, and to those observed during abiotic dehalogenation of CF. Amino acid differences occur at four locations within the two different RDases' active sites, and this study examines whether these differences potentially affect the observed εC, εCl, and ΛC/Cl. Structural protein models approximating the locations of the residues elucidate possible controls on reaction mechanisms and/or substrate binding efficiency. These four locations are not conserved among other chloroalkane reducing RDases with high amino acid similarity (>90%), suggesting that these locations may be important in determining isotope fractionation within this homologous group of RDases.

Molecular cloning of cholecystokinin (CCK) and CCK-A receptor and mechanism of CCK-induced gastrointestinal motility in Suncus murinus

Cholecystokinin (CCK) is a peptide hormone mainly secreted by small intestinal endocrine I-cells and functions as a regulator of gallbladder contraction, gastric emptying, gastrointestinal (GI) motility, and satiety. The cellular effects of CCK in these peripheral tissues are predominantly mediated via CCK-A receptors which are found in smooth muscles, enteric neurons, and vagal afferent neurons in humans and animal models. Although various functions of CCK have been reported to be neurally mediated, it can also stimulate contraction via the CCK receptor on the smooth muscle. However, the entire underlying neural and cellular mechanisms involved in CCK-induced GI contractions are not clearly understood. Here, we first determined the cDNA and amino acid sequences of CCK and CCK-A receptor along with the distributions of cck mRNA and CCK-producing cells in house musk shrew (Suncus murinus, the laboratory strain named as suncus) and examined the mechanism of CCK-induced contraction in the GI tract. Mature suncus CCK-8 was identical to other mammalian species tested here, and suncus CCK-A receptor presented high nucleotide and amino acid homology with that of human, dog, mouse, and rat, respectively. Suncus CCK mRNA and CCK-producing cells were found mainly in small intestine and colon. In the organ bath study, CCK-8 induced dose-dependent contractions in the suncus stomach, duodenum, and jejunum, and these contractions were inhibited by atropine and CCK-A receptor antagonist. These results suggest that CCK-8-induced contraction is mediated in the myenteric cholinergic neural network and that CCK-A receptor is partly responsible for CCK-8-induced contractions. This study indicates that suncus is a useful animal model to study the functions of CCK involved in GI motility.

Discovering B/T Cell-Cancer Antigen Affinity for Targeted Cancer Drug Therapy and Diagnosis Through Deep Neural Network Models

Cancer, one of the leading causes of death worldwide, derives from an uncontrolled division of abnormal cells in a given part of the body. Targeted immunotherapy is a promising avenue of cancer treatments, galvanizing the body's own immune system to marshal B and T cells against abnormal cell growth, pathologically inhibiting antigen function and proliferation. In the current landscape of cancer immunotherapy, however, medicine industries are belabored with the need to use painstaking trial and error and numerous wet-lab investigations to test amino acid sequences' affinities to cancer antigens. Furthermore, most cancer antigens and the structures of their epitopes are unknown, and although most malignancies can be cured when diagnosed early, organ-specific assessments cannot be used for early-stage cancers. To bridge this gap, I innovate a deep convolutional neural network (CNN) model pipeline, which analyzes the complex amino acid makeup of tumor infiltrating B/T cell receptors based on the relationships of biochemical properties among adjacent amino acids predictive of how these receptor polypeptides fold in three-dimensional space, computing high affinity amino acid sequences to revolutionize both targeted drug discovery and early diagnosis.

Tetraploid genes of leptin (leptin-AI, -AII, -BI and -BII) in goldfish: Molecular cloning, bioinformatics analysis, tissue distribution and differential regulation of transcript expression by glucocorticoids

In mammals, leptin is an endocrine hormone involved in the regulation of feeding and energy metabolism, and glucocorticoids were previously reported to induce leptin gene expression. However, the link between glucocorticoids and leptin in fish has not been investigated. Goldfish are tetraploid teleosts because of genomic replication events (FSGD or 3-R), and some specific genes have quadrupled. In this study, two goldfish leptin-B genes, namely, leptin-BI (GenBank: ON468243) and leptin-BII (GenBank: ON468244), were cloned. The cDNA sequences of leptin-AI (GenBank: FJ534535.1) and leptin-AII (GenBank: FJ854572.1) have been reported by our team previously. Goldfish leptin-AI, -AII, -BI and -BII genes are located on chromosomes 43, 18, 29 and 4, respectively. In goldfish, leptin-B proteins shared approximately 30 % amino acid identities with leptin-A proteins and only 20 % with human leptin; however, either goldfish leptin-A or leptin-B showed high amino acid homology between themselves, 79.29 % and 82.74 %, respectively. The four leptin genes in goldfish are all highly conserved in three-dimensional (3-D) protein structures and gene structures. A tissue expression study showed that both the A and B forms of goldfish leptins are predominantly in the brain and liver. The effects of glucocorticoids on four leptin transcripts expression in goldfish were investigated by in vivo intraperitoneal injection and in vitro cell incubation approaches. The results showed that cortisol inhibited hepatic leptin-A transcript expression in a dose-dependent manner, while cortisol stimulated hepatic leptin-B mRNA expression, and RU486 (glucocorticoid receptor inhibitor) abolished cortisol’s effect on hepatic leptin-A and leptin-B mRNA expression. These results reflected the differential regulation of leptin-A and leptin-B by glucocorticoids in goldfish, which implied that leptin-A and leptin-B may mediate different functions in goldfish.