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Objective This study aimed to find global mechanisms related to carnosine synthesis in slow-growing Korat chickens (KRC) using a proteomic approach. Methods M. pectoralis major samples were collected from 10-week-old female KRC including low-carnosine (LC, 2,756.6±82.88 μg/g; n = 5) and high-carnosine (HC, 4,212.5 ±82.88 μg/g; n = 5). Results We identified 152 common proteins, and 8 of these proteins showed differential expression between the LC and HC groups (p

ABSTRACT: Carnosine is a physiologically important molecule in normal human body functions. Chicken meat is an excellent source of carnosine; especially slow-growing Korat chicken (KR) females have a high carnosine content in their meat. The carnosine content of chicken meat can be increased by dietary supplementation of β-alanine (βA) and L-histidine (L-His). Our objective was to reveal the pathways and genes through jejunal transcriptomic profiling related to βA and L-His absorption and transportation. We collected whole jejunum samples from 5 control and 5 experimental KR chicken, fed with 1% βA and 0.5% L-His supplementation. A total of 407 differentially expressed genes (P < 0.05, log2 fold change ≥2) were identified, 272 of which were down-regulated and 135 up-regulated in the group with dietary supplementation compared to the control group. Based on the integrated analysis of the protein–protein interaction network and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway maps, 87 gene ontology terms were identified and 6 KEGG pathways were significantly (P < 0.05) enriched in the jejunum. The analyses revealed 6 key genes, KCND3, OPRM1, CCK, GCG, TRH, and GABBR2, that are related to neuroactive ligand-receptor interaction and the calcium signaling pathway. These findings give insight regarding the molecular mechanism related to carnosine precursor absorption and transportation in the jejunum and help to identify useful molecular markers for improving the carnosine content in slow-growing KR chicken meat.

ABSTRACT: The slow-growing Korat chicken (KR) has been developed to provide an alternative breed for smallholder farmers in Thailand. Carnosine enrichment in the meat can distinguish KR from other chicken breeds. Therefore, our aim was to investigate the effect of enriched carnosine synthesis, obtained by the β-alanine and L-histidine precursor supplementation in the diet, on changes to metabolomic profiles and biochemical compounds in slow-growing KR jejunum tissue. Four hundred 21-day-old female KR chickens were divided into 4 experimental groups: a group with a basal diet, a group with a basal diet supplemented with 1.0% β-alanine, 0.5% L-histidine, and a mix of 1.0% β-alanine and 0.5% L-histidine. The feeding trial lasted 70 d. Ten randomly selected chickens from each group were slaughtered. Metabolic profiles were analyzed using proton nuclear magnetic resonance spectroscopy. In total, 28 metabolites were identified. Significant changes in the concentrations of these metabolites were detected between the groups. Partial least squares discriminant analysis was used to distinguish the metabolites between the experimental groups. Based on the discovered metabolites, 34 potential metabolic pathways showed differentiation between groups, and 8 pathways (with impact values higher than 0.05, P < 0.05, and FDR < 0.05) were affected by metabolite content. In addition, biochemical changes were monitored using synchrotron radiation-based Fourier transform infrared microspectroscopy. Supplementation of β-alanine alone in the diet increased the β-sheets and decreased the α-helix content in the amide I region, and supplementation of L-histidine alone in the diet also increased the β-sheets. Furthermore, the relationship between metabolite contents and biochemical compounds were confirmed using principal component analysis (PCA). Results from the PCA indicated that β-alanine and L-histidine precursor group was highly positively correlated with amide I, amide II, creatine, tyrosine, valine, isoleucine, and aspartate. These findings can help to understand the relationships and patterns between the spectral and metabolic processes related to carnosine synthesis.

Objective: This study aimed to find global mechanisms related to carnosine synthesis in slow-growing Korat chickens (KRC) using a proteomic approach. Methods: M. pectoralis major samples were collected from 10-week-old female KRC including low-carnosine (LC, 2,756.6±82.88 μg/g; n = 5) and high-carnosine (HC, 4,212.5 ±82.88 μg/g; n = 5). Results: We identified 152 common proteins, and 8 of these proteins showed differential expression between the LC and HC groups (p<0.05). Heat shock 70 kDa protein 8, Heat shock 70 kDa protein 2, protein disulfide isomerase family A, member 6, and endoplasmic reticulum resident protein 29 were significantly involved in protein processing in the endoplasmic reticulum pathway (false discovery rate<0.05), suggesting that the pathway is related to differential carnosine concentration in the M. pectoralis major of KRC. A high concentration of carnosine in the meat is mainly involved in low abundances of Titin isoform Ch12 and Connectin and high abundances of M-protein to maintain homeostasis during muscle contraction. These consequences improve meat characteristics, which were confirmed by the principal component analysis. Conclusion: Carnosine synthesis may occur when muscle cells need to recover homeostasis after being interfered with carnosine synthesis precursors, leading to improved muscle function. To the best of our knowledge, this is the first study to describe in detail the global molecular mechanisms in divergent carnosine contents in meat based on the proteomic approach. [ABSTRACT FROM AUTHOR]

ABSTRACT: The Korat chicken (KR) is a slow-growing Thai chicken breed with relatively poor feed efficiency (FE) but very tasty meat with high protein and low fat contents, and a unique texture. To enhance the competitiveness of KR, its FE should be improved. However, selecting for FE has an unknown effect on meat characteristics. Thus, understanding the genetic basis underlying FE traits and meat characteristics is needed. In this study, 75 male KR birds were raised up to 10 wk of age. For each bird, the feed conversion ratio (FCR), residual feed intake (RFI), and physicochemical properties, flavor precursors, and biological compounds in the thigh meat were evaluated. At 10 wk of age, thigh muscle samples from 6 birds (3 with high FCR and 3 with low FCR values) were selected, and their proteomes were investigated using a label-free proteomic method. Weighted gene coexpression network analysis (WGCNA) was used to screen the key protein modules and pathways. The WGCNA results revealed that FE and meat characteristics significantly correlated with the same protein module. However, the correlation was unfavorable; improving FE may result in a decrease in meat quality through the alteration in biological processes including glycolysis/gluconeogenesis, metabolic pathway, carbon metabolism, biosynthesis of amino acids, pyruvate metabolism, and protein processing in the endoplasmic reticulum. The hub proteins of the significant module (TNNT1, TNNT3, TNNI2, TNNC2, MYLPF, MYH10, GADPH, PGK1, LDHA, and GPI) were also identified to be associated with energy metabolism, and muscle growth and development. Given that the same proteins and pathways are present in FE and meat characteristics but in opposite directions, selection practices for KR should simultaneously consider both trait groups to maintain the high meat quality of KR while improving FE.

Objective: Defining the regulators of cell metabolism and signaling is essential to design new therapeutic strategies in obesity and NAFLD/NASH. E3 ubiquitin ligases control diverse cellular functions by ubiquitination-mediated regulation of protein targets, and thus their functional aberration is associated with many diseases. The E3 ligase Ube4A has been implicated in human obesity, inflammation, and cancer. However, its in vivo function is unknown, and no animal models are available to study this novel protein. Methods: A whole-body Ube4A knockout (UKO) mouse model was generated, and various metabolic parameters were compared in chow- and high fat diet (HFD)-fed WT and UKO mice, and in their liver, adipose tissue, and serum. Lipidomics and RNA-Seq studies were performed in the liver samples of HFD-fed WT and UKO mice. Proteomic studies were conducted to identify Ube4A's targets in metabolism. Furthermore, a mechanism by which Ube4A regulates metabolism was identified. Results: Although the body weight and composition of young, chow-fed WT and UKO mice are similar, the knockouts exhibit mild hyperinsulinemia and insulin resistance. HFD feeding substantially augments obesity, hyperinsulinemia, and insulin resistance in both sexes of UKO mice. HFD-fed white and brown adipose tissue depots of UKO mice have increased insulin resistance and inflammation and reduced energy metabolism. Moreover, Ube4A deletion exacerbates hepatic steatosis, inflammation, and liver injury in HFD-fed mice with increased lipid uptake and lipogenesis in hepatocytes. Acute insulin treatment resulted in impaired activation of the insulin effector protein kinase Akt in liver and adipose tissue of chow-fed UKO mice. We identified the Akt activator protein APPL1 as a Ube4A interactor. The K63-linked ubiquitination (K63-Ub) of Akt and APPL1, known to facilitate insulin-induced Akt activation, is impaired in UKO mice. Furthermore, Ube4A K63-ubiquitinates Akt in vitro. Conclusion: Ube4A is a novel regulator of obesity, insulin resistance, adipose tissue dysfunction and NAFLD, and preventing its downregulation may ameliorate these diseases.

ABSTRACT: This study determined the effect of water bath cooking (70°C and 90°C for 40 min) and the extreme heat treatment by an autoclave (121°C for 40 min) on the quality of breast meat of a fast-growing chicken, commercial broiler (CB), and slow-growing chickens, Korat chicken (KC), and Thai native chicken (NC) (Leung Hang Khao), by vibrational spectroscopic techniques, including synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy and Fourier transform Raman (FT-Raman) spectroscopy. Taste-enhancing compounds, including inosine-5ˊ-monophosphate (IMP) and guanosine-5ˊ-monophosphate (GMP), were better retained in cooked KC and NC meats than in cooked CB meat (P < 0.05). The high heat treatment at 121°C depleted the amount of insoluble collagen in all breeds (P < 0.05). Shear force values of slow-growing chicken meat were not affected by high heating temperatures (P > 0.05). In addition, the high heat treatment increased protein carbonyl (P < 0.05), while no effect on in vitro protein digestibility (P > 0.05). SR-FTIR microspectroscopy performed better in differentiating the meat quality of different chicken breeds, whereas FT-Raman spectroscopy clearly revealed differences in meat qualities induced by heating temperature. Based on principal component analysis (PCA), distinct characteristics of chicken meat cooked at 70°C were high water-holding capacity, lightness (L*), moisture content, and predominant α-helix structure, correlating with Raman spectra at 3,217 cm−1 (O–H stretching of water) and 1,651 cm−1 (amide I; α-helix). The high heating temperature at 90°C and 121°C exposed protein structure to a greater extent, as evidenced by an increase in β-sheets, which was well correlated with the Raman spectra at 2,968 and 2,893 cm−1 (C–H stretching), tryptophan (880 cm−1), tyrosine (858 cm−1), and 1,042, 1,020, and 990 cm−1 (C–C stretching; β-sheet). SR-FTIR and FT-Raman spectroscopy show potential for differentiation of chicken meat quality with respect to breeds and cooking temperatures. The marked differences in wavenumbers would be beneficial as markers for determining the quality of cooked meats from slow- and fast-growing chickens.

Objective Growth hormone (GH) and insulin-like growth factor I (IGF-I) play a critical role in animal growth rates. We aimed to investigate the effect of GH and IGF-I genotypes on body weight (BW), dominance, and gene expression in slow-growing chickens at different ages. Methods A total of 613 Korat chickens (KRs) were bred and divided into three groups by genotype – A1A1, A1A3, and A3A3 for GH and AA, AC, and CC for IGF-I. Chickens were weighed every two weeks, and liver and breast muscle tissues were collected at 10 weeks of age. Genetic parameters of KRs were estimated using ASReml software. The GH and IGF-I mRNA levels were measured by quantitative polymerase chain reaction. Significant differences between traits were analyzed using the generalized linear model. Results A significant effect of GH genotypes on BW was found at most ages, and the A1A1 genotype had the highest value of BW. Compared with the A3A3 genotype, the A1A1 and A1A3 genotypes showed a higher dominance effect at 0 and 2 weeks, and genotype A1A1 had the highest value of dominance at 8 weeks of age. A difference in GH mRNA levels between genotypes was detected in breast muscle at 6 weeks and in the liver tissue at 2 weeks. In the case of IGF-I gene, the AA genotype had the highest BW at the beginning of life. Significant differences in BW dominance were found at 2 weeks. However, IGF-I mRNA levels were not different among genotypes in both breast muscles and liver tissues. Conclusion Our results revealed that GH and IGF-I influence growth, but may not be involved in heterosis. GH can be used as a marker gene in selection programs for growth because the homozygous genotype (A1A1) had the highest BW at all ages. The IGF-I is not a useful marker gene for selection programs.

In ovo feeding (IOF) of nutrients is a viable method for increasing muscle mass through hyperplasia and hypertrophy. The objective of this study was to evaluate the effects of IOF of L-arginine (Arg) on breast muscle weight, muscle morphology, amino acid profile, and gene expression of muscle development in slow-growing chickens. Four hundred eighty fertilized eggs were randomly divided into two groups: the first group was the non-injected control group, and the second group was the Arg group, injected with 1% Arg (0.5 mL) into the amnion on day 18 of incubation. After hatching, 160 birds from each group were randomly divided into four replicates of 40 birds each. This experiment lasted for 63 days. The results showed that IOF of Arg did not affect (P > 0.05) breast muscle weight, muscle morphology, and mRNA expression of mammalian target of rapamycin (mTOR) signaling pathway in slow-growing chickens. However, the amino acid profile of breast muscle was altered (P < 0.05) on the day of hatching (DOH), day 21 (D21), and day 42 (D42) post-hatch, respectively. Myogenic factor 5 (Myf5) mRNA expression was upregulated (P < 0.05) on D21 post-hatch. Myogenic regulator 4 (MRF4) mRNA expression was increased (P < 0.05) on DOH. And myogenin (MyoG) was increased (P < 0.05) on DOH and D21 post-hatch, in the Arg group compared to the control group. Overall, IOF of 1% Arg improved the expression of myogenic genes but did not influence muscle morphology and BMW. These results indicate that in ovo Arg dosage (0.5 mL/egg) has no adverse effect on breast muscle development of slow-growing chickens.

Objective The present study was to investigate the extraction conditions of dietary fiber from dried cassava pulp (DCP) and cassava distiller’s dried grains (CDG) under different NaOH concentrations, and the Fourier transform infrared (FTIR) was used to determine the dietary fiber components. Methods The dried samples (DCP and CDG) were treated with various concentrations of NaOH at levels of 2%, 4%, 6%, and 8% using a completely randomized design with 4 replications of each. After extraction, the residual DCP and CDG dietary fiber were dried in a hot air oven at 55°C to 60°C. Finally, the oven dried extracted dietary fiber was powdered to a particle size of 1 mm. Both extracted dietary fibers were analyzed for their chemical composition and determined by FTIR. Results The DCP and CDG treated with NaOH linearly or quadratically or cubically (p< 0.05) increased the total dietary fiber (TDF) and insoluble fiber (IDF). The optimal conditions for extracting dietary fiber from DCP and CDG were under treatment with 6% and 4% NaOH, respectively, as these conditions yielded the highest TDF and IDF contents. These results were associated with the FTIR spectra integration for a semi-quantitative analysis, which obtained the highest cellulose content in dietary fiber extracted from DCP and CDG with 6% and 4% NaOH solution, respectively. The principal component analysis illustrated clear separation of spectral distribution in cassava pulp extracted dietary fiber (DFCP) and cassava distiller’s dried grains extracted dietary fiber (DFCDG) when treated with 6% and 4% NaOH, respectively. Conclusion The optimal conditions for the extraction of dietary fiber from DCP and CDG were treatment with 6% and 4% NaOH solution, respectively. In addition, FTIR spectroscopy proved itself to be a powerful tool for fiber identification.

Objective This study investigated the association between feed efficiency, physicochemical properties, flavor precursors and biomolecules in the thigh meat of Korat (KR) chickens. Methods The feed intake and body weight of individual male KR chickens were recorded from 1 to 10 weeks old to calculate the individual residual feed intake (RFI) of 75 birds. At 10 weeks of age, chickens with the 10 highest (HRFI) and lowest RFI (LRFI) were slaughtered to provide thigh meat samples. The physicochemical properties (ultimate pH, water holding capacity [WHC], drip loss) and flavor precursors (guanosine monophosphate, inosine monophosphate (IMP), adenosine monophosphate and inosine) were analyzed conventionally, and Fourier transform infrared spectroscopy was used to identify the composition of biomolecules (lipids, ester lipids, amide I, amide II, amide III, and carbohydrates) and the secondary structure of the proteins. A group t-test was used to determine significant differences between mean values and principal component analysis to classify thigh meat samples into LRFI and HRFI KR chickens. Results The physicochemical properties of thigh meat samples from LRFI and HRFI KR chickens were not significantly different but the IMP content, ratios of lipid, lipid ester, protein (amide I, amide II) were significantly different (p0.5) while the HRFI group was positively correlated with protein (amide I, amide II), alpha helix protein, IMP content, carbohydrate, ultimate pH and WHC (rg loading >0.5). Conclusion The thigh meat from chickens with different RFI differed in physiochemical properties affecting meat texture, and in the contents of flavor precursors and biomolecules affecting the nutritional value of meat. This information can help animal breeders to make genetic improvements by taking more account of traits related to RFI.

ABSTRACT: The Korat chicken (KR), developed in Thailand, is a slow-growing breed developed as an alternative breed for Thai chicken producers. The growing interest in slow-growing chicken meat, due to its unique taste, distinct texture, health benefits, and higher broiler welfare have led to higher market demand for KR. However, its low feed efficiency (FE) has a significant negative impact on farm profitability. Understanding the molecular mechanism regulating FE allows for designing a suitable selection program and contributing to breeding more efficient chicken for poultry production. Thus, the objective of our study was to investigate the proteome differences and possible pathways associated with FE in male KR using a label-free quantitative proteomic approach. Seventy-five KR males were individually evaluated for FE, and duodenum samples from 6 animals (3 high-FE and 3 low-FE chickens) were collected at 10 wk of age for differential abundant proteins (DAPs), protein networks, functional enrichment, and pathway analyses. In this study, we found 40 DAPs significantly associated with FE pathways, including glycolysis/gluconeogenesis, peroxisome, oxidative phosphorylation, tight junction, and cysteine and methionine metabolism. Thus, variations in observed DAPs or genes related to DAPs could be interesting biomarker candidates for selection for higher feed utilization efficiency in chicken.

ABSTRACT: Carnosine enrichment of slow-growing Korat chicken (KRC) meat helps differentiate KRC from mainstream chicken. We aimed to investigate the effects of β-alanine and L-histidine supplementation on the carnosine synthesis in and quality and secondary structure of proteins in slow-growing KRC meat. Four hundred 21-day-old female KRC were used, and a completely randomized design was applied. The chickens were divided into 4 experimental groups: basal diet (A), basal diet supplemented with 1.0% β-alanine (B), 0.5% L-histidine (C), and 1.0% β-alanine combined with 0.5% L-histidine (D). Each group consisted of 5 replicates (20 chickens per replicate). On d 70, 2 chickens per replicate were slaughtered, and the levels of carnosine, anserine, and thiobarbituric acid reactive substances were analyzed. Biochemical changes were monitored using synchrotron radiation-based Fourier transform infrared microspectroscopy; 5 chickens per replicate were slaughtered, and the meat quality was analyzed. Statistical analysis was performed using ANOVA and principal component analysis (PCA). Group D chickens exhibited the highest carnosine meat content, followed by those in groups B and C. However, amino acid supplementation did not affect anserine content and growth performance. Higher carnosine levels correlated with increasing pH45 min and decreasing drip loss, cooking loss, shear force, and lipid oxidation. PCA revealed that supplementation with only β-alanine or L-histidine was related to increased content of β-sheets, β-turns, and aliphatic bending groups and decreased content of α-helix groups. This study is the first to report such findings in slow-growing chicken. Our findings suggest that KRC can synthesize the highest carnosine levels after both β-alanine and L-histidine supplementation. Higher carnosine contents do not adversely affect meat quality, improve meat texture, and alter the secondary structures of proteins. The molecular mechanism underlying carnosine synthesis in chickens needs further study to better understand and reveal markers that facilitate the development of nutrient selection programs.

Obesity and nonalcoholic fatty liver disease (NAFLD) are global health concerns, and thus, drugs for the long-term treatment of these diseases are urgently needed. We previously discovered that the inositol pyrophosphate biosynthetic enzyme IP6K1 is a target in diet-induced obesity (DIO), insulin resistance, and NAFLD. Moreover, high-throughput screening (HTS) assays and structure−activity relationship (SAR) studies identified LI-2242 as a potent IP6K inhibitor compound. Here, we tested the efficacy of LI-2242 in DIO WT C57/BL6J mice. LI-2242 (20 mg/kg/BW daily, i.p.) reduced body weight in DIO mice by specifically reducing the accumulation of body fat. It also improved glycemic parameters and reduced hyperinsulinemia. LI-2242-treated mice displayed reduced the weight of various adipose tissue depots and an increased expression of metabolism- and mitochondrial-energy-oxidation-inducing genes in these tissues. LI-2242 also ameliorated hepatic steatosis by reducing the expression of genes that enhance lipid uptake, lipid stabilization, and lipogenesis. Furthermore, LI-2242 enhances the mitochondrial oxygen consumption rate (OCR) and insulin signaling in adipocytes and hepatocytes in vitro. In conclusion, the pharmacologic inhibition of the inositol pyrophosphate pathway by LI-2242 has therapeutic potential in obesity and NAFLD.

ABSTRACT: Chicken breed is one of the key factors that influence meat quality. The quality attributes of breast meat from commercial broiler (CB), Thai native chicken (NC, Leung Hang Khao), and the crossbred Korat chicken (KC) were investigated via synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy, Fourier transform Raman (FT-Raman) spectroscopy, and physicochemical analysis. The protein and carbonyl contents of KC and NC meats were higher than that of CB meat, but the lipid content was lower (P < 0.05). CB meat was characterized by high moisture, lightness (L*), and presence of taste-active nucleotides, namely, inosine 5′-monophosphate (IMP) and guanosine 5′-monophosphate (GMP). Moreover, NC meat had the highest insoluble collagen and inosine contents (P < 0.05). The predominant protein secondary structures of KC and NC meats were β-turns and random coils, whereas α-helices were mainly found in CB meat. Based on principal component analysis, the meat quality and spectra were clearly separated by breeds. The high moisture and lipid content of meat corresponded to O–H stretching (3,203 cm−1) and C–H stretching (2,854 cm−1) in the FT-Raman spectra, whereas PO2− stretching (1,240 cm−1), measured via SR-FTIR, was well correlated with the IMP content. In addition, the FT-Raman wavenumber of 934 cm−1, indicating C–C stretching, was correlated with high water-holding capacity (WHC) in KC meat. The quality of meat from slow- and fast-growing chickens significantly varies. Vibrational spectroscopy is a powerful technique that provides insightful molecular information correlated with various meat attributes.

Meat quality attributes vary with chicken age. Understanding the relationship between poultry age and the quality of the meat would be beneficial for efficient poultry farming to meet market needs. The Korat hybrid chicken (KC) is a new crossbred chicken whose meat quality is distinct from that of commercial broiler (CB) chickens and has not been well characterized. In this study, we characterized the physico-chemical properties of KC meat and correlate the findings with Raman spectral data. The protein content of KC breast and thigh meat increased with age. The pH of thigh meat decreased, while the water-holding capacity of breast meat increased as the age of the chickens increased. The amount of cholesterol in breast meat decreased as the rearing period was extended. Inosine 5′-monophosphate and guanosine 5′-monophosphate of breast meat decreased as KC grew older. The shear force values of meat from older birds increased concomitantly with an increase in total collagen. Principle component analysis revealed that the meat quality of CB was greatly different from that of KC meat. High shear force values of KC meat at 20 wk of age were well correlated with an increase in the β-sheet structure (amide I) and amide III of collagen. Raman spectra at 3,207 cm−1 and relative α-helical content were negatively correlated with shear force values of KC breast meat. These could be used as markers to evaluate KC meat quality.

Objective: Obesity and insulin resistance greatly increase the risk of nonalcoholic fatty liver disease and steatohepatitis (NAFLD/NASH). We have previously discovered that whole-body and adipocyte-specific Ip6k1deletion protects mice from high-fat-diet-induced obesity and insulin resistance due to improved adipocyte thermogenesis and insulin signaling. Here, we aimed to determine the impact of hepatocyte-specific and whole-body Ip6k1 deletion (HKO and Ip6k1-KO or KO) on liver metabolism and NAFLD/NASH. Methods: Body weight and composition; energy expenditure; glycemic profiles; and serum and liver metabolic, inflammatory, fibrotic and toxicity parameters were assessed in mice fed Western and high-fructose diet (HFrD) (WD: 40% kcal fat, 1.25% cholesterol, no added choline and HFrD: 60% kcal fructose). Mitochondrial oxidative capacity was evaluated in isolated hepatocytes. RNA-Seq was performed in liver samples. Livers from human NASH patients were analyzed by immunoblotting and mass spectrometry. Results: HKO mice displayed increased hepatocyte mitochondrial oxidative capacity and improved insulin sensitivity but were not resistant to body weight gain. Improved hepatocyte metabolism partially protected HKO mice from NAFLD/NASH. In contrast, enhanced whole-body metabolism and reduced body fat accumulation significantly protected whole-body Ip6k1-KO mice from NAFLD/NASH. Mitochondrial oxidative pathways were upregulated, whereas gluconeogenic and fibrogenic pathways were downregulated in Ip6k1-KO livers. Furthermore, IP6K1 was upregulated in human NASH livers and interacted with the enzyme O-GlcNAcase that reduces protein O-GlcNAcylation. Protein O-GlcNAcylation was found to be reduced in Ip6k1-KO and HKO mouse livers. Conclusion: Pleiotropic actions of IP6K1 in the liver and other metabolic tissues mediate hepatic metabolic dysfunction and NAFLD/NASH, and thus IP6K1 deletion may be a potential treatment target for this disease.

The aim of this study was to evaluate the effects of in ovo feeding (IOF) of L-arginine (Arg) on the hatchability, growth performance, antioxidant capacity, and meat quality of slow-growing chickens. A total of 480 eggs were randomly divided into a non-injected control group (NC group) and a 1% Arg-injected group (Arg group). On day 18 of incubation, 0.5 mL of Arg solution was injected into the embryonic amnion in the Arg group. Upon hatching, 160 mixed-sex chickens were randomly assigned to two groups, with four replicates per group. This experiment lasted for 63 days. The results showed that the hatchability, growth performance, carcass traits, and meat quality were not significantly different (p > 0.05) between the two groups. However, the malondialdehyde (MDA) content was lower (p < 0.05), and the glutathione (GSH) level was higher (p < 0.05) on day of hatching in the Arg group. The total antioxidant capacity (T-AOC) activity was increased (p < 0.05) on day 21 post-hatch in the Arg group compared to that in the NC group. In conclusion, IOF of Arg increased the antioxidant capacity of the breast muscle in the starter period, which may have a positive effect on health status of slow-growing chickens post-hatch.

This study was to determine the effect of the organic raising system (OR) on growth performance, meat quality, and physicochemical properties of slow-growing chickens. Three hundred and sixty (one-day-old) Korat chickens (KRC) were randomly assigned to control (CO) and OR groups. The groups comprised six replicates of thirty chickens each. The chickens were housed in indoor pens (5 birds/m2), wherein those in OR had free access to Ruzi pasture (1 bird/4 m2) from d 21 to d 84 of age. In the CO group, chickens were fed with a mixed feed derived from commercial feedstuffs, while those in the OR group were fed with mixed feed derived from organic feedstuffs. The results revealed a lower feed intake (p < 0.0001) and feed conversion ratio (p = 0.004) in the OR. The OR increased total collagen, protein, shear force, color of skin and meat, and decreased abdominal fat (p < 0.05). The OR improved fatty acid with increased DHA, n-3 PUFA, and decreased the ratio of n-6 to n-3 PUFA in KRC meat (p < 0.05). The synchrotron radiation-based Fourier transform infrared spectroscopy and correlation loading analyses confirmed these results. In conclusion, our results proved that OR could improve growth performance and meat quality and suggested the raising system be adopted commercially. In addition, the observed differences in biochemical molecules could also serve as markers for monitoring meat quality.

The evolutionarily conserved Elongator Complex associates with RNA polymerase II for transcriptional elongation. Elp3 is the catalytic subunit, contains histone acetyltransferase activity, and is associated with neurodegeneration in humans. Elp1 is a scaffolding subunit and when mutated causes familial dysautonomia. Here, we show that "elp3" and "elp1" are required for aversive long-term olfactory memory in "Drosophila." RNAi knockdown of "elp3" in adult mushroom bodies impairs long-term memory (LTM) without affecting earlier forms of memory. RNAi knockdown with coexpression of "elp3" cDNA reverses the impairment. Similarly, RNAi knockdown of "elp1" impairs LTM and coexpression of "elp1" cDNA reverses this phenotype. The LTM deficit in "elp3" and "elp1" knockdown flies is accompanied by the abolishment of a LTM trace, which is registered as increased calcium influx in response to the CS+ odor in the a-branch of mushroom body neurons. Coexpression of "elp1" or "elp3" cDNA rescues the memory trace in parallel with LTM. These data show that the Elongator complex is required in adult mushroom body neurons for long-term behavioral memory and the associated long-term memory trace.

The emergence of the lumpy skin disease virus (LSDV) was first detected in north-eastern Thailand in March 2021. Since then, the abrupt increase of LSD cases was observed throughout the country as outbreaks have spread rapidly to 64 out of a total of 77 provinces within four months. Blood, milk, and nodular skin samples collected from affected animals have been diagnosed by real-time PCR targeting the p32 gene. LSDV was isolated by primary lamb testis (PLT) cells, followed by Madin-Darby bovine kidney (MDBK) cells, and confirmed by immunoperoxidase monolayer assay (IPMA). Histopathology and immunohistochemistry (IHC) of a skin lesion showed inclusion bodies in keratinocytes and skin epithelial cells. Phylogenetic analyses of RPO30 and GPCR genes, and the whole genome revealed that Thai viruses were closely related to the vaccine-derived recombinant LSDV strains found previously in China and Vietnam. Recombination analysis confirmed that the Thai LSDV possesses a mosaic hybrid genome containing the vaccine virus DNA as the backbone and a field strain DNA as the minor donor. This is an inclusive report on the disease distributions, complete diagnoses, and genetic characterisation of LSDV during the first wave of LSD outbreaks in Thailand.

Objective: To investigate the prevalence of trypanosome infection and their phylogeny in Bandicota indica rats from the cadmium-contaminated area of Mae Sot and the Myanmar border. Methods: Blood samples were taken from 100 animals, and parasite infection was examined by light microscopy observation and polymerase chain reaction (PCR) studies. Results: Trypanosoma spp. infection was found in 20% of the thin blood smear samples. PCR showed positive 623 bp DNA bands in 21 samples (21%). The sequencing analysis showed that all of the samples (100%) had the Trypanasoma lewisi 18S ribosomal RNA gene. Phylogenetic analysis confirmed that these 16 isolates of Trypanosoma spp. were closely related to Trypanasoma lewisi. Conclusions: Molecular detection using PCR is as effective as conventional light microscopy analysis. This study confirms that trypanosomal infection in rodents is still high; therefore, fleas as their vectors need to be controlled in order to prevent transmission to humans.

Here, molecular pathways and genes involved in the feed efficiency (FE) and thigh-meat quality of slow-growing Korat chickens were investigated. Individual feed intake values and body weights were collected weekly to the calculate feed conversion ratios (FCR) and residual feed intake. The biochemical composition and meat quality parameters were also measured. On the basis of extreme FCR values at 10 weeks of age, 9 and 12 birds from the high and the low FCR groups, respectively, were selected, and their transcriptomes were investigated using the 8 × 60 K Agilent chicken microarray. A weighted gene co-expression network analysis was performed to determine the correlations between co-expressed gene modules and FE, thigh-meat quality, or both. Groups of birds with different FE values also had different nucleotide, lipid, and protein contents in their thigh muscles. In total, 38 modules of co-expressed genes were identified, and 12 were correlated with FE and some meat quality traits. A functional analysis highlighted several enriched functions, such as biological processes, metabolic processes, nucleotide metabolism, and immune responses. Several molecular factors were involved in the interactions between FE and meat quality, including the assembly competence domain, baculoviral IAP repeat containing 5, cytochrome c oxidase assembly factor 3, and myosin light chain 9 genes.

Korat chicken (KRC) is a slow-growing chicken bred in Thailand, whose meat exhibits a unique toughness. A previous study produced KRC breast meat containing high carnosine content through dietary supplementation with β-alanine or L-histidine; however, the KRC that were fed an L-histidine-supplemented diet produced meat that was significantly more tender. Herein, we performed RNA-Seq to identify candidate genes involved in the regulation of carnosine content and meat toughness. Total RNA was isolated from five female KRC breast muscles in each treatment group that KRC fed diets without supplementation, supplemented with β-alanine or L-histidine. Compared to the non-supplemented group, we identified 118 and 198 differentially expressed genes (DEGs) in the β-alanine or L-histidine supplementation groups, respectively. Genes potentially related to meat tenderness—i.e., those regulating myosin, collagen, intramuscular fat, and calpain—were upregulated (LOC107051274, ACSBG1, and CAPNS2) and downregulated (MYO7B, MYBPH, SERPINH1, and PGAM1). However, carnosine synthase gene was not identified. Functional enrichment analysis identified pathways affected by dietary supplementation, including the insulin signaling pathway (β-alanine supplementation) and the insulin resistance and adipocytokine signaling pathways (L-histidine supplementation). The FoxO signaling pathway was identified as a regulatory network for both supplementation groups. The identified genes can be used as molecular markers of meat tenderness in slow-growing chickens.

Improving feed efficiency is an important breeding target for the poultry industry; to achieve this, it is necessary to understand the molecular basis of feed efficiency. We compared the jejunal transcriptomes of low- and high-feed conversion ratio (FCR) slow-growing Korat chickens (KRs). Using an original sample of 75 isolated 10-week-old KR males, we took jejunal samples from six individuals in two groups: those with extremely low FCR (n = 3; FCR = 1.93 ± 0.05) and those with extremely high FCR (n = 3; FCR = 3.29 ± 0.06). Jejunal transcriptome profiling via RNA sequencing revealed 56 genes that were differentially expressed (p < 0.01, FC > 2): 31 were upregulated, and 25 were downregulated, in the low-FCR group relative to the high-FCR group. Functional annotation revealed that these differentially expressed genes were enriched in biological processes related to immune response, glutathione metabolism, vitamin transport and metabolism, lipid metabolism, and neuronal and cardiac maturation, development, and growth, suggesting that these are important mechanisms governing jejunal feed conversion. These findings provide an important molecular basis for future breeding strategies to improve slow-growing chicken feed efficiency.

(1) Background: We previously demonstrated that disruption of IP6K1 improves metabolism, protecting mice from high-fat diet-induced obesity, insulin resistance, and non-alcoholic fatty liver disease and steatohepatitis. Age-induced metabolic dysfunction is a major risk factor for metabolic diseases. The involvement of IP6K1 in this process is unknown. (2) Methods: Here, we compared body and fat mass, insulin sensitivity, energy expenditure and serum-, adipose tissue- and liver-metabolic parameters of chow-fed, aged, wild type (aWT) and whole body Ip6k1 knockout (aKO) mice. (3) Results: IP6K1 was upregulated in the adipose tissue and liver of aWT mice compared to young WT mice. Moreover, Ip6k1 deletion blocked age-induced increase in body- and fat-weight and insulin resistance in mice. aKO mice oxidized carbohydrates more efficiently. The knockouts displayed reduced levels of serum insulin, triglycerides, and non-esterified fatty acids. Ip6k1 deletion partly protected age-induced decline of the thermogenic uncoupling protein UCP1 in inguinal white adipose tissue. Targets inhibited by IP6K1 activity such as the insulin sensitivity- and energy expenditure-inducing protein kinases, protein kinase B (PKB/Akt) and AMP-activated protein kinase (AMPK), were activated in the adipose tissue and liver of aKO mice. (4) Conclusions: Ip6k1 deletion maintains healthy metabolism in aging and thus, targeting this kinase may delay the development of age-induced metabolic dysfunction.

The selection of rapidly growing animals in breeding programs has had inadvertent detrimental effects on meat quality. Thus, the aim of the present study was to investigate the relationship between body weight (BW) and meat quality traits, and the effects of genes encoding insulin-like growth factor I (IGF-I), insulin-like growth factor II (IGF-II), melanocortin-4 receptor (MC4R), and calpain 1 (CAPN1) on BW, carcass yield, and meat quality of the Thai indigenous chicken, Leung Hang Khao. Five hundred and ten chickens were used for genotyping. PCR-restriction fragment length polymorphism and PCR-single strand conformation polymorphism were used to determine the genotypes of IGF-I, IGF-II, MC4R, and CAPN1. BWs were collected from 0–16 weeks of age. The chickens were sacrificed at 16 weeks and individual carcass yields and meat qualities (drip loss, cooking loss, and shear force) were recorded. The correlations between BW and meat qualities were determined. Significant correlation between BW and cooking loss and shear force of breast meat and between BW and drip loss of thigh meat were detected (P

The aim of the present study was to investigate the effect of single nucleotide polymorphisms in the major histocompatibility complex (MHC) class II gene on resistance to Newcastle disease virus and body weight of the Thai indigenous chicken, Leung Hang Khao (Gallus gallus domesticus). Blood samples were collected for single nucleotide polymorphism analysis from 485 chickens. Polymerase chain reaction sequencing was used to classify single nucleotide polymorphisms of class II MHC. Body weights were measured at the ages of 3, 4, 5, and 7 months. Titres of Newcastle disease virus at 2 weeks to 7 months were determined and the correlation between body weight and titre was analysed. The association between single nucleotide polymorphisms and body weight and titre were analysed by a generalized linear model. Seven single nucleotide polymorphisms were identified: C125T, A126T, C209G, C242T, A243T, C244T, and A254T. Significant correlations between log titre and body weight were found at 2 and 4 weeks. Associations between single nucleotide polymorphisms and titre were found for C209G and A254T, and between all single nucleotide polymorphisms (except A243T) and body weight. The results showed that class II MHC is associated with both titre of Newcastle disease virus and body weight in Leung Hang Khao chickens. This is of concern because improved growth traits are the main goal of breeding selection. Moreover, the results suggested that MHC has a pleiotropic effect on the titre and growth performance. This mechanism should be investigated in a future study.

This study proposes a strategy to manipulate the fatty acid (FA) content in slow-growing Korat chicken (KRC) meat using tuna oil (TO). To determine the optimal level and feeding period of TO supplementation, we conducted a study investigating the effects of dietary TO levels and feeding periods on meat quality, omega-3 polyunsaturated fatty acid (n-3 PUFA) composition, and gene expression related to FA metabolism in KRC breast meat. At 3 wk of age, 700 mixed-sex KRC were assigned to seven augmented factorial treatments with a completely randomized design, each consisting of four replicate pens containing 25 chickens per pen. The control group received a corn-soybean-based diet with 4.5% rice bran oil (RBO), while varying amounts of TO (1.5%, 3.0%, or 4.5%) replaced a portion of the RBO content in the experimental diets. The chickens were fed these diets for 3 and 6 wk, respectively, before being slaughtered at 9 wk. Our results indicated no significant interactions between TO levels and feeding periods on the growth performance or meat quality of KRC (P > 0.05). However, the liver fatty acid-binding protein gene (L-FABP, also known as FABP1), responsible for FA transport and accumulation, showed significantly higher expression in the chickens supplemented with 4.5% TO (P < 0.05). The chickens supplemented with 4.5% TO for a longer period (3 to 9 wk of age) exhibited the lowest levels of n-6 PUFA and n-6 to n-3 ratio, along with the highest levels of eicosapentaenoic acid, docosahexaenoic acid, and n-3 PUFA in the breast meat (P < 0.05). However, even a short period of supplementation with 4.5% TO (6 to 9 wk of age) was adequate to enrich slow-growing chicken meat with high levels of n-3 PUFA, as recommended previously. Our findings indicated that even a short period of tuna oil supplementation could lead to desirable levels of omega-3 enrichment in slow-growing chicken meat. This finding has practical implications for the poultry industry, providing insights into optimal supplementation strategies for achieving desired FA profiles without adversely affecting growth performance or meat quality. [ABSTRACT FROM AUTHOR]

Obesity and nonalcoholic fatty liver disease (NAFLD) are global health concerns, and thus, drugs for the long-term treatment of these diseases are urgently needed. We previously discovered that the inositol pyrophosphate biosynthetic enzyme IP6K1 is a target in diet-induced obesity (DIO), insulin resistance, and NAFLD. Moreover, high-throughput screening (HTS) assays and structure−activity relationship (SAR) studies identified LI-2242 as a potent IP6K inhibitor compound. Here, we tested the efficacy of LI-2242 in DIO WT C57/BL6J mice. LI-2242 (20 mg/kg/BW daily, i.p.) reduced body weight in DIO mice by specifically reducing the accumulation of body fat. It also improved glycemic parameters and reduced hyperinsulinemia. LI-2242-treated mice displayed reduced the weight of various adipose tissue depots and an increased expression of metabolism- and mitochondrial-energy-oxidation-inducing genes in these tissues. LI-2242 also ameliorated hepatic steatosis by reducing the expression of genes that enhance lipid uptake, lipid stabilization, and lipogenesis. Furthermore, LI-2242 enhances the mitochondrial oxygen consumption rate (OCR) and insulin signaling in adipocytes and hepatocytes in vitro. In conclusion, the pharmacologic inhibition of the inositol pyrophosphate pathway by LI-2242 has therapeutic potential in obesity and NAFLD. [ABSTRACT FROM AUTHOR]

Current thought envisions dopamine neurons conveying the reinforcing effect of the unconditioned stimulus during associative learning to the axons of Drosophila mushroom body Kenyon cells for normal olfactory learning. Here, we show using functional GFP reconstitution experiments that Kenyon cells and dopamine neurons from axoaxonic reciprocal synapses. The dopamine neurons receive cholinergic input via nicotinic acetylcholine receptors from the Kenyon cells; knocking down these receptors impairs olfactory learning revealing the importance of these receptors at the synapse. Blocking the synaptic output of Kenyon cells during olfactory conditioning reduces presynaptic calcium transients in dopamine neurons, a finding consistent with reciprocal communication. Moreover, silencing Kenyon cells decreases the normal chronic activity of the dopamine neurons. Our results reveal a new and critical role for positive feedback onto dopamine neurons through reciprocal connections with Kenyon cells for normal olfactory learning.

Cassava pulp contains a lot of starch, but low amounts of protein and high fiber content which limits its use as a feedstuff for broilers. However, fermentation of this pulp with Aspergillus oryzae (A. oryzae) to improve its protein content may increase its usefulness in broiler diets. Therefore, two experiments were conducted to evaluate the potential use of fermented cassava pulp (FCP) in broilers. In experiment 1 the effects of FCP on nutrient digestibility and retention were studied. FCP was prepared using cassava pulp fermented with A. oryzae and urea for 4 days. Forty-nine fifteen-day old male chickens were placed in individual cages and assigned randomly to one of 7 dietary treatment groups (one control and six FCP: 40, 80, 120, 160, 200 and 240 g/kg) for 10 days. The results indicate that nutrient digestibility and retention decreased with increasing levels of FCP (P>0.05), but the decrease was not significant at dietary levels below 160 g/kg. Experiment 2 studied the effect of FCP in broiler diets on growth performance, carcass quality and blood biochemistry. Two hundred and seventy one-day old male chicks were randomly distributed to 6 dietary groups (one control and five FCP: 40, 80, 120, 160 and 200 g/kg) for 42 days. The results show that FCP could be used as an energy source with inclusion levels up to 160 g/kg in broiler diets having no effect on growth performance, carcass composition, meat color or blood biochemistry (P>0.05). Moreover, it was found that FCP had no detrimental effects on the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of broilers (P>0.05). In conclusion, FCP can be used in broiler diets up to 160 g/kg without detrimental effects on nutrient digestibility and retention, growth performance, carcass quality or blood biochemistry.

The purpose of this study was to describe and examine a model for assessing student learning through reflection in service-learning courses. This model utilized a course-embedded process to frame, facilitate, support, and assess students' depth of learning and critical thinking. Student reflection products in two service-learning courses (a freshman course and an upper-level course) at a public university were examined at two times for depth of academic, personal, and civic learning and for level of critical thinking. Depth of learning and levels of critical thinking between freshmen and upperclassmen were compared. Results suggest that the model and associated rubrics were useful in documenting student learning. Students could identify, describe, and apply their learning. They had difficulty, however, evaluating their learning and thinking critically. There was some enhancement in depth of learning and critical thinking over time with upperclassmen achieving greater depth of learning and higher levels of critical thinking in some areas. Findings indicate that the model is a rigorous tool that can be used to document and assess student learning in service-learning courses. (Contains 6 tables.)

1. Korat (KR) chicken is a new meat-type chicken breed established with the purpose of developing Small and Micro Community Enterprise Production. This slow-growing chicken has been recognised for its good texture and flavour, but its low feed efficiency is associated with high cost of production which can hamper the development of local production. This has highlighted the importance of studying the trade-off between feed efficiency and meat quality in KR chicken. 2. This study investigated the phenotypic correlations between feed efficiency, growth performance, and meat quality in KR chicken. Individual body weight and feed intake were recorded weekly for 75 male KR chickens for the calculation of feed conversion ratio (FCR) and residual feed intake (RFI). The growth curve was modelled by Gompertz function and meat quality evaluated at 10 weeks of age, through the measurement of pH (pHu), water-holding capacity (WHC) and drip loss (DL) in breast and thigh muscle. 3. Faster growth rate at young age appeared favourable, regarding feed efficiency, since a moderate negative correlation was found between FCR and initial growth rate. Chickens with a higher initial growth and maturation rates were characterised by lower water-holding capacity in meat. This was not associated with acidic meat, since a higher speed of growth (especially in the first weeks) was positively associated with final pH in breast meat. Relationships between RFI and meat quality traits appeared to be weak. While better FCR could be achieved by increased initial growth rate, this seemed to negatively impact water-holding capacity of the meat. 4. RFI appeared as an interesting alternative as a compromise between FCR and meat quality. [ABSTRACT FROM AUTHOR]

Objective: This study investigated the association between feed efficiency, physicochemical properties, flavor precursors and biomolecules in the thigh meat of Korat (KR) chickens. Methods: The feed intake and body weight of individual male KR chickens were recorded from 1 to 10 weeks old to calculate the individual residual feed intake (RFI) of 75 birds. At 10 weeks of age, chickens with the 10 highest (HRFI) and lowest RFI (LRFI) were slaughtered to provide thigh meat samples. The physicochemical properties (ultimate pH, water holding capacity [WHC], drip loss) and flavor precursors (guanosine monophosphate, inosine monophosphate (IMP), adenosine monophosphate and inosine) were analyzed conventionally, and Fourier transform infrared spectroscopy was used to identify the composition of biomolecules (lipids, ester lipids, amide I, amide II, amide III, and carbohydrates) and the secondary structure of the proteins. A group t-test was used to determine significant differences between mean values and principal component analysis to classify thigh meat samples into LRFI and HRFI KR chickens. Results: The physicochemical properties of thigh meat samples from LRFI and HRFI KR chickens were not significantly different but the IMP content, ratios of lipid, lipid ester, protein (amide I, amide II) were significantly different (p<0.05). The correlation loading results showed that the LRFI group was correlated with high ratios of lipids, lipid esters, collagen content (amide III) and beta sheet protein (rg loading >0.5) while the HRFI group was positively correlated with protein (amide I, amide II), alpha helix protein, IMP content, carbohydrate, ultimate pH and WHC (rg loading >0.5). Conclusion: The thigh meat from chickens with different RFI differed in physiochemical properties affecting meat texture, and in the contents of flavor precursors and biomolecules affecting the nutritional value of meat. This information can help animal breeders to make genetic improvements by taking more account of traits related to RFI. [ABSTRACT FROM AUTHOR]