Rare nonclassic type of Congenital adrenal hyperplasia due to 21-hydroxylase deficiency and genotype-phenotypic correlation.

Objective: To explore the correlation between different CYP21A2 pathogenic gene mutations and clinical phenotypes in Congenital adrenal hyperplasia (CAH) patients. Moreover, combined with the specific phenotypes of patients in the clinic, diagnosis and treatment suggestions should be made for CAH patients. Methods: In this study, a genetic status of a Chinese family in three generations of 21-hydroxylase deficiency was comprehensively presented, and the pathogenic genes in the family were found and traced in detail. We measured CYP21A2 gene in this family by Sanger sequencing and MLPA. The trophoblast cells of female proband's embryos were detected by PGT-M which used Copy-Number Variations of a Single Human Cell and high throughput sequencing. The CYP21A2 gene mutation site in each embryo were detected by Sanger sequencing, whole genome sequencing and single nucleotide polymorphism (SNP). Results: There are many related pathogenic genes of CAH in this family. The female proband showed a compound heterozygous mutation in the CYP21A2 gene, including a CYP21A1P/A2 fusion gene (CH-8) (classical phenotype) and a new mutation c.1034T > C (p. L354S) (unknown clinical significance). In the proband's family, a heterozygous gene mutation of c.1034T > C and a CYP21A1P/A2 fusion gene (CH-8) was carried by her father and mother, respectively. Meanwhile, the husband of the proband also has a genetic family with related disease. Both the husband and his father carried the CYP21A2 gene c.844G > T heterozygous mutation, while his mother had no related mutation in the CYP21A2 gene. Furthermore, PGTM gene detection was carried out on the four blastocysts of the proband's offspring through IVF. The results showed that embryos T1, T2 and T4 all carried CYP21A1P/A2 fusion gene (CH-8), as well as embryo T3 carried c.1034T > C heterozygous mutation of maternal origin. Conclusion: This case is a family report showing a complete genetic map of the proband and her family, describing the genetic process of different pathogenic genes in detail and clearly corresponding to the patient's different phenotypes. It is speculated that the pathogenesis of CAH is caused by different mutations in the CYP21A2 gene and their interactions, which may affect the different phenotypes of CAH patients.

The impact of freezing methods on the quality, moisture distribution, microstructure, and flavor profile of hand-grabbed mutton during long-term frozen storage.

The aim of present study was to investigate the influence of conventional freezing (CF, -18 °C), low-temperaturefreezing (LF, -40 °C), and ultra-low-temperature freezing (ULF, -80 °C) on the quality, moisture distribution, microstructure, and flavor profile of hand-grabbed mutton (HGM) during frozen storage (0, 30, 60, 90, 120, 150 and 180 days). The TPC, TVB-N, and TBARS values increased significantly with prolonged storage, while the moisture content decreased (P < 0.05). Additionally, the concentrations of aldehydes, alcohols, ketones, acids, and alkenes decreased significantly as the storage duration increased. However, the concentrations of esters and heterocyclics increased (P < 0.05). Notably, at 30-180 days of storage, the TBARS and TVB-N values in ULF samples were significantly lower than those in CF and LF samples, while the moisture content was significantly higher (P < 0.05). Low field-nuclear magnetic resonance (LF-NMR) analysis showed that ULF decreased water migration and maintained the original texture characteristics of HGM during frozen storage. The ULF and LF groups had significantly higher levels of volatiles than the CF group (P < 0.05). The findings show that ULF, with its relatively rapid freezing rates, can still maintain the high quality of HGM after 180 days of frozen storage, contributing to quality control.

Investigation Tracing the Origin of Tan Sheep Visceral Tissues through Mineral Elements.

The traceability of quality mineral fingerprints in the viscera of Tan sheep from northwest China was studied. Twenty-five mineral elements in the heart and liver samples of Tan sheep were determined using an inductively coupled plasma mass spectrometer (ICP-MS), and the characteristics of the mineral elements in the visceral tissues of the Tan sheep were further analyzed in combination with a principal component analysis (PCA), hierarchical cluster analysis (HCA), and linear discriminant analysis (LDA) to establish a discriminant model and verify it. The results show that 11 elements (137Ba, 43Ca, 63Cu, 56Fe, 39K, 31P, 60Ni, 78Se, 118Sn, 125Te, and 66Zn) in the Tan sheep heart samples had significant differences among different regions (p < 0.05), and the results of the LDA show that the accuracy rate of the return-generation examination was 85.70%, and the accuracy rate of the hand-over-fork examination was 87.50%; 10 elements (111Cd, 59Co, 52Cr, 56Fe, 39K, 55Mn, 95Mo, 23Na, 121Sb, and 78Se) in the Tan sheep liver samples had significant differences among different regions (p < 0.05), and the results of the LDA showed that the accuracy rate of the return-generation examination was 96.30%, and the accuracy rate of the hand-over-fork examination was 86.25%. This indicates that the multi-element analysis has potential for determining the origin of Tan sheep viscera in certain regions.

Serum levels of anti-Müllerian hormone influence pregnancy outcomes associated with gonadotropin-releasing hormone antagonist treatment: a retrospective cohort study.

As a specific predictor of ovarian reserve, serum anti-Müllerian hormone (AMH) has become an area of intense research interest in the field of assisted reproductive technology. We assessed the relationship between AMH levels and pregnancy outcomes in Chinese patients and investigate the influencing factors of cumulative live birth in patients with high AMH levels. A total of 1379 patients starting their IVF/ICSI cycle were divided into normal (Group A, 1.1-4.0 ng/ml, n = 639) and high (Group B, > 4.0 ng/ml, n = 740) groups by serum AMH levels. Live birth rate (LBR), cumulative live birth rate (CLBR) and cumulative clinical pregnancy rate (CCPR) were also investigated. Compared with Group A, Group B had a significantly higher CLBR (65.80% vs. 43.95%) and CCPR (76.77% vs. 57.14%), respectively. Binomial logistic regression analysis showed that age over 40 years, LH/FSH > 2.5, total Gn dose and Gn duration, and greater than 4000 ng/ml serum E2 levels on HCG day were significantly associated with CLBR in Group B. The AUC value of CLBR averaged 0.664 (ranging from 0.621 to 0.706) (p < 0.001). The patients with high AMH levels had higher CPR, higher LBR, and lower MR with no statistically significant differences, although there were significant improvements in CLBR. Advanced age (> 40 years) still impacted CLBR, even in women with good ovarian reserves. Consequently, it is still recommended that patients over 40 years old with high AMH levels actively receive IVF treatment if they seek to become pregnant. PCOS diagnoses did not influence the CLBR. In summary, this study showed that serum AMH levels could positively predict patient ovarian responses and further affect pregnancy outcomes.

Dietary linseed supplementation improves meat quality and flavor of sheep by altering muscle fiber characteristics and antioxidant capacity.

Linseed as a common oil crop that has been implicated in regulating the meat quality of animals. The aim of this investigation was to establish the effects of dietary linseed supplementation on carcass traits, meat quality, muscle fiber characteristics, volatile compounds and antioxidant capacity of sheep. Twenty-four Sunit sheep were distributed into 2 groups with different diet: control (CO) fed a standard diet and linseed (LS, 8% of extruded linseed). The results showed that linseed not only increased M. longissimus thoracis (LT) increased the loin-eye area but also decreased pH24h, L* and shear force. Meanwhile, linseed increased mRNA expression of MyHCI and MyHCIIx and decreased in cross-sectional area (CSA) and muscle fiber diameter. In addition, linseed altered the composition of meat volatile flavor compounds, such as Z-10-Pentadecen-1-ol, pentanal, 2-Octenal, (E)-, decanal, butane, and 2-heptanone. Moreover, linseed increased total antioxidative capacity (T-AOC) activity, the mRNA expression of glutathione peroxidase (GSH-Px) and catalase (CAT), decreased malondialdehyde (MDA) content. Overall, the results suggest that linseed is an effective feed additive in improving meat quality and flavor. The underlying mechanisms for its effectiveness may be partly due to a change in muscle fiber characteristics and antioxidant capacity.

Effect of dietary probiotics supplementation on meat quality, volatile flavor compounds, muscle fiber characteristics, and antioxidant capacity in lambs.

This study investigated the effects of probiotics on growth performance, meat quality, muscle fiber characteristics, volatile compounds, and antioxidant capacity in lambs. A total of 24 Sunit lambs were randomly allocated into two groups, each consisting of three replicates of four lambs. Throughout the experiment period, the lambs were fed with based diet (CON) and 10 g probiotics/d supplemented diet (PRO). Compared with the CON group, the number of lactic acid bacteria in fecal samples of PRO group was significantly increased (p < .05) and the coliforms were significantly decreased (p < .05). Dietary probiotics supplementation decreased pH24h, L*, and shear force (p < .05). The muscle fibers were switched from type IIB to type I, with a decrease in the mean cross-sectional area (CSA) (p < .05) of longissimus thoracis (LT) muscle. Also, probiotics altered the composition of meat volatile flavor compounds, such as nonanal, undecanal, 1-pentanol, 1-hexanol, and 2,3-octanedione. In addition, probiotics increased the total antioxidative capacity (T-AOC) and catalase (CAT) activity of LT muscle, while it decreased superoxide dismutase (SOD) activity (p < .05). Overall, these results indicated that probiotics could be used as an effective feed additive by improving meat tenderness and flavor.

Impact of dietary Lactobacillus supplementation on intramuscular fat deposition and meat quality of Sunit sheep.

To investigate the impacts of dietary Lactobacillus supplementation on meat quality such as edible quality and nutritional value of Sunit sheep, a 90-day feeding experiment (Lactobacillus dietary group, R group; non-Lactobacillus dietary group, C group) using twelve 3-month-old Sunit sheep was conducted. The deposition of intramuscular fat (IMF) was increased (p < .05) while the share force and cooking loss were decreased (p < .05) in the R group compared with the C group. The proportions of seven kinds of fatty acids (FAs) have changed significantly (p < .05), especially with higher functional FAs and lower trans-FA in the R group. Metabonomics analysis showed that the metabolites and pathway-related lipid syntheses, such as carnitine cycle, tricarboxylic acid cycle, and glycerophosphocholine metabolic pathway, have significantly changed in the R group. The Lactobacillus dietary supplements impacted the variation of IMF deposition and FAs composition by altering the lipid metabolism pathways of Sunit sheep and then changed the edible quality and nutritional value. PRACTICAL APPLICATIONS: It is well known that the intramuscular fat (IMF) and fatty acids composition in livestock is positively correlated with various aspects of meat quality such as edible quality and nutritional value, which are related to consumer preference. The present study analyzed the effects of Lactobacillus supplement on the intramuscular fat deposition and meat quality of Sunit sheep, which resulted in the increase of IMF, and the differences of fatty acids composition, especially the functional fatty acids. It was explored the mechanism of Lactobacillus affect the variation of lipid metabolism pathways and key metabolites in sheep, which suggested that altering the feeding regimen could improve the meat quality of agri-animals.

Effects of oxidative stability variation on lamb meat quality and flavor during postmortem aging.

This study evaluated the effects of oxidative stability variation on meat quality and flavor in biceps femoris (BF) and longissimus dorsi (LD) muscles of lambs during postmortem aging. The samples were stored at 4±1℃; the meat quality, flavor and muscle oxidative stability were measured on day 0, 1, 2, 3, and 4 of postmortem aging. The results showed that malondialdehyde (MDA) content increased in both muscle types; superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities decreased, as did total antioxidative capacity (T-AOC). LD muscle showed lower MDA content and antioxidative activity than BF muscle. Meat quality analyses indicated that aging increased the lightness (L*), redness (a*), and yellowness (b*) values of meat while improving its tenderness. Gas chromatography-mass spectrometry results showed that volatile flavor compounds were more abundant in LD muscle than in BF muscle. As the aging time extended, relative contents of aldehyde and alcohol increased in both muscles. The key flavor compounds during postmortem aging including heptanal, octanal, nonanal, and decanal were screened by relative odor activity value (ROAV), and the content of key flavor compounds showed the trend of increasing, which were usually associated with fresh green grass, nutty, and fat descriptors. In conclusion, the oxidative muscles displayed better antioxidative capacity, and postmortem aging altered the oxidative stability of lamb muscle, which affected the meat quality and flavor. PRACTICAL APPLICATION: Meat aging is an important strategy to improve the quality of various meat traits (including flavor). The results of this work could be of interest to meat professionals who will be able to apply in actual production by choosing the best aging time based on flavor and meat quality for different muscle parts.

Biological function, mediate cell death pathway and their potential regulated mechanisms for post-mortem muscle tenderization of PARP1: A review.

Tenderness is a key attribute of meat quality that affects consumers' willingness to purchase meat. Changes in the physiological environment of skeletal muscles following slaughter can disrupt the balance of redox homeostasis and may lead to cell death. Excessive accumulation of reactive oxygen species (ROS) in the myocytes causes DNA damage and activates poly ADP-ribose polymerase 1 (PARP1), which is involved in different intracellular metabolic pathways and is known to affect muscle tenderness during post-slaughter maturation. There is an urgent requirement to summarize the related research findings. Thus, this paper reviews the current research on the protein structure of PARP1 and its metabolism and activation, outlines the mechanisms underlying the function of PARP1 in regulating muscle tenderness through cysteine protease 3 (Caspase-3), oxidative stress, heat shock proteins (HSPs), and energy metabolism. In addition, we describe the mechanisms of PARP1 in apoptosis and necrosis pathways to provide a theoretical reference for enhancing the mature technology of post-mortem muscle tenderization.

Angiopoietin-like protein 4 regulates breast muscle lipid metabolism in broilers.

The objective of this study was to determine the effects of angiopoietin-like protein 4 (ANGPTL4) on breast muscle lipid metabolism in broilers. In experiment 1, 36 thirty-five-day-old male Arbor Acres broilers were randomly allocated into 6 treatment groups with 6 birds in a completely randomized design. The broilers were subjected to intravenous injection of His-SUMO-ANGPTL4 at the dose of 0 (injection of normal saline [NS]), 20, 100, 500, 2,500, or 12,500 ng/kg BW, respectively. The results showed that broilers at 30 min after His-SUMO-ANGPTL4 at the level of 12,500 ng/kg BW intravenous injection had higher (P < 0.05) concentrations of triglyceride and non-esterified fatty acid in the serum, higher (P < 0.05) adipose triglyceride lipase and carnitine palmitoyltransferase 1 mRNA expression in the breast muscle, but lower (P < 0.05) lipoprotein lipase (LPL) mRNA expression in the breast muscle. In experiment 2, 18 thirty-five-day-old male Arbor Acres broilers were randomly allocated into 3 treatment groups with 6 birds in a completely randomized design. The broilers were subjected to intravenous injection of NS, His-SUMO, or His-SUMO-ANGPTL4 (12,500 ng/kg BW) in order to rule out the effect of His-SUMO tag. It's confirmed that ANGPTL4 could increase (P < 0.05) concentrations of triglyceride and non-esterified fatty acid in the serum, enhance (P < 0.05) adipose triglyceride lipase mRNA expression in the breast muscle, and decrease (P < 0.05) LPL mRNA expression in the breast muscle. In experiment 3 and 4, co-culture experiments of chicken primary myoblasts and NS, His-SUMO, or His-SUMO-ANGPTL4 (250 pg/mL, physiological dose) were set up to monitor the cytotoxicity of ANGPTL4 and the changes of lipid metabolism-related genes expression. It was found that cell viability was not affected but LPL mRNA expression in chicken primary myoblasts was highly reduced (P < 0.05) by ANGPTL4. In conclusion, ANGPTL4 could promote lipodieresis and inhibit LPL in the breast muscle of broilers.

Rumen bacteria and meat fatty acid composition of Sunit sheep reared under different feeding regimens in China.

BACKGROUND: Rumen bacteria play a critical role in feed degradation and productivity. This study evaluated the impact of feeding regimen on the rumen microbial populations and fatty acid composition of the meat of sheep. Twenty-four Sunit sheep were raised on a grass pasture from birth to 9 months of age, at which time they were randomly divided into two feeding groups: pasture feeding (PF) and barn feeding (BF). Sheep in the PF group were allowed to graze freely on wild grassland for 3 months. Sheep in the BF group were confined for 3 months to a dry barn, in which they roamed freely with corn straw and corn. RESULTS: Sheep in the PF group had greater rumen bacteria diversity. The relative abundances of the genera Butyrivibrio_2, Saccharofermentans and Succiniclasticum were increased, and that of the genus RC9_gut_group was decreased, in the PF compared to the BF sheep. The n-3 polyunsaturated fatty acid contents were greater in meat from PF sheep than from BF sheep. In addition, the α-linolenic acid (C18:3 n-3, ALA) and conjugated linoleic acid (CLA) contents were positively correlated with the abundance of Butyrivibrio_2. CONCLUSION: Grazing may improve the diversity of rumen bacteria and increase the proportion of ALA and CLA in sheep meat. © 2020 Society of Chemical Industry.

Effect of feeding regimen on meat quality, MyHC isoforms, AMPK, and PGC-1α genes expression in the biceps femoris muscle of Mongolia sheep.

The effects of two feeding regimens on meat quality, myosin heavy chain (MyHC) types, and key factors regulating muscle fiber type (AMP-activated protein kinase [AMPK] and peroxisome proliferator-activated receptor-coactivator-1α [PGC-1α]) in the biceps femoris muscle of Mongolia sheep were investigated. A total of 20 Mongolia sheep were weaning for 90 days and divided into two groups (pasture group (P) and confinement group (C)) at 10.36 ± 0.35 kg of weaning weight. After weaning, sheep were pasture fed or confinement fed for 9 months. The results showed that live weights, carcass weight, intramuscular fat (IMF), and Warner-Bratzler shear force (WBSF) in P group were significantly lower (p < .05) than that in C group. Compared with P group, color evaluations with respect to L* and b* values were significantly higher (p < .05) in C group. Expression of the MyHC I gene in the P group was significantly higher, while MyHC IIa and MyHC IIb genes expression was significantly lower (p < .05) than that in C group. Also, AMPK activity and expression of AMPKα2 and PGC-1α genes were significantly higher (p < .05) in P group compared with C group. The present study indicated that muscle fiber composition was one of the key differences leading to the differences of meat quality in different feeding regimens. AMPK, particularly AMPKα2, and PGC-1α were considered to be two key factors regulating muscle fiber types in Mongolia sheep. The results support that AMPK activity and the expression of AMPKα2 and PGC-1α genes may affect the composition of muscle fibers; thus, AMPK activity and the expression of AMPKα2 and PGC-1α genes had an effect on meat quality by changed composition of muscle fibers.

Impact of feeding regimens on the composition of gut microbiota and metabolite profiles of plasma and feces from Mongolian sheep.

Mongolian sheep are an indigenous ruminant raised for wool and meat production in China. The gut microbial community plays an important role in animal performance and metabolism. The objective of this study was to investigate the effects of two feeding regimens on the diversity and composition of gut microbiota and metabolite profiles of feces and plasma from Mongolian sheep. A total of 20 Mongolian sheep were assigned to one of two feeding regimens: free grazing (FG) and barn confinement (BC). When samples were collected, the average live weights of the sheep were 31.28 ± 1.56 kg and 34.18 ± 1.87 kg for the FG and BC groups, respectively. At the genus level, the FG group showed higher levels of Bacteroides, RC9_gut_group, Alistipes, Phocaeicola, Barnesiella, and Oscillibacter, and lower levels of Succinivibrio, Treponema, and Prevotella, compared to the BC group. The butyric acid content in feces was lower in the FG group (P > 0.05). Higher levels of palmitic acid, oleic acid, alpha-linolenic acid, L-carnitine, L-citrulline, and L-histidine, and lower levels of L-tyrosine, L-phenylalanine, and L-kynurenine were found in the plasma of the FG sheep. Moreover, there were substantial associations between several gut microbiota genera and alterations in feces and plasma metabolites especially those involved in the metabolism of butyric acid, linolenic acid, and L-tyrosine. Feeding regimens can not only influence the composition of gut microbiota, but also alter metabolic homeaostasis in sheep.

Meat quality, fatty acids, volatile compounds, and antioxidant properties of lambs fed pasture versus mixed diet.

The aim of the present work was to investigate the effects of feeding regimens (pasture vs. mixed diet) on meat quality, fatty acids, volatile compounds, and antioxidant properties in lamb meat. In total, 24 lambs were allotted into two feeding regimens at 10.23 kg live weight. Lambs were fed on pasture grass (PG group, n = 12) or mixed diet (M group, n = 12). Longissimus thoracis (LT) muscle samples from the M group had a higher intramuscular fat (IMF) (p < 0.05), pH45minvalue (p < 0.01), and ash (p < 0.05) than the PG group. In contrast, the shear force (p < 0.05), L*(p < 0.05), and b* (p < 0.001) in M group were lower than in PG group. Analyses indicated that PG group contained higher linolenic acid (C18:3n3) and docosatrienoic acid (C22:3n6) (p < 0.05) than the M group. Major volatile compounds in the muscles included hexanal, heptanal, nonanal, octanal, 1-pentanol, 1-hexanol, 1-octen-3-ol, and 2,3-octanedione. The levels of hexanal, nonanal, and 2,3-octanedione were significantly lower in PG lamb muscle (p < 0.01). In contrast, 1-pentanol and 1-hexanol levels were higher in M lamb muscle (p < 0.01). Muscle from PG lamb exhibited higher catalase (CAT) and glutathione peroxidase (GPx) activity (p < 0.05). PG muscle also contained a higher radical-scavenging ability (RSA; p < 0.001) and cupric-reducing antioxidant capacity (CUPRAC; p < 0.05). Overall, the improved antioxidant status in PG muscle inhibited lipid peroxidation (aldehydes and ketones), thereby improving the meat quality.

Primary peritoneal serous carcinoma with signet ring cells.

► The case is unique as the signet ring-like tumor cells in the ascites occur in primary peritoneal serous tumors. ► The tumor cells may have degenerated, which led to the formation of cytoplasmic vacuoles and resembled signet ring-like cells.