Rh(III)-Catalyzed Dual C-H Functionalization/Cyclization Cascade by a Removable Directing Group: A Method for Synthesis of Polycyclic Fused Pyrano[de]Isochromenes.

An interesting Rh(III)-catalyzed dual C-H functionalization/cyclization cascade of azomethine imine with diazophosphonate by a removable directing group for the synthesis of highly fused pyrano[de]isochromene has been achieved. The transformation shows that the desired pyrano[de]isochromenes with two oxygen atoms on its core scaffold could be constructed with good to excellent yields (up to 86%) via a facile one-pot, multiple-step cascade reaction, along with broad generality and versatility.

Greener liquid-phase synthesis and the ACE inhibitory structure-activity relationship of an anti-SARS octapeptide.

A high-efficiency strategy for resin-free and large scale liquid phase synthesis of the anti-SARS octapeptide AVLQSGFR is described. Herein, tri(4'-diphenylphosphonyloxylbenzoyl phenyl)phosphate (TDPBP) derivatives were designed as C-terminal supports to aid octapeptide intermediate purification without the need for chromatographic separation. Furthermore, the ACE inhibitory structure-activity relationship (SAR) of the anti-SARS octapeptide and its alanine-scanning analogues was systematically studied by in vitro assay and 3D-QSAR via molecular docking. This paper provides a new strategy for the development of peptide-based drugs. Simultaneously, a study on the ACE inhibition and structure-activity relationship of the anti-SARS octapeptide also lays a foundation for further understanding how the anti-SARS octapeptide acts as an ACE inhibitor.

Chronic Inflammatory and Proliferative Lesions of the Gallbladder in Aged Pigs.

Primary epithelial tumors of the gallbladder are rarely reported in animals. In this study, 9 aged pigs (6-12 years old) were histopathologically examined for gallbladder proliferative lesions. At necropsy, a large gallstone occupied the lumen of the gallbladder of 3 pigs. Histopathological examination revealed chronic cholecystitis in all 9 pigs, mucosal hyperplasia in 2 pigs, adenoma in 1 pig, and adenocarcinoma in 2 pigs. Bacilli were detected in the gallbladder lumen of 6 pigs by Warthin-Starry stain. Mucosal hyperplasia, adenoma, and adenocarcinoma were characterized by papillary projections of the mucosa with occasional acinar structures. Tumor invasion of the surrounding tissue was observed in the cases of adenocarcinoma. On Alcian blue and periodic acid-Schiff double-stained sections, the acinar structure of gallbladder mucosa in chronic cholecystitis and mucosal hyperplasia was stained in a mosaic pattern, indicating pyloric gland metaplasia. The results of immunohistochemistry revealed a CD10-positive epithelial brush border and mucin (MUC) 2-positive goblet cells in chronic cholecystitis, adenoma, and adenocarcinomas, indicating intestinal metaplasia. Immunoreactivity of MUC5 AC and cytokeratin 19 was weaker in adenoma and adenocarcinomas compared with the normal and hyperplastic gallbladder mucosa. The number of p53-positive nuclei and the Ki-67 index were higher in adenocarcinomas compared with benign lesions. These results suggest that chronic cholecystitis associated with gallstones and/or bacterial infections may contribute to metaplastic changes and development of gallbladder tumors in aged pigs. Alteration of mucin, cytokeratin, and p53 profiles in gallbladder proliferative lesions in pigs were similar to that in humans, suggesting a common pathogenesis in tumor development.

Melatonin rescues the aneuploidy in mice vitrified oocytes by regulating mitochondrial heat product.

Vitrification of germinal vesicle (GV) stage oocytes has been shown to be closely associated with decreased rates of meiosis maturation and increased rates of aneuploidy. However, little is known about the effects of melatonin on these events in mice vitrified GV oocytes. In this study, the effects of melatonin on meiosis maturation potential and the incidence rate of aneuploidy in mouse vitrified oocytes were analyzed by supplementing in vitro maturation (IVM) solution with melatonin at different concentrations. This study, for the first time, showed that the mitochondrial heat production was markedly increased in vitrified oocytes (P < 0.05), which compromised the first polar body extrusion (PBE) of vitrified oocytes (73.3% vs. 85.1%, P < 0.05). However, 10-11 mol/L melatonin could significantly decrease mitochondrial heat production and ROS level (9.1 vs. 12.0 pixels, P < 0.05), meanwhile increase ATP level (1.1 vs. 0.88 pmol, P < 0.05) and mtDNA copies (107438 vs. 67869, P < 0.05), which rescued the abnormal chromosome alignment (32% vs. 69%, P < 0.05) and reduced the incidence of aneuploidy (15.6% vs. 38.5%, P < 0.05) in vitrified oocytes. The meiosis maturation ability of vitrified oocytes with melatonin supplementation was similar to that of fresh ones (83.4% vs. 85.1%, P > 0.05). Collectively, our data revealed that melatonin has a protective action against vitrification-induced injuries of oocytes meiosis maturation.

Expression of uterine lipocalin 2 and its receptor during early- to mid-pregnancy period in mares.

From previous cDNA subtraction studies analyzing gene expression in equine endometrium, high lipocalin 2 (LCN2) mRNA expression was found in the gravid endometrium. In the uterus, LCN2 may transport hydrophobic molecules and siderophores with iron, or may form a complex with another protein, however, the expression of uterine LCN2 beyond day 20 of equine pregnancy and its receptor has not been characterized. To study the expression and potential roles of uterine LCN2 from pre-implantation to mid-gestation period, stage-specific endometrial samples were obtained from day 13 (day 0 = ovulation) cyclic and days 13, 19, 25, and 60 to 131 pregnant mares. Expression of LCN2 mRNA increased in day 19 gravid endometrium and was abundant from day 60 onward. The expression of LCN2 mRNA was localized to the glandular epithelium. LCN2 protein was detected in day 25 gravid endometrium and luminal fluid, and the protein was localized to the glandular epithelium and luminal cavity, whereas LCN2 receptor expression was found in luminal and glandular epithelium and trophectoderm throughout the experimental period. The presence of matrix metalloproteinase-9 (MMP9) was also examined because MMP9 is known to form a complex with LCN2. Although MMP9 and LCN2 were both found in luminal fluid from day 25 pregnant uterus, the complex of these proteins was not detected. Localization of the receptor in the trophectoderm suggests that endometrial LCN2 could play a role in carrying small substances from the mother to fetus in the equine species.

Current status and prospects for in-feed antibiotics in the different stages of pork production - A review.

Antibiotics have long been of great benefit for people, both in the medical treatment of human disease and in animal food where they improve the growth performance and feed utilization during animal production. Antibiotics as in-feed supplements affect all stages of pork production, including the gestation, nursing, growing, and finishing stages, although the effects show stage-dependent differences. However, the use of antibiotics in animal feed has become a worldwide concern. This review describes why sub-therapeutic levels of antibiotic additives in animal feed have become an integral part of animal feeding programs for more than 70 years, particularly in pork production. It also discusses the threat of the long-term use of sub-therapeutic levels of antibiotics in pork production. In recent years, the effectiveness of in-feed antibiotics has tended to decrease. This review analyzes this change from various perspectives. First, the equipment used at pig farms has improved dramatically and is more sanitary. Worldwide, more pig farms use pig farrowing crates, gestation crates, piglet nursery crates, flooring devices, piggery ventilation and cooler systems, automatic pig feeders, piggery heating equipment, and artificial insemination systems. In addition, scientists have replaced the use of antibiotics with organic acids, fermented mash, probiotics, prebiotics, minerals, oligosaccharides, enzymes, herbs/flavors, and protein/amino acids, and have improved management and husbandry techniques. In addition, animal welfare legislation has been aimed at improving the quality of the floors and living space, ensuring that animals have permanent access to fresh water, and setting a minimum weaning age. Finally, the prospects and the possibility of replacing antibiotics in pork production are described, in line with recent research results.

N6-Methyladenosine in Cell-Fate Determination of BMSCs: From Mechanism to Applications.

The methylation of adenosine base at the nitrogen-6 position is referred to as "N6-methyladenosine (m6A)" and is one of the most prevalent epigenetic modifications in eukaryotic mRNA and noncoding RNA (ncRNA). Various m6A complex components known as "writers," "erasers," and "readers" are involved in the function of m6A. Numerous studies have demonstrated that m6A plays a crucial role in facilitating communication between different cell types, hence influencing the progression of diverse physiological and pathological phenomena. In recent years, a multitude of functions and molecular pathways linked to m6A have been identified in the osteogenic, adipogenic, and chondrogenic differentiation of bone mesenchymal stem cells (BMSCs). Nevertheless, a comprehensive summary of these findings has yet to be provided. In this review, we primarily examined the m6A alteration of transcripts associated with transcription factors (TFs), as well as other crucial genes and pathways that are involved in the differentiation of BMSCs. Meanwhile, the mutual interactive network between m6A modification, miRNAs, and lncRNAs was intensively elucidated. In the last section, given the beneficial effect of m6A modification in osteogenesis and chondrogenesis of BMSCs, we expounded upon the potential utility of m6A-related therapeutic interventions in the identification and management of human musculoskeletal disorders manifesting bone and cartilage destruction, such as osteoporosis, osteomyelitis, osteoarthritis, and bone defect.

Ultrasensitive miRNA Detection by AuNP-based 3D DNA Walker and Catalytic Hairpin Assembly (CHA) Cascade Amplification for Early Cancer Diagnosis.

By combining exquisitely designed hairpins with the catalytic hairpin assembly (CHA) to form tripedal DNA walkers driven by enzyme, we constructed a 3D DNA walker with accordingly complementary hairpins attached on gold nanoparticles (AuNPs) and sensitive fluorescence sensing system for the sensitive detection of target miRNA-21 (miR-21). The presence of miR-21 triggers the CHA among three hairpins (HP1, HP2, and HP3), which lead to the formation of the tripedal DNA walkers. For the walking trajectories, FAM-labeled hairpins (HP4) were attached to the surface of AuNPs, the fluorescence of which was initially quenched due to its close proximity to AuNPs. After the binding/cleaving/moving process of tripedal DNA walkers with HP4 driven by Exonuclease III (Exo III), a number of single-stranded DNAs (ssDNAs) will be released with FAM fluorescence recovered. Benefiting from the DNA walker and CHA cascade amplification, the proposed sensing strategy showed remarkable improvement in sensitivity with the LOD of 42 aM. Owing to the precise design of the system, this method exhibited excellent specificity to distinguish miR-21 from its single-, double-mismatched sequences and non-complementary sequences, showing great versatility and potential for the biological analysis and early disease diagnosis.

Genome-Wide Analysis of lncRNA and mRNA Expression in the Uterus of Laying Hens during Aging.

Eggshell plays an essential role in preventing physical damage and microbial invasions. Therefore, the analysis of genetic regulatory mechanisms of eggshell quality deterioration during aging in laying hens is important for the biosecurity and economic performance of poultry egg production worldwide. This study aimed to compare the differences in the expression profiles of long non-coding RNAs (lncRNAs) and mRNAs between old and young laying hens by the method of high-throughput RNA sequencing to identify candidate genes associated with aging in the uterus of laying hens. Overall, we detected 176 and 383 differentially expressed (DE) lncRNAs and mRNAs, respectively. Moreover, functional annotation analysis based on the Gene Ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) databases revealed that DE-lncRNAs and DE-mRNAs were significantly enriched in "phosphate-containing compound metabolic process", "mitochondrial proton-transporting ATP synthase complex", "inorganic anion transport", and other terms related to eggshell calcification and cuticularization. Through integrated analysis, we found that some important genes such as FGF14, COL25A1, GPX8, and GRXCR1 and their corresponding lncRNAs were expressed differentially between two groups, and the results of quantitative real-time polymerase chain reaction (qPCR) among these genes were also in excellent agreement with the sequencing data. In addition, our study found that TCONS_00181492, TCONS_03234147, and TCONS_03123639 in the uterus of laying hens caused deterioration of eggshell quality in the late laying period by up-regulating their corresponding target genes FGF14, COL25A1, and GRXCR1 as well as down-regulating the target gene GPX8 by TCONS_01464392. Our findings will provide a valuable reference for the development of breeding programs aimed at breeding excellent poultry with high eggshell quality or regulating dietary nutrient levels to improve eggshell quality.

MetaRF: attention-based random forest for reaction yield prediction with a few trails.

Artificial intelligence has deeply revolutionized the field of medicinal chemistry with many impressive applications, but the success of these applications requires a massive amount of training samples with high-quality annotations, which seriously limits the wide usage of data-driven methods. In this paper, we focus on the reaction yield prediction problem, which assists chemists in selecting high-yield reactions in a new chemical space only with a few experimental trials. To attack this challenge, we first put forth MetaRF, an attention-based random forest model specially designed for the few-shot yield prediction, where the attention weight of a random forest is automatically optimized by the meta-learning framework and can be quickly adapted to predict the performance of new reagents while given a few additional samples. To improve the few-shot learning performance, we further introduce a dimension-reduction based sampling method to determine valuable samples to be experimentally tested and then learned. Our methodology is evaluated on three different datasets and acquires satisfactory performance on few-shot prediction. In high-throughput experimentation (HTE) datasets, the average yield of our methodology's top 10 high-yield reactions is relatively close to the results of ideal yield selection.

Activatable fluorescent probes for imaging and diagnosis of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that is primarily manifested as synovitis and polyarticular opacity and typically leads to serious joint damage and irreversible disability, thus adversely affecting locomotion ability and life quality. Consequently, good prognosis heavily relies on the early diagnosis and effective therapeutic monitoring of RA. Activatable fluorescent probes play vital roles in the detection and imaging of biomarkers for disease diagnosis and in vivo imaging. Herein, we review the fluorescent probes developed for the detection and imaging of RA biomarkers, namely reactive oxygen/nitrogen species (hypochlorous acid, peroxynitrite, hydroxyl radical, nitroxyl), pH, and cysteine, and address the related challenges and prospects to inspire the design of novel fluorescent probes and the improvement of their performance in RA studies.

Quantitative proteomics provides insights into the mechanism of the differences in heat-induced gel properties for egg white proteins with different interior quality during ageing in laying hens.

The purpose of this study was to investigate the mechanism for the differences in heat-induced gel properties of egg white proteins with different interior quality during ageing in laying hens. Quantitative proteomic analysis revealed that the abundance of ovotransferrin, avidin, mucin 5B, and clusterin increased with decreasing Haugh units (HU), leading to the transition from disorder to order in the secondary and tertiary structure of egg white proteins, with the burial of hydrophobic groups and a reduction in the negative charge on the protein surface, rendering the egg white protein solution aggregated. These changes would accelerate the rate of aggregation of egg white proteins during heating, resulting in the loss of orientation of the molecular chains, forming coarse and porous gel structures and poor gel properties. This research provides a new idea for improving the gelling properties of egg whites from lower interior quality during ageing in laying hens.

The ionome and proteome landscape of aging in laying hens and relation to egg white quality.

The ionome is essential for maintaining body function and health status by participating in diverse key biological processes. Nevertheless, the distribution and utilization of ionome among different organs and how aging impacts the ionome leading to a decline in egg white quality remain unknown. Thus, we used inductively coupled plasma mass spectrometry (ICP-MS) to analyze 35 elements and their isotopic contents in eight organs of laying hens at 35, 72, and 100 weeks. Moreover, the magnum proteome, amino acids in egg white, and egg white quality were analyzed in laying hens at three different ages using 4D proteomics techniques, an amino acid analyzer, and an egg quality analyzer. Across the organs, we identified varying distribution patterns among macroelements (Mg24, Ca43/44, K39, and P31), transition metals (Zn64/66, Cu63/65, Fe56/57, and Mn55), and toxic elements (Pb208, Ba137, and Sr86). We observed an organ-specific aging pattern characterized by the accumulation of toxic elements (Pb208, Ba137, and Sr86) and calcification in the small intestine. Additionally, a decrease in the utilization of essential trace elements selenium (Se78/82) and manganese (Mn55) was noted in the oviduct. By analyzing ionome in tandem with egg quality, egg white amino acids, and proteome, we unveiled that the reduction of selenium and manganese concentrations in the magnum during the aging process affected amino acid metabolism, particularly tryptophan metabolism, thereby inhibiting the amino acid synthesis in the magnum. Furthermore, it accelerated the senescence of magnum cells through necroptosis activation, leading to a decline in the albumen secretion function of the magnum and subsequently reducing egg white quality. Overall, this study provides insights into the evolution of 35 elements and their isotopes across 8 organs of laying hens with age. It also reveals the elemental composition, interactions, and utilization patterns of these organs, as well as their correlation with egg white quality. The present study highlights the significance of ionome and offers a comprehensive perspective on the selection of ionome for regulating the aging of laying hens.

Effect of RNA interference of BID and BAX mRNAs on apoptosis in granulosa cell-derived KGN cells.

In mitochondrion-dependent type II apoptosis, BH3-interacting domain death agonist (BID) and BCL-2-associated X protein (BAX) promote death ligand and receptor-mediated cell death. In porcine ovaries, the levels of BID and BAX increase in follicular granulosa cells during atresia. In the present study, to confirm the pro-apoptotic activity of BID and BAX in granulosa cells, we examined the effect of RNA interference of BID or BAX on apoptosis using a human ovarian granulosa tumor cell line, KGN. By reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, expression of BID and BAX was detected in KGN cells. Then, we suppressed BID and BAX mRNA expression in KGN cells using small interfering RNA (siRNA). When BID or BAX was suppressed, a significant decrease in the apoptotic cell rate was noted. In granulosa-derived cells, BID and BAX showed pro-apoptotic activity. These results suggest that BID and BAX act as signal-transducing factors in mitochondrion-dependent type II apoptosis.

Intestinal Microbiota Regulate Certain Meat Quality Parameters in Chicken.

Growing evidence of intestinal microbiota-muscle axis provides a possibility to improve meat quality of broilers through regulating intestinal microbiota. Water-holding capacity is a crucial factor to evaluate the meat quality. High quality of water-holding capacity is usually described as a low drip-losing rate. This study aimed to explore the relationship between intestinal microbiota and water-holding capacity of muscle in broilers. According to our results, two native breeds of broilers (the Arbor Acres broilers and the Beijing-You broilers) exhibited remarkable differences in microbiota composition. However, the regular of gut bacteria compositions gradually became similar when the two breeds of broiler were raised in a same feeding environment. Therefore, this similar regular of intestinal microbiota induced similar water-holding capacity of the muscle from the two breeds. In subsequent fecal microbiota transplantation (FMT) experiments, the intestinal microbiota community of the Arbor Acres broilers was remodeling by oral gavage of bacterial suspension that was derived from the Beijing-You broilers. Then, not only body weight and abdominal fat rate were increased, but also drip loss of muscle was decreased in the Arbor Acres broilers. Additionally, muscle fiber diameter of biceps femoris muscle and expression of MyoD1 were notably enlarged. Muscle fiber diameter and related genes were deemed as important elements for water-holding capacity of muscle. Simultaneously, we screened typical intestinal bacteria in both the two native breeds of broilers by 16S rDNA sequencing. Lachnoclostridium was the only bacteria genus associated with drip-losing rate, meat fiber diameter, body weight, and abdominal fat rate. Importance: Higher body weight and superior meat quality in livestock imply an adequate source of protein and substantial commercial value. Regulating the intestinal microbiota of broilers is a promising approach to optimize commercial phenotypes. Our results indicate that the intestinal microbiota profile could be reconstructed by external factors, leading to advantageous changes in muscle characteristics. The cecum microbiota of native broilers have the ability to improve certain meat quality and production performance. The population of Lachnoclostridium spp. could be used to regulate body weight and drip-losing rate in broilers, but more study is needed.

Energy values evaluation and improvement of soybean meal in broiler chickens through supplemental mutienzyme.

This study measured the metabolizable energy of soybean meal (SBM) and evaluated effects of soybean meal specific enzymes supplementation in corn-soybean diets on growth performance, intestinal digestion properties and energy values of 28-day-old broilers. A total of 336 one-day-old male AA broiler chickens were distributed to 7 groups in a completely random design. The birds were given 7 diets containing 6 diets with different combined soybean meals and a fasting treatment, 8 replicates per treatment and 6 birds per replicate (Trial 1). A total of 672 one-day-old male AA broiler chickens were randomly allocated to 7 dietary treatments including a control diet and 6 diets supplemented with 300 mg/kg α-galactosidase, 200 mg/kg ß-mannanase, and 300 mg/kg protease individually or in combination (Trial 2). Apparent metabolizable energy (AME) of broilers was measured from d 25 to 27 in both trial 1 and trial 2. The results showed that AME values of combined soybean meals averaged 2,894 kcal/kg. Dietary ß-mannanase and protease supplementation increased body weight gain (P < 0.05) during d 0 to 14, whereas did not affect the growth performance (P > 0.05) during d 14 to 28. Addition of ß-mannanase in combination with other enzymes significantly increased lipase and trypsin content (P < 0.05) in ileum. In addition, dietary ß-mannanase and protease supplementation individually or in combination enhanced trypsin enzyme content in jejunum (P < 0.05). The ß-mannanase enzyme enhanced villus height and villus height to crypt depth ratio (P < 0.05) of ileum compared with control diet. Moreover, supplementation of enzyme except for protease enhanced raffinose and stachyose degradation ratio (P < 0.05). Dietary ß-mannanase supplementation individually or in combination enhanced AME and AMEn values (P < 0.05). This study demonstrated that dietary enzyme supplementation especially ß-mannanase improved intestinal digestion properties and contributed to high energy values.

Bid and Bax are involved in granulosa cell apoptosis during follicular atresia in porcine ovaries.

More than 99% of follicles undergo "atresia" during follicular development and growth. Follicular atresia is predominantly regulated by granulosa cell apoptosis. However, the intracellular signaling pathway of apoptosis in granulosa cells has not been revealed. In the present study, we examined changes in the expression of BH3-interacting domain death agonist (Bid) and Bcl-2-associated X protein (Bax), which are considered to promote the cell death ligand/receptor-mediated process in mitochondrion-dependent type II apoptosis, in porcine granulosa cells during atresia. Levels of mRNA and protein of Bid and Bax were determined by the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting techniques, respectively. Levels of Bid and Bax mRNA and protein were markedly increased in granulosa cells of early atretic follicles compared with those of healthy follicles. In situ hybridization and immunohistochemical staining revealed that mRNA and protein of Bid and Bax were present in the granulosa cells, though only traces were found in healthy follicles; however, strong staining was noted in atretic follicles. These results indicate that Bid and Bax appear to be signal transduction factors in granulosa cells during follicular atresia and appear to play proapoptotic roles and confirm that the porcine granulosa cell is a mitochondrion-dependent type II apoptotic cell.

Effects of Supplementation with a Quebracho Tannin Product as an Alternative to Antibiotics on Growth Performance, Diarrhea, and Overall Health in Early-Weaned Piglets.

This study assessed the feasibility of using a vegetable extract, MGM-P (quebracho tannin product), as an alternative to antibiotics for weaned piglets; it investigated MGM-P effects on growth performance, diarrhea, and overall health in early-weaned piglets. In total, 24 piglets were allocated to three treatment groups fed basal diets supplemented with 0, 0.2%, or 0.3% MGM-P for 20 days. The addition of 0.3% MGM-P to the diet of early-weaned piglets improved diarrhea incidence, hematological parameters, and intestinal mucosa structure. Furthermore, the addition of 0.2% or 0.3% MGM-P to the diet of early-weaned piglets did not affect their overall health. Importantly, MGM-P had no effects on average daily gain (ADG), average daily feed intake (ADFI), or feed conversion ratio (FCR). Gut morphology analysis showed that treatment with 0.3% MGM-P enhanced the jejunal villus height (p < 0.05) while reducing the ileal crypt depth (p < 0.05) and colon mucosal thickness (p < 0.05). Collectively, the findings suggested that the use of MGM-P as an alternative to dietary antibiotics could improve diarrhea incidence in early-weaned piglets without negative effects on growth performance or overall health.

Metabolic conversion of zearalenone to alpha-zearalenol by goat tissues.

Zearalenone (ZEA), an estrogenic mycotoxin produced by several Fusarium species, is converted into a more active metabolite, alpha-zearalenol (alpha-ZOL), and a less active metabolite, beta-zearalenol (beta-ZOL), by liver subcellular fractions, but evidence of this reaction in other tissues is limited. In order to clarify the role of various tissues in ZEA metabolism in ruminant, we investigated the in vitro metabolic conversion of ZEA by various tissues of adult male and female goats. The results indicate that in the liver, alpha-ZOL was a major metabolite in cytosolic fractions, whereas beta-ZOL was a predominant metabolite in microsome fractions. Such a feature of ZEA metabolism was confirmed by the K(m) and V(max) values from an enzyme kinetics experiment. Post-mitochondrial fractions of the liver converted ZEA predominantly to alpha-ZOL, indicating that the goat liver may function as an activation organ rather than as an inactivation organ, for ZEA metabolism in goats. In most other tissues including rumen tissue, the activity converting ZEA to alpha-ZOL was higher than that to beta-ZOL. The amount of alpha-ZOL formed by gastrointestinal tissues was 1/8-1/3 of that by the liver tissue in terms of the amount per mg protein, but the contribution of all gastrointestinal tissues to production of alpha-ZOL was estimated to be comparable to that of the liver because of the large mass of gastrointestinal tissues in ruminants. Overall the results show the importance of not only the liver tissue, but also other tissues, especially gastrointestinal ones, in the formation of a potent estrogenic metabolite, alpha-ZOL.

The Error-Prone Kinetochore-Microtubule Attachments During Meiosis I in Vitrified Oocytes.

Oocytes vitrification is frequently applied in assisted reproductive technologies. However, chromosomes segregation was error-prone during meiosis maturation of vitrified oocytes. The fidelity of chromosomes segregation depends on the correct kinetochore-microtubule attachments (KT-MTs). In meiosis I, the Aurora B/C would not spatially separate from the attachment sites upon bivalents stretched. Oocytes lack a mechanism for coordinating bivalent stretching and Aurora B/C inhibition in meiosis I. Thus, the KT-MTs are unstable in oocytes. In this study, we firstly found the incorrect KT-MTs were markedly increased in vitrified oocytes. The Aurora B/C activity in vitrified oocytes was significantly increased when the bivalents were stretched. This Aurora B/C activity could not induce a SAC response, as the SAC protein Mad2 was significantly decreased during MI stage in vitrified oocytes. Thus, the KT-MTs in vitrified oocytes were error-prone. This study, for the first time, revealed the mechanism of the incorrect KT-MTs occurred in vitrified oocytes and provided a theoretical basis for further improvement of oocytes vitrification.