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Preeclampsia (PE) is a complex multisystem disease characterized by hypertension of sudden onset (>20 weeks' gestation) coupled with the presence of at least one additional complication, such as proteinuria, maternal organ dysfunction, or uteroplacental dysfunction. Hypertensive states during pregnancy carry life-threatening risks for both mother and baby. The pathogenesis of PE develops due to a dysfunctional placenta with aberrant architecture that releases factors contributing to endothelial dysfunction, an antiangiogenic state, increased oxidative stress, and maternal inflammatory responses. Previous studies have shown a correlation between grade 3 placental calcifications and an elevated risk of developing PE at term. However, little is known about the molecular pathways leading to placental calcification. In this work, we studied the gene and protein expression of c-Jun N-terminal kinase (JNK), Runt-related transcription factor 2 (RUNX2), osteocalcin (OSC), osteopontin (OSP), pigment epithelium-derived factor (PEDF), MSX-2/HOX8, SOX-9, WNT-1, and β-catenin in placental tissue from women with late-onset PE (LO-PE). In addition, we employed von Kossa staining to detect mineral deposits in placental tissues. Our results show a significant increase of all these components in placentas from women with LO-PE. Therefore, our study suggests that LO-PE may be associated with the activation of molecular pathways of placental calcification. These results could be the starting point for future research to describe the molecular mechanisms that promote placental calcification in PE and the development of therapeutic strategies directed against it. [ABSTRACT FROM AUTHOR]

Tumour necrosis factor (TNF) superfamily member 11 (TNFSF11), also known as RANKL, plays a crucial role in regulating several physiological and pathological activities. Additionally, it is a vital factor in bone physiology, and the sex hormone progesterone regulates the expansion of stem cells and the proliferation of mammary epithelial cells. It is essential for animal growth and reproductive physiological processes. This study aimed to evaluate the tissue-specific expression characteristics and promoter activity of the TNFSF11 gene in pigs. As a result, the study examined the presence of TNFSF11 expression in the tissues of Xiangsu pigs at 0.6 and 12 months of age. Moreover, the core promoter region of TNFSF11 was also identified by utilizing a combination of bioinformatic prediction and dual-luciferase activity tests. Finally, the effect of transcription factors on the transcriptional activity of the core promoter region was determined using site-directed mutagenesis. TNFSF11 was uniformly expressed in all tissues; however, its expression in muscles was comparatively low. The core promoter region of TNFSF11 was located in the −555 to −1 region. The prediction of the transcription start site of TNFSF11 gene-2000 ∼ + 500bp showed that there was a CpG site in 17 ∼ + 487bp. Analysis of mutations in the transcription factor binding sites revealed that mutations in the Stat5b , Myog , Trl , and EN1 binding sites had significant effects on the transcriptional activity of the TNFSF11 gene, particularly following the EN1 binding site mutation (P < 0.001). This study provides insights into both the tissue-specific expression patterns of TNFSF11 in the tissues of Xiangsu pigs and the potential regulatory effects of transcription factors on its promoter activity. These results may be helpful for future research aimed at clarifying the expression and role of the porcine TNFSF11 gene. [ABSTRACT FROM AUTHOR]

Background/Objectives: Stearoyl-coenzyme A desaturase 1 (SCD1) plays a crucial role in fatty acid metabolism. However, its roles in the feeding habit transformation of mandarin fish (Siniperca chuatsi) remain largely unknown. Methods: Juvenile mandarin fish (10.37 ± 0.54)g were trained to feed on an artificial diet and then divided into artificial diet feeders and nonfeeders according to their feed preference. Afterwards, the scd1 gene of mandarin fish (Sc-scd1) was identified and characterized, and its transcription difference was determined between S. chuatsi fed live artificial diets and those fed prey fish. Results: Our results show that Sc-scd1 coding sequence is 1002 bp long, encoding 333 amino acids. The assumed Sc-SCD1 protein lacks a signal peptide, and it contains 1 N-linked glycosylation site, 24 phosphorylation sites, 4 transmembrane structures, and 3 conserved histidine elements. We found that Sc-SCD1 exhibits a high similarity with its counterparts in other fish by multiple alignments and phylogenetic analysis. The expression level of Sc-scd1 was detected with different expression levels in all tested tissues between male and female individuals fed either live prey fish or artificial diets. Conclusions: In particular, the Sc-scd1 expression level was the highest in the liver of both male and female mandarin fish fed artificial diets, indicating that scd1 genes may be associated with feed adaption of mandarin fish. Taken together, our findings offer novel perspectives on the potential roles of scd1 in specific domestication, and they provide valuable genetic information on feeding habits for the domestication of mandarin fish. [ABSTRACT FROM AUTHOR]

Abstract Background Endometriosis is a gynecological disease characterized by the presence of endometrial tissue in abnormal locations, leading to severe symptoms, inflammation, pain, organ dysfunction, and infertility. Surgical removal of endometriosis lesions is crucial for improving pain and fertility outcomes, with the goal of complete lesion removal. This study aimed to analyze the location and expression patterns of poly (ADP-ribose) polymerase 1 (PARP-1), epithelial cell adhesion molecule (EpCAM), and folate receptor alpha (FRα) in endometriosis lesions and evaluate their potential for targeted imaging. Methods Gene expression analysis was performed using the Turku endometriosis database (EndometDB). By immunohistochemistry, we investigated the presence and distribution of PARP-1, EpCAM, and FRα in endometriosis foci and adjacent tissue. We also applied an ad hoc platform for the analysis of images to perform a quantitative immunolocalization analysis. Double immunofluorescence analysis was carried out for PARP-1 and EpCAM, as well as for PARP-1 and FRα, to explore the expression of these combined markers within endometriosis foci and their potential simultaneous utilization in surgical treatment. Results Gene expression analysis revealed that PARP-1, EpCAM, and FOLR1 (FRα gene) are more highly expressed in endometriotic lesions than in the peritoneum, which served as the control tissue. The results of the immunohistochemical study revealed a significant increase in the expression levels of all three biomarkers inside the endometriosis foci compared to the adjacent tissues. Additionally, the double immunofluorescence analysis consistently demonstrated the presence of PARP-1 in the nucleus and the expression of EpCAM and FRα in the cell membrane and cytoplasm. Conclusion Overall, these three markers demonstrate significant potential for effective imaging of endometriosis. In particular, the results emphasize the importance of PARP-1 expression as a possible indicator for distinguishing endometriotic lesions from adjacent tissue. PARP-1, as a potential biomarker for endometriosis, offers promising avenues for further investigation in terms of both pathophysiology and diagnostic-therapeutic approaches.

Maize is a significant food and feed product, and abiotic stress significantly impacts its growth and development. Arabidopsis Toxicosa en Levadura (ATL), a member of the RING-H2 E3 subfamily, modulates various physiological processes and stress responses in Arabidopsis. However, the role of ATL in maize remains unexplored. In this study, we systematically identified the genes encoding ATL in the maize genome. The results showed that the maize ATL family consists of 77 members, all predicted to be located in the cell membrane and cytoplasm, with a highly conserved RING domain. Tissue-specific expression analysis revealed that the expression levels of ATL family genes were significantly different in different tissues. Examination of the abiotic stress data revealed that the expression levels of ATL genes fluctuated significantly under different stress conditions. To further understand the biological functions of maize ATL family genes under high-temperature stress, we studied the high-temperature phenotypes of the maize ZmATL family gene ZmATL10 and its homologous gene AtATL27 in Arabidopsis. The results showed that overexpression of the ZmATL10 and AtATL27 genes enhanced resistance to high-temperature stress. [ABSTRACT FROM AUTHOR]

Background: Endometriosis is a gynecological disease characterized by the presence of endometrial tissue in abnormal locations, leading to severe symptoms, inflammation, pain, organ dysfunction, and infertility. Surgical removal of endometriosis lesions is crucial for improving pain and fertility outcomes, with the goal of complete lesion removal. This study aimed to analyze the location and expression patterns of poly (ADP-ribose) polymerase 1 (PARP-1), epithelial cell adhesion molecule (EpCAM), and folate receptor alpha (FRα) in endometriosis lesions and evaluate their potential for targeted imaging. Methods: Gene expression analysis was performed using the Turku endometriosis database (EndometDB). By immunohistochemistry, we investigated the presence and distribution of PARP-1, EpCAM, and FRα in endometriosis foci and adjacent tissue. We also applied an ad hoc platform for the analysis of images to perform a quantitative immunolocalization analysis. Double immunofluorescence analysis was carried out for PARP-1 and EpCAM, as well as for PARP-1 and FRα, to explore the expression of these combined markers within endometriosis foci and their potential simultaneous utilization in surgical treatment. Results: Gene expression analysis revealed that PARP-1, EpCAM, and FOLR1 (FRα gene) are more highly expressed in endometriotic lesions than in the peritoneum, which served as the control tissue. The results of the immunohistochemical study revealed a significant increase in the expression levels of all three biomarkers inside the endometriosis foci compared to the adjacent tissues. Additionally, the double immunofluorescence analysis consistently demonstrated the presence of PARP-1 in the nucleus and the expression of EpCAM and FRα in the cell membrane and cytoplasm. Conclusion: Overall, these three markers demonstrate significant potential for effective imaging of endometriosis. In particular, the results emphasize the importance of PARP-1 expression as a possible indicator for distinguishing endometriotic lesions from adjacent tissue. PARP-1, as a potential biomarker for endometriosis, offers promising avenues for further investigation in terms of both pathophysiology and diagnostic-therapeutic approaches. [ABSTRACT FROM AUTHOR]

The insulin-like growth factor (IGF) system plays a key regulatory role in biological processes, such as growth, reproduction, early development, cell differentiation, and proliferation, in fish and other vertebrates. The IGF family system consists of two IGFs (IGF1 and IGF2), two IGF receptors (IGF-1R and IGF-2R), and six IGF-binding proteins (IGF-binding proteins IGFBP1–6) in mammals. By binding to the cell surface-specific receptor (IGF-Rs), IGFs activate the activity of related protein kinases, initiate the signaling cascade in the cell, and participate in the regulation of various biological reaction processes. The research on fish IGFs mainly focuses on IGF1 and IGF2, and several studies have confirmed that they are widely involved in multiple biological processes, such as growth regulation, immune response, gonadal development, and maturation. IGF3 is a fish-specific sub-type; it was first detected in zebrafish (Danio rerio) and Nile tilapia (Oreochromis niloticus) in 2008 and plays an important role in gender differentiation in many reports. However, it is unclear whether it affects ovarian development.Turbot (Scophthalmus maximus) is an economically valuable flatfish species that is widely farmed in Europe and China. It cannot spawn eggs naturally in captivity and needs to be hand-stripped and artificially fertilized. The IGF system plays an important role during turbot ovary maturation. In this study, the full-length turbot IGF3 sequence was obtained via RACE cloning technology, its bioinformatics characteristics were analyzed, and the tertiary structure and docking situation were predicted. The relative expression levels during the whole tissue and ovarian development were detected using qRT-PCR. Hematoxylin-eosin staining was used to determine the gonadal development stage. The results showed that the full-length cDNA sequence of turbot IGF3 was 1 255 bp, encoding 259 amino acids, and displayed the highest homology with halibut (Hippoglossus stenolepis). A total of 33 amino acid signal peptides and 59 amino acid IGF-specific domains were found. It was a hydrophilic protein with a molecular mass of 26.39 kDa and a hydrophobic isoelectric point of 8.13. The turbot IGF3 is highly homologous with other fish. Turbot IGF3 is a hydrophobic protein, and its tertiary structure consists of three α spirals connected in series, closely integrated with IGF-1R and IGF-2R. igf3 is expressed differently between female and male turbot. In the liver, kidney, stomach, gonads, and brain, the male IGF3 expression was significantly higher than in females. In the heart, intestines, and pituitary gland, female IGF3 expression was significantly higher than males; however, the difference was not significant in the spleen. In male turbot, igf3 is mostly expressed in the brain, while in female turbot, igf3 is mostly expressed in the brain and intestine. During the five stages of development of turbot ovaries (previtellogenesis, early vitellogenesis, late vitellogenesis, migratory nucleus, and atresia), igf3 expression increased significantly in late vitellogenesis and maintained high expression in the migratory nucleus stage and decreased in the atresia stage. The results showed that turbot IGF3 has typical IGF family structure characteristics: local endocrine factors, widely distributed in tissues, significant sex dimorphism, and participates in regulating the growth and maturation of oocytes. Therefore, these findings lay a foundation for an in-depth research on the effects of IGF3 on ovarian development and oocyte maturation in turbot and provides crucial ideas for exploring the novel functions of IGF3 in fish.

The grass carp (Ctenopharyngodon idella) holds significant economic value worldwide. However, intensive farming practices often result in lipid metabolic disorder of fish. It is essential to explore the influence of pivotal genes in the process of fat accumulation, particularly from the perspective of fat transport. Fatty acid binding proteins (FABPs) are crucial for selectively binding and transporting free fatty acids (FAs) within cells. Nevertheless, the types of fabp genes in the grass carp genome and their responses to various FAs remain unclear. In the current study, exhaustive bioinformatic analysis identified nine fabp genes in the grass carp genome: fabp1a, fabp1b.1, fabp2, fabp3, fabp6, fabp7a, fabp10a, fabp11a, and fabp11b. Although these genes showed widespread expression across various tissues, each gene displayed distinct preferences for specific primary expression tissues. Notably, fabp1a exhibited prominent expression in muscle, fabp1b.1 in abdominal fat, fabp2 and fabp6 in the intestine, fabp7a in the brain, fabp10a in the hepatopancreas, and fabp11a showed maximal expression in the eye. Findings from primary grass carp hepatocytes revealed that the addition of palmitic acid (PA), high concentrations oleic acid (OA) and docosahexaenoic acid (DHA) (400 µM), significantly increased intracellular lipid accumulation. Intriguingly, the addition of PA resulted in increased expression of fabp2 and fabp11a, while DHA significantly enhanced the expression of fabp7a (P < 0.01). Given that the accumulated FAs are primarily saturated fatty acids, these results suggest that regulating fabps, especially fabp2 and fabp11a, could be a strategy to control fat deposition in grass carp.

Simple Summary: The research explores the potential of the Wnt receptor ROR1 as a target for cancer treatment, an area gaining significant attention. Despite ongoing therapeutic investigations, the understanding of ROR1's presence in different tissues remains limited. To address this, the study conducted immunohistochemistry analyses on a large variety of tumor types, including sarcomas and carcinomas, revealing a diverse ROR1 expression pattern. Notably, high ROR1 prevalence was observed in mesothelioma, liposarcoma, gastrointestinal stromal tumors, and uterine endometrioid carcinoma. Conversely, other cancers showed lower expression. These findings highlight the potential for ROR1-targeted therapies, particularly in mesothelioma, which exhibits frequent and high ROR1 expression, suggesting a novel therapeutic avenue. This research may inform the development of more effective cancer treatments targeting ROR1. The Wnt receptor ROR1 has generated increased interest as a cancer therapeutic target. Research on several therapeutic approaches involving this receptor is ongoing; however, ROR1 tissue expression remains understudied. We performed an immunohistochemistry analysis of ROR1 protein expression in a large cohort of multiple tumor and histologic types. We analyzed 12 anonymized multi-tumor tissue microarrays (TMAs), including mesothelioma, esophageal and upper gastrointestinal carcinomas, and uterine endometrioid carcinoma, among other tumor types. Additionally, we studied 5 different sarcoma types of TMAs and 6 patient-derived xenografts (PDX) TMAs developed from 19 different anatomic sites and tumor histologic types. A total of 1142 patient cases from different histologic types and 140 PDXs placed in TMAs were evaluated. Pathologists assessed the percentage of tumor cells in each case that were positive for ROR1 and the intensity of staining. For determining the prevalence of staining for each tumor type, a case was considered positive if >1% of its tumor cells showed ROR1 staining. Our immunohistochemistry assays revealed a heterogeneous ROR1 expression profile. A high prevalence of ROR1 expression was found in mesothelioma (84.6%), liposarcoma (36.1%), gastrointestinal stromal tumors (33.3%), and uterine endometrioid carcinoma (28.9%). Other histologic types such as breast, lung, renal cell, hepatocellular, urothelial carcinoma, and colon carcinomas; glioblastoma; cholangiocarcinoma; and leiomyosarcoma showed less ROR1 overall expression, ranging between 0.9 and 13%. No ROR1 expression was seen in mesenchymal chondrosarcoma, rhabdomyosarcoma, or gastric adenocarcinoma cases. Overall, ROR1 expression was relatively infrequent and low in most tumor types investigated; however, ROR1 expression was infrequent but high in selected tumor types, such as gastroesophageal GIST, suggesting that ROR1 prescreening may be preferable for those indications. Further, mesothelioma exhibited frequent and high levels of ROR1 expression, which represents a previously unrecognized therapeutic opportunity. These findings can contribute to the development of ROR1-targeted therapies. [ABSTRACT FROM AUTHOR]

[Objective]Pholidota chinensis Lindl is a rare and endangered epiphytic orchid. The epiphytes have evolved many special morphological structures and physiological and biochemical characteristics compared to geophytes. The transcription factors play a role in regulating plant growth and development. The aim of this paper is to explore and discover transcription factors in P. chinensis that regulate its growth and development.[Method]This study used PacBio and Illumina sequencing techniques to obtain the full-length transcriptome data of P. chinensis and the gene expressions in the roots, rhizomes, pseudobulbs and leaves. After further bioinformatics analysis, the four transcription factor family members, physicochemical properties and tissue expression specificity of P. chinensis were revealed. The expression of C2H2 family and bHLH family was analyzed by RT-qPCR using sodium chloride(NaCl)and methyl jasmonate(MeJA)induced adversity stresses.[Result] The four identified transcription factors and their families members in P. chinensis were: 21 members in C2H2 family, 27 members in bHLH family, 30 members in WRKY family, and 19 members in bZIP family. These four families and their members were mainly hydrophilic unstable proteins, and their subcellular localization was mainly located in the nucleus. Some members of bHLH families were located in cytoplasm, chloroplast and other organelles. The gene family had typical conserved domain characteristics, and showed obvious tissue specificity in the roots, rhizomes, pseudobulbs and leaves of P. chinensis. Expressions of all genes in C2H2 families and bHLH families were affected by NaCl and MeJA, but family members differed in whether they were promoted or repressed in the expression and in the amount of their expression.[Conclusion] The number of four transcription factor family members in P. chinensis was identified, and the members of each family are mainly hydrophilic unstable proteins located in the nucleus with conserved structural domain features and tissue expression specificity. [ABSTRACT FROM AUTHOR]

[Objectives]The paper aimed to provide a basis for studying the function of 14-3-3 protein and breeding of Jerusalem artichoke, we identified the 14-3-3 gene family of Jerusalem artichoke and analyzed their expression patterns in response to high temperature, cold, salt and drought stresses. [Methods]Cloning and bioinformatics analysis were used to study gene properties, and RNA-seq data analysis and RT-qPCR were used to study gene response patterns to abiotic stress. [Results]Ten members of the 14-3-3 gene family, namely HtGRF1-HtGRF10(GenBank accession No.OP132618-OP132627), were cloned from Jerusalem artichoke. According to the evolutionary relationship, these genes were divided into two subfamilies, HtGRF1-HtGRF7 belonged to the non-ε group, and HtGRF8-HtGRF10 belonged to the ε group, which usually formed homologous or heterodimers. The expression level of HtGRF2/3/6/9 in sprouts, roots, stems and leaves was higher than those of other genes. The expression level of HtGRF3/5/9 was higher in stolon and tuber formation stages than that in the tuber rapid intamescentia and maturity stages during the tuber development. Comprehensive analysis of HtGRF in response to abiotic stress in roots and leaves showed that HtGRF6 expression level decreased under high temperature, salt and drought stresses, while HtGRF2/3/7/8/9 expression level decreased under salt stress, but increased under drought stress. HtGRF1 expression level increased under cold stress, HtGRF4/5 expression level decreased under drought stress, and HtGRF10 expression level did not change significantly. [Conclusions]The 14-3-3 protein family in Jerusalem artichoke was highly conserved and encoded by multiple genes, and played an important role in the growth and development and in coping with complex environments. [ABSTRACT FROM AUTHOR]

Copyright of Xinjiang Agricultural Sciences is the property of Xinjiang Agricultural Sciences Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The Transporter 1/Peptide Transporter Family (NPF) is essential for the uptake and transport of nitrate nitrogen. Significant increases in nitrogen have been increasingly reported for many mycorrhizal plants, but there are few reports on maize. Here, we have identified the maize NPF family and screened for arbuscular mycorrhiza fungi (AMF) induced NPFs. In this study, a systematic analysis of the maize NPF gene family was performed. A total of 82 NPF genes were identified in maize. ZmNPF4.5 was strongly induced by AMF in both low and high nitrogen. Lotus japonicus hairy root-induced transformation experiments showed that ZmNPF4.5 promoter-driven GUS activity was restricted to cells containing tufts. Yeast backfill experiments indicate that ZmNPF4.5 functions in nitrate uptake. Therefore, we speculate that ZmNPF4.5 is a key gene for nitrate-nitrogen uptake in maize through the mycorrhizal pathway. This is a reference value for further exploring the acquisition of nitrate-nitrogen by maize through AMF pathway. [ABSTRACT FROM AUTHOR]

Lysosomes-associated membrane proteins (LAMPs), a family of glycosylated proteins andmajor constituents of the lysosomal membranes, play a dominant role in various cellular processes, including phagocytosis, autophagy and immunity in mammals. However, their roles in aquatic species remain poorly known. In the present study, three lamp genes were cloned and characterized from Micropterus salmoides. Subsequently, their transcriptional levels in response to different nutritional status were investigated. The full-length coding sequences of lamp1, lamp2 and lamp3 were 1251bp, 1224bp and 771bp, encoding 416, 407 and 256 amino acids, respectively. Multiple sequence alignment showed that LAMP1-3 were highly conserved among the different fish species, respectively. 3-D structure prediction, genomic survey, and phylogenetic analysis were further confirmed that these genes are widely existed in vertebrates. The mRNA expression of the three genes was ubiquitously expressed in all selected tissues, including liver, brain, gill, heart, muscle, spleen, kidney, stomach, adipose and intestine, lamp1 shows highly transcript levels in brain and muscle, lamp2 displays highly expression level in heart, muscle and spleen, but lamp3 shows highly transcript level in spleen, liver and kidney. To analyze the function of the three genes under starvation stress in largemouth bass, three experimental treatment groups (fasted group and refeeding group, control group) were established in the current study. The results indicated that the expression of lamp1 was significant induced after starvation, and then returned to normal levels after refeeding in the liver. The expression of lamp2 and lamp3 exhibited the same trend in the liver. In addition, in the spleen and the kidney, the transcript level of lamp1 and lamp2 was remarkably increased in the fasted treatment group and slightly decreased in the refed treatment group, respectively. Collectively, our findings suggest that three lamp genes may have differential function in the immune and energetic organism in largemouth bass, which is helpful in understanding roles of lamps in aquatic species. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Northwest A & F University - Natural Science Edition is the property of Editorial Department of Journal of Northwest A&F University (Natural Science Edition) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Corticotropin-releasing hormone (CRH) mediates various physiological/pathological roles such as stress and reproduction in vertebrates by specifically activating its receptors (CRHRs), and the regulation is intricately intertwined with the CRH binding protein (CRHBP). Although numerous research on mammalian CRHRs and CRHBP has been conducted, reports regarding the teleost CRHRs and CRHBP are limited. Herein, using Nile tilapia (ti-) as a model, we conducted the characterization, functional analysis, and examination of tissue distribution of tiCRHRs and tiCRHBPs. Our study showed that: (1) tiCRHR1a, tiCRHR1b and tiCRHR2 encoded proteins of 432, 436 and 414 amino acids (-aa), while tiCRHBP1 and tiCRHBP2 were predicated to generate a 323- and 322-aa precursor respectively; (2) All tiCRHRs could be activated by tiCRH and subsequently coupled to cAMP/PKA signaling pathways, while only tiCRHR1a and tiCRHR1b could trigger MAPK/ERK signaling cascade; (3) tiCRHBP1 and tiCRHBP2 could does-dependently suppress tiCRH-upregulated luciferase activity in HEK293 cells expressing tiCRHRs; (4) All tiCRHRs mRNA were considerably detected in the central nervous system (CNS), but they exhibited distinct expression pattern in examined peripheral tissues; (5) tiCRHBP1 exhibited an exclusive expression pattern in the pituitary, whereas tiCRHBP2 mRNA was abundantly expressed in the telencephalon. Collectively, our data will help to facilitate a better understanding of the members and functions of CRHRs and CRHBP in teleost.

Crustacyanin (CRCN), a family of lipocalin proteins specific to crustaceans, was primarily found in the exoskeleton of crustaceans. By binding with astaxanthin to form the astaxanthin-binding protein complex, it resets the proton at the end of astaxanthin and modifies the acidic site in astaxanthin, thereby regulating crustacean shell color. In addition to regulating shell colors, crustacyanin is involved in the transport of small molecule lipids such as steroid hormones and pheromones and has important biological functions in moult growth, gonadal and nervous system development, resistance to heavy metal lipid metabolism, and hypoxic stress. It was first discovered that the pigment in the blue shell of lobster could be extracted with ammonium chloride via a complex combination of organic bases and lipid pigments. It was later proposed that the pigment was a combination of astaxanthin and multimeric protein complex known as α-CRCN. This complex is composed of an octomer of dimeric β-CRCN subunits, with this dimer formed by two types of CRCN A and C in association with two astaxanthin molecules. Natural crustacyanins are all α-CRCN, comprising a total of 16 molecules. Procambarus clarkii, a member of the order Crustacea (Decapoda, Crayfish), is native to North America and is an important freshwater crustacean in China. It had been established that P. clarkii is susceptible to low oxygen stress during the culture process. The low oxygen environment inhibits the metabolic rate of P. clarkii, leading to increased susceptibility to pathogens; therefore, the ability of P. clarkii to tolerate and physiologically regulate stress due to other environmental factors was also affected, which can lead to irreversible damage and even death in severe cases. To understand the role of crustacyanin-like genes in gonadal development and hypoxic-reoxygenation stress in P. clarkii, a cDNA sequence of the PcCRCN-L gene was isolated from the hepatopancreas of this organism. The structural characteristics and evolutionary patterns of the PcCRCN-L gene were analyzed, and the expression characteristics of the PcCRCN-L gene in different tissues and gonad development stages were investigated. The expression response pattern of PcCRCN-L under hypoxic-reoxygenation stress was investigated. The DNA sequence of the PcCRCN-L gene was 6 130 bp long and located on chromosome 12 of the P. clarkii genome. The cDNA sequence was 2 700 bp, and its open reading frame (ORF) length was 1 587 bp. It contained five exons and four introns, encoded 528 amino acid residues, had a theoretical isoelectric point of 5.71 and a relative molecular weight of 55 613.55, and was a hydrophilic protein. The intron/exon splicing pattern was in accordance with the GT-AG rule. The PcCRCN-L protein had a complete lipocalin domain, which included the typical sequences G-X-W of conserved region Ⅰ (SCR1), T-D-Y of conserved region Ⅱ (SCR2), and arginine R of conserved region Ⅲ (SCR3). The results of multiple sequence alignments and phylogenetic analyses showed that PcCRCN-L, as well as the crustacyanin A and crustacyanin C subgroups, were separately clustered into a branch. The expression characteristics of the PcCRCN-L gene in different tissues showed that PcCRCN-L was expressed in all the tissues; however, the highest expression level was found in the hepatopancreas. The expression trend of the PcCRCN-L gene in the ovary and the hepatopancreas was similar in different stages of gonadal development. The expression level of the PcCRCN-L gene was significantly decreased in stages Ⅰ~Ⅱof ovarian development (P < 0.05), but no significant difference was found in stages Ⅱ~Ⅴ (P > 0.05). The expression of the PcCRCN-L gene was significantly decreased under hypoxia stress for 1 h (P < 0.05), but there was no significant difference between hypoxia stress for 1 h and hypoxia stress for 6 h. After reoxygenation for 1 h, the expression of the PcCRCN-L gene was significantly up-regulated (P < 0.05). Compared with reoxygenation for 1 h, the expression of the PcCRCN-L gene was significantly increased at 12 h (P < 0.05). The results showed that PcCRCN-L should be closely involved in the regulation of gonadal development and hypoxia-reoxygenation stress in P. clarkii.

Preeclampsia (PE) is a complex multisystem disease characterized by hypertension of sudden onset (>20 weeks’ gestation) coupled with the presence of at least one additional complication, such as proteinuria, maternal organ dysfunction, or uteroplacental dysfunction. Hypertensive states during pregnancy carry life-threatening risks for both mother and baby. The pathogenesis of PE develops due to a dysfunctional placenta with aberrant architecture that releases factors contributing to endothelial dysfunction, an antiangiogenic state, increased oxidative stress, and maternal inflammatory responses. Previous studies have shown a correlation between grade 3 placental calcifications and an elevated risk of developing PE at term. However, little is known about the molecular pathways leading to placental calcification. In this work, we studied the gene and protein expression of c-Jun N-terminal kinase (JNK), Runt-related transcription factor 2 (RUNX2), osteocalcin (OSC), osteopontin (OSP), pigment epithelium-derived factor (PEDF), MSX-2/HOX8, SOX-9, WNT-1, and β-catenin in placental tissue from women with late-onset PE (LO-PE). In addition, we employed von Kossa staining to detect mineral deposits in placental tissues. Our results show a significant increase of all these components in placentas from women with LO-PE. Therefore, our study suggests that LO-PE may be associated with the activation of molecular pathways of placental calcification. These results could be the starting point for future research to describe the molecular mechanisms that promote placental calcification in PE and the development of therapeutic strategies directed against it.

[Objective] The Larix kaempferi flavanone 3-hydroxylase 2 (LkF3H2) gene in Larix kaempferi plays a crucial role in flavonoid biosynthesis. This study is aimed to investigate the function of the LkF3H2 in L. kaempferi and plant flavonoid biometabolism. [Method] The LkF3H2 gene of L. kaempferi was cloned based on preliminary transcriptome data, and bioinformatics analysis and tissue expression analysis were performed. The gene was then stably transformed into tobacco to examine the relationship between gene expression and flavonoid content using tissue expression analysis and flavonoid content determination. [Result] The cDNA sequence length of LkF3H2 gene is 1 074 bp, and its protein encodes 358 amino acids with a molecular formula of C1785H2807N481O541S18 and a molecular weight of 40.24 kD, indicating that it is an unstable hydrophilic protein lacking a signal peptide. Moreover, phylogenetic analysis revealed that LkF3H2 is closely related to the F3H genes of Pinus taeda, Pinus radiata, Picea abies, and Picea glauca. In addition, the LkF3H2 gene had the highest expression in the leaves of one-month-old L. kaempferi seedlings and the stems of one-year L. kaempferi seedlings, as determined by tissue expression analysis. Moreover, it had the highest expression in the transgenic tobacco leaves; notably, transgenic tobacco leaves had the highest flavonoid content, followed by stems and roots, indicating a positive relationship between flavonoid content and LkF3H2 gene expression in transgenic tobacco. [Conclusion] The LkF3H2 gene of L. kaempfer belongs to the 2-oxoglutarate-dependent dioxygenase family and is pivotal in flavonoid biosynthesis. [ABSTRACT FROM AUTHOR]

Odorant binding protein(OBP) is an important protein in the process of insect olfactory communication. OBPs are involved in the first step of insect odor recognition, and play an important role in insect activities such as foraging, aggregation, courtship, and looking for spawning places. In order to determine the expression characteristics of OBP genes in Agasicles hygrophila and to preliminarily understand its functions, in this study, three OBP genes were cloned and analyzed using bioinformatics. The relative expression levels of OBPs in different tissues were analyzed by quantitative real-time PCR. The results showed that the open reading frame of AhygOBP27, AhygOBP32, and AhygOBP44 were 351, 408, 423 bp, respectively. The proteins encoded by the three OBPs were acidic soluble proteins with small molecular weight. AhygOBP27 was mainly expressed in the antennae, head, feet and wings of both sexes. AhygOBP32 was expressed in antennae, head, leg and wings of the male and in antennae, feet and wings of the female. AhygOBP44 was expressed in the antennae, heads and feet of both sexes. The expression levels of above three genes in the antennae of male and female were significantly higher than that of other tissues. The expression level of AhygOBP27 and AhygOBP44 in female antennae were significantly higher than that in male antennae, and the expression level of AhygOBP32 in the antennae of males was significantly higher than that of females. The three genes might play an important role in the olfactory recognition of Agasicles hygrophila, AhypOBP27, and AhygOBP44 might be related to the recognition of host plant volatiles and the search for spawning ground; AhygOBP32 might be involved in the recognition of sex pheromone emitted by females. [ABSTRACT FROM AUTHOR]

To elucidate the function and reproductive regulation mechanism of the Vitellogenin(GmVg) gene in Grapholita molesta, a significant pest of fruit trees, and to provide a theoretical basis for further research on the function of the GmVg gene, in this study, GmVg was cloned and identified and a comprehensive bioinformatics analysis was performed. The expression patterns of GmVg at various developmental stages, across sexes, and in different tissues were examined using quantitative reverse transcription PCR(qRT-PCR). The results revealed that the full-length of GmVg gene spanned 5 573 base pairs(bp), with an open reading frame(ORF) of 5 364 bp that encoded 1 787 amino acids. The signal peptide consisted of 15 amino acids, and the molecular formula was C9041H14090N2544O2743S57. The percentage of the amino acids alanine(Ala), glutamine(Gln), and serine (Ser) was large and they comprised 8.2%, 7.9%, and 7.8% of the protein, respectively. The protein contained 187 acidic residues(Asp+Glu) and 204 basic residues(Arg+Lys). Bioinformatics predictions indicated that the molecular weight of the GmVg protein was 204.1 ku, with an isoelectric point of 8.79, and it featured three characteristic conserved domains: Vitellogenin-N, DUF1943, and VWD. The expression of GmVg was markedly higher in adults compared to that in eggs, larvae, and pupae, reaching its peak 24 h post-emergence. The gene displayed sex-specific expression, with negligible levels in males. Within female adults, tissue-specific patterns were noted in head, ovary, and fat body; the expression in the fat body was substantially higher than in the head and ovary-21.49-fold and 16.4-fold greater, respectively. [ABSTRACT FROM AUTHOR]

Background: Genomic imprinting refers to expressing parent-specific genes in mammalian diploid cells. The NDN gene is maternally imprinted in humans and mice and correlates with the timing of puberty. This study aimed to investigate its imprinting status and its relationship with the onset of puberty in Dolang sheep. Methods and results: In this study, cloning and sequencing obtained the NDN gene cDNA sequence of 1082 bp of Dolang sheep, coding for 325 amino acids. Similarity analysis and phylogenetic tree showed that the NDN gene conformed to the law of speciation and was highly conserved among mammals. RT-qPCR results showed the highest expression of NDN mRNA was found in the hypothalamus at puberty, and the expression was significantly increased and then significantly decreased from prepuberty to postpuberty in the hypothalamus, pituitary, and ovary and oviduct. Based on expressed single nucleotide polymorphism (SNP), the NDN gene was expressed monoallelically in the tissues of adult and neonatal umbilical cords, and the expressed allele was paternally inherited. The NDN promoter region of 3400 bp was obtained by cloning and identified in monoallelic-expressing tissues (hypothalamus, ovary, spleen) as a differentially methylated region (DMR). Conclusion: These findings will enrich the number of imprinted genes in sheep and suggest that the NDN gene could be a candidate gene for studying puberty initiation in Dolang sheep. [ABSTRACT FROM AUTHOR]

To explore the function and expression characteristics of the short neuropeptide receptor AhsNPFR in Agasicles hygrophila, and to lay a theoretical foundation for exploring the biological control role of Agasicles hygrophila, in this study, the AhsNPFR gene was cloned using PCR technology and analyzed with bioinformatics software. The spatiotemporal expression profiles of AhsNPFR across different developmental stages and tissues were investigated by quantitative real-time PCR(qPCR). The results showed that the complete sequence of 1 669 bp was obtained, which was found to contain a 1 257 bp open reading frame(ORF), encoding 418 amino acids. The predicted molecular weight was 48.11 ku and the theoretical isoelectric point(pI) was 8.21. AhsNPFR had 7 typical conserved transmembrane domains and belonged to the GPCRs family. Phylogenetic tree analysis showed that AhsNPFR was closely related to Diabrotica virgifera virgifera sNPFR. AhsNPFR gene was expressed throughout the developmental stages tested with the highest expression levels at the first instar larva which was 9.06 times higher than those eggs, the stage with the lowest expression level. The expression level in female adult was significantly higher than that in male adult. The AhsNPFR gene was expressed in different tissues and was significantly highly expressed in the hindgut of the 3rd instar larvae, which was 12.21 times higher than those fat body. The AhsNPFR gene was significantly highly expressed in the hindgut of male and female adults, with no difference expression between the sexes across all tissues. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The purpose of this study was to examine STC-1's structure, function, and differential expression in large and miniature pigs. We cloned the Hezuo pig's coding sequence, compared its homology, and used bioinformatics to assess the structure. RT-qPCR and Western blot were used to detect the expression in ten tissues of Hezuo pig and Landrace pig. The results showed that Hezuo pig was most closely related to Capra hircus and most distantly related to Danio rerio. The protein STC-1 has a signal peptide and its secondary structure is dominated by the alpha helix. The mRNA expression in the spleen, duodenum, jejunum, and stomach of Hezuo pigs was higher than that of Landrace pigs. And except for heart and duodenum, expression of the protein in Hezuo pig was higher than in another. In conclusion, STC-1 is highly conserved among different breeds of pigs, and the expression and distribution of its mRNA and protein are different in large and miniature pigs. This work can lay the foundation for future study into the mechanism of action of STC-1 in Hezuo pigs and the enhancement of breeding in miniature pigs. [ABSTRACT FROM AUTHOR]

Copyright of South China Fisheries Science is the property of South China Fisheries Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Insulin-like growth factor binding proteins (IGFBPs) play crucial roles in regulating biological activities of insulin-like growth factors (IGFs) and growth performance in vertebrates. IGFBP is a six-member protein family (IGFBP1–6) with a high affinity for IGF. It affects the distribution, stability, and biological activity of IGF by regulating the interaction between IGF ligands and receptors. Recently, IGFBP-1 was identified as a regulator of growth, reproduction, and development in bony fish, such as juvenile Atlantic salmon, where IGFBP-1 interacts with cortisol to regulate growth. IGFBP-1 regulates cell metabolism and growth through interaction with insulin in primary hepatocytes incubated in vitro in orange-spotted grouper (Epinephelus coioides). IGFBP-2 has a wide range of tissue expression characteristics in bony fish, and IGF regulation may occur through autocrine or paracrine pathways. For example, long-term fasting induced increased igfbp-2 mRNA expression in the liver of zebrafish. In addition, IGFBP-2 can inhibit the activity of IGF ligand through its high affinity with IGF to play an inhibitory and regulatory role in zebrafish growth. igfbp-2 mRNA expression was significantly up-regulated in Carassius auratus (goldfish) liver after fasting, and quickly recovered to normal levels after feeding. This indicated that igfbp-2 mRNA expression may be related to the anabolism and catabolism of goldfish, and is regulated by metabolic factors.Yellowtail kingfish (Seriola aureovittata) of the Carangidae family has high economic value and nutrition value. It is a warm and temperate oceanic fish with long distance migration characteristics that is distributed in the middle and upper oceans globally. This species is large with a fast growth rate, and is a promising candidate for aquaculture in land-based industrial circulating water system, deep-water cages, net pen systems, and aquaculture craft, etc. Our laboratory studied the regulatory mechanism of the rapid growth of yellowtail kingfish and cloned gh, igf-1, igf-2, ghr, and other growth-related functional genes to reveal their molecular regulation in early growth and development. igfbp genes regulate the growth, development, and nutrient metabolism of fish through their interaction with growth axis. The mechanism of IGFBP and how it influences the regulation of yellowtail kingfish growth is unreported. This paper further studied the growth of yellowtail kingfish by analyzing the key growth axis factors.RNAiso Plus reagent (TaKaRa) was used to extract total RNA from tissues. RNA integrity was detected by agarose gel electrophoresis, and RNA quality was determined by a NanoDrop 2000C spectrophotometer (Thermo Fisher Scientific, USA). First, cDNA strand was synthesized using a PrimeScript™ RT reagent kit with a gDNA Eraser (Perfect Real-Time) reverse transcription kit (TaKaRa). Primers were designed to clone the predicted sequence of yellowtail kingfish igfbp gene according to the NCBI GenBank database. The product was amplified by polymerase chain reaction, purified from agarose gel electrophoresis, linked with T4 Ligases, transformed into Trans1-T1 Phage Resistant Chemically Competent Cell, and positive clones were selected and tested. Quantitative real time polymerase chain reaction (qRT-PCR) was used to analyze the distribution of yellowtail kingfish tissues and the expression patterns of liver tissues under different densities of industrial culture.The lengths of igfbp-1, igfbp-2a, igfbp-2b open reading frame (ORF) domains were 741 bp, 882 bp, and 801 bp, and encoded 246 amino acids, 293 amino acids, and 269 amino acids, respectively. The conserved domain of insulin growth factor-binding protein homologues (IB) was present in the N-terminus of the three igfbps, and the conserved domain of thyroglobulin type-1 repeat (Ty-1) was present in the C-terminus. They had a wide range of tissue expression characteristics and were highly expressed in the liver. There were differences between the expression of the same gene in the same tissues of male and female fish. For example, the expression of igfbp-1 and igfbp-2b genes was significantly higher in the liver of male fish compared with female fish, while igfbp-2a expression was significantly higher in the liver of female fish compared with male fish. The tissue differential expression of genes between sexes indicates that igfbp may have sex dimorphism when it plays a physiological role. However, the specific characteristics of this difference between males and females and the possible signaling pathway are unclear. The fish in the low-density group showed the greatest growth rate and the highest expression levels of igfbp-1, igfbp-2a, igfbp-2b, igf-1, and igf-2 under industrial culture conditions. These expression levels were significantly different from those of the medium and high-density groups (P < 0.05). However, there were no significant differences in the expression levels of growth and liver genes between the middle and high-density groups.igfbp-1, igfbp-2a, and igfbp-2b participated in the growth regulation of yellowtail kingfish. The expression regulation of igf-1 and igf-2 had a positive synergistic effect with growth regulation of yellowtail kingfish. The ORF regions of igfbp-1, igfbp-2a, and igfbp-2b genes of yellowtail kingfish were cloned, and the structural characteristics, tissue expression characteristics, and relationships with growth performance were analyzed under different culture densities. The content of serum IGF-1, GH, cortisol, and glucose concentration was detected in the early stage. The expression trend of igfbp-1, igfbp-2, igf-1, and igf-2 was the same as that of serum IGF-1 and GH in the liver of yellowtail kingfish rapidly growing at the low-density group, but contrary to the expression trend of cortisol content and glucose concentration. This indicated that the key factors of growth axis, cortisol, and glucose participated in the growth regulation of yellowtail kingfish at different densities. However, the specific regulatory mechanism requires further study. The results provide a theoretical basis for interpreting the molecular mechanism of the growth of yellowtail kingfish and the regulation of suitable culture densities under industrial conditions.

Yellowtail kingfish (Seriola aureovittata), a pelagic marine finfish species with a worldwide distribution, is regarded as an emerging candidate for the aquaculture industry owing to its fast growth, superior flesh quality, and farming suitability in both sea cages and land-based facilities in China. The species has high economic value and is the second most produced Seriola species in the world following Japanese yellowtail Seriola quinqueradiata. Researchers worldwide have studied the role of regulatory factors neuropeptide Y (NPY) in fish feeding regulation. In recent years, there has been great progress in research on food intake in fish, however, very little attention has been paid to the endocrine regulation mechanism of food intake. Methods on strengthening the production performance of fish through appetite regulation is still a hot research topic.The control of food intake and energy metabolism in vertebrates are complex processes involving several neural pathways. Some hypothalamic signals are released by peripheral tissues that are associated with energy homeostasis or nutrient availability. Among the signaling molecules involved, NPY plays a key role. NPY is recognized as one of the most effective appetite regulators, which primarily function as a signaling factor to regulate a variety of biological processes such as food intake and glucose homeostasis. The orexigenic actions of NPY have been well investigated thoroughly over the past decades. Much evidence supports that NPY´s functional role as a regulator of energy homeostasis and appetite control is conserved across vertebrates, including teleosts. In several species, including rainbow trout, Nile tilapia, and grass carp, NPY injections increase food intake, supporting an orexigenic role. In line with this, food deprivation increased npy mRNA expression in the brain, such as seen for goldfish and Johnny carp. Moreover, refeeding normalized npy mRNA abundance following food deprivation. As S. aureovittata feeds heavily and fiercely, the breeding industries need to understand its feeding control mechanism. To make real-time adjustment to feeding strategy, it is necessary to obtain high-quality and high-yield aquatic products with the least input to maximize economic benefits. As a potent appetite stimulating factor, npy has been proven to promote feeding, but this gene has not been identified in S. aureovittata. Therefore, it is necessary to explore the variable rules of the npy gene in feeding and starvation compensation mechanism, to provide the special compound feed for breeding.To gain insight into the existence of npy in S. aureovittata, we used homologous cloning, RNA extraction and reverse-transcription to obtain the ORF sequence of npy. npy belongs to the pancreatic polypeptide (PP) family, which plays an important role in appetite regulation and energy expenditure in mammals and fish. The ORF of S. aureovittata npy is 300 nucleotides in size and encodes a 99-amino-acid precusor, with a calculated molecular mass and isoelectric point of 11.24 kDa and 5.02, respectively. The precursor protein is composed of a predicted signal peptide of 28-aa in size, 36-aa putative mature peptide, a GKR protein proteolytic site, and a 32-aa C terminus of unknown function. Bioinformatics analysis on the amino acid sequence identities and evolutionary relationships of the npy was performed. Comparison of homology of the precursor peptide sequences of npy analysis revealed that S. aureovittata npy displayed a high degree of identity with the counterparts of Seriola dumerili (99.0%), Morone saxatilis (98.0%), Micropterus salmoides (96.0%), and Scophthalmus maximus (94.9%), followed by Cynoglossus semilaevis (93.9%) and Oryzias latipes (92.9%). Phylogenetic analysis highly supported that the npy of S. aureovittata was closely related to that of S. dumerili. Furthermore, using real-time quantitative PCR, we found that the npy mRNA is widely expressed in 12 tissues, with abundant expression in the brain, followed by the pituitary and stomach. In addition, except for the intestine and gonad, npy was found to have no significant difference in all other detected tissues of both sexes. To establish the functional link between npy and feeding, the expression profiles of npy mRNA during food deprivation and refeeding were examined in S. aureovittata. We detected the 7 d, 14 d, and 21 d starvation and 7 d refeeding effect on npy mRNA levels. Results showed that fasting induced an increase of npy mRNA levels in brain, pituitary, and stomach when compared to the control groups. Interestingly, the pituitary npy transcripts significantly increased after 21 d of starvation compared with the control group. In addition, refeeding normalized npy mRNA abundance following food deprivation in the brain, pituitary, and stomach. These results indicated that npy is involved in the regulation of feeding and energy homeostasis in S. aureovittata. Collectively, we provided initial evidence for the existence of npy in S. aureovittata and suggested its involvement in the regulation of feeding, which plays an important role in the starvation compensation mechanism.In summary, we obtained the ORF sequence of npy and clarified its role as a potent orexigenic peptide in feeding regulation of S. aureovittata, which would be beneficial for specific feed for this species.

Genomic imprinting refers to the expression of parent-specific genes in diploid mammalian cells. MAGEL2 gene is a maternally imprinted gene that has been identified in mice and humans and is associated with the onset of puberty. The purpose of this study was to investigate its imprinting status and its relationship with the onset of puberty in Dolang sheep. The sequence of 3734 bp cDNA of MAGEL2 in Dolang sheep was obtained by cloning and sequencing, encoding 1173 amino acids. The results of the nucleotide and amino acid similarity analysis showed that it was highly conserved among different mammalian species. The MAGEL2 gene was expressed monoallelically in the tissues of adult and neonatal umbilical cords, and the expressed allele was paternally inherited. Real Time quantitative PCR (RT-qPCR) results showed that the MAGEL2 gene was highly expressed in the hypothalamus and pituitary gland, increased significantly from prepuberty to puberty, and decreased significantly after puberty. This study suggests that MAGEL2 is a paternally expressed and maternally imprinted gene in Dolang sheep, which may be involved in the initiation of puberty in Dolang sheep. This study provides a theoretical basis for further research on the mechanism of the imprinted gene MAGEL2 regulating the onset of puberty in sheep, and provides a new idea for the future research on the mechanism of onset of puberty in sheep. [ABSTRACT FROM AUTHOR]

[Objective] The study aimed to systematically identify and analyze DBB gene family in rice, and to lay a theoretical foundation for the functional study of DBB family in rice. [Method] Based on the genome-wide data of rice, the chromosome location, physicochemical properties, evolutionary relationships, conserved motifs, cis-regulatory elements and expression patterns of DBB gene family in rice were comprehensively analyzed by bioinformatics methods. [Result] A total of 13 DBB genes were identified from rice, named OSDBBl - OSDBB13, and these OsDBB genes were unevenly distributed on 7 of 12 chromosomes of rice. According to phylogenetic analysis, rice DBB gene family can be divided into four subfamilies: Group I - Group 4, and the number and arrangement of introns and exons of DBB gene in the same subfamily were similar. The 13 OsDBB genes in rice contain 10 conserved motifs, among which Motif 1 was the common conserved motifs of these genes, which is speculated to be related to conserved domain. The promoter sequence of OsDBB gene contained regulatory elements related to growth and development, hormones, stress response, and light response. Different OsDBB genes were expressed differently in different tissues and have certain specificity. [ Conclusion] Rice OsDBB gene plays an important role in plant growth and development and response to abiotic stress. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Dalian Ocean University is the property of Journal of Dalian Ocean University Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Odorant-binding proteins (OBPs) are expressed at extremely high concentrations in the chemo-sensilla lymph of insects and have long been thought to be crucial for delivering the semiochemicals to the odorant receptors. They are represented by multiple classes: general odorant-binding proteins (GOBP1 and GOBP2) and pheromone-binding proteins. In the current study, we identified a total of 35 OBPs in the antennal transcriptome of Peridroma saucia, a worldwide pest that causes serious damage to various crops. A gene expression value (TPM, transcripts per million) analysis revealed that seven OBPs (PsauPBP1/2/3, PsauGOBP1/2, PsauOBP6, and PsauOBP8) were highly abundant in the antennae. Next, we focused on the expression and functional characterization of PsauGOBP2. Real-time quantitative-PCR analysis demonstrated that PsauGOBP2 was predominantly expressed in the antennae of both sexes. Fluorescence binding assays showed that the recombinant PsauGOBP2 strongly binds to the female sex pheromone components Z11-16: Ac (Ki = 4.2 μM) and Z9-14: Ac (Ki = 4.9 µM) and binds moderately (6 µM ≤ Ki ≤ 13 µM) to the host plant volatiles phenylethyl acetate, β-myrcene, and dodecanol. Further 3D structural modeling and molecular docking revealed that several crucial amino acid residues are involved in ligand binding. The results not only increase our understanding of the olfactory system of P. saucia but also provide insights into the function of PsauGOBP2 that has implications for developing sustainable approaches for P. saucia management. [ABSTRACT FROM AUTHOR]

The growth traits are economically important traits in sheep. Improving growth rates will increase the profitability of producers. The aim of this study was to identify alleles of CD8B (encoding T-cell surface glycoprotein CD8 beta chain) that are aberrantly expressed in different tissues and to assess the effects and associations of its different genotypes on weight and size traits in sheep. Using quantitative real-time reverse transcription PCR arrays, expression profiling of CD8B was performed in various organs and tissues. CD8B was ubiquitously expressed, with very high expression in the lung, spleen, lymph, duodenum, and liver. One intronic mutation (chr3:62,718,030 (Oar_rambouillet_v1.0, same below) G > A) was identified using pooled DNA sequencing. Subsequently, the variants (AA, AG, and GG) were genotyped using the KASPar® PCR single nucleotide polymorphism (SNP) genotyping system. The results of association analysis with body weight and body size traits in 1304 sheep showed that increases in multiple phenotypic traits correlated with the AA genotype (body weight, p < 0.05; body length, p < 0.05). Thus, SNP chr3:62,718,030 G > A is a promising molecular marker for marker-assisted selection in sheep breeding. [ABSTRACT FROM AUTHOR]

[ Objective]The study aimed to clone the CDS region of EGR2 gene transcript XI (EGR2-XI) of bovine, detect the expression patterns of EGR2-XI in different tissues and preadipocytes of different differentiation degrees, and predict the structure and function of its encoded protein. [Method] CDS region of EGR2-XI gene was cloned by PCR and sequencing techniques. Then, preadipocytes were isolated by collagenase digestion. In addition, the expression level of EGR2-XI gene was detected in 7 tissues ( heart, liver, spleen, lung, kidney, muscle and back fat) and during preadipocytes differentiation by RT-qPCR. At the same time, the physicochemical properties and protein structures of EGR2-XI were analyzed by ProtParam, Phyre2, NetPhos 3.1 and other software. [Result]The CDS region of bovine EGR2-XI was successfully cloned and it was 1458 hp in length. It encoded an unstable 485 amino acid hydrophilic protein without transmembrane structures and signal peptides. The protein contained three typical zinc finger domains, and its amino acids were mainly composed of irregular coiled and extended chains, which mainly played roles in the nucleus. At the same time, the prediction of protein interaction network found that EGR2-Xl may interact with SOXI0, EGRl, EGR3 and other proteins. Phylogenetic tree analysis showed that the genetic relationship between bovine and buffalo was the closest, and zebrafish was the farthest. It was indicated that EGR2-Xl protein was highly conserved among different species. EGR2-Xl gene was expressed in heart, liver, spleen, lung, kidney, muscle and back fat, with the highest expression in the lung, followed by back fat and liver, and the lowest in kidney. The expression level of EGR2-Xl gene was the highest on the 3nl day, then gradually decreased, and the lowest on the 7th day. [ Conclusion]EGR2-XJ plays an important role in regulating the proliferation, differentiation and lipid metabolism of bovine adipocytes. [ABSTRACT FROM AUTHOR]

In order to explore the role of Isthmin-1 (Ism-1) in glucose and lipid metabolism of grass carp, we cloned the open reading frame (ORF) of Ism-1 by RT-PCR, and the sequence was analyzed by bioinformatics technology. The distribution characteristics of Ism-1 in different tissues of grass carp were detected by RT-qPCR, and the expression of Ism-1 under different nutritional conditions were analyzed in vitro and in vivo. In this study, ORF region of Ism-1 was successfully cloned. Sequence analysis showed that the ORF region of Ism-1 was 1380 bp, encoding 459 amino acids, and the relative molecular weight of protein was 50.96 kD. Amino acid multiple sequence alignment and phylogenetic tree analysis showed that the evolutionary relationship of Ism-1 between grass carp and fathead minnow was the closest (amino acid similarity was up to 96.51%). Ism-1 was widely expressed in multiple tissues, and had higher expression in red muscle, next were gill, brain and white muscle of grass carp. The fasting and refeeding experiment indicated the expression of Ism-1 in hepatopancreas was significantly upregulated after 14 days of starvation treatment (P<0.05), and the expression decreased after refeeding, but was still significantly higher than that of control group (P<0.05). The expression of Ism-1 in white muscle was significantly up-regulated after starvation and refeeding (P<0.05). The results of intraperitoneal injection of glucagon experiment eventuated that the expression of Ism-1 in hepatopancreas was significantly downregulated compared with control group (P<0.05). In addition, the expression of Ism-1 was significantly decreased (P<0.001) in hepatocytes after treatment with 80 μg/mL oleic acid for 24h. Overall, this research confirmed Ism-1 may be involved in the regulation of glucose and lipid metabolism of grass carp, which would provide essential data for the following study of the function of Ism-1. [ABSTRACT FROM AUTHOR]

Simple Summary: Since existing control strategies are not yet effective, in order to elucidate the molecular mechanisms of protein–ligand binding for effective pest control, three ligands that bind best to chemosensory protein 2 of Sirex nitobei were screened in this study. The host plant volatile (+)-α-pinene, symbiotic fungal volatiles terpene and (−)-globulol were found to bind most stably by molecular docking and dynamic simulations, and their free binding energies were calculated by the molecular mechanics Poisson–Boltzmann surface area method. Furthermore, some key amino acid residues were deeply explored, providing a favorable molecular basis for regulating the behavioral interactions of insects and developing new pest control strategies. Sirex nitobei is an important wood-boring wasp to conifers native to Asia, causing considerable economic and ecological damage. However, the current control means cannot achieve better efficiency, and it is expected to clarify the molecular mechanism of protein–ligand binding for effective pest control. This study analyzed the expression pattern of CSP2 in S. nitobei (SnitCSP2) and its features of binding to the screened ligands using molecular docking and dynamic simulations. The results showed that SnitCSP2 was significantly expressed in female antennae. Molecular docking and dynamic simulations revealed that SnitCSP2 bound better to the host plant volatile (+)-α-pinene and symbiotic fungal volatiles terpene and (−)-globulol than other target ligands. By the molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) method, the free binding energies of the three complexes were calculated as −44.813 ± 0.189 kJ/mol, −50.446 ± 0.396 kJ/mol, and −56.418 ± 0.368 kJ/mol, and the van der Waals energy was found to contribute significantly to the stability of the complexes. Some key amino acid residues were also identified: VAL13, GLY14, LYS61, MET65, and LYS68 were important for the stable binding of (+)-α-pinene by SnitCSP2, while for terpenes, ILE16, ALA25, TYR26, CYS29, GLU39, THR37, and GLY40 were vital for a stable binding system. We identified three potential ligands and analyzed the interaction patterns of the proteins with them to provide a favorable molecular basis for regulating insect behavioral interactions and developing new pest control strategies. [ABSTRACT FROM AUTHOR]

In order to explore the mRNA expression level of cyclin-dependent protein kinase 8 gene (CDK8) in various tissues of red deer (Cervus elaphus) antler velvet and the biological characteristics of the protein encoded by the gene, the full-length coding sequence (CDS) of cyclin-dependent protein kinase 8 gene of red deer was cloned by PCR and connected to pMD-18T vector to measure nucleotide sequences. Thereafter, similarity alignment and phylogenetic tree construction could be carried out, and the resulting nucleotide sequences was further used for similarity comparison and phylogenetic tree construction. The composition, structure and physicochemical properties of CDK8 protein were analyzed by bioinformatics methods. Real-time PCR was used to detect the expression level of CDK8 gene mRNA in antler tissues. The results showed that the coding region of red deer CDK8 was 1, 254 bp in length, and a total of 417 amino acids were encoded. The encoded protein has a relative molecular mass of 47,744. 11 and a theoretical isoelectric point pI of 7.28, which is a hydrophilic unstable protein with no signal peptide and localized to the nucleus with a total of 38 phosphorylation sites, and the secondary structure of the protein is dominated by irregular curls and α helixes. The expression level of CDK8 mRNA is the highest in furry tissue, followed by mesenchymal, then cartilage, and the lowest is in precartilage. CDK8 is an important part of the transcription machine the mediator transcriptional regulatory complex. In this study, the full-length coding sequence of red deer CDK8 was cloned for the first time, which can provide important reference data for us to understand the mechanism and function of CDK8 during antler regeneration. [ABSTRACT FROM AUTHOR]

nARs are ligand-activated transcription factors associated with gonadal development and reproductive regulation. In this study, two nuclear androgen receptor genes (ara and arb) were cloned from an ovoviviparous teleost, black rockfish (Sebastes schlegelii). The phylogenetic analysis of nARs showed that nARα and nARβ clustered into teleost nARα and nARβ, respectively, but differed from the nARs of tetrapods. Four module domains of the nuclear receptor superfamily are present in the black rockfish nARs, an N-terminal domain (NTD), a DNA-binding domain (DBD), a hinge region (HR) and a ligand-binding domain (LBD). Among the four domains, only the DBD and LBD of nARα and nARβ are relatively conserved. Tissue distribution analysis revealed that ara was mainly expressed in the gonad, muscle, intestine and kidney in males and in female, while arb was mainly detected in the gonad, followed by the intestine, kidney and head kidney. During the gonadal development process, the expression levels of ara and arb were significantly decreased from the regenerating to late stages of spermatogenesis and significantly increased in the degeneration stage in the testis (P

Simple Summary: This article is mainly about that molecular characterization and expression pattern of leptin in yellow cheek carp (Elopichthys bambusa) and its transcriptional changes in response to fasting and refeeding. In this study, the authors used PCR to clone the CDS of leptin in yellow cheek carp, and analyzed the sequence differences of the gene with other species, constructed the phylogenetic tree, used real-time PCR for analyzing the expression of leptin in different tissues, including the expression of leptin in the brain and liver after fasting–refeeding of yellow cheek carp. This paper found that the full-length cDNA sequence of Eblep was 1140 bp and the length of the open reading frame (ORF), which can encode a protein of 174 amino acids, was 525 bp. The Eblep mRNA transcript was detected in all tested tissues, with the highest expression in the liver and lowest expression in the spleen. It was found that the change in the mRNA expression of EbLep may be an adaptive strategy for different energy levels by studying the expression of EbLep mRNA in the brain and liver under fasting and refeeding. Leptin, a secretory protein encoded by obese genes, plays an important role in regulating feeding and energy metabolism in fish. To study the structure and function of the Leptin gene in yellow cheek carp (Elopichthys bambusa), the full-length cDNA sequence of leptin was cloned, named EbLep. The full-length cDNA of Eblep was 1140 bp, and the length of the open reading frame (ORF), which can encode a protein of 174 amino acids, was 525 bp. The signal peptide was predicted to contain 33 amino acids. Sequence alignment showed that the amino acid sequence of Leptin was conserved in cyprinid fish. Despite large differences between primary structures, the tertiary structure of the EbLep protein was similar to that of the human protein and had four α-helices. The EbLep mRNA transcript was detected in all tested tissues, with the highest expression in the liver and lowest expression in the spleen. In this study, short-term fasting significantly increased the mRNA expression of EbLep in the liver, which returned to a normal level after 6 days of refeeding and was significantly lower than the normal level after 28 days of refeeding. In the brain, the mRNA expression of EbLep significantly decreased during short-term fasting and significantly increased to a higher value than the control group after 1 h of refeeding. It then rapidly decreased to a lower value than the control group after 6 h of refeeding, returning to the normal level after 1 day of refeeding, and significantly decreasing to a lower value than the control group after 28 days of refeeding. To sum up, the change in the mRNA expression of EbLep in the brain and liver may be an adaptive strategy for different energy levels. [ABSTRACT FROM AUTHOR]

Janus kinase-signal transducers and activators of transcription (JAK-STAT) signaling pathway is one of the most important ubiquitous cell signaling pathways in animals. In vertebrates, it is mainly involved in regulating the expression and secretion of many cytokines. The members of the suppressor of cytokine signaling (SOCS) family are the important JAK-STAT pathway inhibitory factors. Cytokine-induced SH-2 containing protein (CISH) is one known family members of SOCS, with its main function involved in regulating the signaling of interleukin-2. In this study, the full-length cDNA sequence of Cyprinus carpio CISH with the length of 1 685 bp was cloned. The open reading frame (ORF) was 666 bp in length and encoded 221 amino acids. The molecular weight of the deduced protein was 24.91 ku. C. carpio CISH possesses typical SOCS family structure, with three functional domains including N region, central SH2 domain and C terminal SOCS box domain, the latter two being conservatively present in CISH proteins of other bony fish. The phylogenetic analysis showed that the common carp CISH was the closest to CISHs of grass carp and zebrafish, which also belong to Cyprinidae. The results showed that the mRNA expression level of C. carpio CISH in blood was the highest, and the lowest expression level in liver. After being stimulated by Aeromonas hydrophila, the expression of C. carpio CISH in blood was up-regulated significantly at 24 h, and returned to the normal level at 48 h, suggesting that it may play an important role in the process of the resistance to bacterial infection. [ABSTRACT FROM AUTHOR]

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SHCBP1 (Shc SH2-domain binding protein 1) is a member of the Src and collagen homolog (Shc) protein family and is closely associated with multiple signaling pathways that play important roles during hair follicle induction, morphogenesis, and cycling. The purpose of this study was to investigate SHCBP1 gene expression, polymorphisms, and the association between SHCBP1 and wool quality traits in Chinese Merino sheep. The SHCBP1 gene was shown, by qPCR, to be ubiquitously expressed in sheep tissues and differentially expressed in the adult skin of Chinese Merino and Suffolk sheep. Four SNPs (termed SHCBP1SNPs 1–4) were identified by Sanger sequencing and were located in exon 2, intron 9, intron 12, and exon 13 of the sheep SHCBP1 gene, respectively. SHCBP1SNPs 3 and 4 (rs411176240 and rs160910635) were significantly associated with wool crimp (P < 0.05). The combined polymorphism (SHCBP1SNP3-SHCBP1SNP4) was significantly associated with wool crimp (P < 0.05). Bioinformatics analysis showed that the SNPs associated with wool crimp (SHCBP1SNPs 3 and 4) might affect the pre-mRNA splicing by creating binding sites for serine–arginine-rich proteins and that SHCBP1SNP4 might alter the SHCBP1 mRNA and protein secondary structure. Our results suggest that SHCBP1 influences wool crimp and SHCBP1SNPs 3 and 4 might be useful markers for marker-assisted selection and sheep breeding. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to study VdActin7 gene expression pattern in different tissues of Vaccinium dunalianum Wight.【Method】The cDNA and DNA of VdActin7 gene were successfully cloned by PCR technique based on the transcriptome database of 3 generations of V. dunalianum. The expression of VdActin7 gene in young leaves, mature leaves, flower buds, flowers, green fruits, red fruits, green fruit stems and red fruit stems of V. dunalianum was further detected and analyzed using qRT-PCR technique. 【Result】The full length of VdActin7 gene and cDNA were 1657 and 1387 bp, respectively, which contained 3 introns, namely, intron1 of 93 bp, intron2 of 89 bp and intron3 of 88 bp. It had an open reading frame length of 1134 bp, encoding 377 amino acids. The protein VdActin7 with a relative molecular weight of 41.70 KD and a theoretical isoelectric point of 5.31, was a stable hydrophilic protein, without signal peptide and transmembrane domain. Phylogenetic analysis revealed that the VdActin7 was closest to V. ashei Actin7, which also belonged to the Vaccinium family. The codon usage bias analysis of the VdActin7 gene showed that GC contents of codon(GC) and 3rd nucleotide in codon GC3s were 48.2% and 44.1%, respectively, the effective number of codon(ENC) was 49.28%, a total of high-frequency codons was 31. The codon bias was similar to that of Ziziphus, both with the strongest bias for CUU, followed by GUU, and the bias for CUU was significantly stronger in V. dunalianum(3.43) than that in Ziziphus(2.57). qRT-PCR analysis showed that the VdActin7 gene expression showed obvious tissue specificity, with the highest in young leaves while lower expression in mature leaves and flowers, meanwhile no detectable expression or no expression in fruits, suggesting that the VdActin7 gene might play an important role in the development of young leaves. 【Conclusion】In the paper, we reported the Actin7 gene sequence and its characteristics in V. dunalianum for the first time, and found that the VdActin7 gene had tissue expression specificity. This laid the foundation for further study of VdActin7 gene function, expression regulation and mechanism of action. [ABSTRACT FROM AUTHOR]

Matrix metalloproteinase-12 (MMP12), or macrophage metalloelastase, plays important roles in extracellular matrix (ECM) component degradation. Recent reports show MMP12 has been implicated in the pathogenesis of periodontal diseases. To date, this review represents the latest comprehensive overview of MMP12 in various oral diseases, such as periodontitis, temporomandibular joint dysfunction (TMD), orthodontic tooth movement (OTM), and oral squamous cell carcinoma (OSCC). Furthermore, the current knowledge regarding the distribution of MMP12 in different tissues is also illustrated in this review. Studies have implicated the association of MMP12 expression with the pathogenesis of several representative oral diseases, including periodontitis, TMD, OSCC, OTM, and bone remodelling. Although there may be a potential role of MMP12 in oral diseases, the exact pathophysiological role of MMP12 remains to be elucidated. Understanding the cellular and molecular biology of MMP12 is essential, as MMP12 could be a potential target for developing therapeutic strategies targeting inflammatory and immunologically related oral diseases. [ABSTRACT FROM AUTHOR]

The Dof (DNA binding with one finger) genes encode a family of plant-specific transcription factors and play critical roles in numerous biological processes in plants. Piper nigrum L. (black pepper) is a popular spice crop with high economic value. Phytophthora capsici infection has become a key problem in P. nigrum industry, but the molecular studies about P. capsici have been rarely performed. This study aimed to conduct a genome-wide identification and characterization of the Dof gene family in P. nigrum, and provide some candidates differentially expressed genes under P. capsica infection. A total of 46 Dof genes were identified in the P. nigrum genome, and these genes were unevenly distributed on 20 chromosomes. Afterward, a phylogenetic tree was constructed by the N-J (neighbor-joining) method, and seven groups (not including one group that was not present in P. nigrum but present in Arabidopsis thaliana (L.) Heynh.) were obtained. The Dof genes in the same group had similar gene and protein structures. Selection pressure analysis indicated that duplicated genes underwent purification selection during evolution. Tissue expression analysis showed that P. nigrum Dof genes had significant tissue-specific expression characteristics, and they also expressed significantly different among different groups. In addition, according to the RNA-seq sequence data of Dof genes under the infection of P. capsici, a large number of Dof genes were found with differential expression profiles, and several genes with high response to P. capsici were screened out. This study revealed evolutionary and functional characteristics of P. nigrum Dof genes, and provided important references for the follow-up study of Dof genes in response to the biological stress of P. capsici. [ABSTRACT FROM AUTHOR]

In the present investigation, goat β-defensin (GBD) expression in different tissues of Osmanabadi goat was studied. Goat tongue epithelia had the highest level of GBD expression, followed by reticulum, rumen, omasum, kidney, spleen, liver and uterus. Minimal expression of GBD was observed in Osmanabadi tracheal tissue. To our knowledge, this is the first study to report expression of GBD in goat rumen.

Mucosal immunity plays a critical role in the protection of teleost fish against infection, but mucosal immunoglobulin of important aquaculture species unique to Southeast Asia remained greatly understudied. In this study, the sequence of immunoglobulin T (IgT) from Asian sea bass (ASB) is described for the first time. IgT of ASB possesses the characteristic structure of immunoglobulin with a variable heavy chain and four CH4 domains. The CH2-CH4 domains and fulllength IgT were expressed and CH2-CH4 specific antibody was validated against full-length IgT expressed in Sf9 III cells. Subsequent use of the anti-CH2-CH4 antibody in immunofluorescence staining confirmed the presence of IgTpositive cells in the ASB gill and intestine. The constitutive expression of ASB IgT was characterized in different tissues and in response to red-spotted grouper nervous necrosis virus (RGNNV) infection. The highest basal expression of secretory IgT (sIgT) was observed in the mucosal and lymphoid tissues such as the gills, intestine and head kidney. Following NNV infection, IgT expression was upregulated in the head kidney and mucosal tissues. Moreover, a significant increase in localized IgT was found in gills and intestines of infected fish on day 14 post-infection. Interestingly, a significant increase in NNV-specific IgT secretion was only observed in the gills of the infected group. Our results suggest that ASB IgT may play an important role in the adaptive mucosal immune responses against viral infection and could potentially be adapted as a tool for the evaluation of prospective mucosal vaccines and adjuvants for the species. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Hydrobiology is the property of Editorial Department of Journal of Hydrobiology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)