Syntheses and anti-inflammatory activity of diphenylamine-2,2'-dicarboxylic acid and its metal complexes

Diphenylamine-2,2'-dicarboxylic acid and its Cu(II), Ni(II), Co(II) and Zn(II) complexes have been synthesized and characterized by their elemental analyses, molecular weight determination, molar conductance, infrared and electronic spectra and magnetic measurements. The Zinc complex was tested by different methods for its anti-inflammatory activity and found to be equipotent to naproxen and ibuprofen, though at higher doses.     

Quantitative analysis of retromer complex-related genes during embryo development in the mouse

The retromer complex is a heteropentameric protein unit associated with retrograde transport of cargo proteins from endosomes to the trans-Golgi network. Functional silencing study of the Vps26a gene indicated the important role of the retromer complex during early developmental stages in the mouse. However, individual expression patterns and quantitative analysis of individual members of the retromer complex during the early developmental stages has not been investigated. In this study, we conducted quantitative expression analysis of six retromer complex genes (Vps26a, Vps26b, Vps29, Vps35, Snx1, and Snx2) and one related receptor gene (Ci-mpr) during the eleven embryonic stages with normal MEF (mouse embryonic fibroblast) and Vps26a(-/-) MEF cells. Remarkably, except for Vps26a (maternal expression pattern), all tested genes showed maternal-zygotic expression patterns. And five genes (Vps26b, Vps29, Vps35, Snx2, and Ci-mpr) showed a pattern of decreased expression in Vps26a(-/-) MEF cells by comparative analysis between normal MEF and Vps26a(-/-) MEF cells. However, the Snx1 gene showed a pattern of increased expression in Vps26a(-/-) MEF cells. From our results, we could assume that retromer complex-related genes have important roles during oocyte development. However, in the preimplantation stage, they did not have significant roles.     

Role of host switching in the development of pesticide tolerance in Helopeltis theivora (Hemiptera: Miridae), the major pest of tea in India

The polyphagous pest Helopeltis theivora Waterhouse (Heteroptera: Miridae) is one of the major pests of tea [Camellia sinensis, (Theaceae)]. Duranta repens (Verbenaceae), an ornamental plant producing various allelochemicals, is used in many tea estates as hedge plant. This plant was earlier reported to be an alternative host of H. theivora. To gain an insight into the pest and host–plant relationships, the present work reports studies on host-based life-cycle traits, pesticide tolerance status and levels of detoxification enzymes, such as general esterases (GEs), glutathione S-transferases (GSTs) and cytochrome P450 (CYP450). Host switching in the development of pesticide tolerance in H. theivora was evident. While comparing the biology of this pest on the primary host (tea) and the alternative host (D. repens) it was found that total developmental period was higher in the Duranta-reared population, along with significantly higher nymphal mortality and significantly less fecundity. However, there was no significant difference in incubation period, hatchability, sex ratio, pre-oviposition period and adult longevity. Susceptibility of the Duranta-reared population against quinalphos (a commonly used insecticide) was 1.41 times less than the tea-reared H. theivora population. The host-based variation in the relative susceptibility against insecticide was corroborated by differential activity of the three major xenobiotic detoxifying enzymes mentioned above. GE, GST and CYP450 activities were 2.67-fold, 1.43-fold and 1.37-fold higher in the Duranta-reared population than in the tea-reared population, possibly signifying that host switching can play a role in the development of tolerance of H. theivora against insecticides.     

Genomic structure and expression analyses of the PYGM gene in the thoroughbred horse

Muscle glycogen Phosphorylase (PYGM) has been shown to catalyze the degradation of glycogen to glucose-1-phosphate. The PYGM gene can contribute to providing energy to the body by disassembling the glycogen in muscle. Here, we analyzed the genomic structure and expression of the PYGM gene in the thoroughbred horse. The PYGM gene, containing several transposable elements (MIRs, LINEs, and MERs), was highly conserved in mammalian genomes. In order to understand the expression of the horse PYGM gene, we performed quantitative RT-PCR using 11 thoroughbred horse tissue samples. The horse PYGM gene was broadly expressed in all tissues tested. In particular, the highest expression of the horse PYGM gene was observed in skeletal muscle tissue relative to the other tissues. Interestingly, the horse PYGM gene contains fewer mobile elements than its human ortholog, resulting in an increase in the structural stability of the PYGM gene sequence. This study provides insights into the genomic structure of the horse PYGM gene that may be useful in future studies of its association with exercise capability.     

Genetic structure and variability of the working dog inferred from microsatellite marker analysis

Working dogs serve as military watch dogs, search dogs, rescue dogs, and guide dogs with un-come-at-able character. They are drafted by in-training examination including concentration, capacity for locomotion, boldness and earthly desires. In this study, genetic diversity and relationships among two groups of working dogs (pass and fail group in-training examination) were assessed based on 15 microsatellite markers in 25 individuals of working dogs (military watch dogs and Korean search dogs). For the 15 microsatellite markers, the values of allelic richness (A _R ) ranged from 2.21 (pass group) to 1.60 (fail group) in military watch dogs, while A _R ranged from 2.79 (pass group) to 2.72 (fail group) in Korean search dogs. Among 52 different alleles of military watch dogs, 22 alleles were detected in pass group only, while 8 alleles in fail group only. In case of Korean search dogs, 3 alleles were observed in pass group only, while 13 alleles in fail group only. These group-specific unique alleles reflect good biomarker for selecting working dogs (military watch dogs and Korean search dogs), indicating that those group specific microsatellite alleles could separate working dogs to be pass or fail group in out-training dog population. Taken together, this study demonstrates the feasibility of microsatellite analyses for the selection of superior working dogs objectively. Furthermore, this approach could be used for the proper selection of working dogs in combination with in-training examination.     

Selection of internal reference genes for SYBR green qRT-PCR studies of rhesus monkey (<Emphasis Type="Italic">Macaca mulatta</Emphasis>) tissues

Background  

The rhesus monkey (Macaca mulatta) is a valuable and widely used model animal for biomedical research. However, quantitative analyses of rhesus gene expression profiles under diverse experimental conditions are limited by a shortage of suitable internal controls for the normalization of mRNA levels. In this study, we used a systematic approach for the selection of potential reference genes in the rhesus monkey and compared their suitability to that of the corresponding genes in humans.     

Long interspersed element-1 open reading frame 1 protein expression profiles in ovarian cancers

Long interspersed element-1 (LINE-1) retrotransposons are autonomous mobile DNA elements with unique activity that account for about one-fifth of the human genome. Recently, it has been reported that the expression of LINE-1 is closely related to cancer prognosis, and LINE-1 hypomethylation might contribute to the acquisition of aggressive tumor behavior. Despite the importance of LINE-1 expression in cancers, research on the expression of LINE-1 open reading frame (ORF) proteins is very limited. Here, we investigated the expression profiles of LINE-1 ORF1p in ovarian cancer tissue microarrays containing 100 surgical specimens including adjacent normal ovary tissue, primary ovarian cancers, and metastatic ovarian cancers in lymph node. The tissue microarray was stained with mouse monoclonal antibody to LINE-1 ORFp1 for immunofluorescence analysis, and expression levels were evaluated by image analysis. LINE-1 ORFp was significantly overexpressed in ovarian cancers compared with normal tissues and especially upregulated in metastatic ovarian cancers. In addition, the expression of LINE-1 ORF1p was significantly higher in older ovarian cancer patients compared with young patients. These results indicate that expression of LINE-1 ORF1p is related to the progression of ovarian cancers and, in particular, to the age of the patient and the metastatic potential of the cancer.     

Molecular cloning, gene expression, and tissue distribution of adiponectin and its receptors in the Japanese monkey, Macaca fuscata

Background  Adiponectin is an adipocyte-derived hormone that affects regulation of metabolic syndrome such as insulin resistance, type-2 diabetes, and obesity. It functions via seven transmembrane domain receptors [i.e., adiponectin receptors 1 (AdipoR1) and 2 (AdipoR2)] that have been scarcely investigated in non-human primates.
Methods  Molecular cloning of cDNAs for adiponectin, AdipoR1, and AdipoR2 that included the whole protein-coding region in the Japanese monkey, Macaca fuscata , was carried out. Tissue-specific expression of respective genes was analyzed with Northern blot hybridization.
Results  The essential Cys36 and four lysine residues in adiponectin, and transmembrane-spanning domains in AdipoR1 and AdipoR2 appear well conserved. While adiponectin mRNA is expressed only in adipose tissues, AdipoR1 mRNA was found to be expressed in various tissues including the brain.
Conclusions  These results significantly add to the understanding of the molecular basis of obesity-related adipokines and their receptors in non-human primates.