Cell-cycle-dependent structural transitions in the human CENP-A nucleosome in vivo

In eukaryotes, DNA is packaged into chromatin by canonical histone proteins. The specialized histone H3 variant CENP-A provides an epigenetic and structural basis for chromosome segregation by replacing H3 at centromeres. Unlike exclusively octameric canonical H3 nucleosomes, CENP-A nucleosomes have been shown to exist as octamers, hexamers, and tetramers. An intriguing possibility reconciling these observations is that CENP-A nucleosomes cycle between octamers and tetramers in?vivo. We tested this hypothesis by tracking CENP-A nucleosomal components, structure, chromatin folding, and covalent modifications across the human cell cycle. We report that CENP-A nucleosomes alter from tetramers to octamers before replication and revert to tetramers after replication. These structural transitions are accompanied by reversible chaperone binding, chromatin fiber folding changes, and previously undescribed modifications within the histone fold domains of CENP-A and H4. Our results reveal a cyclical nature to CENP-A nucleosome structure and have implications for the maintenance of epigenetic memory after centromere replication.     

Polymorphisms in the promoter regions for human MMP-1 and MMP-13 lead to differential responses to the alpha and beta isoforms of estrogen receptor and their ligand in vitro

Estrogen receptors (ER) are present in connective tissues and therefore it is possible that the loss of estrogen after menopause influences the integrity of these tissues, contributing to development of degenerative conditions such as osteoporosis and osteoarthritis in a subset of women. Aberrant expression of matrix metalloproteinases (e.g. MMP-1 and MMP-13) has been implicated in the progression of these diseases. The present study investigated potential molecular mechanisms involved in the regulation of expression of MMP-1 and MMP-13 promoter variants by ER-alpha and ER-beta (+/-estrogen) in a transient transfection system. The results demonstrate that the activity of human MMP-1 and MMP-13 polymorphic variants is elevated in the presence of ER-alpha and ER-beta, and the single nucleotide polymorphisms present in the promoters of MMP-1 and MMP-13 variants leads to differential activities in response to the ER isoforms. Furthermore, the influence of 17-beta estradiol also varies depending upon whether the alpha or the beta isoform of ER is the modulator of these polymorphic variants. These findings support the conclusion that ER isoforms may be contributing to disease development and/or progression in genetically distinct subsets of women following menopause, and provide mechanistic insights into how such contributions are manifested.     

Molecular dissection of QTL governing grain size traits employing association and linkage mapping in Basmati rice

Grain size is one of the key traits that determines the quality of Basmati rice from the consumers’ as well as the traders’ point of view. Though many genes governing grain size have been identified in indica and japonica, little work has been done in Basmati rice. The present study aims at dissection of a QTL region governing grain size traits in Basmati employing association and linkage mapping approaches. Association mapping revealed that three markers, i.e., RM 6024 (grain breadth), RM1237 and RM18582 (grain length-breadth ratio), which cover 889 kb in the targeted QTL region have been significantly associated with grain size traits. Using linkage mapping, the targeted QTL region has been further delimited to a physical distance of 268 kb that comprises 24 annotated genes. The gene expression analysis of parents, revealed 19 genes differentially expressing within the QTL. Of them, 15 genes showed high expression in Basmati370, while four were expressed in Jaya, and whereas five genes did not show any differential expression between parents. Among differentially expressed genes, a highly expressed gene in Basmati370, Os05g0374200 (Cytokinin dehydrogenase 1 precursor) seems to be involved in accumulation of cytokinins, thus affecting the grain size. Therefore, our findings demonstrated that by complimenting association and linkage mapping, it is likely to dissect a QTL governing grain size traits in Basmati rice and also the QTL could be a potential target for marker-assisted breeding and map-based cloning studies.     

An outbreak of fetal and neonatal losses associated with the diagnosis of bovine viral diarrhea virus

A severe outbreak of dairy herd pregnancy wastage was investigated. At the beginning of the outbreak, a total of 121 lactating cattle were pregnant and considered to be at risk. Overall, 33.1% of the population at risk aborted, while 25.6% gave birth to calves that either died during the early neonatal period or demonstrated signs compatible with congenital defects (abnormal births). A laboratory diagnosis of bovine viral diarrhea virus (BVDV) infection was made in two surviving neonatal calves with symptoms of cerebellar hypoplasia and blindness. An on-farm investigation was conducted to determine if the abortions and abnormal births were associated with BVDV infection. The rate of abortions versus abnormal births was biphasic when graphed by the date of occurrence. The cases of abortion occurred early in the outbreak and were followed by the neonatal losses. Within the population at risk, the mean values for gestational age at the beginning of the outbreak were different between the subpopulations described by gestational outcome. The outcome of each pregnancy that existed at the beginning of the outbreak was determined. Classifications included normal birth (birth of a normal calf), abnormal birth (a neonatal loss of the type described above), abortion, and continued gestation (normal, uncompleted pregnancy). The average gestational age at the time of the index case (the first cases of pregnancy wastage) for these four pregnancy outcome classfications was 142.0, 106.2, 86.7 and 31.3 days, respectively. Reasons for assuming that this outbreak was related to BVDV are discussed.     

Cytotoxicity studies of chromium(III) complexes on human dermal fibroblasts

The cytotoxicity of certain Cr(III) complexes, such as [Cr(salen)(H(2)O)(2)](+), [Cr(edta)(H(2)O)](-), [Cr(en)(3)](3+), [Cr(ox)(3)](3-), [Cr(pic)(3)], and CrCl(3), which differ in ionic character and ligand environment in human dermal skin fibroblasts, has been studied. After 72 h of exposure to 100 microM doses of chromium(III) complexes, the order in which the complexes had an inhibitory effect on cell viability was [Cr(en)(3)](3+) > [Cr(salen)(H(2)O)(2)](+) > [Cr(ox)(3)](3-) > [Cr(edta)(H(2)O)](-) > [Cr(pic)(3)] > CrCl(3). Based on viability studies it was confirmed that [Cr(en)(3)](3+), a triply charged cation, inhibits cell proliferation, and therefore, it was chosen to carry out further investigations. [Cr(en)(3)](3+), at a dose of 50 microM, was found to bring about surface morphological changes, evidenced by cellular blebbing and spike formation accompanied by nuclear damage. TEM analysis revealed substantial intracellular damage to fibroblasts in terms of the formation of apoptotic bodies and chromatin condensation, thus reflecting cell death. FACS analysis further revealed DNA damage by formation of a sub-G(1) peak with 84.2% DNA as aneuploid DNA and arrest of the G(2) / M phase of the cell cycle. Cellular DNA damage was confirmed by agarose gel electrophoresis with the characteristic appearance of a DNA streak in DNA isolated from [Cr(en)(3)](3+)-treated fibroblasts. The proposed mechanism suggests the plausible role of Cr(V), formed as a result of oxidation of Cr(III) by cellular oxidative enzymes, in the cytotoxic response. Consequently, any Cr(III) complex that is absorbed by cells and can be oxidized to Cr(V) must be considered a potential carcinogen. This has potential implications for the increased use of Cr(III) complexes as dietary supplements and highlights the need to consider the cytotoxicity and genotoxicity of a variety of Cr(III) complexes and to understand the potential hazards of Cr(III) complexes encountered in research laboratories.     

Diagnosis and management of a malignant granulosa cell tumor in a holstein nulligravida: a case study

Transrectal palpation of an 18-mo-old, anestrous Holstein nulligravida revealed an enlarged left ovary. Ultrasonographically, the mass resembled known equine granulosa cell tumor (GCT) images. Slowly, virilism developed. Fifty-five days after initial diagnosis, unilateral ovariectomy was performed The presurgical serum concentration of testosterone was equal to that of a mature bull. During the 3 mo period following surgery, estrus expression resumed, the serum testosterone concentration returned to baseline, and the heifer was diagnosed pregnant following AI. A low grade, malignant GCT was the final histopathologically diagnosis. Clinical diagnosis of GCT should involve ultrasonography and steroid hormone assay. An ultrasound image, similar to the one shown, may be characteristic of GCT in the bovine.     

The role of Haemophilus somnus in early embryonic death. I. The effect of the organism on embryos by day 8 postbreeding

A study using 23 healthy, mature, virgin Holstein-Friesian heifers was designed to determine if H. somnus caused detrimental effects in early bovine embryos and the mechanism(s) that induced these effects. Superovulated heifers were artificially inseminated 12 and 24 h after standing estrus using highquality, Haemophilus-free semen from a single ejaculate of one bull. Treatment heifers (n=12) were exposed by intrauterine infusion 12 h after the second insemination to approximately 1.5 x 10(9)H. somnus organisms (Iowa strain 1229) suspended in 10 ml of sterile 0.85% phosphate buffered saline (PBS). Control heifers (n=11) were inseminated and the infused with sterile PBS. Embryos were recovered 8 d after the second insemination using non-surgical technique and evaluated microscopically and graded on their estimated survivability. Representative embryos were also examined for in vitro culture survival time, histopathological changes, vital stain uptake and bacterial contamination. Following embryo recovery, uterine flush solution was centrifuged at 10,000 x G. Sediment was submitted for bacteriologic examination and supernatant preserved for quantitation of H. somnus immunoglobulins. Results to date indicate that H. somnus had a detrimental effect on early bovine embryos. H. somnus was recovered from the tissues of one treated animal. Significantly more (P 相似文献   

Next‐generation sequencing for identification of candidate genes for Fusarium wilt and sterility mosaic disease in pigeonpea (Cajanus cajan)

To map resistance genes for Fusarium wilt (FW) and sterility mosaic disease (SMD) in pigeonpea, sequencing‐based bulked segregant analysis (Seq‐BSA) was used. Resistant (R) and susceptible (S) bulks from the extreme recombinant inbred lines of ICPL 20096 × ICPL 332 were sequenced. Subsequently, SNP index was calculated between R‐ and S‐bulks with the help of draft genome sequence and reference‐guided assembly of ICPL 20096 (resistant parent). Seq‐BSA has provided seven candidate SNPs for FW and SMD resistance in pigeonpea. In parallel, four additional genotypes were re‐sequenced and their combined analysis with R‐ and S‐bulks has provided a total of 8362 nonsynonymous (ns) SNPs. Of 8362 nsSNPs, 60 were found within the 2‐Mb flanking regions of seven candidate SNPs identified through Seq‐BSA. Haplotype analysis narrowed down to eight nsSNPs in seven genes. These eight nsSNPs were further validated by re‐sequencing 11 genotypes that are resistant and susceptible to FW and SMD. This analysis revealed association of four candidate nsSNPs in four genes with FW resistance and four candidate nsSNPs in three genes with SMD resistance. Further, In silico protein analysis and expression profiling identified two most promising candidate genes namely C.cajan_01839 for SMD resistance and C.cajan_03203 for FW resistance. Identified candidate genomic regions/SNPs will be useful for genomics‐assisted breeding in pigeonpea.