Disruption of POF1B binding to nonmuscle actin filaments is associated with premature ovarian failure

Premature ovarian failure (POF) is characterized by elevated gonadotropins and amenorrhea in women aged <40 years. In a Lebanese family with five sisters who received the diagnosis of POF, we established linkage to the long arm of the X chromosome (between Xq21.1 and Xq21.3.3), using whole-genome SNP typing and homozygosity-by-descent mapping. By sequencing one candidate gene within that region, POF1B, we identified a point mutation localized in exon 10. This substitution of a nucleotide (G-->A), at position 1123, results in an arginine-->glutamine mutation of the protein sequence at position 329 (mutation R329Q). All the affected family members were homozygous for the mutation, whereas the unaffected members were heterozygous. Because POF1B shares high homology with the tail portion of the human myosin, we assessed the ability of both wild-type and mutant POF1B proteins to bind nonmuscle actin filaments in vitro. We found that the capacity of the mutant protein to bind nonmuscle actin filaments was diminished fourfold compared with the wild type, suggesting a function of POF1B in germ-cell division. Our study suggests that a homozygous point mutation in POF1B influences the pathogenesis of POF by altering POF1B binding to nonmuscle actin filaments.     

A method for estimating age of Danish medieval sub-adults based on long bone length

The preferred method for aging archaeological sub-adult skeletons is by dental examination. In cases where no dental records are available, age estimation may be performed according to epiphyseal union, skeletal elements or diaphyseal lengths. Currently no data have been produced specifically for aging archaeological Danish sub-adults from the medieval period based on diaphyseal lengths. The problem with using data on Danish samples, which have been derived from a different population, is the possibility of skewing age estimates. In this study 58 Danish archaeological sub-adults were examined, aged from approximately six years to twenty-one years. The samples were aged according to two dental methods: Haavikko and Ubelaker. Regression formulae were constructed for aging according to their diaphyseal lengths both for individual long bones and combinations of upper and lower long bones. This study indicated that with the regression formulae developed, estimation of age can be done with reasonable results on Danish sub-adults. The Danish data were then compared to data from a different archaeological sample and a modern sample. It showed that the modern data indicated a consistently lower age compared to this sample which increased until reaching a maximum of nearly five years and six months. When comparing the archaeological data to this study, the growth profile crossed over at 12.5 years with a maximum age difference before the cross point of two years and three months lower for the archaeological data. After the cross point there was a maximum difference of three years and four months higher for the archaeological data. This study has shown the importance of using data for age estimation for archaeological material which has been developed specifically for that population. In addition it has presented a possible solution for Danish sub-adult material when dental material is not available.     

Diacylglycerol stimulates acrosomal exocytosis by feeding into a PKC- and PLD1-dependent positive loop that continuously supplies phosphatidylinositol 4,5-bisphosphate

Acrosomal exocytosis involves a massive fusion between the outer acrosomal and the plasma membranes of the spermatozoon triggered by stimuli that open calcium channels at the plasma membrane. Diacylglycerol has been implicated in the activation of these calcium channels. Here we report that this lipid promotes the efflux of intraacrosomal calcium and triggers exocytosis in permeabilized human sperm, implying that diacylglycerol activates events downstream of the opening of plasma membrane channels. Furthermore, we show that calcium and diacylglycerol converge in a signaling pathway leading to the production of phosphatidylinositol 4,5-bisphosphate (PIP(2)). Addition of diacylglycerol promotes the PKC-dependent activation of PLD1. Rescue experiments adding phosphatidic acid or PIP(2) and direct measurement of lipid production suggest that both PKC and PLD1 promote PIP(2) synthesis. Inhibition of different steps of the pathway was reverted by adenophostin, an agonist of IP(3)-sensitive calcium channels, indicating that PIP(2) is necessary to keep these channels opened. However, phosphatidic acid, PIP(2), or adenophostin could not trigger exocytosis by themselves, indicating that diacylglycerol must also activate another factor. We found that diacylglycerol and phorbol ester stimulate the accumulation of the GTP-bound form of Rab3A. Together our results indicate that diacylglycerol promotes acrosomal exocytosis by i) maintaining high levels of IP(3) - an effect that depends on a positive feedback loop leading to the production of PIP(2) - and ii) stimulating the activation of Rab3A, which in turn initiates a cascade of protein interactions leading to the assembly of SNARE complexes and membrane fusion.     

Brassinosteroid and brassinosteroid-mimic differentially modulate Arabidopsis thaliana fitness under drought

Plant Growth Regulation - Brassinosteroids (BRs) are widely used to promote plant growth/development and alleviate environmental stresses’ adverse effects. However, its low stability in the...     

A Monte Carlo study to investigate the feasibility to use the Moroccan panoramic irradiator in sterile insect technique programs

Radiation and Environmental Biophysics - Mediterranean fly pest (Ceratitis) is one of the most destructive pests of fruit species in Morocco. The sterile insect technique (SIT) is an...     

MicroRNA-145 Is Downregulated in Glial Tumors and Regulates Glioma Cell Migration by Targeting Connective Tissue Growth Factor

Glioblastomas (GBM), the most common and aggressive type of malignant glioma, are characterized by increased invasion into the surrounding brain tissues. Despite intensive therapeutic strategies, the median survival of GBM patients has remained dismal over the last decades. In this study we examined the expression of miR-145 in glial tumors and its function in glioma cells. Using TCGA analysis and real-time PCR we found that the expression of miR-145/143 cluster was downregulated in astrocytic tumors compared to normal brain specimens and in glioma cells and glioma stem cells (GSCs) compared to normal astrocytes and neural stem cells. Moreover, the low expression of both miR-145 and miR-143 in GBM was correlated with poor patient prognosis. Transfection of glioma cells with miR-145 mimic or transduction with a lentivirus vector expressing pre-miR 145 significantly decreased the migration and invasion of glioma cells. We identified connective tissue growth factor (CTGF) as a novel target of miR-145 in glioma cells; transfection of the cells with this miRNA decreased the expression of CTGF as determined by Western blot analysis and the expression of its 3′-UTR fused to luciferase. Overexpression of a CTGF plasmid lacking the 3′-UTR and administration of recombinant CTGF protein abrogated the inhibitory effect of miR-145 on glioma cell migration. Similarly, we found that silencing of CTGF decreased the migration of glioma cells. CTGF silencing also decreased the expression of SPARC, phospho-FAK and FAK and overexpression of SPARC abrogated the inhibitory effect of CTGF silencing on cell migration. These results demonstrate that miR-145 is downregulated in glial tumors and its low expression in GBM predicts poor patient prognosis. In addition miR-145 regulates glioma cell migration by targeting CTGF which downregulates SPARC expression. Therefore, miR-145 is an attractive therapeutic target for anti-invasive treatment of astrocytic tumors.     

Final Disposition and Quality Auditing of the Rehabilitation Process in Wild Raptors Admitted to a Wildlife Rehabilitation Centre in Catalonia,Spain, during a Twelve Year Period (1995–2007)

Background

Variability in reporting and classification methods in previous published data of the final dispositions in the rehabilitation of wild raptors makes use of this data limited in trying to audit the quality of the rehabilitation process. Crude as well as stratified disposition rates are needed if quality auditing of the rehabilitation process is to be adequately performed.

Methodology

Final dispositions of 6221 hospitalized wild raptors admitted at a wildlife rehabilitation centre (WRC) of Catalonia during 1995–2007 were analyzed. These dispositions were calculated as the euthanasia (Er), unassisted mortality (Mr), release (Rr) and captivity rates (Cr)., time to death (Td) for dead and euthanized raptors, and length of stay for released (Tr) raptors was estimated. Stratified analyses by main causes of admission and clinical signs were performed.

Results

The disposition for the total population were: Er  = 30.6%, Mr = 19.1%, Rr  = 47.2%, and Cr  = 3%. By main causes of admission, Er was higher in the trauma category (34.2%), whereas Mr was found similar between trauma (37.4%) and non-trauma categories (34.8%). The highest Rr was observed for the orphaned group (77.9%). Furthermore, Cr was low in all the categories (<4%). By clinical signs, the highest Er was found in animals suffering musculoskeletal (37.9%) or skin (32.3%) lesions; Mr was high in infectious/parasitic diseases (66.7%) and in case of neurological symptoms (64.5%). The euthanized birds had a median Td  = 1 day (P₁₀ = 0-P₉₀ = 59) for both trauma and non-trauma categories, and Td  = 36 days for the orphaned young group (P₁₀ = 0; P₉₀ = 596). The median Td in the unassisted dead birds was 2 days for all the categories (P₁₀ = 0-P₉₀ = 31). Finally, the median Tr in the centre was variable among categories.

Conclusions/Significance

Reporting of final dispositions in wildlife rehabilitation should include the crude and stratified rates (Er, Mr, Rr, and Cr), by causes and clinical presentation, as well as Td and Tr, to allow meaningful auditing of the rehabilitation process quality.     

Evolution of the Portulaca oleracea L. aggregate in Egypt on molecular and phenotypic levels revealed by morphology,inter-simple sequence repeat (ISSR) and 18S rDNA gene sequence markers

Molecular markers including inter simple sequence repeats (ISSR) and 18S rDNA gene sequence markers were combined with a detailed morphological analysis to seek characters that discriminate four taxa of Portulaca oleracea s.l. These taxa were identified as the microspecies Portulaca nitida, Portulaca oleracea, Portulaca rausii and Portulaca granulatostellulata. Morphological characters did not provide a clear distinction among the four taxa of P. oleracea s.l. It was found that mixed populations of the taxa occur in several locations and morphological similarities between the microspecies were detected. ISSR analysis indicates that gene flow among populations of different taxa was high and most of the genetic variation (61.9%) occurs within population. These results are inconsistent with the general characteristics of P. oleracea as a self- or 5% cross-pollinated species. A close relationship between P. oleracea, P. rausii and P. granulatostellulata was supported by ISSR and 18S rDNA gene sequence. The ISSR and 18s data support the specific status of P. nitida as a recently evolved taxon. We conclude that P. oleracea exists as a polymorphic species and is not divisible into microspecies based on seed surface as the primary morphological trait, and inter simple sequence repeats (ISSR) and 18S rDNA gene sequence markers. We suggest that the taxa do not merit specific rank as morphological characters and absence of a breeding barrier fail to separate the different populations when they become sympatric.     

Asymmetry and steric hindrance in tripodal ligands: Reaching the limit for octahedral geometry with the newly synthesized [(6-bromo 2-pyridylmethyl) (6-fluoro 2-pyridylmethyl) (2-pyridylmethyl)] amine tripod in FeCl2 complexes

We have synthesized the fully unsymmetrical [(6-bromo 2-pyridylmethyl) (6-fluoro 2-pyridylmethyl) (2-pyridylmethyl)] amine tripod FBrTPA. The synthesis involves preparation of the already known [(6-bromo 2-pyridylmethyl) (2-pyridylmethyl)] amine BrDPA, which is obtained either by classical condensation of α-substituted pyridine carboxaldehyde with aminomethyl pyridine, or by a pathway involving the protection/deprotection sequence of this primary amine. This second way is useful for syntheses of monosubstituted DPAs in the cases where α-substituted pyridine carboxaldehydes are unavailable. The crystal structure of the FeCl₂ complex shows that the ligand binds in a κ⁴-N fashion, with however relatively long ligand-to-metal distances. The spectroscopic and electrochemical studies support decoordination of the bromopyridyl substituent in solution, with κ³-N coordination mode of the tripod within a complex displaying a trigonal bipyramidal geometry at the metal centre. This complex reacts easily with dry oxygen to yield an unsymmetrical μ-oxo diferric complex, the structure of which is also reported.     

Neuroprotective effect of pyruvate and oxaloacetate during pilocarpine induced status epilepticus in rats

Recent research data have shown that systemic administration of pyruvate and oxaloacetate causes an increased brain-to-blood glutamate efflux. Since increased release of glutamate during epileptic seizures can lead to excitotoxicity and neuronal cell death, we tested the hypothesis that glutamate scavenging mediated by pyruvate and oxaloacetate systemic administration could have a neuroprotective effect in rats subjected to status epilepticus (SE). SE was induced by a single dose of pilocarpine (350mg/kgi.p.). Thirty minutes after SE onset, a single dose of pyruvate (250mg/kgi.p.), oxaloacetate (1.4mg/kgi.p.), or both substances was administrated. Acute neuronal loss in hippocampal regions CA1 and hilus was quantitatively determined five hours after SE onset, using the optical fractionator method for stereological cell counting. Apoptotic cascade in the hippocampus was also investigated seven days after SE using caspase-1 and -3 activity assays. SE-induced neuronal loss in CA1 was completely prevented in rats treated with pyruvate plus oxaloacetate. The SE-induced caspase-1 activation was significantly reduced when rats were treated with oxaloacetate or pyruvate plus oxaloacetate. The treatment with pyruvate and oxaloacetate caused a neuroprotective effect in rats subjected to pilocarpine-induced SE.