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.     

Adaptation to freshwater in the palaemonid shrimp Macrobrachium amazonicum: comparative ontogeny of osmoregulatory organs

The ontogeny of osmoregulatory organs was studied in two geographically isolated populations of the palaemonid shrimp Macrobrachium amazonicum, one originating from the Amazon estuary (A) and the other from inland waters of the Pantanal (P) in northeastern and southwestern Brazil, respectively. A previous investigation had shown that the estuarine population is able to hypo-osmoregulate in seawater, whereas the hololimnetic inland population has lost this physiological function. In the present study, the structural development of the branchial chamber and excretory glands and the presence of Na⁺/K⁺-ATPase (NKA) were compared between populations and between larval and juvenile stages after exposure to two salinities representing hypo- and hypertonic environments. In the newly hatched zoea I stage of both populations, gills were absent and NKA was localized along the inner epithelium of the branchiostegite. In intermediate (zoea V) and late larval stages (decapodids), significant differences between the two populations were observed in gill development and NKA expression. In juveniles, NKA was detected in the gills and branchiostegite, with no differences between populations. At all developmental stages and in both populations, NKA was present in the antennal glands upon hatching. The strong hypo-osmoregulatory capacity of the early developmental stages in population A could be linked to ion transport along the inner side of the branchiostegite; this seemed to be absent or weak in population P. The presence of fully functional gills expressing NKA appears to be essential for efficient hyper-osmoregulation in late developmental stages during successful freshwater adaptation and colonization.     

Two new cave rnicolous genera of Julidae (Diplopoda, Julida), with notes on the tribe Brachyiulini and on julid subanal hooks and anchors

Two remarkable genera and species of the millipede family Julidae, Titanophyllum spiliarumgen. n., sp. n. and Mammamia profugagen. n. sp., n., are described from caves in Greece and Italy, respectively. The presence of a flagellum and the absence of a 'pro-mesomerital forceps' on the gonopods place them in the tribe Brachyiulini Verhoeff, 1909, an unnatural grouping based on plesiomorphic characters. Both are outstanding in being the only hitherto known blind julidans having such gonopodal features. A dichotomous key to the nine valid brachyiulinine genera based on peripheral and gonopodal characters is presented. Moreover, notes on subanal hooks and anchors in Julida are provided with hypotheses on their possible function.     

Ranolazine block of human Na v 1.4 sodium channels and paramyotonia congenita mutants

The antianginal drug ranolazine exerts voltage- and use-dependent block (UDB) of several Na+ channel isoforms, including Na(v) 1.4. We hypothesized that ranolazine will similarly inhibit the paramyotonia congenita Na(v) 1.4 gain-of-function mutations, R1448C, R1448H, and R1448P that are associated with repetitive action potential firing. Whole-cell Na+ current (I(Na)) was recorded from HEK293 cells expressing the hNa(v) 1.4 WT or R1448 mutations. At a holding potential (HP) of -140 mV, ranolazine exerted UDB (10 Hz) of WT and R1448 mutations (IC 50 = 59 - 71 μM). The potency for ranolazine UDB increased when the frequency of stimulation was raised to 30 Hz (IC 50 = 20 - 27 uM). When the HP was changed to -70 mV to mimic the resting potential of an injured skeletal muscle fibre, the potency of ranolazine to block I(Na) further increased; values of ranolazine IC 50 for block of WT, R1448C, R1448H, and R1448P were 3.8, 0.9, 6.3, and 0.9 uM, respectively. Ranolazine (30 uM) also caused a hyperpolarizing shift in the voltage-dependence of inactivation of WT and R1448 mutations. The effects of ranolazine (30 uM) to reduce I(Na) were similar (~35% I(Na) inhibition) when different conditioning pulse durations (2-20 msec) were used. Ranolazine (10 μM) suppressed the abnormal I(Na) induced by slow voltage ramps for R1448C channels. In computer simulations, 3 μM ranolazine inhibited the sustained and excessive firing of skeletal muscle action potentials that are characteristic of myotonia. Taken together, the data indicate that ranolazine interacts with the open state and stabilizes the inactivated state(s) of Na(v)1.4 channels, causes voltage- and use-dependent block of I(Na) and suppresses persistent I(Na). These data further suggest that ranolazine might be useful to reduce the sustained action potential firing seen in paramyotonia congenita.     

Development of NaCl-tolerant line in Chrysanthemum morifolium Ramat. through shoot organogenesis of selected callus line

Plants were regenerated successfully through shoot organogenesis of a NaCl-selected callus line of Chrysanthemum morifolium Ramat. cv. Maghi Yellow (a salt sensitive cultivar), developed through stepwise increase in NaCl concentration (0-100mM) in the MS medium. The stepwise increase in NaCl concentration from a relatively low level to cytotoxic level was found to be a better way to isolate NaCl-tolerant callus line, since direct transfer of callus to high saline medium was detrimental to callus survival and growth. The selected callus line exhibited significant increase in superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) activities compared to control callus (grown in medium devoid of NaCl). Stability of salt tolerance character of the selected callus line was checked by growing the calli in NaCl-free medium for 3 consecutive months followed by re-exposure to higher salinity stress (120mM NaCl). Among different growth regulator treatments, a combination of 5mgl(-1) TDZ (Thidiazuron) along with 0.25mgl(-1) NAA and 0.5mgl(-1) GA(3) was found to be the most effective for shoot organogenesis in selected callus line. The regeneration potential of the NaCl-tolerant callus ranged from 20.8% to 0% against 62.4% to 0% in control callus line. Under elevated stress condition (medium supplemented with 250mM NaCl), selected calli derived regenerants (S1 plants) exhibited significantly higher SOD and APX activities over both PC (positive control: control callus derived plants grown on MS medium devoid of NaCl) and NC (negative control: control callus derived plants subjected to 250mM NaCl stress) plants. In addition, the NC plants showed stunted growth, delayed root initiation, and had lesser number of roots as compared to S1 plants. Based on growth performance and antioxidant capacity, the S1 plants could be considered as NaCl-tolerant line showing all positive adaptive features towards the salinity stress. Further study on agronomic performance of these S1 plants under saline soil condition need to be undertaken to check the genetic stability of the induced salt-tolerance.     

Effect of sport practice and warm-up duration on the morning–evening difference in anaerobic exercise performance and perceptual responses to it

This study was designed to assess the effect of sport practice and warm-up duration on the morning–evening differences in muscle power and fatigue during performance of anaerobic exercise and perceptual responses to it. Twenty-two male physical education students – twelve trained (TG) (21.51 ± 1.25 years, 182.17 ± 4.37 cm and 82.88 ± 11.23 kg) and ten untrained (NTG) (23.89 ± 3.17 years, 176.8 ± 2.2 cm and 82.24 ± 8.43 kg) – participated in a crossover randomized study. They were asked to perform a 30-s Wingate test during six experimental sessions (three at 08:00 and three at 18:00 h) after different active warm-up (AWU) durations (5 min, 15 min, or 20 min). The AWU consisted of pedaling at a constant pace of 60 rpm against 50% of maximal aerobic power. Rate of perceived exertion (RPE) was recorded after the AWU and again immediately after the Wingate test. Heart rate and temperature (T) were recorded during each session at rest, after AWU and immediately at the end of the Wingate test. During the Wingate test, peak power (PP), mean power (MP), and the fatigue index were recorded. While the RPE estimations were higher in NTG, no time-of-day effect was recorded in either experimental group (morning or evening). T, PP, and MP were higher in the afternoon than in the morning (p < 0.001 for PP and MP; p < 0.05 for T). Similarly, PP and MP during the Wingate test were significantly higher in the TG than in the NTG (p < 0.001). The morning–evening difference of PP and MP was affected by AWU duration; AWU₁₅ was best in the morning for improving lower limb power for both the TG and NTG, whereas reducing this period to 5 min in the evening was appropriate for TG only.     

Prediction of auxin response elements based on data fusion in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The plant hormone “auxin” is a key regulator of plant development and environmental responses. Many genes in Arabidopsis thaliana are known to be up-regulated in response to auxin. Auxin response factors activate or repress the expression of genes by binding at their promoter regions within auxin response elements (AuxRE) that are key regulatory cis-acting motives. Therefore, the identification of auxin-response elements is among the most important issues to understand the auxin regulation mechanisms. Thus, searching the TGTCTC motif is an unreliable method to identify AuxRE because many AuxRE variants may also be functional. In the present study, we perform an In-silico prediction of AuxREs in promoters of primary auxin responsive genes. We exploit microarray data of auxin response in Arabidopsis thaliana seedlings, in order to provide biological annotation to AuxRE. We apply a data fusion method based on the combined use of evidence theory and fuzzy sets to scan upstream sequences of response genes.