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[Display omitted] This article addresses the influence of sulfuric acid on the alternating current electrospinning of polyamide solutions aimed at enhancing the efficiency of the production of nanofibrous materials. Sets of three homopolyamides and three heteropolyamides solutions modified with sulfuric acid were prepared accompanied by the measurement of their dynamic viscosities and conductivities. The solutions were then electrospun. The nanofiber layers produced were analyzed using a scanning electron microscope along with the determination of the diameters of the nanofibers. The evaluation of the spinning of the various solutions and the quality of the fibers was followed by the testing of their productivity and their comparison with the productivity of the original non-acidified solutions. The experiment revealed that the minor addition of sulfuric acid to the solutions served to enhance the productivity of the alternating electrospinning process but at the cost of increases in the diameters of the fibers obtained. In addition, the use of less polar polyamides led to the degradation of the spinning process. The method described in the study is thus suitable for enhancing the productivity of polyamides. [ABSTRACT FROM AUTHOR]

Microplastics can promote microbial colonisation and biofilm growth, thus being referred to as “plastispheres”. The global plastic pollution surge is likely to adversely impact ecology and human health by providing a novel habitat for microbial communities. Even though microplastics in marine environments have been the subject of in-depth research, plastispheres have recently received attention. Thus, the current study investigates the prevalence and distribution of plastispheres along the Maharashtra coast of India, considering their plausible implications for ecology and human health. Microplastics were isolated from sediment and water samples obtained from 10 sampling sites. Subsequently, these microplastic particles were subjected to ATR-FTIR and scanning electron microscopy (SEM) analyses to ascertain their chemical composition, surface topography, and presence of attached biofilms. The predominant polymers composing the microplastic particles were polypropylene (42.8%), polyethylene (28.6%), polystyrene (14.3%), and polyvinyl chloride (14.3%). SEM analysis revealed the presence of topographical structures and degradation effects, facilitating microbial attachment on the microplastic surface. About 50% of the microplastic particles tested positive for biofilms, with over 66% of those collected from Girgaon and Malvan beaches exhibiting biofilm presence. These positively screened particles also displayed comparatively rough surface structures, likely enhancing microbial colonisation. Microplastic ageing and polymer type could positively affect microbial colonisation. Diatoms and fungal hyphae exhibit varied interactions with microplastic polymers. Notably, microplastics host various reproductive stages of fungi, as evidenced by filamentous networks, mycelia, and conidiophores.

Psychosis is a state of altered thoughts which often accompanies schizophrenia. It was suggested that changes in energetic metabolism accompany psychosis and post-psychosis states. Here, we use the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 to experimentally induce psychosis-like behavior in rats. We addressed an effect of single and repeated (5×) MK-801 application (0.3 mg/kg; i.p.) on the energy metabolism in homogenates and crude mitochondrial fraction (CMF) of the striatum (STR), prefrontal cortex (PFC), and the hippocampus (HIP) of the adult male Wistar rat (n = 39). In each brain region, we assessed activity of glycolytic (hexokinase (HK) and lactate dehydrogenase (LDH)) and Krebs cycle enzymes (citrate synthase (CS) and malate dehydrogenase (MDH)) 2 h and 3 days (3d) after the last MK-801 application together with relative respiratory rates assessment in tissue homogenate. In STR, a single MK-801 application led to a decrease in the LDH (p = 0.0035) and the increase of the MDH (p = 0.0043) activities following 3d. Therein, repeated MK-801 doses evoked increased LDH (p = 0.0204) and CS (p = 0.0019) activities in the homogenate 2 h and increased HK (p = 0.0007) 3d after the last application. Elevated HK activity within CMF was observed after 3d (p = 0.0054). In PFC, repeated MK-801 application decreased HK activity in the homogenate 3d after the final application (p = 0.0234). Correspondingly, PFC HK activity in CMF of repeated administration samples dropped (p = 0.003). In HIP, repeated MK-801 administration led to increased respiration of SDH (p = 0.0475) only 2 h after the last application and decreased CS activity (p = 0.0160) was observed 3d after the last application. Our results indicate a progressive metabolic dysregulation of glycolytic and Krebs cycle enzymes following repeated inhibition of NMDA receptors activity in a region-specific manner. Energetic alterations may form a basis for persisting cognitive problems during and following a psychosis in schizophrenia patients.

ABSTRACTIntroduction of non-native species is considered a grave threat for global biodiversity. It can negatively affect native biota and cause serious socioeconomic losses. In this study, we confirmed the existence of black-striped pipefish (Syngnathus abaster) in the freshwater Tbilisi Reservoir (the middle Kura Drainage), which had been translocated from the Black Sea Basin. The introduction was in part a result of the local name of the target freshwater reservoir called the “Tbilisi Sea,” which implied a connection with the Black Sea for local hobbyists. S. abasteris expanding its range with or without human help, and introduction of this species is expected in other regions. To address this problem, we made a species distribution model using MaxEnt software to test the species environmental suitability around the globe. Lowland inland waterbodies and shorelines in temperate, Mediterranean, and arid climate zones were indicated as localities where this fish could survive if released. The risk of introduction currently seems low, but exploitation of S. abasterfor ornamental purposes due to expanding local and international pet trade increases the likelihood of future releases. For this reason, it is important to verify new cases of successful establishment of this species.

Hard red spring wheat (Triticum aestivumL.) (HRSW) is used for bread baking while durum wheat (Triticum turgidumL. ssp. durum) is used for pasta production. This study investigated the impact of alleles from durum wheat for yield component traits on HRSW end‐use quality traits. Near‐isogenic lines for four quantitative trait loci, QGw.mst‐3B, QGw.mst‐7A, QTn.mst‐5B, and QYld.mst‐2B, were evaluated in rainfed and irrigated environments. Durum alleles were found to significantly impact end‐use quality at all four loci. The durum allele QGw.mst‐3Bfor increased kernel weight (KWT) positively impacted end‐use quality traits with increases in kernel protein and loaf volume. The durum allele at QTn.mst‐5Bdelayed heading date, impacting productive tiller number and KWT and additional end‐use quality traits. The durum allele at QYld.mst‐2Bfor GY showed both positive and negative effects on end‐use quality. The durum allele for reduced spikelet number per spike at QGw.mst‐7Ahad little to no impact on end‐use quality. Results from the present study indicate that the alleles from durum wheat are potential sources of cultivar improvement; however, interactions between yield component and end‐use quality traits may influence utilization. This study demonstrates the potential utility of introgressed durum yield component alleles for genetic improvement in HRSW.

Hexaploid bread wheat (Triticum aestivumL.) and tetraploid durum wheat (Triticum durumDesf.) have been cultivated in similar geographic areas for ∼10,000 yr. The crossing barrier caused by ploidy difference suggests that different favorable alleles for yield‐related traits may have accumulated in the two crops. Previous work allowed identification of favorable alleles at six quantitative trait loci (QTL) from durum wheat in a recombinant inbred line (RIL) population from a cross of ‘Mountrail’ durum and ‘Choteau’ spring wheat. The purpose of this study was to determine the impact of six durum alleles at yield component QTL in several spring wheat backgrounds. Three spring wheat cultivars were crossed with six hexaploid lines derived from the original Choteau/Mountrail cross to generate RILs. Heterozygous RILs, containing both the durum and the bread wheat alleles, were identified for each of the QTL. The heterozygous RILs were used to develop near‐isogenic lines (NILs) for the six introgressed QTL. The NILs were grown in five environments under irrigated and rainfed conditions in Montana in 2017 and 2018. A durum allele QTL on chromosome 3B resulted in increased kernel weight in all five environments. The introgressed durum QTL alleles caused pleiotropic interactions among yield component traits. Environment and genetic background significantly affected the stability of introgressed QTL on yield components for four of the six QTL. Results suggest that alleles from durum may be useful for yield improvement of hexaploid spring wheat. However, interrelationships of yield components, pleiotropic interactions, and environment will affect the value of durum wheat alleles in hexaploid wheat backgrounds.

Culture media used in assisted reproduction are commonly supplemented with gonadotropin hormones to support the nuclear and cytoplasmic maturation of in vitro matured oocytes. However, the effect of gonadotropins on protein synthesis in oocytes is yet to be fully understood. As published data have previously documented a positive in vitro effect of follicle-stimulating hormone (FSH) on cytoplasmic maturation, we exposed mouse denuded oocytes to FSH in order to evaluate the changes in global protein synthesis. We found that dose-dependent administration of FSH resulted in a decrease of methionine incorporation into de novo synthesized proteins in denuded mouse oocytes and oocytes cultured in cumulus-oocyte complexes. Similarly, FSH influenced methionine incorporation in additional mammalian species including human. Furthermore, we showed the expression of FSH-receptor protein in oocytes. We found that major translational regulators were not affected by FSH treatment; however, the amino acid uptake became impaired. We propose that the effect of FSH treatment on amino acid uptake is influenced by FSH receptor with the effect on oocyte metabolism and physiology.

The β-adrenergic signaling pathways and antioxidant defence mechanisms play important roles in maintaining proper heart function. Here, we examined the effect of chronic normobaric hypoxia (CNH, 10% O2, 3 weeks) on myocardial β-adrenergic signaling and selected components of the antioxidant system in spontaneously hypertensive rats (SHR) and in a conplastic SHR-mtBN strain characterized by the selective replacement of the mitochondrial genome of SHR with that of the more ischemia–resistant Brown Norway strain. Our investigations revealed some intriguing differences between the two strains at the level of β-adrenergic receptors (β-ARs), activity of adenylyl cyclase (AC) and monoamine oxidase A (MAO-A), as well as distinct changes after CNH exposure. The β2-AR/β1-AR ratio was significantly higher in SHR-mtBN than in SHR, apparently due to increased expression of β2-ARs. Adaptation to hypoxia elevated β2-ARs in SHR and decreased the total number of β-ARs in SHR-mtBN. In parallel, the ability of isoprenaline to stimulate AC activity was found to be higher in SHR-mtBN than that in SHR. Interestingly, the activity of MAO-A was notably lower in SHR-mtBN than in SHR, and it was markedly elevated in both strains after exposure to hypoxia. In addition to that, CNH markedly enhanced the expression of catalase and aldehyde dehydrogenase-2 in both strains, and decreased the expression of Cu/Zn superoxide dismutase in SHR. Adaptation to CNH intensified oxidative stress to a similar extent in both strains and elevated the IL-10/TNF-α ratio in SHR-mtBN only. These data indicate that alterations in the mitochondrial genome can result in peculiar changes in myocardial β-adrenergic signaling, MAO-A activity and antioxidant defence and may, thus, affect the adaptive responses to hypoxia.

The study investigated the effect of frequency on the electrospinning of polyamide 6 using an alternating electric field. The study included the investigation of the behavior of two polyamide solutions with respect specifically to the dependence of the spinning behavior on increasing voltage frequency. All the measurements were taken applying a rectangular signal for the spinning. The study demonstrated the fundamental influence of frequency on the electrospinning process and, above all, its efficiency. It was discovered that the time of so-called effective spinning, or the period during which the appearance of a more significant number of Taylor cones and polymer nozzles is observed, remains, in certain cases, basically independent of the selected frequency value. This finding thus indicates the application of higher frequencies than the standard 50 Hz in order to enhance the efficiency of the polyamide 6 spinning process. This conclusion was supported by the measurement of the productivity of the process at various frequencies, as well as SEM images that served to prove the preservation of the fibrous nature of the products via the creation of histograms of the fiber diameters. The nanofibrous layers produced by this method are suitable for application in the fields of chemical analytics or filtration.

A reprint of the article "Disneyland for the Double Reed Player:Tales from the IDRS in Tokyo" by Natasha Kalous, which appeared in the Reeding Matter is presented.

The expanding geographical range of wheat stem sawfly (WSS) damage, coupled with the limited number of effective control measures, calls for a need to better characterize and explore the genetic variability for resistance in wheat (Triticum aestivumL.) germplasm from North America. An association‐mapping analysis for stem solidness and wheat stem sawfly resistance was conducted using a set of 244 elite spring wheat lines from 10 North American breeding programs. Three previously identified quantitative trait loci (QTL) for resistance were confirmed, including QTL for stem solidness, heading date, and stem cutting. Three new QTL on chromosomes 2A, 3A, and 5B were identified to be associated with larval mortality and a QTL associated with early stem solidness was identified on chromosome 5D. Genetic variation for late stem solidness is mainly determined by the solid‐stem QTL on chromosome 3B, while variation for early solidness is affected by chromosomal regions on 1B, 3B, and 5D. Potential sources of a novel resistance mechanism causing larval mortality were identified among the panel lines. Favorable alleles for stem solidness and other resistance traits are available within North American wheat germplasm, but only Montana and South Dakota breeding programs appear to have favorable alleles for all QTLs detected in this study. Improvements in wheat stem sawfly management, for both historically impacted areas and for regions first experiencing damage, may be obtained using alleles that already exist in elite germplasm.

The paper presents the theoretical background and implementation details of a multi-domain symbolic analysis program developed with the emphasis on linearized hybrid systems combining classical electrical circuits, controllers, electro-mechanical converters and mechanical parts of electric drives. The systems analyzed can be composed of both basic elements and more complex behavioral blocks (regulators, converters, drives, etc.). All models are defined in an easily extensible library. Implemented algorithms for equation/matrix-based symbolic simplification allow obtaining symbolic expressions of acceptable complexity even for larger systems.

Distal parts of the fish intestine were analyzed for presence of bifidobacteria and lactobacilli using selective agars. Seventy seven samples from Cyprinus carpio, Oncorhynchus mykiss, Carassius auratus, Tinca tinca, Perca fluviatilis, Rutilus rutilus, Scardinius erythrophthalmus, Oreochromis niloticus, and Squalius cephalus were collected randomly throughout years 2008 and 2009. Bifidobacteria were detected in 5 samples from 4 fish species at counts 2.18–4.29 log CFU/g, lactobacilli were present in 6 fish species at counts 1.21–3.65 log CFU/g. Seven bifidobacterial isolates were identified to the species level using biochemical tests and by sequencing of 16S rRNA gene. Three strains belonged to species B. longum, two isolates were identified as B. dentium, one strain as B. asteroides and one isolate was not determined to the species level by employed methods. As identified bifidobacterial species are considered to be of human, animal or honeybee origin, they probably derived as contamination from sewage or other sources. After further more detail testing, the possible use of isolated bifidobacteria as probiotics is promising since they were able to pass through the digestive tract successfully.

Introduction of genes from outside a primary germplasm base has the potential to impact phenotypic diversity, genetic structure, and patterns of linkage disequilibrium (LD). Our goals were to monitor the introduction of key genes in a hard red spring wheat (Triticum aestivumL.) germplasm set using historical cultivars, to determine the impact of the introduced genes on phenotypic diversity in elite germplasm, and to assess the impact of selection history on LD. A superior allele for bread making was introduced early in the 20th century and quickly became fixed. An allele for solid stems introduced in the mid 20th century is the major source of variation in current germplasm. An introduced allele for photoperiod insensitivity is an important source of variation for heading date in current germplasm. The allelic introductions impacted patterns of LD. The unlinked loci for reduced height, RHT‐B1and RHT‐D1,were in LD. Linkage disequilibrium for physically linked markers near introduced genes did not exceed previous estimates of genome‐wide LD. Our results showed a pattern of gene introductions into the spring wheat germplasm base that continues to have an impact on phenotypic diversity and patterns of LD.

Alterations in cell cycle regulators have been implicated in human malignancies including breast cancer. PD 0332991 is an orally active, highly selective inhibitor of the cyclin D kinases (CDK)4 and CDK6 with ability to block retinoblastoma (Rb) phosphorylation in the low nanomolar range. To identify predictors of response, we determined the in vitrosensitivity to PD 0332991 across a panel of molecularly characterized human breast cancer cell lines. Forty-seven human breast cancer and immortalized cell lines representing the known molecular subgroups of breast cancer were treated with PD 0332991 to determine IC50values. These data were analyzed against baseline gene expression data to identify genes associated with PD 0332991 response. Cell lines representing luminal estrogen receptor-positive (ER+) subtype (including those that are HER2 amplified) were most sensitive to growth inhibition by PD 0332991 while nonluminal/basal subtypes were most resistant. Analysis of variance identified 450 differentially expressed genes between sensitive and resistant cells. pRb and cyclin D1were elevated and CDKN2A (p16) was decreased in the most sensitive lines. Cell cycle analysis showed G0/G1arrest in sensitive cell lines and Western blot analysis demonstrated that Rb phosphorylation is blocked in sensitive lines but not resistant lines. PD 0332991 was synergistic with tamoxifen and trastuzumab in ER+ and HER2-amplified cell lines, respectively. PD 0332991 enhanced sensitivity to tamoxifen in cell lines with conditioned resistance to ER blockade. These studies suggest a role for CDK4/6 inhibition in some breast cancers and identify criteria for patient selection in clinical studies of PD 0332991.

The propagation of Ca2+ waves in a network of microglial cells, after its initiation by glutamate, is mediated by purinergic transmission. In this study, we investigated the mechanisms by which glutamate releases ATP from cultured spinal cord microglia. The 4-fold increase in ATP release from microglia in response to glutamate (0.5 mM) was blocked by alpha-aminohydroxy-5-methyl-isoxazole-4-proprionate (AMPA)/kainate receptor antagonist 6-cyano-7-nitroguinoxaline-2,3-dione and specific AMPA receptor antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466) but not by N-methyl-d-aspartic acid or metabotropic glutamate receptor antagonists. Glutamate acting on AMPA receptors evoked an ATP release that was blocked by antagonizing the rise in intracellular Ca2+ as a result of its release from internal stores as well as by antagonizing protein kinase C with chelerythrine. Glutamate-stimulated ATP release was significantly antagonized by the cystic fibrosis transmembrane conductance regulator (CFTR) blockers flufenamic acid and glibenclamide. A role for the CFTR was further confirmed using microglia from CFTR knockout mice, which released significantly less ATP than microglia from control wild-type mice in response to glutamate. Use of 6-methoxy-1-(3-sulfopropyl)quinolinium fluorescence assay revealed functional CFTR in microglia. These observations suggest that glutamate acted on microglial AMPA receptors to stimulate release of Ca2+ from intracellular stores as well as a Ca2+-dependent isoform of protein kinase C, which then acts to trigger release of ATP with the CFTR acting as a regulator of the ATP release process, perhaps through another channel or transporter.

Objective: Superoxide radicals (O2-) are generated during reoxygenation following asphyxia, possibly more when higher concentrations of O2 are used during resuscitation. Superoxide dismutase (SOD) is an antioxidant enzyme, which scavenges O2-. We tested the hypothesis that a single intravenous dose of recombinant human Cu,Zn SOD (rhSOD) could influence microcirculation and biochemical markers of asphyxia in piglets reoxygenated with 21% or 100% O2 after combined cerebral hypoxemia-ischemia-hypercapnia. Methods: Anesthetized newborn piglets were randomized to asphyxia (n = 40) or control (n = 3). Asphyxia was induced by ventilation with 8% O2, adding CO2, and temporary occlusion of both common carotid arteries. After 20 min, 16 piglets received rhSOD 5 mg/kg intravenously and reoxygenation with 21% O2 (rhSOD, 21%; n = 8) or 100% O2 (rhSOD, 100%; n = 8), and 24 piglets received saline and reoxygenation with 21% O2 (21%, n = 13) or 100% O2 (100%, n = 11). The cortical microcirculation was assessed by laser Doppler flowmetry, and glutamate in the striatum and hypoxanthine in the cortex were measured by in vivo microdialysis. Results and conclusion: rhSOD peaked in plasma after 5 min. No rhSOD was detected in brain tissue. There were no significant differences between rhSOD and non-rhSOD groups in any measured variable.

The effect of the p42/44 mitogen-activated kinase (MAPK) inhibitor, PD98059, on MAPK activation and meiosis resumption in mouse oocytes was studied. When germinal vesicle (GV)-stage denuded oocytes (DOs) were cultured continuously in 50 μM PD98059, germinal vesicle breakdown (GVBD) was postponed for 2-3 h. MAPK phosphorylation and activation was delayed as well. However, PD98059 did not impair histone H1 kinase activation. After 14 h of culture there was no significant difference in the rate of DOs reaching metaphase II (MII) arrest in either control or experimental conditions. The effect of PD98059 on MAPK inhibition was further tested in epidermal growth factor (EGF)-treated oocyte–cumulus complexes (OCCs). Exposure of GV-stage OCCs for 5 min to EGF (10 ng/ml) induced a considerable increase in MAPK phosphorylation. After OCCs were further cultured in 50 μM PD98059 a rapid dephosphorylation of MAPK was induced. Already after 1 min of treatment the non-phosphorylated form of MAPK dominated, indicating the high effectivity of PD98059. This result indicates that short EGF/PD98059 treatment of OCCs induced MAPK phosphorylation/dephosphorylation in cumulus cells only. As only a transient delay in MAPK phosphorylation and activation was observed in PD98059-treated DOs we conclude that there is also another PD98059-nonsensitive pathway(s) leading to MAPK activation in mouse oocytes. The data obtained suggest that meiosis resumption in mouse oocytes is somehow influenced by the MEK/MAPK activation pathway.

In this study a specific inhibitor of cyclin-dependent kinases (cdks), butyrolactone I (BL I), was used for activation of pig and cattle metaphase II (MII) oocytes. BL I at a concentration of 100 μM was able to induce activation of both pig and cattle MII oocytes in a manner dependent on exposure time; however, precise timing of BL I exposure was required for the best activation results. The optimum activation rates were obtained when cattle MII oocytes were treated for 5 h with BL I and subsequently for 3-11 h in control medium, and pig MII oocytes for 8 h in BL I and then for 8-16 h in control medium; the percentage of activated oocytes after such treatment varied between 55% and 74% and between 53% and 81% for cattle and pig oocytes, respectively. Shorter exposures to BL I led to re-entry of the oocytes to the metaphase state in 35-50% of oocytes, the remaining oocytes forming a pronuclear stage; longer exposure to BL I led to increased numbers of oocytes being abnormal or degenerated. The behaviour of histone H1 kinase and mitogen activated protein (MAP) kinase, also measured during the experiment, reflected the morphological changes in the oocytes: both were inactivated after BL I treatment, though the inactivation of histone H1 kinase occurred 2 h ahead of that of MAP kinase. However, in the oocytes treated for a shorter time with BL I, with the reoccurrence of condensed chromatin in proportion of the oocytes cultured in control medium after BL I treatment, both kinases became reactivated. Taken together, these results suggest the possibility of using BL I for activation and cloning experiments in both species.

The sterol 4,4-dimethyl-5-cholesta-8,14,24-trien-3-ol (follicular fluid meiosis-activating sterol [FF-MAS]) isolated from human follicular fluid induces resumption of meiosis in mouse oocytes cultured in vitro. The purpose of this study was to examine the hypothesis that differential signal transduction mechanisms exist for FF-MAS-induced and spontaneous in vitro resumption of meiosis in mouse oocytes. Mouse oocytes were dissected from ovaries originating from mice primed with FSH 48 h before oocyte collection. Mechanically denuded germinal vesicle (GV) oocytes were in vitro matured in medium supplemented with hypoxanthine and FF-MAS or allowed to mature spontaneously; both groups were exposed to individual compounds known to inhibit specific targets in the cell. After 20–22 h of in vitro maturation, resumption of meiosis was assessed as the frequency of oocytes in GV breakdown (GVBD) stage. Pertussis toxin (2.5 μg/ml) did not influence resumption of meiosis in either group. Dibutyryl cyclic GMP (320 μM) inhibited FF-MAS-induced GVBD, but not spontaneous GVBD, whereas the subtype 5 phosphodiesterase-inhibitor zaprinast (50 μM) inhibited GVBD in both groups. Microinjection of the catalytic subunit of cAMP-dependent protein kinase into oocytes inhibited spontaneous GVBD, but not FF-MAS-induced GVBD. An inhibitor of cytoplasmic polyadenylation, cordycepin (80 μM), inhibited or retarded spontaneous GVBD to a further extent than it did FF-MAS-induced GVBD. Spontaneous GVBD was more sensitive to the histone H1 kinase-inhibitor olomoucine (250 μM) than was FF-MAS-induced GVBD. Addition of the mitogen-activated protein kinase (MAPK)-inhibitor PD 98059 (50 μM), phospholipase C-inhibitor U-73122 (10 μM), p21ras-inhibitor lovastatine (250 μM), and the src-like kinase inhibitor PP2 (20 μg/ml) inhibited FF-MAS-induced GVBD, but not spontaneous GVBD. Both MAPKs, extracellular regulated kinase (ERK) 1 and ERK2, were phosphorylated under FF-MAS-induced meiotic resumption, in contrast to spontaneous meiotic resumption, in which ERK1 and ERK2 phosphorylation occurred 2 h after GVBD. In the present study, we show that FF-MAS acts through an MAPK-dependent pathway, and we suggest that src-like kinase, p21ras, and phosphoinositide signaling lie upstream of MAPK in the FF-MAS-activated signaling pathway. Clearly, striking pathway differences are present between spontaneous versus FF-MAS-induced meiotic resumption.

We have investigated pathogenic effects of the tRNALys A8344G mutation associated with the syndrome myoclonus epilepsy with ragged-red fibres (MERRF) by using fibroblasts and fibroblast-derived cytoplasmic hybrid cells harbouring different percentages of mutated mitochondrial DNA (mtDNA). The activity of cytochrome c oxidase (COX) in patient fibroblasts with 89% mutated mtDNA was decreased to 20% of the control levels. COX exhibited altered kinetics, with a decreased Vmax for both the low-affinity and high-affinity phases; however, the Km values were not significantly changed. The substrate-dependent synthesis of ATP was decreased to 50% of the control. Analysis of the mitochondrial membrane potential, δΨ, in digitonin-treated cells with tetramethylrhodamine methyl ester (TMRM) with the use of flow cytometry showed a 80% decrease in δΨ at state 4 and an increased sensitivity of δΨ to an uncoupler in fibroblasts from the patient. The investigation of transmitochondrial cytoplasmic hybrid clones derived from the patient's fibroblasts enabled us to characterize the relationship between heteroplasmy of the MERRF mutation, COX activity and δΨ. Within the range of 87-73% mutated mtDNA, COX activity was decreased to 5-35% and δΨ was decreased to 6-78%. These results demonstrate that the MERRF mutation affects COX activity and δΨ in different proportions with regard to mutation heteroplasmy and indicate that the biochemical manifestation of the MERRF mutation exerts a very steep threshold of δΨ inhibition.

Abstract.: We studied drug-induced cardiotoxic effects in 22 pregnant women having tocolysis with intravenous fenoterol and verapamil. Because CK-MB is released from the uterus and placenta, we used the determination of cardiac troponin T (cTnT) as it is one of the most sensitive and specific indicators of myocardial necrosis. Cardiac troponin T levels were within physiological range (0.08± 0.01 μg/l) in all healthy pregnant women tested between 32 and 36 weeks of gestation (control group). In the pregnant women having tocolysis cTnT levels started to increase slightly during the first day of treatment (0.10±0.03 μg/l) and were significantly higher (p<0.05) during the third day (0.35±0.14 μg/l) of tocolytic therapy. The cTnT levels in cord blood (0.13±0.03 μg/l) did not correspond with maternal cTnT concentrations.

Summary: Analysis of the course of D-glucose binding to insulin has shown that the mechanism of glucose-insulin interaction is a cooperative process. Binding of D-glucose molecules to insulin is facilitated by the dissociation of insulin aggregates caused by insulin-glucose interaction. Thus, insulin behaves as a system with strong positive cooperativity. The results have been treated in accordance with theories for interactions coupled to association equilibria. The data obtained support the idea that insulin monomers are the active species responsible for insulin action.

Polyborate anions were found to inhibit mitochondrial ATPase. Mercapto and chloro derivatives of dicarbononaborates showed full inhibition of the enzyme activity at 0.5–0.8 mM. The inhibitory effect of dodecaborates was lower. The inhibition was of competitive type with respect to ATP. The inhibition of soluble F1-ATPase indicates a direct interaction of the polyborate anion with the catalytic part of the enzyme molecule.

The meiotic competence and meiosis resumption of Blue fox (Alopex lagopus) oocytes from anoestrous animals were followed. Oocyte–cumulus complexes (OCC) were cultured in modified TC 199 medium with or without FSH, recombinant bovine somatotrophin (bST) and okadaic acid (OA). The results showed that oocytes less than 100 μm in diameter did not achieve germinal vesicle breakdown (GFBD) by 72 h of culture, which indicates their meiotic incompetence. Oocytes larger than 100 µm in diameter underwent GVBD after 48 h of culture (27%) and reached metaphase II (MII) after 72 and 96 h (20% and 27%) in control medium. Both bST and OA accelerated resumption of meiosis (bST: 55% GVBD and 42% MII after 48 h; OA: 66% GVBD after 18 h). In contrast, FSH significantly reduced meiosis resumption (only 3% GVBD and MII after 72 h) and induced changes in the shape of cumulus granulosa (CG) cells and F-actin assembly typical for cumulus expansion. However, the innermost layers of CG cells (corona radiata) remained connected with the oocyte via gap junctions until the end of culture. Cumuli of oocytes cultured in control, bST-supplemented or OA-supplemented medium did not expand (changes in cell shape and F-actin redistribution did not occur). Moreover, especially in media with bST and OA an increased detachment and rapid disconnection of their gap junctions with the oocyte were observed. These results suggest that under in vitro conditions FSH stimulates expansion of the CG cells and the attached membrana granulosa cells but in contrast it secures heterologous gap junctions between cytoplasmic processes of the corona radiata cells and oolemma during 3 days of culture. Thus, in agreement with the in vivo situation in which Canidae oocytes are ovulated in the GV stage, the cumulus, mainly corona radiata cells, controls resumption of meiosis in Blue fox oocytes under in vitro conditions also.

Ampholytic surfactants of the betaine type are titrated directly with sodium tetraphenylborate in acidic medium, in which they become cationic forms. Generally, ampholytic surfactants and cationic surfactants that contain hydrophilic groups can be determined by precipitation with sodium tetraphenylborate, the excess of which is back-titrated with thallium(I) nitrate. An aluminium wire electrode coated with a plasticized PVC membrane is used for end-point indication. Characteristic data are given for determinations of nine surfactants and applications to textiles are outlined.

The inhibitory effect of mercapto- and chloro-dicarbanonaborates on mitochondrial, FAD-linked, L-glycerol-3-phosphate dehydrogenase activity was tested. The mercaptodicarbanonaborate was found to be a more potent inhibitor than the chloro derivative. The Kiof mercapto- and chlorodicarbanonaborates were 0.8 M and 480 M, respectively. Whereas the mercaptodicarbanonaborate inhibited the enzyme activity competitively, the inhibitory effect of chloro derivative was noncompetitive. The inhibitory effect of mercaptodicarbanonaborate was completely prevented by addition of bovine serum albumin.

In liver mitochondria isolated from hypothyroid rats, the rate of ATP synthesis is lower than in mitochondria from normal rats. Oligomycin-sensitive ATP hydrolase activity and passive proton permeability were significantly lower in submitochondrial particles from hypothyroid rats compared to those isolated from normal rats. In mitochondria from hypothyroid rats, the changes in catalytic activities of F0F1-ATP synthase are accompanied by a decrease in the amount of immunodetected β-F1, F01-PVP, and OSCP subunits of the complex. Northern blot hybridization shows a decrease in the relative cytosolic content of mRNA for β-F1subunit in liver of hypothyroid rats. Administration of 3,5,3′-triodo-L-thyronine to the hypothyroid rats tends to remedy the functional and structural defects of F0F1-ATP synthase observed in the hypothyroid rats. The results obtained indicate that hypothyroidism leads to a decreased expression of F0F1-ATP synthase complex in liver mitochondria and this contributes to the decrease of the efficiency of oxidative phosphorylation.

Pig and cattle oocytes, when released from the follicle, spontaneously resume first meiotic division within 20 or 8 hr, respectively. In oocytes of both species, the activity of histone H1 kinase increases during maturation, exhibiting a maximum in metaphase I. Treatment of these oocytes with okadaic acid results in acceleration of germinal vesicle breakdown (GVBD) and of histone H1 kinase activation. This effect is more important in pig oocytes, in which the acceleration rises for 6 hr, as compared to 2 hr in cattle. Moreover, under these conditions, H1 kinase activity measured after 12 hr of culture appears higher than that observed in control metaphase I oocytes. When added to prophase oocytes, both cycloheximide and 6-DMAP (6-dimethylaminopurine) block GVBD and histone H1 kinase activation. Okadaic acid, at a concentration of 2.5 μM,is able to release the inhibitory effect exerted by cycloheximide on histone H1 kinase activity; however, GVBD occurred only in two-thirds of pig and one-quarter of cattle oocytes after 20 hr of culture. In addition, okadaic acid fully reverses the effect of 6-DMAP on H1 kinase activity and on GVBD in both species. The opposite effects of 6-DMAP and okadaic acid on MPF activation are discussed, as well as the nature of the protein, which has to be synthesized during the first meiotic division and may be involved in the MPF activation cascade.