Changes in microtubule-associated protein-2 (MAP2) expression during development and after status epilepticus in the immature rat hippocampus

In this study, we analyzed the spatiotemporal expression patterns of the high-molecular weight (MAP2a and b) and low-molecular weight (MAP2c and d) cytoskeletal microtubule-associated protein-2 (MAP2) isoforms with Western blotting, and the cellular localization of the high-molecular weight MAP2 isoforms with immunocytochemistry in the hippocampi of 1- to 21-day-old rats. Moreover, the temporal profile (from 30 min to 1 week) of MAP2 isoform reactivity to kainic acid-induced status epilepticus was studied in P9 rats. During development, the expression of the high-molecular weight MAP2 isoforms significantly increased, while the low-molecular weight isoforms decreased, the most prominent changes occurring during the second postnatal week. This developmental increase in the high-molecular weight MAP2 expression was also confirmed with immunocytochemistry, which showed increased immunoreactivity, particularly in the molecular layers of the dentate gyrus, and in CA1 and CA3 stratum radiatum. In 9-day-old rats, status epilepticus resulted in a rapid transient increase (about 210%) in the high-molecular weight MAP2 expression, without any effect on the low-molecular weight MAP2. Moreover, disturbed dendritic structure in the CA1 and CA3 stratum radiatum was manifested as formation of varicosities 3h after the kainic acid treatment. The strictly developmentally regulated MAP2 isoform expression suggests different functional roles for these proteins during the postnatal development in the rat hippocampus. Moreover, high-molecular weight MAP2s may play a role in nerve cell survival during cell stress.     

Alterations in the ultrastructure of cardiac autonomic nervous system triggered by crotoxin from rattlesnake (Crotalus durissus cumanensis) venom.

This study explored the toxic effects of crotoxin isolated from Crotalus durissus cumanensis venom on the ultrastructure of mice cardiac autonomic nervous system. Mice were intravenously injected with saline (control group) and crotoxin diluted in saline venom (study group) at a dose of 0.107 mg/kg mouse body weight. Samples from the inter-ventricular septum were prepared for electron microscopy after 6 h (G1), 12 h (G2), 24 h (G3) and 48 h (G4). The G1 group showed some cardiomyocyte with pleomorphic mitochondria. Capillary swollen walls, nerve cholinergic endings with depleted acetylcholine vesicles in their interior and other depletions were observed. A space completely lacking in contractile elements was noticed. The G2 group demonstrated a myelinic figure, a subsarcolemic region with few myofibrils and nervous cholinergic terminal with scarce vacuoles in their interior. The G3 group demonstrated a structure with a depleted axonic terminal, mitochondrias varying in size and enhanced electron density. In addition, muscular fibers with myofibrillar structure disorganization, a depleted nervous structure surrounded by a Schwann cell along with an abundance of natriuretic peptides, were seen. An amyelinic terminal with depleted Schwann cell and with scarce vesicles was also observed. Finally, axonic lysis with autophagic vacuoles in their interior and condensed mitochondria was observed in the G4 group. This work describes the first report of ultrastructural damage caused by crotoxin on mice cardiac autonomic nervous system.     

The survey of recent life experiences: A psychometric evaluation

Hitherto, various critics have claimed that the most commonly used measure for daily hassles is confounded with psychological well-being in both content and format. In order to circumvent such contamination, the Survey of Recent Life Experiences (SRLE) was developed by Kohn and MacDonald (1992). In the present study, the SRLE was psychometrically evaluated within a general sample of the Dutch population. Confirmatory factor analysis showed that, with the exception of one item, the original six-factor structure was strongly replicated. For five of six factors, internal consistency reliabilities proved satisfactory. It is argued that the rather low internal consistency reliability of the sixth factor may be improved if additional items are subjoined. In addition, yielded relationships between the SRLE and other variables were in accordance with previous research. It is therefore argued that the results are in support of cross-cultural construct validity of the SRLE. Future use within other Western European societies is recommended.     

Large deletions in the polycystic kidney disease 1 (PKD1) gene

Since identification of the genes mutated in patients with Autosomal Dominant Polycystic Kidney Disease, PKD1 and PKD2, a large number of different germ line mutations in both genes have been found by conventional PCR-based mutation detection methods. Nevertheless, in approximately 40% of the PKD1 families the disease-causing mutation remains to be elucidated. Complex germ line rearrangements are often not detectable by these standard diagnostic techniques. To detect large deletions in the PKD1 gene we performed Field Inversion Gel Electrophoresis (FIGE) followed by Southern blot analysis with probes selected in the unique and in the reiterated region of this gene. Our analysis revealed 4 deletions in 125 patients, indicating that large deletions in PKD1 are rare. Likely, patients with a deletion that also affects the neighbouring Tuberous Sclerosis Complex 2 (TSC2) gene will be diagnosed as patients with tuberous sclerosis. It was speculated that the exceptional polypyrimidine tract located in intron 21 and the small tract in intron 22, might play a role in the pathogenesis of ADPKD. Since this region is extremely difficult to amplify by PCR, we analysed the 5.8 kb BamHI fragment that contains the polypyrimidine tracts. We did not observe a disease-linked alteration although we detected two different rare variants either in PKD1 or in one of its homologues.     

Glucose deprivation depolarizes plasma membrane of cultured astrocytes and collapses transmembrane potassium and glutamate gradients

Primary cultures of astrocytes were used to investigate the effects of glucose deprivation on plasma membrane potential, on the respiration and on the energy status of these cells. Plasma membrane potential, as monitored with a cyanine dye, 3,3'-diethylthiadicarbocyanine, hyperpolarized by about 100% when glucose was added to substrate-deprived cells. The effect of glucose was prevented by iodoacetate or ouabain. In the absence of glucose, cellular adenosine triphosphate/adenosine diphosphate ratio was extensively reduced and pyruvate was unable either to restore energy status or to hyperpolarize the plasma membrane of astrocytes, although it was the preferential substrate for mitochondria within the cells. Glucose deprivation and inhibition of glycolysis or respiration in the presence of glucose caused dramatic decrease in transmembrane potassium ion and L-glutamate gradients. The gradients were not restored in the presence of pyruvate. Thus, aerobic glycolysis, rather than oxidation of pyruvate, is required to maintain maximal plasma membrane potential, adenosine triphosphate/adenosine diphosphate ratios as well as K+ and L-glutamate gradients. This evidence, together with the unresponsiveness of astrocyte respiration to ouabain, indicates a functional dissociation between energy dissipation at the plasma membrane and mitochondrial synthesis of adenosine triphosphate. The results are discussed with regard to the vulnerability of glia at low levels of blood glucose and the contribution of glial dysfunction to development of hypoglycaemic encephalopathy.     

Determination of GABA receptor-linked Cl- fluxes in rat cerebellar granule cells using a fluorescent probe SPQ

Gamma-Aminobutyric acid (GABA)-induced Cl- fluxes in cultured rat cerebellar granule cells were measured using the chloride-sensitive fluorescent probe SPQ (6-methoxy-N-(3-sulphopropyl)quinolinium) incorporated into the cells. The fluorescence of SPQ is quenched by Cl-ions. GABA and pentobarbitone increased the fluorescence of the probe when the Cl- gradient was directed outward by bathing cells, grown in the presence of GABA, in a low Cl- medium. Picrotoxin and bicuculline inhibited the response to GABA. The results suggest that SPQ is a suitable probe for measuring GABA-induced Cl- fluxes in living cells.     

High-intensity interval training can improve hand grip strength,inspiratory muscle,and quality of life in systemic sclerosis subjects

IntroductionSystemic sclerosis/scleroderma (SSc) is a chronic autoimmune disease with connective tissue, multi-organ, and multisystem involvement. The disease has three main characteristics, namely vasculopathy, fibrosis, and autoimmunity. The effect of high-intensity interval training (HIIT) in aerobic exercise on other rheumatic diseases has been studied, for example in patients with rheumatoid arthritis (RA) and juvenile idiopathic arthritis (JIA). The purpose of this work is to investigate the effectiveness of HIIT of aerobics exercise on improving the inspiratory muscle, quality of life and functional ability for systemic sclerosis subjects.Material and methodsThe study was conducted on patients with confirmed systemic sclerosis who met the inclusion criteria. The research was carried out for 12 months in the outpatient clinic and gait laboratory of the Department of Physical Medicine and Rehabilitation.ResultsAfter HIIT in aerobic exercise, we found significant changes in inspiratory muscle (SNIP values 45.67 [30.92] vs. 54.25 [22.71]), handgrip (13.14 [4.42] vs. 15.63 [4.08]), walking speed (184.70 [26.86] vs. 246.6 [12.30]), metabolic equivalent (3.53 [0.30] vs. 4.21 [1.25]) and Scleroderma-Specific Health Assessment Questionnaire Disability Index for all visual analog scale (VAS) domains except Disability Index. Exercise approaches are characterized by repeated cycles of exercise interrupted by rest. For a range of clinical conditions, HIIT in aerobic exercise is known to remedy blood vessel function.ConclusionsOur results suggest that HIIT in aerobic exercise has improved functional ability, respiratory muscle strength, and quality of life in SSc subjects. Training twice a week in a 12-week HIIT program is considered to be safe for this population. We have to consider internal and external factors that influenced the result. A larger sample and further exploration of the feasibility of combined exercise in SSc patients should be the focus for future research.