Functional expression and properties of the human skeletal muscle sodium channel

Full-length deoxyribonucleic acid, complementary (cDNA) constructs encoding the-subunit of the adult human skeletal muscle Na⁺ channel, hSkM1, were prepared. Functional expression was studied by electrophysiological recordings from cRNA-injectedXenopus oocytes and from transiently transfected tsA201 cells. The Na⁺ currents of hSkM1 had abnormally slow inactivation kinetics in oocytes, but relatively normal kinetics when expressed in the mammalian cell line. The inactivation kinetics of Na⁺ currents in oocytes, during a depolarization, were fitted by a weighted sum of two decaying exponentials. The time constant of the fast component was comparable to that of the single component observed in mammalian cells. The block of hSkM1 Na⁺ currents by the extracellular toxins tetrodotoxin (TTX) and -conotoxin (CTX) was measured. The IC₅₀ values were 25 nM (TTX) and 1.2 M (CTX) in oocytes. The potency of TTX is similar to that observed for the rat homolog rSkM1, but the potency of CTX is 22-fold lower in hSkM1, primarily due to a higher rate of toxin dissociation in hSkM1. Single-channel recordings were obtained from outside-out patches of oocytes expressing hSkM1. The single-channel conductance, 24.9 pS, is similar to that observed for rSkM1 expressed in oocytes.     

Molecular mechanisms of virus-induced carcinogenesis: the interaction of viral factors with cellular tumor suppressor proteins

Accumulating evidence indicates that tumor viruses represent a major etiological factor in a significant portion of human cancers. These cancers include human papillomavirus induced anogenital cancers, hepatitis B and C virus associated hepatocellular carcinomas, nasopharyngeal carcinomas and lymphomas linked to Epstein-Barr virus infection, and human T cell leukemia virus associated adult T cell leukemias. This review summarizes the recent progress made in understanding the molecular mechanisms of viral carcinogenesis, with a particular focus on the interaction of viral factors with cellular tumor suppressor proteins. The functional inactivation of tumor suppressor proteins may represent a common strategy by which several tumor viruses contribute to malignant cell transformation.Abbreviations EBV Epstein-Barr virus - E6AP E6-associated protein - HBV Hepatitis B virus - HCC Hepatocellular carcinoma - HPV Human papillomavirus - HTLV Human T cell leukemia virus - pRb Retinoblastoma protein - RB Retinoblastoma - SV40 Simian virus 40     

Responses to eccentric rotation in two space-bound subjects

Summary Two subjects were rotated eccentrically in the manner described previously. In contrast to a normal control group, settings of a luminous line to the subjective vertical were almost unrelated to the gravitoinertial vector before, and totally so shortly after, space flight. Only 3 days postflight did a clear relation to the gravitoinertial vector re-establish itself in the one subject who actually flew. The correspondence became normal 5 days after the flight. Since there were no clinical abnormalities evident in the subjects, it is suggested that both subjects suppressed their vestibular information, presumably as an effect of vestibular deconditioning training before the flight. In addition, as a consequence of the flight experience one subject continued to ignore it several days after the flight.Abbreviations DARA Deutsche Agentur für Raumfahrtangelegenheiten - DLR Deutsche Forschungsanstalt für Luft-und Raumfahrt - ESA European space agency - ESTEC European space research and technology centre - G _z G-Units (9.81 m/s²) in z-axis direction - L-xx Launch minus xx days - R+xx Return - VESTA ESA Neurovestibular experiment facility     

Somatotopic organization along the central sulcus, for pain localization in humans, as revealed by positron emission tomography

Regional cerebral blood flow was measured with positron emission tomography (PET) in six healthy volunteers at rest and during experimentally induced, sustained cutaneous pain on the dorsum of the right hand or on the dorsum of the right foot. Pain was inflicted by intracutaneous injection of capsaicin, providing a mainly C-fibre nociceptive stimulus. Statistical analysis showed significant activations along the central sulcus (SI) area when comparing pain in the hand to pain in the foot. Separate comparison of both pain states to a baseline revealed different locations along the central sulcus for hand pain and foot pain. The encountered differences are consistent with what is previously known about the somatotopics of non-painful stimuli. When comparing painful stimuli to baseline, the contralateral anterior cingulate gyrus, the ipsilateral anterior insular cortex and the ipsilateral prefrontal cortex were implicated. The results are consistent with an involvement of SI in the spatial discrimination of acute cutaneous pain. Received: 17 October 1996 / Accepted: 12 May 1997     

Visual motion perception from stimulation of the human medial parieto-occipital cortex

Summary Visual phenomena evoked by direct electrical stimulation of extrastriate cortex were observed in 30 epileptic patients as part of a presurgical investigation. An incremental sequence of low-level bipolar stimulation trains was delivered at medial and lateral pairs of contacts of stereotaxically-implanted multilead intracerebral electrodes in parietal, occipital and posterior temporal regions. Diffusion of stimulus afterdischarges was monitored by electrodes in temporal and frontal lobes and by the non-stimulated contacts of the stimulated electrode. Localized stimulations evoked few visual phenomena. The strongest anatomo-perceptual correlation was found for stimulation in the medial parieto-occipital fissure which evoked visual motion phenomena in all three patients stimulated in that region. The evoked motion perceptions were not associated with eye movements or any particular localization of the epileptic focus. These perceptions were only evoked once outside of the medial PO region at the 61 sites examined. The results suggest that the medial parieto-occipital region is closely linked to the human visual motion processing system.     

Neuropathology of the Central Nervous System in Acquired Immune Deficiency Syndrome (AIDS) in Japan

The neuropathologiesl features of the central nervous system in IS autopsy cases of Japanese male with AIDS were reported. Nine patients had various histological changes including a variety of opportunistic infections in six patients (40%), primary malignant lymphoma of the brain in two (13%), AIDS encephalopathy in four (27%) and vacuolar myelopathy in one (7%). Usually, these pathological changes were present concomitantly. AIDS encephalopathy was characterized by infiltration of mono and multinucleated cells and myelin pallor with astrogliosis located predominantly in the cerebral white matter and subcortical gray matter. Furthermore, unevenly distributed neuronal loss of the cerebral cortex was apparent in one case. Diffuse astrocytosis of the gray matter out of proportion to neuronal loss was also an outstanding finding in another case. The present study suggested that not only the white matter changes but also gray matter alterations might be the morphological substrates of AIDS encephalopathy.     

The sinusoidal barrier in alcoholic patients without liver fibrosis

Summary Alcohol induces morphological changes in the endothelial and perisinusoidal cells at the fibrotic stage of alcoholic liver diseases. Directly or indirectly, through hemodynamic disturbances linked to the enlargement of steatotic hepatocytes, alcohol may modify this barrier before the onset of fibrosis. Liver biopsies were obtained from control and from alcoholic patients and perfusion-fixed. Volume and surface densities of endothelial cells, perisinusoidal cells and their processes were measured. Liver histology was normal in the 2 groups except for steatosis in the alcoholics. Volume densities represented 8.2%, 4.7% and 3.2% of the sinusoid in controls for endothelial cells, perisinusoidal cells and their processes whereas surface densities represented respectively 0.5, 0.23, 0.21 m²/cm³ of sinusoid. Morphometric values were not significantly different in the alcoholic patients. In none of the alcoholic patients did fine morphological studies of sinusoidal cells give any indication of the possible evolution of the alcoholic disease towards fibrosis. These results indicate that in the group of patients studied, alcohol, before the fibrotic stage, did not significantly alter the sinusoidal barrier.     

Transcranial magnetic stimulation

We investigated the effects of unilateral cold-water vestibular stimulation on healthy subjects' performance in two cognitive tasks known to be differentially mediated by the two cerebral hemispheres. In a first experiment (right-hemisphere task), subjects memorized object-location associations while being stimulated with cold water in the left ear or right ear or not at all (control group). In the second experiment (left-hemisphere task), subjects memorized a list of sequentially presented function words while being stimulated in the same manner as the subjects in the first experiment. A recall phase followed each encoding phase. In the first experiment, subjects who had been stimulated in the left ear recalled the object locations significantly faster than subjects who had been stimulated in the right ear and those in the control group. The second experiment yielded the reverse pattern: correct word recognition was faster for subjects who had been stimulated in the right ear than for subjects stimulated in the left ear and those of the control group. We suggest that unilateral caloric stimulation leads to a selective activation of contralateral cerebral structures and speeds up cognitive processes mediated by these structures. These results are discussed with respect to findings in neglect patients and functional-imaging studies in healthy subjects.     

Response of the human triceps surae muscle to electrical stimulation during varying levels of blood flow restriction

The changes in muscle force associated with varying degrees of lower-limb ischaemia were investigated. Isometric torque production by the triceps surae muscle was measured during a 5-min continuous train of 2-Hz electrical stimulation in six healthy young adults under different thigh cuff occlusion pressures. The reproducibility of this protocol when performed under complete ischaemia (tested five times over a 2-week period) was assessed as having a coefficient of variation (CV) for fatigue (end/initial force) of [mean (SEM) 12 (1)%; n=5]. This compares favourably with that obtained for maximum voluntary contraction torque [CV 9 (1)%]. In six subjects, triceps surae muscle fatigue was assessed under thigh cuff pressures of 0, 6.7 kPa (50 mmHg, venous occlusion) and 28 kPa (210 mmHg, complete ischaemia), as well as two intermediate levels of occlusion that were established by cuff pressures of 13.4 (0.5) and 20.3 (1.1) kPa [103 (4) and 152 (8) mmHg, respectively]. These corresponded to ankle-brachial pressure indices of 1.3 and 0.8, respectively when subjects were seated, or 0.8 and 0.36 when supine. With undisturbed lower-leg circulation, force potentiated steadily over the 5 min of stimulation such that the final force was 135 (8)% of the initial value. With complete ischaemia, force fell to 47 (2)% of the initial value. Stimulation under thigh occlusion pressures of 6.7, 13.4 and 20.3 kPa elicited intermediate levels of reduction in force, graded according to the increasing restriction of perfusion. The results show that low-force twitch contractions, which themselves do not occlude blood flow, are extremely sensitive to impaired perfusion and may represent a viable alternative to established methods of muscle performance assessment in patients with blood flow insufficiency. Accepted: 5 November 1999     

HIV－1Protease在大肠杆菌中的高表达

艾滋病的致病因子为人免疫缺陷病毒。该病毒的蛋白酶在病毒复制和成熟中具有决定性的意义。由于目前国内外尚未获得艾滋病病毒蛋白酶高效表达的重组子及简便的活性检测系统，限制了它的研究与应用。本文将用ＰＣＲ技术修饰的ＨＩＶＰｒ基因克隆入原核高效表达载体ｐＴＴＱ１８的ＥｃｏＲⅠ和ＨｉｎｄⅢ酶切位点之间，并用豆芽核酸酶将ＥｃｏＲⅠ的粘端削平，构建了读框正确的表达载体，ＩＰＴＧ诱导表明，该重组子在大肠杆菌中获得了高表达，激光扫描结果表明：重组的ＨＩＶＰｒ占细菌总蛋白８．９％以上。