Normal values of sudomotor function assessed by electrochemical skin conductance in African population.

BACKGROUND: Dysfunction of small nerve fibers remains a major public health concern. Subjects suspected of having small nerve fiber damage need to undergo reliable tests to confirm the diagnosis. Sudomotor function test is a reliable noninvasive exploration for detecting peripheral neuropathies. Nevertheless, the normal reference values derived from the sudomotor function test are not known in the African population. The objective of this study was therefore to describe the normal values of Electrochemical Skin Conductance (ESC) measured by the sudomotor function test in healthy African subjects. MATERIALS AND METHODS: Between December 1st, 2021 and May 31st, 2022, ESC was measured in 475 healthy subjects (median age: 42 [31-53] years, 46% men) using a sudomotor function test, in the hands and feet. Investigators proposed the examination and received participants' consent; demographic, anthropometric, biological, and clinical data were obtained before the test. Data on 475 healthy study participants who underwent sudomotor function testing was collected and analyzed. The sociodemographic (age, sex), anthropometric (weight, height, waist circumference, body mass index), diastolic blood pressure, systolic blood pressure, heart rate, and electrochemical skin conductances of the hands and feet were assessed. RESULTS: ESC values were statistically higher in men compared to women (right hand ESC: 70 [60-78] versus 63 [53-72], left hand ESC: 72 [61-80] versus 68 [57-75], right foot ESC: 77 [82-99] versus 72 [64-79], ESC left foot: 76 [68-82] versus 72 [62-78] respectively). ESC values were significantly inversely correlated with age (right hand ESC: r=-0.12, P=0.006; left hand ESC: r=-0.11, P=0.01; right foot ESC: r=-0.37, P<0.0001; ESC left foot: r=-0.38, P<0.0001). ESC values measured in feet were significantly inversely correlated with body mass index (right foot r=-0.22, P<0.0001; left foot r=-0.21, P<0.0001). CONCLUSION: This study reports normal reference values for ESCs according to age and gender in the healthy African population. Progressive decrease in ESC with aging is confirmed by our data. The value of ESC seems lower in the African population than in other reported ethnicities. This finding needs to be further explored in additional studies.

T-tubules: a key structure of cardiac function and dysfunction.

The sarcolemmal membrane of mammalian ventricular cardiomyocytes are characterised by the presence of invaginations called transverse tubules (T-tubules) which constitute a network, the transverse tubule system. T-tubules occur at the Z line as transverse elements with also longitudinal extensions. While the existence of T-tubules has been known since a long time, recent studies have suggested their structure and function can be more complex than previously believed. Many of the proteins involved in excitation-contraction coupling process are concentrated within the T-tubular network, suggesting T-tubules are a highly specialized membrane system. Thus, T-tubules are a key determinant of cardiac cell function. The fundamental role of T-tubules is emphasized by changes in their structure and protein expression occurring during pathologies such as cardiac hypertrophy and heart failure. This review summarizes recent studies which highlight the key-role of the T-tubules in the regulation of cardiac function. Changes observed in pathological conditions are also discussed.

Quantification of calcium entry at the T-tubules and surface membrane in rat ventricular myocytes.

The action potential of cardiac ventricular myocytes is characterized by its long duration, mainly due to Ca flux through L-type Ca channels. Ca entry also serves to trigger the release of Ca from the sarcoplasmic reticulum. The aim of this study was to investigate the role of cell membrane invaginations called transverse (T)-tubules in determining Ca influx and action potential duration in cardiac ventricular myocytes. We used the whole cell patch clamp technique to record electrophysiological activity in intact rat ventricular myocytes (i.e., from the T-tubules and surface sarcolemma) and in detubulated myocytes (i.e., from the surface sarcolemma only). Action potentials were significantly shorter in detubulated cells than in control cells. In contrast, resting membrane potential and action potential amplitude were similar in control and detubulated myocytes. Experiments under voltage clamp using action potential waveforms were used to quantify Ca entry via the Ca current. Ca entry after detubulation was reduced by approximately 60%, a value similar to the decrease in action potential duration. We calculated that Ca influx at the T-tubules is 1.3 times that at the cell surface (4.9 vs. 3.8 micromol/L cytosol, respectively) during a square voltage clamp pulse. In contrast, during a cardiac action potential, Ca entry at the T-tubules is 2.2 times that at the cell surface (3.0 vs. 1.4 micromol/L cytosol, respectively). However, more Ca entry occurs per microm(2) of junctional membrane at the cell surface than in the T-tubules (in nM/microm(2): 1.43 vs. 1.06 during a cardiac action potential). This difference is unlikely to be due to a difference in the number of Ca channels/junction at each site because we estimate that the same number of Ca channels is present at cell surface and T-tubule junctions ( approximately 35). This study provides the first evidence that the T-tubules are a key site for the regulation of action potential duration in ventricular cardiac myocytes. Our data also provide the first direct measurements of T-tubular Ca influx, which are consistent with the idea that cardiac excitation-contraction coupling largely occurs at the T-tubule dyadic clefts.

cGMP-mediated inhibition of cardiac L-type Ca(2+) current by a monoclonal antibody against the M(2) ACh receptor.

The effects of a monoclonal antibody (B8E5) directed against the second extracellular loop of the muscarinic M(2) receptor were studied on the L-type Ca(2+) currents (I(Ca,L)) of guinea pig ventricular myocytes using the whole cell patch-clamp technique. Similar to carbachol, B8E5 reduced the isoproterenol (ISO)-stimulated I(Ca,L) but did not significantly affect basal I(Ca,L). Atropine blocked the inhibitory effect of B8E5. The electrophysiological parameters of ISO-stimulated I(Ca,L) were not modified in presence of B8E5. Inhibition of I(Ca,L) by B8E5 was still observed when intracellular cAMP was either enhanced by forskolin or maintained constant by using a hydrolysis-resistant cAMP analog (8-bromoadenosine 3',5'-cyclic monophosphate) or by applying the phosphodiesterase inhibitor IBMX. The effect of B8E5 was mimicked by 8-bromoguanosine 3',5'-cyclic monophosphate, a potent stimulator of cGMP-dependent protein kinase, and prevented by a selective inhibitor of nitric oxide-sensitive guanylyl cyclase [1H-(1,2,4)oxadiazolo[4,3-a]quinoxaline-1-one]. These results indicate that the antibody B8E5 inhibits the beta-adrenergic-stimulated I(Ca,L) through activation of the M(2) muscarinic receptor and further suggest that the antibody acts not via the classical pathway of decreasing intracellular cAMP, but rather by increasing cGMP.

Inhibitory activity of antibodies against the human atrial 5-HT(4)receptor.

Antibodies directed against the second extracellular loop of G protein-coupled receptors have been shown to exert "agonist-like" activities. In order to test the hypothesis that this is a general phenomenon, antibodies were raised in rabbits against a synthetic peptide corresponding to the second extracellular loop of the newly sequenced human cardiac 5-HT(4)receptor. The antibodies were affinity-purified and shown to recognize the 5-HT(4)receptor in immunoblots of Chinese hamster ovary (CHO) cells expressing the receptor. The antibodies had no intrinsic effect but could depress the activation of L -type calcium channel induced by serotonin in human atrial cells. This antagonist-like effect was exerted both by intact IgG and by Fab fragments. These results are physiologically important since it has been shown that the 5-HT(4)receptor could be a target for autoantibodies in mothers at risk of giving birth to children with neonatal atrio-ventricular block.

Anti-SSA/Ro52 autoantibodies blocking the cardiac 5-HT4 serotoninergic receptor could explain neonatal lupus congenital heart block.

The 52-kDa SSA/Ro (Ro52) ribonucleoprotein is an antigenic target strongly associated with the autoimmune response in mothers whose children develop neonatal lupus and congenital heart block. When sera from patients with systemic lupus erythematosus were used as autoimmune controls in an enzyme immunoassay to screen for antibodies against the human serotoninergic 5-HT4-receptor, a high correlation was found between the presence of anti-Ro52 protein antibodies in such sera and antibodies reacting with a synthetic peptide, corresponding to the second extracellular loop of the human 5-HT4 receptor (amino acid residues 165-185). Homology scanning between the 5-HT4 peptide and the sequence of the Ro52 protein indicated two potential common epitopes located between residues 365 and 396 of the Ro52 protein. Cross-reactivity was found between the peptide derived from the 5-HT4 receptor, and a peptide corresponding to residues 365-382 of the Ro52 protein. Autoantibodies, affinity-purified on the 5-HT4 receptor peptide, specifically recognized both the Ro52 protein and the 5-HT4 receptor protein in immunoblots. The affinity-purified antibodies antagonized the serotonin-induced L-type Ca channel activation on human atrial cells. This effect could explain the electrophysiological abnormalities in neonatal lupus.