Autoantibodies against the serotoninergic 5-HT4 receptor and congenital heart block: a reassessment

The specificity of autoantibodies against the serotoninergic 5-HT4 receptor in congenital heart block has led to conflicting observations. In order to clarify the situation, a collaborative effort was undertaken to discover the reasons for these discrepancies and to reassess the importance of such autoantibodies by making use of the Research Registry for Neonatal Lupus. Sera from 128 patients (101 anti-SSA/Ro52 positive mothers among which 74 have children with congenital heart block (CHB), 9 anti-SSA/Ro52 negative patients of which 1 had a child with heart block and 18 healthy donors) were assessed in a single blind test using an ELISA coated with a 5-HT4 receptor-derived peptide. Discrepancies between previous observations in our two groups could be ascribed to small differences in the set up of the assay. Of the 75 sera from mothers of children with CHB, 12 were reactive with the 5-HT4 peptide. Four sera among which three were from 35 Ro52 negative mothers without affected children as well as 2 in the 18 controls were positive. Interestingly, in 1 mother with an isolated child with CHB but who had no detectable anti-SSA/Ro52 antibodies and 1 mother with a child with a structural heart block and no detectable antibodies to any component of SSA/Ro, reactivity with the 5-HT4 receptor was noted. While 5-HT4 receptor autoantibodies do not have the predictive value of anti-Ro52 autoantibodies, the presence of these antibodies in a minor subset of mothers whose children have CHB suggests an additional risk factor which may contribute to the pathogenesis of disease.     

Disturbances of Haemostasis in Diabetes Mellitus

Diabetes mellitus is associated with disturbances in haemostasis that could contribute to the development of thrombotic complications.The present study was undertaken to determine the behavior of coagulation variables and fibrinolytic system in diabetes mellitus. Forty five diabetic patients and forty five matched controls were evaluated by doing the following haemostatic parameter, prothrombin time, partial thromboplastin time, thrombin time, coagulation factors assay II, VII, IX, & plasma fibrinogen, ADP-induced platelet aggregation, protein C, a₂- antiplasmin, PAI and FDPs. Generally diabetic patients have high levels of fibrinogen, a₂- antiplasmin, & PAI and lower level of protein C. Other haemostatic parameters did not show statistically significant difference between diabetic patients and control group. Significantally elevated levels of PAI, a₂- antiplasmin together with low protein C level in diabetic patients may result in the disturbance of haemostatic balance favoring thrombotic events. Conclusion: High levels of plasma fibrinogen, a₂A- antiplasmin with low plasma protein C activity could lead to a prothrombotic tendency in insulin dependent diabetic patients. Moreover, in non-insulin dependent diabetic patients, the above mentioned parameters together with high levels of ADP-induced platelet aggregation and plasminogen activator inhibitor may increase the risk of thrombotic complications. Obesity can be considered as an additional risk factor for development of thrombosis in diabetic patients.     

Chryseomonas luteola identified as the source of serious infections in a Moroccan University Hospital

Chryseomonas luteola has only rarely been reported as a human bacterial pathogen. It has been shown that this organism in particular affects patients with health or indwelling disorders. Most reported cases showed septicemia, meningitis, endocarditis, or peritonitis. Two C. luteola infections observed in Morocco are described in the present study.     

Temperature-mediated heteroduplex analysis for detection of pncA mutations associated with pyrazinamide resistance and differentiation between Mycobacterium tuberculosis and Mycobacterium bovis by denaturing high- performance liquid chromatography

The goal of this study was to apply temperature-mediated heteroduplex analysis using denaturing high-performance liquid chromatography to identify pyrazinamide (PZA) resistance in Mycobacterium tuberculosis isolates and simultaneously differentiate between M. tuberculosis and Mycobacterium bovis. Features that contributed to an optimal assay included the use of two different reference probes for the pncA gene targets from wild-type M. tuberculosis and wild-type M. bovis, optimization of the column temperature, increasing the starting concentration of the elution buffer, and reducing the rate of elution buffer increase (slope). A total of 69 strains were studied, including 48 wild-type M. tuberculosis strains (13 were PZA-resistant strains) and 21 M. bovis strains (8 were BCG strains). In all isolates tested, wild-type M. tuberculosis generated a single-peak pattern when mixed with the M. tuberculosis probe and a double-peak pattern with the M. bovis probe. In contrast, all M. bovis isolates generated a double-peak pattern when mixed with the M. tuberculosis probe and a single-peak pattern with the M. bovis probe. PZA-resistant mutant M. tuberculosis isolates generated characteristic patterns that were easily distinguishable from both wild-type M. tuberculosis and M. bovis isolates. Chromatographic patterns generated by the two reference probes allowed the rapid detection of PZA resistance with the simultaneous ability to distinguish between M. tuberculosis and M. bovis. This approach may allow the detection of drug resistance-associated mutations, with potential application to clinical and epidemiological aspects of tuberculosis control.     

Synthetic organic hard capsule colouring agents: in vitro effect on human true and pseudo-cholinesterases

Hard capsules are made of pure gelatin and small quantities of additives, including colouring agents permitted for use in food. In this study, the effects of three colouring agents (sunset yellow, quinoline yellow and erythrosine) on true and pseudo-cholinesterases (ChE) are assessed in erythrocytes and plasma, respectively. Results indicated that the synthetic compounds affected both true and pseudo ChE activity. The concentration of sunset yellow which caused 50% inhibition (IC50) of true ChE was about 64% that of pseudo-ChE; for erythrosine, IC50 was approximately the same for both true and pseudo-ChE; and for quinoline yellow, IC50 for true ChE was 25% of pseudo-ChE, although its effect on both true and pseudo-ChE was greater than seen with the other two dyes. Inhibitions of both true and pseudo-ChE were of mixed type (competitive and non-competitive). The enzyme-inhibitor dissociation constant (Ki) indicated that quinoline yellow was most potent and erythrosine was least potent out of the three compounds. Inhibition of both true and pseudo-ChE by each of the three dyes was abolished by dialysis, indicating that the effects were reversible.     

Intestinal antilectin immunoglobulin A antibody response and immunity to Entamoeba dispar infection following cure of amebic liver abscess

We followed 93 subjects with amebic liver abscess (ALA) and 963 close associate controls at 3-month intervals for 36 months to characterize intestinal and humoral antibody responses to the amebic galactose-inhibitable lectin and to determine whether immunity developed to Entamoeba histolytica or Entamoeba dispar infection following cure of ALA. We found that ALA subjects had a higher prevalence and level of intestinal antilectin immunoglobulin A (IgA) and serum anti-LC3 (cysteine-rich recombinant lectin protein) IgA and IgG antibodies, P < 0.01 and P < 0.05, respectively, compared to controls. The intestinal antilectin IgA antibody response was sustained over a longer time period in ALA subjects (71.8% remained positive at 18 months and 52.6% at 36 months, P < 0.001 compared to 17.6% and 10.3% of controls, respectively). ALA subjects were highly immune to E. dispar infection throughout the study (0% infected at 6 and 36 months, compared to 6.5% and 4.9% of control subjects, respectively, P < 0.05). Upon entry into the study, 6.3% of ALA subjects were infected with E. histolytica; the incidence of new E. histolytica infections in controls (as determined by culture) was too low (1.4%) to determine whether ALA subjects exhibited immunity to new infections. We found that stool cultures every 3 months markedly underestimated the occurrence of new E. histolytica infections, as 15.3% of controls seroconverted after 12 months of follow-up. Unfortunately, under the field conditions present in Durban, South Africa, enzyme-linked immunosorbent assay for detection of lectin antigen in stool yielded unreliable results. In summary, subjects cured of ALA exhibited sustained mucosal IgA antibody responses to the amebic galactose-inhibitable lectin and a high level of immunity to E. dispar infection. Determination of immunity to E. histolytica following cure of ALA will require the use of more sensitive and reliable diagnostic methods.     

Median frequency of the myoelectric signal

Summary A study was performed to investigate the changes that occur in the median frequency of the myoelectric signal during local ischemia or reduction of intramuscular temperature produced by surface cooling. Data was obtained from experiments which involved the first dorsal interosseous muscle of 10 female and 16 male subjects. These subjects were asked to perform isometric constant-force abduction contractions of the index finger at 20% and 80% of maximal voluntary contraction level. The initial median frequency (IMF) of the myoelectric signal during the first 0.5 s of contraction was calculated. Results showed a significant reduction of the IMF in contractions performed under ischemic conditions; upon release, the IMF recovered quickly. At 80% maximal voluntary level of contraction, a greater decrease of the IMF was recorded. Similar results were demonstrated during reduction of intramuscular temperature with gradual recovery of the IMF after cooling. These results demonstrate that the median frequency of the myoelectric signal displays behavior similar to that reported for conduction velocity and this is consistent with the notion that accumulation of metabolic byproducts in muscle tissue causes a decrease in the conduction velocity of the muscle fibers.Dr. R. Merletti was on a leave of absence from the Institute of Electrical Engineering, Politecnico di Torino, Italy     

Thermal transition and frequency response of the dielectric behaviour of some cellulosic substances

The real part of the relative permittivity ?′_r of some cellulosic materials contaminated with commercial impurities and NaCl (determined as weight percent ash contents), and that of the purified fibres, as well as their hydrolysed residues, free of such contaminations, was measured in the frequency range 10² Hz ≤ f ≤ 10⁵ Hz and in the temperature range 283,15 K ≤ T ≤ 333,15 K. Additional measurements of the imaginary part of relative permittivity ?″_r have been carried out in the same frequency and temperature ranges for hydrocellulose samples free of inorganic impurities and contaminated with NaCl. The results obtained show that the variation with temperature of the relative permittivity of the samples examined exhibits a change in slope in the vicinity of 303,15 K except for hydrocellulose samples free of impurities. They further show that the presence of inorganic contaminations has a significant effect on the magnitude of the relative permittivity and its variation with temperature, particularly at low frequency. In the case of samples contaminated with inorganic impurities, interfacial polarization is responsible for the anomalous dielectric behaviour observed with the examined samples.     

Endocrinology: The effect of growth hormone on granulosa cell function during in-vitro fertilization

The effect of growth hormone addition to human menopausal gonadotrophin(HMG), after pituitary down-regulation, on granulosa cell function,in in-vitro fertilization (IVF) was evaluated. Growth hormoneor placebo were added in a prospective, randomized and double-blindmanner to an existing IVF stimulation protocol. Forty-two normalovulatory women (38 years old) with mechanical factor infertilityand normal male factor were included in the study. Gonadotrophin-releasinghormone agonist (GnRHa) was given from day 21 of the previouscycle until human chorionic gonadotrophin (HCG) administration.Follicular stimulation with HMG was started after pituitarydown-regulation. Growth hormone 12 IU/day or placebo were administeredon alternate days, beginning day 1 until day 7 of HMG treatment.Granulosa cell function was evaluated, in all patients, by follicularfluid levels of ovarian steroids and insulin-like growth factor-I(IGF-I). In 14 patients, chosen arbitrarily granulosa luteincells were cultured in the presence and absence of additionalHCG. Follicular fluid levels of oestradiol, progesterone, testosteroneand IGF-I were similar in both growth hormone and placebo groups.Basal and post-HCG levels of oestradiol and progesterone didnot differ significantly between the two groups of granulosalutein cell cultures. We conclude that after pituitary down-regulation,in-vivo administration of growth hormone with HMG in young ovulatorywomen does not seem to affect granulosa cell function when comparedto the administration of HMG alone.