Extensive depolarization and lack of recovery of leech Retzius neurons caused by 2,4 diaminobutyric acid.

In this paper we present, for the first time, a detailed account of electrophysiological effects of 2,4-diaminobutyric acid (2,4-DABA). 2,4-DABA is a neurotoxic non-protein amino acid produced by Cyanobacteria with a possible link to neurodegenerative disorders in animals and humans. Intracellular recordings were performed on Retzius nerve cells of the leech Haemopis sanguisuga using glass microelectrodes filled with 3 mol/L KCl. Our results show that 2,4-DABA is an excitatory amino acid, causing membrane depolarization in a concentration-dependent manner. The most prominent depolarizations of 39.63±2.22 mV and 47.05±4.33 mV, induced by 5×10-3 and 10-2 mol/L 2,4-DABA respectively, are several times larger than maximal depolarizations induced by either Glutamate, Aspartate, ß-N-methylamino-alanine (BMAA) or ß-N-oxalylamino-alanine (BOAA) on our model. These 2,4-DABA induced depolarizations evolve through two distinct stages, which is a novel phenomenon in electrical cell activity upon application of an excitatory amino acid, at least on our model. Involvement of two separate mechanisms, suggested by the two stage phenomenon, is discussed in the paper. We also provide evidence that 2,4-DABA induces irreversible functional disturbances in neurons in a concentration-dependent manner, since only half of the cells recovered normal electrical activity after application of 5×10-3 mol/L 2,4-DABA, and none recovered after application of 10-2 mol/L 2,4-DABA. Effects of both L-2,4-DABA and DL-2,4-DABA were tested and are not significantly different.

Is lymphocytic thyroiditis a unique type or merely a type of Hashimoto's thyroiditis?

AIM: Objective of the study was to clarify the role of apoptosis in the pathogenesis of lymphocytic thyroiditis (LT) and the existence of difference between Hashimoto's thyroiditis (HT) and LT. METHODS: We evaluated levels of antithyroglobulin and antithyroperoxidase antibodies, the apoptosis by in situ Cell Death Detection-TUNEL and the expression of Bcl2 and Bax by immunohistochemistry in thyroid tissues from 16 patient with HT, 10 with LT and 10 with euthyroid goiter-EG (control group). RESULTS: It was found that apoptosis of thyrocytes in HT (mean 3.05%, SD 1.29%) and LT (mean 2.70%, SD 1.17%) was statistically significantly higher than EG (mean 0.56%, SD 0.23%), but the difference in the percentage of thyrocytes between HT and LT was not statistically significant. In HT the percentage of apoptotic infiltrating lymphocytes (mean 0.59%, SD 0.23%) was smaller than in EG (mean 2.26%, SD 1.42%), but it showed no significant difference in comparison to LT. The expression of Bax in infiltrating lymphocytes in HT (mean 0.72%, SD 0.34%) was statistically significantly higher than LT (mean 0.11%, SD 0.06%). The level of thyroglobulin was lower in HT compared to LT (P<0.01) and compared to EG (P<0.01). The level of antithyroglobulin/antithyroperoxidase antibodies was higher in HT compared to LT (P<0.01) and compared to EG (P<0.01). There was no statistically significant difference in the level of thyroglobulin and level of antibodies between LT and EG. CONCLUSION: These results suppose that apoptosis represents one of significant mechanisms in the pathogenesis of both HT and LT and that LT probably differs from HT.

Peritubular membrane potential in kidney proximal tubular cells of spontaneously hypertensive rats.

Peritubular membrane potential in kidney proximal tubular cells of spontaneously hypertensive rats (SHR-Okamoto strain adult rats) was measured with conventional 3 mol KCl microelectrodes, in vivo. Peritubular cell membrane potential was not different in SHR (-66.5 ± 0.7 mV) as compared with normotensive control Wistar rats (-67.5 ± 1.2 mV). To test the effects of possible altered sodium membrane transport in SHR on proximal tubule peritubular membrane potential, we allowed SHR and control rats to drink 1% NaCl for two weeks. Again, proximal tubule peritubular membrane potential was not different in SHR on 1% NaCl (-67.0 ± 1.0 mV) as compared with control rats on 1% NaCl (-64.7 ± 1.3 mV). From these results we concluded that peritubular membrane potential in kidney proximal tubular cells of SHR was not different from normotensive Wistar control rats, and if some alteration of sodium transport in kidney proximal tubular cells of SHR could exist, that was not possible to evaluate from the measurements of peritubular membrane potential in kidney proximal tubular cells.

Differential expression of KCNQ1 K+ channel in tubular cells of frog kidney.

The aim of this study was to evaluate KCNQ1 K+ channel expression in the frog kidney of Rana esculenta. KCNQ1 K+ channel, also known as KvLQT1, is the pore forming a-subunit of the IKs K+ channel, a delayed rectifier voltage-gated K+ channel, which has an important role in water and salt transport in the kidney and gastrointestinal tract. The expression of KCNQ1 K+ channel along tubular epithelium differs from species to species. In the present study the expression of KCNQ1 K+ channel in the frog kidney has been demonstrated by immunohistochemistry. The presence of KCNQ1 K+ channel was demonstrated in the epithelial cells of distal convoluted tubule and collecting duct. However, the pattern of expression of KCNQ1 K+ channel differs between distal convoluted tubules and collecting duct. All epithelial cells of distal convoluted tubules revealed basolateral expression of KCNQ1 K+ channel. On the contrary, only the single cells of collecting duct, probably intercalated cells, showed diffuse cell surface staining with antibodies against KCNQ1 K+ channel. These findings suggest that KCNQ1 K+ channel has cell-specific roles in renal potassium ion transport.

Serum beta2-microglobulin as a marker of congenital toxoplasmosis and cytomegalovirus infection in preterm neonates.

BACKGROUND: Fetal serum beta(2)-microglobulin (beta(2)M) has been reported as a reliable indicator of fetal infectious diseases. OBJECTIVES: To evaluate serum beta(2)M as a marker of congenital toxoplasmosis or cytomegalovirus (CMV) infection in neonates. METHODS: beta(2)M was retrospectively measured in 72 neonatal serum samples from preterm neonates. Of these, 32 originated from neonates with serological evidence of congenital toxoplasmosis (n = 12) and CMV infection (n = 20), while 40 samples from neonates in which both infections were serologically excluded served as controls. beta(2)M levels were compared between the infection and control groups. RESULTS: Mean (+/-SEM) beta(2)M levels were significantly higher in the groups of neonates infected with Toxoplasma (5.64 +/- 0.61 mg/l) (p = 0.014) and CMV (6.06 +/- 0.66 mg/l) (p < 0.0001) than in the control group (3.80 +/- 0.2). Against the cut-off level of 5 mg/l, beta(2)M was normal in 36 of the 40 uninfected neonates examined, indicating a specificity of 90%. In contrast, it was elevated in 66.7% (8/12) and 65% (13/20) of the Toxoplasma and CMV-infected neonates, respectively, indicating an overall sensitivity of 66%. CONCLUSIONS: In the absence of urogenital disorders, an increase in beta(2)M in neonates is likely to be infection-induced. We showed that serum beta(2)M is increased in congenital toxoplasmosis and CMV infection in the first weeks of life.