Effects of histamine and related compounds on the release of immunoreactive thyrotropin-releasing hormone from the rat stomach in vitro

Effects of histamine and related compounds on the release of immunoreactive thyrotropin-releasing hormone (ir-TRH) from the rat stomach in vitro were studied. The rat stomach was incubated in medium 199 with 1.0 mg/ml of bacitracin and 100 micrograms/ml of ascorbic acid (pH 7.4) for 20 min. The amount of TRH release into the medium was measured by radioimmunoassay. The ir-TRH release from the rat stomach was enhanced significantly in a dose-related manner with the addition of histamine and inhibited with the addition of famotidine, but not with mepyramine. The stimulatory effect of histamine on ir-TRH release from the stomach was partially blocked with the addition of famotidine, but not with mepyramine. The elution profile of acid-methanol-extracted rat stomach on Sephadex G-10 was identical to that of synthetic TRH. These findings suggest that histamine stimulated ir-TRH release from the rat stomach in vitro, and that histamine's effects may be mediated via a H2-receptor.     

Expression of neurotrophin receptors trkB and trkC and their ligands in rat adrenal gland and the intermediolateral column of the spinal cord

Neurotrophins and their trk receptors constitute major classes of signaling molecules with important actions in the developing and adult nervous system. With regard to the sympathoadrenal cell lineage, which gives rise to sympathetic neurons and chromaffin cells, neurotrophin-3 (NT-3) and nerve growth factor (NGF) are thought to influence developing sympathetic neurons. Neurotrophin requirements of chromaffin cells of the adrenal medulla are less well understood than those for NGF. In order to provide the bases for understanding of putative functions of neurotrophins for the development and maintenance of chromaffin cells and their preganglionic innervation, in situ hybridization has been used to study the expression of brain-derived neurotrophic factor (BDNF) and NT-3, together with their cognate receptors trkB and trkC, in the adrenal gland and in the intermediolateral column (IML) of the spinal cord. BDNF is highly expressed in the embryonic adrenal cortex and later in cells of the cortical reticularis zone. Adrenal medullary chromaffin cells fail to express detectable levels of mRNAs for BDNF, NT-3, and their cognate receptors trkB and trkC. Neurons in the IML express BDNF and trkB, and low levels of NT-3 and trkC. Our data make it unlikely that BDNF and NT-3 serve as retrograde trophic factors for IML neurons but suggest roles of BDNF and NT-3 locally within the spinal cord and possibly for sensory nerves of the adrenal cortex.     

Distinctive gene expression profile in women with history of postpartum depression

Postpartum depression (PPD) is a disease which incorporates a variety of depressive states differing in nature and severity. To assist in the understanding of the pathogenesis of the disease, we aimed to ascertain a molecular mechanism underlying PPD development. We applied microarray technology to characterize gene expression of euthymic women with a history of PPD and compared the results with healthy controls. Our study demonstrated that women who considered euthymic on a clinical level, in fact, had an altered molecular profile when compared to participants with no PPD history. We identified nine genes significantly distinguished expression in post- depressive women; they may serve as a diagnostic tool for the detection of a predisposition to PPD. Our findings contribute significantly to the understanding of PPD etiology and its pathogenesis, offer a plausible explanation for the risk of the PPD recurrence, and may also contribute to clinical treatment.     

CEREBRAL LIPIDS IN A CASE OF SYSTEMIC GM2-GANGLIOSIDOSIS OF A LATE INFANTILE TYPE1

—Quantitative analyses performed on the lipids of cerebral grey matter from brains of a normal child and a child with Tay-Sachs (T-S) disease were compared with such analyses on the brain of a 6-year-old, non-Jewish male with systemic G_M2-gangliosidosis of a late infantile type (G_M2-LI). Analysis of gangliosides showed a 3·5-fold increase of total gangliosides in the G_M2-LI brain and a six-fold increase in the T-S brain, compared to normal brain. Both pathological brains had similar distribution patterns for gangliosides, with the G_M2-ganglioside component constituting more than 80 per cent of the total. Lipid components in the T-S brain were below normal values except for lecithin and cholesterol, while in the G_M2-LI brain there were increases in sulphatides, cerebrosides, sphingomyelin and cholesterol. Approximately twice as much ceramide trihexoside was present in the T-S brain as in the G_M2-LI brain, and none could be detected in the normal brain. The clinical, pathological and biochemical data support the conclusion that this case represents a new variant of systemic late-infantile gangliosidosis in which there is an accumulation of the G_M2-ganglioside like that in Tay-Sachs disease.     

Roles of ferritin and iron in ischemic preconditioning of the heart

Iron and copper play major roles in biological systems, catalyzing free radical production and consequently causing damage. The relatively high levels of these metals, which are mobilized into the coronary flow following prolonged ischemia, have been incriminated as key players in reperfusion injury to the heart. In the present communication we investigated other roles of iron – providing protection to the ischemic heart via preconditioning (PC).PC was accomplished by subjecting isolated rat hearts to three episodes of 2 min ischemia separated by 3 min of reperfusion. Prolonged ischemia followed the PC phase. PC hearts (group I) were compared to hearts subjected to normal perfusion (group II, no ischemia) and to ischemia without PC (group III). Group I showed a marked improvement in the recovery of hemodynamic function vs. group III. Biochemical parameters further substantiated the PC protection provided to group I against prolonged ischemia. Correspondingly, group I presented markedly lower re-distribution and mobilization of iron and copper into the coronary flow, following prolonged ischemia, as evinced from the decrease in total levels, and in the 'free' fraction of iron and copper.During the PC phase no loss of cardiac function was observed. A small wave of re-distribution and mobilization of iron (typically less than 4–8% of the value of 35 min ischemia) was recorded. The cellular content of ferritin (Ft) measured in the heart was significantly higher in group I than in group III (0.90 and 0.54 g/mg, respectively). Also, iron-saturation of Ft was significantly lower for PC hearts, compared to both groups II and III (0.22 vs. 0.32 and 0.31 g/mg, for 35 min ischemia, respectively). These findings are in accord with the proposal that intracellular re-distribution and mobilization of small levels of iron, during PC, cause rapid accumulation of ferritin – the major iron-storage protein.It is proposed that iron play a dual role: (i) It serves as a signaling pathway for the accumulation of Ft following the PC phase. This iron is not involved in cardiac injury, but rather prepares the heart against future high levels of 'free' iron, thus reducing the degree of myocardial damage after prolonged ischemia. (ii) High levels of iron (and copper) are mobilized following prolonged ischemia and cause tissue damage.     

Ferritin-bearing lymphocytes and T-cell levels in peripheral blood of patients with breast cancer

Summary Peripheral blood lymphocytes bearing surface ferritin and thymus-dependent lymphocyte (T cell) levels were determined in 15 breast cancer patients in stage I–II, 5 in stage III, 10 with benign breast disease, 4 with Thalassaemia, and 25 normal controls. The results of this study demonstrate that a subpopulation of lymphocytes (16.6%) bearing surface ferritin was found in patients with breast cancer in stage I–II. None were demonstrated in patients with either benign breast disease, or with Thalassaemia, the latter known to have high serum ferritin levels, and almost none (1.7%) in normal individuals. A significant decrease in the percentage of ERFC as compared with the percentage of T cells, determined with anti-T cell antiserum (P<0.01), was observed in patients with breast cancer in stage I–II. Yet, the mean T-cell percentage in this group of patients was significantly higher than the mean percentage of T cells in normal controls (P<0.01). In patients with benign breast disease, the percentage of T cells corresponded to the percentage of ERFC and did not significantly differ from those in normals. Stage III breast cancer patients seem to constitute a biologically distinct group, since the ferritin-positive lymphocyte subpopulation disappeared and the percentage of ERFC and T cells returned to the values of normal controls.Overnight incubation of lymphocytes from patients exhibiting a ferritin-positive lymphocyte subpopulation in culture media containing 20% FCS resulted in the removal of ferritin from the surface of the cells and in restoration of the percentage of ERFC.     

Antibiotic effects on mitochondrial translation and in patients with mitochondrial translational defects

The infantile presentation of mitochondrial respiratory chain defects frequently simulates acute bacterial infection and sepsis. Consequently, broad spectrum antibiotic therapy is often initiated before definitive diagnosis is reached and without taking into consideration the potential harm of antibiotics affecting mitochondrial translation. Here, we demonstrate that some commonly used translation-targeted antibiotics adversely affect the growth of fibroblasts from patients with defective mitochondrial translation systems. In addition, we show that these antibiotics inhibit mitochondrial translation in vitro. Our results suggest that patients with mitochondrial translation defects may be more vulnerable to toxic-side-effects following the administration of certain translation-targeted antibiotics.     

Regulation by Thyroid Hormones of Glucose Transport in Cultured Rat Myotubes

Primary cultures of skeletal muscle obtained from neonatal rats possess a saturable process for active glucose uptake, the myotubes having a relatively high affinity for the substrate with a Km of 1 mM. The expression of the glucose transport system was most apparent after fusion of single myoblasts to multinucleated myotubes [3-4 days in vitro (DIV)], at which time glucose uptake increased sharply to reach plateau values at about 6-8 DIV. Treatment of the cells at age 6 DIV with triiodothyronine or thyroxine caused a marked increase in glucose uptake beginning 4 h after treatment and reaching a maximum at 24 h. Thyroid hormone-induced increase in glucose uptake was not reduced by either tetrodotoxin or verapamil, thus indicating that the effect was not secondary to the ability of the hormone to increase contractile activity. The effect of thyroid hormones was eliminated completely by inhibition of protein synthesis. The results indicate that thyroid hormones play an important role in regulation of glucose transport in skeletal muscle.     

Understanding the origins of time-dependent inhibition by polypeptide deformylase inhibitors

The continual bacterial adaptation to antibiotics creates an ongoing medical need for the development of novel therapeutics. Polypeptide deformylase (PDF) is a highly conserved bacterial enzyme, which is essential for viability. It has previously been shown that PDF inhibitors represent a promising new area for the development of antimicrobial agents, and that many of the best PDF inhibitors demonstrate slow, time-dependent binding. To improve our understanding of the mechanistic origin of this time-dependent inhibition, we examined in detail the kinetics of PDF catalysis and inhibition by several different PDF inhibitors. Varying pH and solvent isotope led to clear changes in time-dependent inhibition parameters, as did inclusion of NaCl, which binds to the active site metal of PDF. Quantitative analysis of these results demonstrated that the observed time dependence arises from slow binding of the inhibitors to the active site metal. However, we also found several metal binding inhibitors that exhibited rapid, non-time-dependent onset of inhibition. By a combination of structural and chemical modification studies, we show that metal binding is only slow when the rest of the inhibitor makes optimal hydrogen bonds within the subsites of PDF. Both of these interactions between the inhibitor and enzyme were found to be necessary to observe time-dependent inhibition, as elimination of either leads to its loss.     

Regulation of trunk myogenesis by the neural crest: a new facet of neural crest-somite interactions

It is well established that the somitic mesoderm regulates early stages of neural crest development and further segmentation of crest-derived peripheral ganglia. The possibility that neural crest progenitors feed back on the somites was, however, not explored. Two recent studies provide evidence that the neural crest regulates somite-derived myogenesis by distinct mechanisms.