In situ optical monitoring of the movpe growth of ZnSe on GaAs

In situ optical reflection measurement was employed to study surface processes during the MOVPE growth of ZnSe films under an alternate supply of diethylzinc (DEZn) and dimethylselenide (DMSe) using H₂ and/or N₂ as carrier gases. We have found that the time-dependent reflection signal exhibits a unique saw-toothed pattern during the DEZn supply, which is attributed to the adsorption and structural change of the DEZn. In contrast, the influence of DMSe on the time-dependent signal appears to be rather marginal. A growth mechanism is proposed based on these experimental results, through which the important role of ambient hydrogen is discussed.     

Reversal of multidrug resistance (MDR) by aspidofractinine-type indole alkaloids

A series of indole alkaloids of the aspidofractinine-type was assessed for their potential in reversing MDR in vincristine-resistant KB cells. Of the compounds tested, kopsiflorine, kopsamine, pleiocarpine, 11-methoxykopsilongine, lahadinine A and N-methoxycarbonyl-11,12-methylenedioxy-delta 16,17-kopsinine were found to show appreciable activity.     

Differential expression of SHP2, a protein-tyrosine phosphatase with SRC homology-2 domains, in various types of renal tumour

The synthesis of DNA was studied in the proximal tibial growth plate of 25-day-old healthy NMRI mice by using the thymidine analog bromodeoxyuridine (BrdUrd), which is incorporated into cells in the S-phase. Such cells were found only in the upper three fifths of the morphologically defined proliferating zone. This zone was therefore subdivided into a functional proliferating zone (the S-phase zone) where most, if not all, chondrocytes proliferate, and a remaining maturation zone. The BrdUrd containing immunoreactive cells could then be followed at different intervals and they were found at the chondro-osseous junction after only 36 h. By using double-labeling with BrdUrd and iododeoxyuridine (IdUrd) the duration of cell cycle components could be estimated; that is, the time for DNA synthesis (S-phase), second gap and mitosis (G2 + M-phase), and remaining first gap (G1). We determined an S-phase time of 7.1 h and an average cell-cycle duration of 36 h. The G2 + M-phase was estimated as 3.5-4 h, leaving an average G1-phase time of 25 h, which probably varies considerably between chondrocytes. By combining these data with morphometrical data regarding distances between cells, we calculated a total growth rate of 9.0 microm/h. Of this rate, 80% was entirely related to the process of hypertrophy--that is, longitudinal expansion without any corresponding increase in cell number--and 75 % was the result of processes outside the S-phase zone. Five percent of the growth was due to the expansion of cell distances within the S-phase zone. In this way longitudinal expansion can be studied at different levels in the growth plate and the data permit calculation of changes in volumes of the extracellular matrix. The largest increases in matrix volume occurred in the hypertrophic zone. These data may serve as a basis for further studies on matrix turnover in relation to growth.     

Higher sensitivity in LEC rat cells to a topoisomerase II inhibitor, ellipticine

A concentration of ellipticine, an inhibitor of topoisomerase II, required to reduce cell survival to 37% (D37) is used as an index to compare the cellular sensitivity. D37 values of LEC and WKAH rat cells were 1.2 and 2.2 microM, respectively. Thus, LEC rat cells were approximately 1.8-fold more sensitive than WKAH rat cells to ellipticine. There was no significant difference between the topoisomerase II activities in nuclear extracts of LEC and WKAH rat cells. These results suggested that the high sensitivity of LEC rat cells to ellipticine is not associated with the level of topoisomerase II activity.     

Maternal stress induces synaptic loss and developmental disabilities of offspring

Mild prenatal stress affects the serotonergic system in the hippocampus of rat offspring. Pregnant rats were daily exposed to mild stress treatments (consisting of crowding and saline injection) during days 15 to 21 of pregnancy. Their offspring were assessed by a series of biochemical, histological and behavioral tests. On 35 days after birth, 5-hydroxytryptamine (5-HT) level was decreased by 17% (P < 0.05), whereas 5-hydroxyindolacetic acid (5-HIAA) level was increased by 18% (P < 0.05) in the offspring of prenatally stressed rats. The metabolic rate (5-HIAA/5-HT) was increased by 49% (P < 0.01). Synaptic density in the hippocampus of prenatally stressed offspring was also decreased by 32% (P < 0.0001) on postnatal day 35. There was no significant group difference in the spatial learning acquisition test of the Morris water maze; however, in the reversal task, prenatally stressed 5-week old rats spent more time than control animals searching for the platform of the pool. Escape latency in the cued test showed no significant difference. Together with data in our previous studies, that have shown 5-HT to facilitate synapse formation and maintenance in the central nervous system, synaptic loss is suggested to occur in relation to changes of 5-HT system in the hippocampus of prenatally stressed offspring. This may be associated with reported changes in behavior and learning ability in prenatally stressed offspring.     

Effects of thyroid hormone on the sorbitol pathway in streptozotocin-induced diabetic rats

Sorbitol accumulation plays an important role in diabetic complications involving the kidney, nerves, retina, lens and cardiac muscle. To investigate the influence of thyroid hormone on the sorbitol pathway, we studied the effects of thyroid hormone on polyol metabolism in normal and diabetic rats. Rats were divided into three groups: controls, streptozotocin (STZ)-induced diabetic euthyroid rats (DM) and STZ-induced diabetic hyperthyroid (thyroxine-injected) rats (DM+HT). The sorbitol (Sor) concentrations in the kidney, liver and sciatic nerve (2.53+/-0.74, 0.97+/-0.16 and 24.0+/-5.1 nmol/mg protein, respectively) of the DM rats were significantly higher than those (1.48+/-0.31, 0.58+/-0.13 and 3. 1+/-0.6 nmol/mg protein) of the control rats. The Sor concentrations in the kidney and sciatic nerve of the DM+HT rats (1.26+/-0.29 and 9. 40+/-1.2 nmol/mg protein) were significantly lower than those in the DM rats. These values were reduced in the liver, unchanged in the kidney, and increased in the sciatic nerve from the hyperthyroid rats without diabetes. Thyroid hormone reduced the aldose reductase (AR) activities in the kidney, liver and sciatic nerve of the DM rats, and similarly reduced AR in the kidney and liver, but not in the sciatic nerve, of the non-diabetic rats. The sorbitol dehydrogenase (SDH) activities were decreased by thyroid hormone in the kidney and liver but not the sciatic nerve of DM rats. In the non-diabetic rats, this enzyme activity was decreased in liver, but not in kidney or sciatic nerve. A positive correlation between the Sor concentration and AR activity was observed in the kidney and liver but not in the sciatic nerve from control, DM and DM+HT rats. A negative correlation was observed between the Sor concentration and SDH activities in the same organs. These data suggest that thyroid hormone affects the sorbitol pathway, but the detailed mechanism whereby this hormone reduces the sorbitol content (especially in diabetic rats) remains to be clarified.     

A possible role of RGS9 in phototransduction. A bridge between the cGMP-phosphodiesterase system and the guanylyl cyclase system

In the current concept of phototransduction, the concentration of cGMP in retinal rod outer segments is controlled by the balance of two enzyme activities: cGMP phosphodiesterase (PDE) and guanylyl cyclase (GC). However, no protein directly mediates these two enzyme systems. Here we show that RGS9, which is suggested to control PDE activity through regulation of transducin GTPase activity (He, W., Cowan, C. W., and Wensel, T. G. (1998) Neuron 20, 95-102), directly interacts with GC. When proteins in the Triton X-100-insoluble fraction of bovine rod outer segments were isolated by two-dimensional gel electrophoresis and binding of GC to these proteins was examined using a GC-specific antibody, proteins (55 and 32 kDa) were found to interact with GC. However, the activity of GC bound to the 55-kDa protein was not detected. This observation was elucidated by the finding that the 55-kDa protein inhibited GC activity in a dose-dependent manner. Amino acid sequence showed that five peptides derived from the 55-kDa protein were identical to corresponding peptides of RGS9. Together with other biochemical characterization of the 55-kDa protein, these observations indicate that the 55-kDa protein is RGS9 and that RGS9 inhibits GC. RGS9 may serve as a mediator between the PDE and GC systems.     

Synthesis and evaluation of oligopeptide RGDS exhibiting cell-attachment activity

Summary Tetrapeptide Arg-Gly-Asp-Ser (RGDS), an amino acid sequence existing in the cell-attachment domain of fibronectin, was synthesized using improved solid-phase procedure. Cell-attachment activity of the RGDS toward L-929 fibroblast cells originating in mouse epithelia was examined by measuring (a) the number of cells attached onto RGDS-immobilized polyvinyl alcohol (PVA) film, and (b) the % inhibition of cell-attachment onto polystyrene substrate from suspension of the cells in the presence of RGDS molecules. It was found that (a) a number of cells attached to the RGDS-immobilized PVA films, and (b) RGDS molecules remarkably attached to the cells, and as a result, RGDS inhibited the cells to adhere onto the substrate.