Cutaneous Warm and Cool Sensation Thresholds and the Inter-threshold Zone in Malaysian and Japanese Males

The purpose of this study was to investigate ethnic differences in cutaneous thermal sensation thresholds and the inter-threshold sensory zone between tropical (Malaysians) and temperate natives (Japanese). The results showed that (1) Malaysian males perceived warmth on the forehead at a higher skin temperature (T_sk) than Japanese males (p<0.05), whereas cool sensations on the hand and foot were perceived at a lower T_sk in Malaysians (p<0.05); (2) Overall, the sensitivity to detect warmth was greater in Japanese than in Malaysian males; (3) The most thermally sensitive body region of Japanese was the forehead for both warming and cooling, while the regional thermal sensitivity of Malaysians had a smaller differential than that of Japanese; (4) The ethnic difference in the inter-threshold sensory zone was particularly noticeable on the forehead (1.9±1.2 C for Japanese, 3.2±1.6 °C for Malaysians, p<0.05). In conclusion, tropical natives had a tendency to perceive warmth at a higher T_sk and slower at an identical speed of warming, and had a wider range of the inter-threshold sensory zone than temperate natives.     

Multicenter evaluation of prototype real‐time PCR assays for Epstein‐Barr virus and cytomegalovirus DNA in whole blood samples from transplant recipients

Quantitative PCR is becoming widespread for diagnosing and monitoring post‐transplantation diseases associated with EBV and CMV. These assays need to be standardized to manage patients in different facilities. Five independent laboratories in Japan compared home‐brew assays and a prototype assay system to establish a standard quantitative procedure for measuring EBV and CMV. Reference standards and a total of 816 (642 EBV and 174 CMV) whole blood samples from post‐transplantation recipients were used for this multicenter evaluation. The prototype reference standard for EBV was compared to a panel of samples, with a theoretical expected value made using EBV‐positive cells containing two virus genome copies per cell. The mean ratio of the reference standard at each site to the standard of the prototype assay was ≤4.15 for EBV among three different sites and ≤3.0 for CMV between two laboratories. The mean of the theoretical expected number of the EBV genome: prototype reference was close to 1.0. The correlation coefficients between the viral copy numbers determined using the prototype assay and those using each home‐brew assay were high (EBV, 0.73–0.83, median = 0.78; CMV, 0.54–0.60, median = 0.57). The dynamics of the EBV and CMV loads in transplant recipients were similar between the assay types. There was an inter‐laboratory difference among the quantification results, indicating that a unified protocol and kit are favorable for standardizing the quantification of EBV and CMV. Such standardization will help to standardize the diagnosis and monitoring of diseases associated with EBV and CMV.     

New aminopropandiol derivatives as orally available and short-acting calcium-sensing receptor antagonists

Synthesis and structure–activity relationship studies on a new aminopropandiol class of derivatives as calcium-sensing receptor antagonists are described. Modification of the phenolic moiety of a calcilytic compound NPS 2143 led to the identification of an orally available compound (R,R)-31 which demonstrated a rapid and transient stimulation of PTH release in rats.     

Novel Phorbol Ester-binding Motif Mediates Hormonal Activation of Na+/H+ Exchanger

Protein kinase C (PKC) is considered crucial for hormonal Na⁺/H⁺ exchanger (NHE1) activation because phorbol esters (PEs) strongly activate NHE1. However, here we report that rather than PKC, direct binding of PEs/diacylglycerol to the NHE1 lipid-interacting domain (LID) and the subsequent tighter association of LID with the plasma membrane mainly underlies NHE1 activation. We show that (i) PEs directly interact with the LID of NHE1 in vitro, (ii) like PKC, green fluorescent protein (GFP)-labeled LID translocates to the plasma membrane in response to PEs and receptor agonists, (iii) LID mutations markedly inhibit these interactions and PE/receptor agonist-induced NHE1 activation, and (iv) PKC inhibitors ineffectively block NHE1 activation, except staurosporin, which itself inhibits NHE1 via LID. Thus, we propose a PKC-independent mechanism of NHE1 regulation via a PE-binding motif previously unrecognized.     

Comparative analysis of the reactive oxygen species‐producing enzymatic activity of Arabidopsis NADPH oxidases

Reactive oxygen species (ROS) produced by NADPH oxidases, called respiratory burst oxidase homologs (Rbohs), play crucial roles in development as well as biotic and abiotic stress responses in plants. Arabidopsis has 10 Rboh genes, AtRbohA to AtRbohJ. Five AtRbohs (AtRbohC, ‐D, ‐F, ‐H and ‐J) are synergistically activated by Ca²⁺‐binding and protein phosphorylation to produce ROS that play various roles in planta, although the activities of the other Rbohs remain unknown. With a heterologous expression system, we found a range of ROS‐producing activity among the AtRbohs with differences up to 100 times, indicating that the required amounts of ROS are different in each situation where AtRbohs act. To specify the functions of AtRbohs involved in cell growth, we focused on AtRbohC, ‐H and ‐J, which are involved in tip growth of root hairs or pollen tubes. Ectopic expression of the root hair factor AtRbohC/ROOT HAIR DEFECTIVE 2 (RHD2) in pollen tubes restored the atrbohH atrbohJ defects in tip growth of pollen tubes. However, expression of AtRbohH or ‐J in root hairs did not complement the tip growth defect in the atrbohC/rhd2 mutant. Our data indicate that Rbohs possess different ranges of enzymatic activity, and that some Rbohs have evolved to carry specific functions in cell growth.     

Reproducing bursting interspike interval statistics of the gustatory cortex

Cortical neurons in vivo generate highly irregular spike sequences. Recently, it was experimentally found that the local variation of interspike intervals, LV, is nearly constant for every spike sequence for the same neurons. On the contrary, the coefficient of variation, CV, varies over different spike sequences. Here, we first show that these characteristic features are also applicable in bursting spike sequences that are obtained from the rat gustatory cortex. Next, we show that the conventional leaky integrate-and-fire model does not fully account for reproducing these statistical features in data of real bursting spike sequences. We resolve this difficulty by proposing an alternative neuron model which is a reduction of the bursting neuron model involving the persistent sodium current. Our study implies that (1) the characteristic features of CV and LV are the results of the endogenous bursting and (2) the bursting behavior in the gustatory cortex is caused mainly by the persistent sodium current.     

Effect of iodoacetic acid on maintenance of pancreatic endocrine cells of the neonatal rat

This report describes the specific cytotoxicity of iodoacetic acid (IAA) in selectively destroying the fibroblastoid cells and stimulating the in vitro function of neonatal B cells prepared from rat pancreases. Under culture conditions with a basal medium containing 5.5 mM D-glucose alone, the responses to insulin secretagogues tested were abolished by day 7 of culture. In contrast, the addition of 10 microM IAA enhanced either insulin release evoked by D-glucose (16.7 mM), L-leucine (10 mM) and 2-ketoisocaproate (10 mM) or the cellular insulin content to approximately twice the initial levels (day 0). L-Glutamine (10 mM) augmented the stimulatory effect of L-leucine or 2-ketoisocaproate. Moreover, the continuous application of IAA significantly increased the rates of glutamine oxidation in endocrine cells after 7 days of culture. On the other hand, the IAA-supplemented medium did not preserve the function of A cells. The phase-contrast microphotograph examination revealed the selective removal of fibroblasts from the monolayer cultures. This corresponded very closely with a remarkable reduction in culture DNA content.     

Ebselen prevents mitochondrial ageing due to oxidative stress: in vitro study of fish erythrocytes

Nucleated trout erythrocytes under oxidative stress suffer DNA membrane damage and inactivation of glutathione peroxidase. In addition, oxidative damage increases with the age of the cell. In the present paper, we evaluate the effects of oxidative stress and ageing on mitochondrial functionality by means of transmission electron microscopy and cytofluorimetric determination of mitochondrial membrane potential and intracellular levels of reactive oxygen species. The protective activity of the antioxidant organoselenium compound ebselen, a mimic of glutathione peroxidase, is also evaluated. Ebselen prevents the drastic structural and functional changes in mitochondria in aged RBCs induced by oxidative stress. However, the antioxidant does not prevent swelling of the mitochondria.