Hypertension aggravates glomerular dysfunction with oxidative stress in a rat model of diabetic nephropathy

Oxidative stress may contribute to the pathogenesis of diabetic nephropathy (DN), although the detailed mechanism of reactive oxygen species (ROS) regulation is still unclear. This study examined the effect of high-salt diet on ROS production and expression of antioxidant enzymes in control and experimentally diabetic rats. Wistar fatty rats (WFR) as a type 2 diabetes mellitus model and Wistar lean rats (WLR) as a control were fed a normal-salt diet (NS) and high-salt diet (HS) from the age of 6 to 14 weeks. We then examined the blood pressure, urinary albumin excretion (UAE), and urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels. The expression of antioxidant enzymes including α-catalase (CAT), Cu-Zn superoxide dismutase (SOD), Mn SOD, and glutathione peroxidase (GPx) were analyzed in the glomeruli of the rats using Western blotting. The expression of NAD(P)H oxidase p47^phox and NFκB p65 was evaluated using immunohistochemical staining. By 14 weeks of age, the WFR-HS group exhibited hypertension and markedly increased UAE. The level of 8-OHdG, a marker of oxidative damage, in the WFR-HS group was also higher than that in the WLR groups or WFR-NS group. The expression of α-CAT and Mn SOD proteins was significantly decreased in isolated glomeruli in the WFR-HS group. GPx and Cu-Zn SOD expression did not differ between the WFR and WLR groups. High expression of ROS and decreases in antioxidants were seen in the glomeruli of diabetic rats with hypertension, suggesting that oxidative stress may be involved in the development of DN.     

Age-dependent increases of calcium and phosphorus in human epiglottal cartilage

To elucidate compositional changes of the elastic cartilage with aging, the authors investigated age-related changes of elements in the epiglottal cartilages by inductively coupled plasma-atomic emission spectrometry. After the ordinary dissection by medical students at Chiang Mai University was finished, the epiglottises were resected from the subjects. The epiglottal cartilages were isolated and the element contents were determined. The subjects consisted of 11 men and 14 women, ranging in age from 39 to 92 yr old. It was found that although the extent of accumulation of calcium and phosphorus was slight, calcium and phosphorus increased progressively in the epiglottal cartilages with aging. In contrast, sulfur, magnesium, zinc, iron and sodium did not change significantly in them. Regarding the relationships among elements, it was found that there were significant correlations among calcium, phosphorus, magnesium, and sodium in the epiglottal cartilages, with one exception between calcium and sodium contents. In comparison between men and women, no significant differences were found in the predominant elements such as calcium, sulfur, and phosphorus in the epiglottal cartilages.     

Isolation and Structure of a Novel Isoflavone Derivative in Red Clover

Absolute configurations of C–2 and C–3 in “Acid III” (IIa), a degradation product of piericidin A diacetate, was determined as R and S, respectively. Consequently both C–9 and C–10 in piericidin A were confirmed to have S configurations. On the basis of UV and NMR specta, configurations of the double bonds in piericidin A were assigned as shown in Ia.     

Sea resident Japanese eel collected from Kozushima Island,Japan

A Japanese eel Anguilla japonica was collected off Kozushima Island, one of the volcanic Izu Islands in Japan. The external morphology corresponded to a silvering stage (Silvering Index: S1) with a melanized body coloration of the head, and the tip of the lower jaw to the anus. However, the value of the gonado-somatic index of the female eel was low, and the oocytes were not matured (peri-nucleolus stage). Otolith analyses indicated that the individual had exclusively inhabited the sea for five years without entering freshwater. This record is the first collection of silver-phased Japanese eel in the Kuroshio region, which could be on the spawning migration route of the species, although it was not determined if the eel was settling around the Izu Islands or migrating to the spawning ground.     

Evidence for a role of tight junctions in regulating sodium permeability in zebrafish (Danio rerio) acclimated to ion-poor water

Freshwater teleosts are challenged by diffusive ion loss across permeable epithelia including gills and skin. Although the mechanisms regulating ion loss are poorly understood, a significant component is thought to involve paracellular efflux through pathways formed via tight junction proteins. The mammalian orthologue (claudin-4) of zebrafish (Danio rerio) tight junction protein, claudin-b, has been proposed to form a cation-selective barrier regulating the paracellular loss of Na⁺. The present study investigated the cellular localization and regulation of claudin-b, as well as its potential contribution to Na⁺ homeostasis in adult zebrafish acclimated to ion-poor water. Using a green fluorescent protein-expressing line of transgenic zebrafish, we found that claudin-b was expressed along the lamellar epithelium as well as on the filament in the inter-lamellar regions. Co-localization of claudin-b and Na⁺/K⁺-ATPase was observed, suggesting its interaction with mitochondrion-rich cells. Claudin-b also appeared to be associated with other cell types, including the pavement cells. In the kidney, claudin-b was expressed predominantly in the collecting tubules. In addition, exposure to ion-poor water caused a significant increase in claudin-b abundance as well as a decrease in Na⁺ efflux, suggesting a possible role for claudin-b in regulating paracellular Na⁺ loss. Interestingly, the whole-body uptake of a paracellular permeability marker, polyethylene glycol-400, increased significantly after prolonged exposure to ion-poor water, indicating that an increase in epithelial permeability is not necessarily coupled with an increase in passive Na⁺ loss. Overall, our study suggests that in ion-poor conditions, claudin-b may contribute to a selective reduction in passive Na⁺ loss in zebrafish.     

CYP3A is responsible for N-dealkylation of haloperidol and bromperidol and oxidation of their reduced forms by human liver microsomes

We studied the biotransformation of haloperidol, bromperidol and their reduced forms by human liver microsomes. Nifedipine oxidation (CYP3A) activity correlated significantly with N-dealkylation rates of haloperidol and bromperidol and oxidation rates of their reduced forms, while neither ethoxyresorufin O-deethylation (CYP1A2) activity nor dextromethorphan O-deethylation (CYP2D6) activity did. In chemical and immunoinhibition studies, only troleandomycin and anti-CYP3A4 serum inhibited both formation rates of 4-fluorobenzoylpropionic acid, a metabolite of haloperidol and bromperidol, and back oxidation rates. Among 10 recombinant isoforms examined, only CYP3A4 showed catalytic activity. The Vmax and Km values of N-dealkylation of bromperidol and reoxidation of reduced bromperidol were similar to those of haloperidol and reduced haloperidol, respectively. The present study indicates that CYP3A plays a major role in N-dealkylation of and oxidation back to bromperidol as well as haloperidol and suggests that modification of in vivo CYP3A activity by inhibition or induction may affect the pharmacokinetics and therapeutic effects of haloperidol and bromperidol.     

Differences in neurogenic potential in floor plate cells along an anteroposterior location: midbrain dopaminergic neurons originate from mesencephalic floor plate cells

Directed differentiation and purification of mesencephalic dopaminergic (mesDA) neurons from stem cells are crucial issues for realizing safe and efficient cell transplantation therapies for Parkinson's disease. Although recent studies have identified the factors that regulate mesDA neuron development, the mechanisms underlying mesDA neuron specification are not fully understood. Recently, it has been suggested that mesencephalic floor plate (FP) cells acquire neural progenitor characteristics to generate mesDA neurons. Here, we directly examined this in a fate mapping experiment using fluorescence-activated cell sorting (FACS) with an FP cell-specific surface marker, and demonstrate that mesencephalic FP cells have neurogenic activity and generate mesDA neurons in vitro. By contrast, sorted caudal FP cells have no neurogenic potential, as previously thought. Analysis of dreher mutant mice carrying a mutation in the Lmx1a locus and transgenic mice ectopically expressing Otx2 in caudal FP cells demonstrated that Otx2 determines anterior identity that confers neurogenic activity to FP cells and specifies a mesDA fate, at least in part through the induction of Lmx1a. We further show that FACS can isolate mesDA progenitors, a suitable transplantation material, from embryonic stem cell-derived neural cells. Our data provide insights into the mechanisms of specification and generation of mesDA neurons, and illustrate a useful cell replacement approach for Parkinson's disease.