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Notes: The purpose of the present study was to evaluate the role of angiotensinogen (AGT) gene polymorphisms in the progression of IgA nephropathy (IgAN). The association of the haplotypes defined by A-20C and M235T, which correspond to G-6 A polymorphisms, with the severity of glomerular or interstitial lesions and renal prognosis were investigated.Patients with histologically proven IgAN were recruited after informed consent was obtained for the genetic study, which was approved by the ethics committee of our institute. Genomic DNA from each patient was prepared from peripheral blood leucocytes using an automatic DNA isolation system (NA-100, Kurabo, Osaka, Japan). A–C transition at nucleotide −20, G–A transition at −6, C–T transition at +68 and M235T variant at exon two in the AGT gene were determined as described previously.1,2The renal survival rate was analysed for 114 IgAN patients with a creatinine clearance (Ccr) level of 70 mL/min or greater and followed up more than 24 months. The patients with haplotype T235 and C-20 (n = 63) were compared with those without T235 and C-20 (n = 51) for age, sex, blood pressure, proteinuria, serum Cr, Ccr at the time of renal biopsy, drugs administered, and severity of renal histopathologic lesion.Light microscopic evaluation of all specimens was performed in a double blind fashion according to the grading classification described previously, including glomerular cellular proliferation, mesangial matrix increase, global or segmental sclerosis, endocapillary proliferation, leucocyte exudation, duplication of glomerular basement membrane, crescent formation, and tufts adhesion to Bowman’s capsule as well as tubulointerstitial lesions.3The Kaplan-Meier method and the Cox proportional hazards regression model analysed the time course from renal biopsy to end point (initiation of dialysis or sCr level doubled after the time of diagnosis). Several covariates were selected by a stepwise backward method and the effects of these covariates were expressed by a hazard ratio. P 〈 0.05 was considered statistically significant.The genotype distributions in the present study were not different from that in Hardy–Weinberg equilibrium. The genotype and allele frequencies of AGT variants were compatible with previous studies for Japanese population.1,4 Haplotype analysis showed a complete linkage disequilibrium between M235T and A-20C alleles (linkage-disequilibrium coefficient: D′, 1.00). Because the AGT gene variant at −20 was observed only in a subset of the 235T alleles, the following haplotypes were determined: T235 and C-20; T235 and A-20; and M235 and A-20. The incidence of hypertension was not different between each haplotype of AGT.An association study of histopathological findings revealed that patients with haplotype T235 and C-20 had significantly higher grading scores in crescent formation (P = 0.017) than those without haplotype T235 and C-20. Renal survival rate was significantly lower in the patients with haplotype T235 and C-20 (〈link href="#f1"〉Fig. 1, P = 0.003). The Cox proportional hazards regression model showed an increased hazard ratio (HR) for haplotype T235 and C–20 of 5.9 from multivariate analysis (〈link href="#f2"〉Fig. 2; 95% CI, 2.1–16.7; P 〈 0.001). Furthermore, in IgAN patients without administration of angiotensin I converting enzyme inhibitors (n = 77), renal survival rate was significantly lower in those with haplotype T235 and C–20 (P = 0.011).〈figure xml:id="f1"〉1〈mediaResource alt="image" href="urn:x-wiley:13205358:NEP19:NEP_19_f1"/〉Effect of AGT haplotype on the renal survival rate in IgAN patients. Solid and dashed lines represent the renal survival rate in the patients with and without haplotype T235 and C–20, respectively.〈figure xml:id="f2"〉2〈mediaResource alt="image" href="urn:x-wiley:13205358:NEP19:NEP_19_f2"/〉Cox proportional hazards regression model to test the predictors for renal survival time in multivariate analysis. Covariates were selected by stepwise backward analysis. U-protein, urinary protein; HT at renal biopsy, hypertensives at the time of renal biopsy; ACEI, angiotensin I converting enzyme inhibitor. The bars represent the 95% confidence intervals of hazard ratio.This work provides the evidence that the C–20 gene polymorphism of AGT, a subset of 235T alleles, is associated with histopathological severity of glomerular injury, and progression of renal dysfunction in Japanese patient with IgAN.

Notes: Abstract The main objective of this study was to assess the blood-brain barrier (BBB) permeability in periventricular areas of the normal mouse brain to test the hypothesis that the fragility of the BBB in periventricular areas may play a role in periventricular white matter lesions. Vascular permeability to intravenously injected horseradish peroxidase (HRP) was examined in the periventricular areas of adult mouse brain using light and electron microscopy. Staining for HRP appeared in the periventricular area adjacent to medial side of the lateral ventricle as well as in BBB-free areas, in the lateral septal nucleus, in the medial portion of the hippocampus and in the dorsal portion of the thalamus. In addition, the staining for HRP appeared in ependymal cell layer located near the choroid plexus and was found early after HRP injection in the wall of some vessels located at medial side of the optic tract. Ultrastructural examination of the vessel wall revealed that staining for HRP in the perfusion-fixed mice after circulation of the tracer for 5 min appeared in the perivascular space, in the basal lamina, in several vesicular profiles of the endothelial cell cytoplasm including abluminal pits, in vesicular profiles of perivascular cells and in the adjacent extracellular space. In the mice perfusion-fixed after HRP circulation for 90 min, staining for HRP in the vessels at medial side of the optic tract appeared in the cytoplasm of the perivascular cells, in vesicular structures of the endothelial cell cytoplasm such as plasmalemmal vesicles, endosomes and multivesicular bodies and occasionally in the vascular basal lamina. No clear staining reaction for HRP was found in the periventricular areas adjacent to lateral side of the lateral ventricles. These findings indicate that the BBB in the periventricular area adjacent to medial side of the lateral ventricle near the root of the choroid plexus is not so tight as it is in the cortex or in the lateral periventricular areas, and suggest that the perivascular cells play a scavenger role in the periventricular area as a component of the BBB. In addition, they indicate that blood-borne macromolecules can also invade the areas adjacent to the ventricles such as the lateral septal nucleus, the medial portion of the hippocampus and the dorsal portion of the thalamus.

Notes: Summary To determine whether E-643, a new α-blocking agent, would reduce the blood pressure, regardless of the posture, a 1 mg dose was given 3 times daily for 7 consecutive days, to 8 male and 7 female inpatients, aged 37–73 years, with essential hypertension. Blood pressure and pulse rate were measured daily in the supine, sitting and standing positions. Before and after the treatment with E-643, plasma levels of noradrenaline, adrenaline, dopamine-β-hydroxylase, renin and aldosterone were determined, samples being obtained with the subjects recumbent and after standing upright for 60 min. A significant reduction in the systolic and diastolic blood pressures was evident in the supine (172±31/100±12 → 151±28/89±14 mmHg), sitting (158±22/101±11 → 138±28/89±15 mmHg) and standing (153±32/103±21 → 129±31/89±20 mmHg) positions. The reduction in blood pressure remained unchanged throughout the period of administration of E-643. Pulse rate was not affected when the subjects were supine (67±10 → 69±10 beats/min), but was increased in the sitting (68±10 → 73±9 beats/min) and standing (73±10 → 81±11 beats/min) positions. The increased pulse rate tended to decline during continued administration of E-643. Treatment with E-643 produced no significant change in plasma levels of adrenaline, noradrenaline, dopamine-β-hydroxylase, renin and aldosterone. The antihypertensive effect of treatment was more prominent in the patients with higher levels of plasma catecholamines and dopamine-β-hydroxylase, and was less prominent in those with higher plasma renin and aldosterone. Two patients had temporary bouts of dizziness and visual disturbances, but there were no subjective complaints during treatment.

Notes: Summary Periodical changes in Ca2+-ATPase and Mg2+-ATPase activity were observed cytochemically in the crayfish gastrolith epithelium during the molting cycle in relation to the calcium transport mechanism. The ATPase activity was demonstrated by a new one-step lead citrate method. The reaction products were mainly restricted to the matrix of type II cell mitochondria. The Ca2+-ATPase activity was intensely observed in two calcium moving stages, the small gastrolith period which indicates the beginning of gastrolith formation, and the aftermolt, when the calcified gastrolith has been dissolved in the stomach and then reabsorbed from the stomach epithelium into the newly formed soft exoskeleton through the blood. Although the intensity of reaction products of Mg2+-ATPase varied in each stage, the enzymatic activity was observed throughout all molting stages. Reaction products were observed in all mitochondria, basement membranes, apical cytoplasmic membranes, and in some lysosomes. In conclusion, periodical changes in the two types of ATPase activity were seen in the mitochondria of gastrolith epithelium during the molting cycle, but Ca2+-ATPase activity seemed to be more prominently synchronized to the calcium movement in the gastrolith epithelium than Mg2+-ATPase activity. These results provide the strong evidence that Ca2+-ATPase may act strongly in the calcium transport system of crayfish molting.

Notes: Summary Ester derivatives (PCA-nED) of dodecyl alcohol, ethyleneglycol dodecyl ether, and diethyleneglycol dodecyl ether with DL-2-pyrrolidone-5-carboxylic acid were prepared, and their properties as emulsion stabilizer were estimated by using a surface-tension meter and multiple drop method in the presence of SDS and compared to those of a series of the corresponding polyethyleneglycol dodecyl ether (nED) as stabilizers. PCA-nED system remarkably reduced the interfacial tension compared tonED system, and the interfacial tension for both PCA-nED andnED systems decreased with the increase of ethylene oxide chain length. The stability of oil drop remarkably increased by addition of PCA-nED compared tonED system, and increased with the increase of ethylene oxide chain length. A series of PCA-nED was found to be utilized as emulsion stabilizers.

Notes: Summary Surface and micellar properties of a homologous series of Octaethylene glycol-n-alkyl ethers (C n E8;n = 9 to 15) have been studied in aqueous solutions by the surface tension measurements. The effects of the alkyl chain length comprising even and carbon numbers have been examined in order to evaluate the surface free energy ΔG A-W and the standard free energy ΔG m for the micellization obtained from their surface tension data. The areas per molecule and the equilibrium surface tension values at the CMC decreased with an increasing carbon number and they showed zigzag curves by the difference in even and odd carbon numbers. These findings may be attributed to the differences in the molecular orientation between the molecules with even carbon number and ones with odd carbon number on the air-water interface at CMC. ΔG A-W values decreased linearly with an increasing alkyl chain length but did not show a zigzag line by the differences in even and odd carbon numbers. This suggests that the molecular orientation is not influenced by the difference between their even and odd carbon numbers in the alkyl chain on the surface of the very diluted solution, and their molecules form some stable adsorbed films with an increase of the alkyl chain length. A division of ΔG A-W into the contribution made both by the hydrophilic group ΔG A-W (-W) and by the hydrophobic group ΔG A-W (-CH2-) was attempted as follows; ΔG A-W (-CH2-) = − 0.80 kcal/mol and ΔG a-W (-W) = + 0.15 kcal/mol. The free energy changes ΔG m of micellization were discussed on the basis of the CMC data obtained from the surface tension measurements by treating the formation of micelles as analogous to phase separation, and the contribution from the each moieties in the molecule were calculated as follows; ΔG m (-CH2-) = − 0.68 kcal/mol and ΔG m (-W) = + 1.54 kcal/mol. The difference between ΔG A-W and ΔG m is discussed using their data.