Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats.

AIMS: Mineralocorticoid receptor (MR) blockade is an effective treatment for hypertension and diabetic nephropathy. There are no data on the effects of MR blockade on diabetic peripheral neuropathy (DPN). The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin (STZ)-induced diabetic rats. METHODS: Expression of MR protein and messenger RNA (mRNA) was examined in the peripheral nerves using Western blot analysis and RT-PCR. We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity (NCV), morphometric changes and cyclooxygenase-2 (COX-2) gene and NF-κB protein expression in the peripheral nerves of STZ-induced diabetic rats. RESULTS: Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney. Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone (39.1 ± 1.2 m/s) or candesartan (46.4 ± 6.8 m/s) compared with control diabetic rats (33.7 ± 2.0 m/s) (p < 0.05). Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats. Eplerenone and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats (p < 0.05). COX-2 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats, and treatment with eplerenone or candesartan reduced these changes in gene expression (p < 0.05). CONCLUSION: MR blockade may have neuroprotective effects on DPN.

Planned release of contaminated water from the Fukushima storage tanks into the ocean: Simulation scenarios of radiological impact for aquatic biota and human from seafood consumption.

The radiological impact for human and aquatic biota as a result of a planned release of contaminated water stored in tanks near the Fukushima Dai-ichi Nuclear Power Plant to the Pacific Ocean is assessed. The total activity for 10 dominant radionuclides (3H, 14C, 60Co, 90Sr, 99Tc, 106Ru, 125Sb, 129I, 134Cs, 137Cs) in tanks is estimated. The compartment model POSEIDON-R is applied to compute the concentration of activity for each radionuclide in water, bottom sediments, and biota, and corresponding doses to marine organisms and humans from seafood consumption. Predicted concentrations of activity in marine products in future will not exceed food safety limits in Japan. The computed maximum committed effective dose to humans is less than 1 µSv per year with the highest contribution from 129I and 14C. Maximum absorbed doses to non-human biota are in the order of 0.05 to 20 µGy per year, meaning that no deleterious effects are expected.

Concentration ratios of stable elements for selected biota in Japanese estuarine areas.

For the estimation of radiation doses to organisms, concentration ratios (C ( R )s) of radionuclides are required. In the present study, C(R)s of various elements were obtained as analogues of radionuclides for algae, molluscs, and crustaceans, in eight estuarine areas around Japan. The elements measured were Na, Mg, K, Ca, V, Mn, Fe, Co, Ni, Cu, Rb, Sr, Y, Mo, Cd, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, Pb, and U. The geometric mean (GM) values of C(R)s (GM-C(R)s) for alkali and alkaline earth elements, Mo, and U for all biota, as well as V for crustaceans, were less than 100 L/kg, while GM-C(R)s for the other elements were higher. When the obtained GM-C(R)s were compared with the C(R)s recommended in IAEA Technical Report Series 422 for marine organisms, no big differences between them were found; however, several elements (i.e. Cd and U for algae, Mn for molluscs, and Pb for crustaceans) were lower than the recommended C(R)s. In the present study, conversion factors (the ratio of C(R) for the whole body to that for muscle) for molluscs and crustaceans were also calculated, since data on edible parts of these organisms are generally available in the literature. For crustaceans, GM conversion factors of all the elements were more than one. For molluscs, GM conversion factors of rare earth elements and U were slightly higher than those for crustaceans, while GM conversion factors of the other elements were almost the same and less than 10. These results indicate that some elements tend to be concentrated in the internal organs of biota collected in the estuarine areas. For environmental radiological assessment, conversion factors from tissue to whole-body C(R) values are useful parameters.

Temporal and spatial variations of 134Cs and 137Cs levels in the Sea of Japan and Pacific coastal region: Implications for dispersion of FDNPP-derived radiocesium.

To investigate the dispersion of Fukushima Dai-ichi Nuclear Power Plant (FDNPP)-derived radiocesium in the Sea of Japan and western Pacific coastal region and determine the sources of radiocesium in these areas, we examined the temporal and spatial variations of 134Cs and 137Cs concentrations (activities) during 2011-2016 in seawaters around the western Japanese Archipelago, particularly in the Sea of Japan. In May 2013, the surface concentration of 134Cs was ∼0.5 mBq/L (decay-corrected to March 11, 2011), and that of 137Cs exceeded the pre-accident level in this study area, where the effects of radiocesium depositions just after the FDNPP accident disappeared in surface waters in October 2011. Subsequently, radiocesium concentrations gradually increased during 2013-2016 (∼0.5-1 mBq/L for 134Cs), exhibiting approximately homogeneous distributions in each year. The temporal and spatial variations of 134Cs and 137Cs concentrations indicated that FDNPP-derived radiocesium around the western Japanese Archipelago, including the Sea of Japan, has been supported by the Kuroshio Current and its branch, Tsushima Warm Current, during 2013-2016. However, in the Sea of Japan, the penetration of 134Cs was limited to depths of less than ∼200 m during three years following the re-delivery of FDNPP-derived radiocesium.

Protein composition of human metaphase chromosomes analyzed by two-dimensional electrophoreses.

A large amount of metaphase chromosomes were isolated from synchronized human cell lines by a polyamine procedure. All the chromosomal proteins extracted by an acetic acid extraction method were fully dissolved into the sample solutions for isoelectric focusing (IEF) or radical free and highly reduced (RFHR) two-dimensional electrophoreses (2-DEs). As a result, well-separated and highly reproducible 2-DE patterns were obtained. This could not be attained by an ordinary acetone precipitation method. The 2-DE patterns visualized using Coomassie Brilliant Blue (CBB) staining indicated that more than one hundred proteins were involved in the isolated metaphase chromosomes, although the most abundant proteins, histones, occupied a greater part of the chromosomal proteins. It was also shown that colcemid treatment for cell cycle synchronization had little effect on the 2-DE pattern compared to that obtained without the treatment. Furthermore, no significant differences were observed in the 2-DE patterns among the chromosomal proteins prepared from two different human cell lines, BALL-1 and K562. However, 2-DE analysis of isolated metaphase chromosomes from HeLa cells apparently showed a smaller number of proteins than the BALL-1 and K562 cell lines at a neutral pI range. The present study paves the way for elucidating protein composition of human metaphase chromosomes.

Identification of alloantisera reacting with HLA-C blank (Cx52) using a mouse L-cell transfected with the HLA-Cw*1201 allele.

The HLA-C locus frequently has a serologically undefined "blank" (CwBL) specificity. A cDNA clone derived from the HLA-C gene with a blank specificity (Cx52) strongly associated with the most common haplotype in a Japanese population, A24-CwBL-B52-DR2-DQ6-DP9, has been recently cloned and sequenced in our laboratory and officially designated Cw*1201 as an allelic name, indicating that the inability to define the HLA-C antigen serologically in this haplotype is not due to an HLA-C antigen gene deletion or mutation, but to the absence of typing sera. In this paper, a mouse L-cell transfectant expressing this Cw*1201 gene product was constructed and employed for screening of alloantisera recognizing the HLA-C antigen with the Cw*1201 specificity. Two alloantisera against Cw*1201 were thus identified and characterized using HLA homozygous B-cell lines and local panel PBL cells, indicating that a transfectant expressing a single HLA provides an efficient screening system for collection of HLA-typing sera, especially reacting with serologically unclassifiable "blank" antigens.

HLA class II antigens are associated with Japanese pemphigus patients.

We investigated the HLA class II antigens in 30 Japanese cases of pemphigus, 17 cases of pemphigus vulgaris (PV) and 13 cases of pemphigus foliaceus (PF), by both serologic and restriction fragment length polymorphism (RFLP) analyses. We detected two major haplotypes susceptible to PV, i.e., DRw12-DQw7 and DRw6-DQw5. In contrast, DR2 was absent in PV. RFLP analyses showed that DRw6 PV patients had a disease-associated restriction fragment representing DQw5, the same association as that found in DRw6 Jewish PV patients. However, DRw12 Japanese PV patients had DQw7, whereas DR4 Jewish PV patients had DQw8. On the other hand, all 13 PF patients were serologically typed for DQw1, which could not be further subdivided into DQw5 by RFLP analyses. These results suggest that Japanese and Jewish PV patients may be immunogenetically closely related to each other, but Japanese PV patients appear to be immunogenetically different from Japanese PF patients.

The potency of substituted benzimidazoles such as E3810, omeprazole, Ro 18-5364 to inhibit gastric H+, K(+)-ATPase is correlatedwith the rate of acid-activation of the inhibitor.

The half maximal inhibitory concentrations (IC50) of substituted benzimidazoles for the H+, K(+)-ATPase in hog gastric vesicles were measured by using the pyruvate kinase-lactate dehydrogenase-linked system in which hydrolysis of ATP was coupled with the oxidation of NADH. The vesicles were incubated in a solution containing a high concentration of KCl, valinomycin and Mg-ATP, and the intravesicular medium was acidified. The inhibitor was activated in the acidic medium and reacted with SH groups on the luminal (intravesicular) side of the ATPase. The active compound formed in the extravesicular medium (pH 6.11) was quenched by GSH. Under these conditions, IC50 of new compound E3810, 2[(4-(3-methoxypropoxy)-3-methylpyridine-2-yl)methyl-sulfinyl]-1H- benzimidazole sodium salt, was 0.072 microM and that of omeprazole was 0.47 microM at 25 degrees. On the other hand, the rates of formation of active compounds, tetracyclic sulfenamide derivatives, from original substituted benzimidazoles in 0.1 N HCl (k) were determined by measuring optical density at the characteristic wavelengths of the active compounds. There was a good correlation between IC50 and k for various substituted benzimidazoles including E3810, methoxy derivative of E3810, omeprazole, Ro 18-5364, H compound, picoprazole and timoprazole. This fact suggest that the rate of the formation of the acid-activated compound is a main factor determining the potency of the inhibitor.

Infection of cardiac suture line after ventricular aneurysmectomy. Report of two cases.

Two cases of sepsis following ventricular aneurysmectomy are presented. In both, the source of sepsis was an infected caridac suture line in which Teflon felt strips were used to reinforce the closure. One patient had a pseudoaneurysm and a chronic empyema of the left side of the chest, and the second had a ventriculocutaneous fistula. Early operation with removal of the foreign body is warranted in this major complication of a ventricular aneurysmectomy.

Histomorphometric analysis of the response of rat tibiae to shape memory alloy (nitinol).

The bone reaction to nitinol (Ni-Ti), a metal with shape memory, and other materials inserted transcortically and extending into the medullary canal of rat tibiae was quantitatively assessed using an image processing system. The materials examined were implants, all of the same shape and size, composed of nitinol, pure titanium (Ti), anodic oxidized Ti (AO-Ti), a titanium alloy (Ti-6Al-4V) and pure nickel (Ni). While the other four implant materials were progressively encapsulated with bone tissues, Ni was encapsulated with connective tissues through the 168-day experimental period, and the Ni implants showed no bone contact at any time during the experimental period. Histometric analysis revealed no significant difference among the tissue reactions to Ti, AO-Ti and Ti-6Al-4V, but Ni-Ti implants showed significantly (P < 0.01) lower percentage bone contact and bone contact area than any of the other titanium or titanium alloy materials.

Effects of chronic exercise on blood pressure in Dahl salt-sensitive rats.

We tested the hypothesis that daily exercise would reduce directly measured arterial blood pressure (BP) and sympathetic nervous system support of BP in conscious, unrestrained, female Dahl salt-sensitive rats consuming 4.0% NaCl. Dahl S/Jr inbred rats were assigned to daily exercise (EX) or sedentary (SED) treatment conditions (n = 12/group) at 4 weeks of age. Rats in the EX group were housed in cages with attached running wheels. After 5 weeks of exercise, rats were running 10.3 +/- 1.7 km/day. After at least 5 wks of treatment, all rats in both groups were placed on a 4.0% NaCl diet for 2 weeks to produce sodium-induced hypertension. Rats continued to either exercise daily or remain sedentary for an additional 2 weeks while consuming the high sodium diet. Carotid and jugular catheters were then implanted for measurements in conscious, resting, unrestrained rats on two separate days. Daily wheel running exercise for 7 to 9 weeks did not alter BP or HR in Dahl S/Jr rats consuming a 4.0% NaCl diet. However, acute arterial depressor responses to ganglionic blockade were less in EX rats. Furthermore, greater alpha-adrenergic (phenylephrine-induced) pressor responses were observed in the EX group while under ganglionic blockade. The findings suggest that overall resting sympathetic neural activity or cardiac beta-adrenergic responsiveness to sympathetic activity is reduced in this model of hypertension by daily wheel running exercise.

Mechanism of inhibitory action of mesaconitine in acute inflammations.

Mesaconitine (MA) inhibited carrageenin-induced hind-paw edema in sham-operated mice as well as adrenalectomized mice. Hind-paw edema produced by subplantar injection of histamine, serotonin and prostaglandin E1 was suppressed by MA, indicating that it elicits the antiinflammatory activity at the early exudative stage of inflammations. However, MA did not affect the biosynthesis of the prostaglandins. Trazoline and propranolol had no effect on the inhibitory activity of MA on carrageenin-induced hind-paw edema. MA when administered i.c. at the doses where it shows marked analgesic activity produced dose-dependent antiinflammatory responses on paw edema produced by carrageenin and on vascular permeability accelerated by acetic acid and agar. The inhibitory activity of morphine on carrageenin-induced paw edema failed to be potentiated by the concurrent administration of MA, demonstrating that the mechanism of the antiinflammatory activity of MA involves the central nervous system.

HLA antigens as immunogenetic markers of alcoholism and alcoholic liver disease.

HLA antigens were studied among 94 chronic alcoholics. Concerning A and B-loci, there was no significant change of phenotype frequency (PF) in the HLA typing between the patients and controls (80 healthy subjects). However, there was a significant difference in the PF of CW3 between chronic alcoholics and controls (58.5% in alcoholics vs 30.0% in controls). The corrected p value was less than 0.05 with relative risk value being 3.29 HLA-DR loci were also detected in 26 patients, but there was no significant difference between the patients and controls. All alcoholics were subdivided according to the hepatic morphology, and the PF of HLA was examined. A significant high frequency of HLA CW3 was found in patients with hepatitis (64%) compared to controls (30%). There was also an increased PF of CW3 in the liver cirrhosis group (59% in cirrhosis group vs 30% in controls). In conclusion, chronic alcoholics have a significantly higher PF of HLA-CW3 as compared to controls. This characteristic feature becomes even more distinct in alcoholics with severe hepatic lesions.

Association of HLA-B40 and DRW9 with Japanese alcoholic liver cirrhosis.

Seventy-seven chronic alcoholics with liver disease were studied to evaluate the HLA antigen association. There were no significant differences of HLA antigen phenotype frequencies (PF) between the patients and controls regarding A and C loci, (62 healthy Japanese). Prevalences of HLA-B40 complex (B40 . 48 . 13) and DRW9 tended to increase among chronic alcoholics. When chronic alcoholics were divided according to whether they had liver cirrhosis or not, the cirrhosis group (42 cases) revealed a significantly higher frequency of HLA-DRW9 (chi 2 = 10.88, p less than 0.001, corrected p less than 0.05, relative risk (R.R.) = 4.17) as compared to controls. There was also a tendency of B40 complex to increase in frequency (chi 2 = 5.51, p less than 0.05, R.R. = 2.65) in the cirrhosis group. Haplotype frequency and linkage disequilibrium parameters of HLA-B40 . 48-DRW9 were significantly higher than those of controls. Moreover, the increased frequency of DRW9 in the cirrhosis group was similar to that in autoimmune disease like ulcerative colitis or SLE. These data suggest that HLA-DRW9 and/or HLA-B40-DRW9 might be closely associated with susceptibility to developing alcoholic cirrhosis and that autoimmune mechanisms might be involved partly in its etiology.

[Lymphocytes in transplantation immunity--in the relation to HLA antigens].

Allograft rejection, in kidney transplantation, is initiated by recipient's lymphocytes with the recognition of genetically determined antigens, histocompatibility antigens, which are expressed on the graft, followed by killer T cell induction and antibody production against the graft. Recent advances in our knowledge of the human major histcompatibility complex antigens-HLA antigens, which have an important role in transplantation immunity, are herein reviewed briefly and the relation to allograft rejection is discussed.

Structure of the cyclic glucan produced from amylopectin by Bacillus stearothermophilus branching enzyme.

The thermostable branching enzyme (BE, EC 2.4.1.18) from Bacillus stearothermophilus TRBE14 produces large cyclic glucans from waxy rice amylopectin similar to those obtained from amylose as described elsewhere [H. Takata, T. Takaha, S. Okada. M. Takagi, and T. Imanaka, J. Bacteriol., 178 (1996) 1600-1606]. The structure of the product (P-1) from the late-stage reaction was analyzed in detail. The weight-average degree of polymerization (dpw) of P-1 was 900. Its chain-length distribution was not significantly changed compared with that of amylopectin, although the amount of long chains (dp > 38) was slightly decreased. The cyclic component of P-1, which was isolated by the extensive action of glucoamylase, had dpw of 49. Three point five alpha-1,6 linkages were directly involved in the formation of the ring structure with several non-cyclic side chains linked to the ring. Based on these results, the action and new roles of BE are discussed.

Cellular basis of gastrulation in the sand dollar Scaphechinus mirabilis.

The processes of gastrulation in the sand dollar Scaphechinus mirabilis are quite different from those in regular echinoids. In this study, we explored the cellular basis of gastrulation in this species with several methods. Cell-tracing experiments revealed that the prospective endodermal cells were convoluted throughout the invagination processes. Histological observation showed that the ectodermal layer remained thickened, and the vegetal cells retained an elongated shape until the last step of invagination. Further, most of the vegetal ectodermal cells were skewed or distorted. Wedge-shaped cells were common in the vegetal ectoderm, especially at the subequatorial region. In these embryos, unlike the embryos of regular echinoids, secondary mesenchyme cells did not seem to exert the force to pull up the archenteron toward the inner surface of the apical plate. In fact, the archenteron cells were not stretched along the axis of elongation and were in close contact with each other. Here we found that gastrulation was completely blocked when the embryos were attached to a glass dish coated with poly-L-lysine, in which the movement of the ectodermal layer was inhibited. These results suggest that a force generated by the thickened ectoderm, rather than rearrangement of the archenteron cells, may play a key role in the archenteron elongation in S. mirabilis embryos.

Anabolic principles of Aconitum roots.

The methanol extracts of raw and processed roots of Aconitum carmichaeli were shown to stimulate amino acid incorporation into mouse liver protein after approx. 10 h of ingestion. The extract of the raw roots was fractionated by monitoring the anabolic activity to furnish the aconitine alkaloids as active principles, among which mesaconitine exhibited the strongest activity. Amino acid incorporation into liver protein in the mesaconitine treated mice was inhibited by actinomycin D to the level of normal mice. Long-term administration of mesaconitine induced no reinforcement of the anabolic activity in mouse liver. Examination of the anabolic activity in liver, spleen, kidney, testis and serum revealed that mesaconitine potentiated protein synthesis only in liver and rather reduced it in kidney.

Establishment and characterization of a cell line, KaMi, from human lung large cell carcinoma.

A cell line of human lung large cell carcinoma (LCC) was established directly from the metastatic skin tumor tissue. The clinical course of the patient who carried this carcinoma was peculiar; generalized lymphadenopathy, histologically resembling Hodgkin's disease, was found as the first clinical symptom. The lung tumor was not discovered until the time of autopsy. This cell line (KaMi) grew adherent to culture vessels with the population doubling time of 20.6h, formed colonies in soft agars with efficiency of 22.6%, and formed tumors in athymic nude mice. The authenticity of KaMi was confirmed by chromosomal analysis and isoenzyme patterns. KaMi cells bore a strong resemblance to the original tumor cells which were composed of small spindle cells, large polygonal cells, and multinucleated giant cells. Immunohistochemically, KaMi cells showed a weak tendency to differentiate to squamous cells, and these immunohistochemical reactivities were almost compatible to those of the original tumor cells, but ultrastructurally, KaMi cells were more immature than the original ones. Treatment with several reagents could not augment a differentiation of KaMi cells. Cytokeratin profiles showed a tendency of squamous cell differentiation. KaMi cells may aid in elucidating the pathogenesis and biology of LCC and its relationship to other lung tumors.

Immortalization of rat spleen and thymus T cells by human T-cell leukemia virus type I.

Co-cultivation of thymus and spleen cells of Fisher and Lewis rats with lethally irradiated MT-2 cells harboring human T-cell leukemia virus type I (HTLV-I) resulted in the establishment of lymphoid cell lines, FIRT-1, FIRS-1, LERT-1, and LERS-1, respectively. Cells of these cell lines had rat T-cell characters as demonstrated by the positive reaction to monoclonal antibodies (MAbs) to rat T cell antigens (Thy 1 and pan T). They lacked surface immunoglobulins and strongly expressed rat interleukin-2 receptor antigen (Tac) and Ia antigen. Karyotypic analysis revealed that they had the normal rat karyotype in early cultures, but showed marked aneuploidy after long cultivation. None of them expressed HTLV gag proteins (p19 and p24) or virus particles, but they contained HTLV-I proviral DNA monoclonally and weakly expressed pX gene products (p40x). They were not transplantable into syngeneic newborn rats.