Involvement of oxidative stress in atherosclerosis development in subjects with sarcopenic obesity.

INTRODUCTION: Co­existing decreased muscle mass and increased visceral fat, an age­associated change called sarcopenic obesity, results in fragility and cardiovascular disease. To assess the pathogenesis of sarcopenic obesity, we assessed the associations of clinical parameters with psoas muscle mass in elderly male subjects with obesity and type 2 diabetes. METHODS: The subjects were 55 patients, over 65 years of age and with a visceral fat area exceeding 100 cm2, with type 2 diabetes. The cross­sectional area of the psoas muscle is considered to provide an estimation of overall muscle mass. Sarcopenia was considered to be present when the total psoas muscle area was low, defined as a value below 500 mm2 m−2 on a computed tomographic scan. RESULTS: The maximum intima­media thickness (max IMT) and urinary 8­isoprostane values were significantly higher in the sarcopenic group. Multiple linear regression analysis revealed max IMT to be an independent variable related to muscle mass decline. In addition, logistic analysis showed max IMT and urinary 8­isoprostane to be variables independently contributing to total psoas muscle area <500 mm2 m−2. CONCLUSION: Worsening surrogate markers for systemic oxidative stress and atherosclerosis were associated with declining muscle mass in elderly subjects with obesity and type 2 diabetes. These results indicate that systemic oxidative stress is among the mechanisms underlying atherosclerosis development in subjects with sarcopenic obesity.

Positive association of free triiodothyronine with pancreatic ß-cell function in people with prediabetes.

AIM: To analyse the effects of thyroid hormones on ß-cell function and glucose metabolism in people with prediabetes who are euthyroid. METHODS: A total of 111 people who were euthyroid underwent 75-g oral glucose tolerance tests, of whom 52 were assigned to the normal glucose tolerance and 59 to the prediabetes groups. Homeostatic model assessment of ß-cell function, insulinogenic index and areas under the curve for insulin and glucose were evaluated as indices of pancreatic ß-cell function. RESULTS: In both groups, BMI, fasting insulin, homeostasis model assessment ratio and HDL cholesterol correlated significantly with all indices of pancreatic ß-cell function. Free triiodothyronine correlated positively with all insulin secretion indices in the prediabetes group. Multiple linear regression analysis showed that free triiodothyronine was an independent variable that had a positive correlation with all indices of ß-cell function in the prediabetes group. By contrast, no such correlation was found in the normal glucose tolerance group. CONCLUSIONS: Free triiodothyronine is associated with both basal and glucose-stimulated insulin secretion in people with prediabetes who are euthyroid; therefore, the regulation of insulin secretion by thyroid hormones is a potentially novel therapeutic target for the treatment of diabetes.

Expression pattern of Galectin 4 in rat placentation.

Galectin 4 (Gal4) is abundantly expressed in the epithelium of the gastrointestinal tract, and functional analysis has concentrated on its roles associated with polarized membrane trafficking. This study aimed to investigate the expression of Gal4 in placentation. The expression level of Gal4 was revealed to be lower in differentiated Rcho-1 cells (a model system of rat trophoblast differentiation) than in proliferative cells. In the rat placenta, immunohistochemical analysis showed that Gal4 is preferentially located in the maternal-fetal junctional zone. These results suggest that down-regulation of Gal4 may be involved in the promotion of trophoblast cell differentiation.

Lecithin-cholesterol acyltransferase (LCAT) deficiency without mutations in the coding sequence: a case report and literature review.

Familial lecithin-cholesterol acyltransferase (LCAT) deficiency (FLD) is a rare genetic disease characterized by corneal opacities, normocytic anemia, dyslipidemia, and proteinuria progressing to chronic renal failure. In all FLD cases, a mutation has been found in the coding sequence of the LCAT gene. FLD is clinically distinguished from an acquired form of LCAT deficiency by the presence of corneal opacities. Here we describe a 36-year-old woman presenting with clinical, pathological, and laboratory data compatible with FLD. Her mother and elder sister had corneal opacities. However, genetic analysis revealed there were no mutations in the LCAT coding sequences and no alterations in LCAT mRNA expression. Furthermore, we were unable to find any underlying conditions that may lead to LCAT deficiency. The present case therefore demonstrates that LCAT deficiency may be caused by factors other than mutations in the coding sequence and we suggest that a translational or posttranslational mechanism may be involved.

Evidence for a pioneer round of mRNA translation: mRNAs subject to nonsense-mediated decay in mammalian cells are bound by CBP80 and CBP20.

Nonsense-mediated decay (NMD) eliminates mRNAs that prematurely terminate translation. We used antibody to the nuclear cap binding protein CBP80 or its cytoplasmic counterpart eIF4E to immunopurify RNP containing nonsense-free or nonsense-containing transcripts. Data indicate that NMD takes place in association with CBP80. We defined other components of NMD-susceptible mRNP as CBP20, PABP2, eIF4G, and the NMD factors Upf2 and Upf3. Consistent with the dependence of NMD on translation, the NMD of CBP80-bound mRNA is blocked by cycloheximide or suppressor tRNA. These findings provide evidence that translation can take place in association with CBP80. They also indicate that CBP80-bound mRNA undergoes a "pioneer" round of translation, before CBP80-CBP20 are replaced by eIF4E, and Upf2 and Upf3 proteins dissociate from upstream of exon-exon junctions.

Evidence that phosphorylation of human Upfl protein varies with intracellular location and is mediated by a wortmannin-sensitive and rapamycin-sensitive PI 3-kinase-related kinase signaling pathway.

Human Upf1 protein (p), a group 1 RNA helicase, has recently been shown to function in nonsense-mediated mRNA decay (NMD) in mammalian cells. Here, we demonstrate that the estimated 3 x 10(6) copies of hUpf1 p per exponentially growing HeLa cell are essentially equally distributed among polysomal, subpolysomal, and ribosome-free fractions. We also demonstrate that hUpf1p binds RNA and is a phosphoprotein harboring phosphoserine and phosphothreonine. hUpf1p is phosphorylated to the highest extent when polysome-associated and to the lowest extent when ribosome free. We find that serum-induced phosphorylation of hUpf1p is inhibited by wortmannin at a concentration that selectively inhibits PI 3-kinase related kinases and, to a lesser extent, by rapamycin. These and other data suggest that phosphorylation is mediated by a wortmannin-sensitive and rapamycin-sensitive PI 3-kinase-related kinase signaling pathway. Comparisons are made of hUpf1p to Upf1p and SMG-2, which are the orthologs to hUpf1p in Saccharomyces cerevisiae and Caenorhabditis elegans, respectively.

Virus-mediated transduction of apolipoprotein E (ApoE)-sendai develops lipoprotein glomerulopathy in ApoE-deficient mice.

Lipoprotein glomerulopathy (LPG) is a unique renal disease characterized by thrombus-like substances in markedly dilated glomerular capillaries, dysbetalipoproteinemia, and elevated plasma concentrations of apoE. Recent studies identified several apoE mutations in patients with LPG, including apoE2(R145P) Sendai (apoE-Sendai). Virus-mediated transduction of apoE-Sendai in apoE-deficient hypercholesterolemic mice resulted in insufficient correction of hypercholesterolemia and a marked and temporal induction of plasma triglyceride levels. In vitro binding studies showed that apoE-Sendai has a reduced affinity for the low density lipoprotein receptor, suggesting that dysbetalipoproteinemia in LPG is caused by the apoE mutation. Furthermore, histological examination revealed marked intraglomerular depositions of apoE-containing lipoproteins in mice injected with apoE-Sendai virus. These LPG-like depositions were detected 6 days after virus injection and were sustained for at least 60 days. Our results demonstrated that apoE-Sendai is an etiological cause of LPG.

Development and characterization of a DNA solar dosimeter.

In this paper, we report the development and characterization of a solar ultraviolet (UV) dosimetry system that can be used as a film badge for radiation monitoring. DNA molecules are coated on a thin nylon membrane as a UV dosimeter. The membrane is sealed in a polyethylene filter envelope with silica gel to keep the humidity low. After exposure to UV or solar light, induced DNA damage is measured by an immunochemical reaction. The intensity of color developed during the immunological reaction can be correlated linearly with the irradiated UV dose delivered by an Oriel solar simulator within a limited dose range. We observe no effects of temperature on the level of damage induction. The membrane is proficient for measuring DNA damage for more than 21 days when stored at either 37 or 4 degrees C. The induced damage remains stable on the membrane for at least 22 days at both 37 and 4 degrees C. In addition to these indoor experiments, we report measurements of solar UV dose in outdoor experiments.

Enhanced human tumor cell transplantability in a new congenic immunodeficient mouse; KSN-BNX.

We introduced two mutant genes (beige; bg that induces the deficiency of natural killer (NK) activity and xid that decreases the production of immunoglobulin) into KSN nude mice with high reproductive performances. We produced KSN bg/bg(nu/nu) (KSN-bg), KSN-xid/xid(nu/nn) (KSN-xid), KSN xid/xid,bg/bg(nu/nu) (KSN-BNX) and KSN-nu/+ (KS) mice by back-cross (cross-intercross method). All strains showed as high a reproductivity rate as the parental KSN mice. KSN-xid and KSN-BNX mice had a reduced percentage of B220 positive cells in the spleens compared to KSN and KSN-bg mice, but they showed increased percentages of Thy-1 and asialo GM1 positive cells. The serum immunoglobulin concentrations of KSN-BNX were as low as KSN-xid. Both KSN-bg and KSN-BNX mice showed deficient NK activity in spleens, whereas KSN-xid mice showed an elevated NK activity. Compared to nude mice, the growth of both human tumor cell TCO-1 and BxPc-3 transplanted subcutaneously was enhanced in KSN-BNX mice. However Panc-1 cells that was rejected in nude mice was not accepted in KSN-BNX mice. Liver metastasis of human pancreatic tumor cells; Capan-1, BxPc-3 and MIAPaCa-2 were studied. No significant difference was observed in the percentage of metastasis formed mice between nude and KSN-BNX mice.

Anti-angiogenic effect of TGFbeta in aqueous humor.

Neovascularization is mediated by various factors in ocular tissues. Recent studies have emphasized the role of vascular endothelial growth factor in the induction of angiogenesis. We have previously reported that aqueous humor (AH) suppressed vascular endothelial cell growth and angiogenesis. We speculated that the anti-angiogenic effect of AH is mediated by transforming growth factor beta (TGFbeta). In order to clarify the presence of TGFbeta in bovine AH, we applied it on the heparin-sepharose affinity column and prepared two fractions (bound and unbound fractions). We measured TGFbeta concentration in each fraction and examined how the anti-TGFbeta antibody decreased the inhibitory effect of AH on human umbilical vein endothelial cell growth and on in vitro angiogenesis. We found the presence of TGFbeta2, but not TGFbeta1, in the heparin bound fraction, and the inhibitory effect was detected in the heparin-bound fraction. Anti-TGFbeta antibody completely and dose-dependently extinguished the inhibitory effect of AH. We propose that the inhibitory effect of AH on endothelial cell growth and in vitro angiogenesis are both mediated by TGFbeta2. Our results indicate TGFbeta2 is normally present in AH and protects the eye tissue against abnormal neovascularization.

A comparison of UVB-carcinogenesis between nude mice and nude beige mice.

To gain an insight into the relationship between UVB-carcinogenesis and natural killer activity, we examined ultraviolet light-induced carcinogenesis in mice with high natural killer, activity (KSN) and mice with natural killer deficiency (KSN-bg). We exposed mice six times a week to three levels of daily ultraviolet B (UVB) doses; 320, 160 and 0 J/m2/day. During the latency period of skin tumor development in KSN mice, we detected no suppression of the natural killer activity at both 320 and 160 J/m2/day. Even at 1340 J/m2/day, we could not detect any significant suppression of NK activity in KSN mice. When we irradiated spleen cells in vitro, we observed NK activity suppression. Next, we compared the carcinogenic effects of UVB-irradiation on KSN and KSN-bg mice. At 320 J/m2/day, we detected no significant differences between them. In contrast, at 160 J/m2/day, KSN-bg mice showed a significantly higher rate of skin tumor induction than KSN mice (p < 0.05). Most UVB-induced tumors were squamous cell carcinoma, the rest were spindle cell carcinoma, papilloma and mixed type. Our results suggest that NK activity plays a protective role against UVB-carcinogenesis from low daily-doses of UVB-irradiation.

An UVB-carcinogenesis model with KSN nude mice.

We established and characterized a systematic ultraviolet light-induced carcinogenesis model using KSN nude mice. We prepared five groups of KSN mice and exposed them six times a week to five levels of daily ultraviolet B (UVB) doses; 1340, 670, 320, 160 and 0 J/m2/day. In 670, 320 and 160 J/m2/day, the latency period tended to become shorter in proportion to the daily doses and prevalence data fitted well to log-normal distribution. In the log-log plot of days till 50% prevalence versus daily dose, we saw a linear relationship for 1 mm tumor diameter. From this analysis, we determined that days necessary to reach 50% prevalence is in proportion to the -0.49 power of daily dose. The average number of tumors per survivor correlated with prevalence data. Direct measured rates of tumor growth were independent of daily UVB-dose. Therefore we speculated that UV-irradiation did not affect tumor growth after its appearance. Most UVB-induced tumors were squamous cell carcinoma, the rest were spindle cell carcinoma, papilloma and mixed type. We concluded that our experimental data with nude mice was in accordance with data with hairless mice in nature.

Supranuclear melanin caps reduce ultraviolet induced DNA photoproducts in human epidermis.

Melanin can form supranuclear caps in human epidermis, suggesting that intracellular melanin reduces ultraviolet transmission to underlying cell nuclei and inhibits the formation of ultraviolet induced DNA photoproducts. The purpose of this study was to determine the photoprotective effect of epidermal melanin. We irradiated normal human skin explants with ultraviolet B and determined the formation of cyclobutane pyrimidine dimers and (6-4)photoproducts in individual epidermal cells by indirect immunofluorescence and by laser cytometry using monoclonal antibodies specific for cyclobutane dimers or for (6-4)photoproducts. We found that epidermal cells with supranuclear melanin caps had significantly less DNA photoproducts (both types) than epidermal cells without supranuclear melanin caps. Moreover, the protection factor against both types of photolesions correlated with melanin concentration in epidermal cells. These results indicate that melanin reduces ultraviolet induced DNA photoproducts in human epidermis in a concentration dependent manner.

Monitoring the free radial forearm flap in pharyngo-oesophageal reconstruction.

Monitoring the viability of the free radial forearm flap after pharyngo-oesophageal reconstruction poses technical problems. A method of monitoring using a small island flap elevated distal to the main forearm flap is presented. This monitor flap is easy to elevate. It is placed externally and provides information about the perfusion of the buried reconstruction flap. Postoperative management requires only assessment of tissue colour, turgor, capillary refill and bleeding, as for standard skin flap monitoring.

New immunodeficient mouse strains bred by introducing beige and xid mutations into the KSN nude strain.

We introduced two mutant genes (beige or bg, which induces a deficiency of natural killer activity, and xid, which decreases the production of immunoglobulins) into KSN nude mice with high reproductive performance. At first we produced the KSN-bg/bg(nu/nu) (KSN-bg) and KSN-xid/xid(nu/nu) (KSN-xid) congenic strain by backcross (cross-intercross method). After we identified homozygosity at the biochemical locus and polymorphic microsatellite loci, we mated KSN-bg and KSN-xid mice, and selected the KSN-xid/xid;bg/bg(nu/nu) (KSN-BNX) mice from the progeny. Furthermore we introduced the non-nude gene, which originated from CBA/N, into the KSN strain and produced KSN-nu/+ (KS) mice that have the same genetic background except for the nu locus. All strains had as high a reproductivity rate as the parental KSN mice. The KSN-xid and KSN-BNX mice had a reduced percentage of B220-positive cells in the spleen compared with KSN and KSN-bg mice, but they had increased percentages of Thy-1 and asialo GM1-positive cells. The serum immunoglobulin concentrations of BNX were as low as those of KSN-xid mice. Both KSN-bg and KSN-BNX mice had deficient natural killer activity in the spleen, whereas KSN-xid mice had increased natural killer activity. Compared with nude mice, the growth of the human thyroid tumor cell line transplanted subcutaneously was enhanced in BNX mice. These KSN, KSN-bg, KSN-xid, KSN-BNX, and KS nice not only are of value for use in various fields as the hosts of xenograft but also are good models of the combination effect of multiple immunodeficient genes.

[The effectiveness of discharge planning (DSP) in home care--comparison of conditions of cancer patient shifted to home treatment before and after DSP introduction].

We developed a discharge planning (DSP) procedure as a method to assess the family and cancer patient situation when making the shift to home care, and we herewith report its significance. DSP is intended to sustain the patient's quality of life (QOL) and improve it; for a smooth transition to treatment at home, the family and patient situation must be assessed from various perspectives. The prospective illness conditions, needed medical treatment and care should be appropriately planned, with due recognition of the roles to be played and the requisite pooling of professional information in order to handle emergencies and the like when such arise. In the present study, in order to clarify issues dealing with DSP effectiveness, we compared 34 patients on home care, 17 before DSP and 17 thereafter. The results indicated that following the introduction of DSP, the home care increased for patients with gastroenterological problems such as pain, general debilitation, poor appetite, constipation, diarrhea and the like and those impulsiveness, vomiting and mental symptoms overlapping. With the proper assessment at the time of the transition to home care, this care could be continued. Moreover, with patient satisfaction, there was the need for assessment over the course in order to evaluate treatment and care. In the days to come, there is the need to improve this approach with a scoring system in the evaluation.

[Factors affecting continuation of home medical care--based on the relationship between patient and caregiver status].

In recent years it has gradually become possible to have some control over the medical condition of the end-stage cancer patient. However, there are various problems involved in the continuation of home care. In order to clarify how the mutual relationship between patient and caregiver effects the continuation of home care, we conducted a study on 28 end-stage cancer patients who made the transition from our hospital to home care by means of our HOme Care Service. Using the Kaneda Assessment Score, we obtained information from patients' charts, added reasons for rehospitalizations and analyzed the data. The results showed that the group of patients who remained on home care, despite the fact that their condition was significantly poorer than that of the group who discontinued home care, showed a significantly higher proportion of "wishing to be cared for at home to the last," in the view of the caregiver, a joint commitment of the caregiver together with the family, and a suitable response to changes in the patient. As a direction for forthcoming support in this area, there is a need for sufficient assessment in order to evaluate the balance between caregiver and family. It was suggested that guidance in each case and the use of community resources will be very important.

[Treatment of multiple sclerosis with lymphocytapheresis using a leukocyte adsorption column].

Twenty four patients with multiple sclerosis and four with retrobulbar neuritis were treated with lymphocytapheresis using leukocyte adsorption column. Although intravenous pulsed methylpredonine therapy followed by oral prednisolone therapy was used to treat these patients, they still fail to respond to treatment. Lymphocytapheresis was carried using a "Cellsorba" leukocyte adsorption column (Asahi Medical Co., Tokyo). About 3000 ml of whole blood was treated with the anticoagulant nafamostat mesilate and passed through the column. Eight of the 24 patients who underwent lymphocytapheresis therapy showed marked to moderate improvement. Prior to treat with lymphocytapheresis, their EDSS grades varied 3.0 to 8.5. Within 4-12 hours after therapy each of the eight patients showed EDSS grade improvement of 2 or more steps. Although about 50% of all lymphocytes were removed by lymphocytapheresis, Tal-positive activated T cells were removed more efficiently. None of the patients experienced any side effects following lymphocytapheresis. Treatment efficacy seems to be correlated with the removal of certain pathologically-activated lymphocytes, in particular, Tal-positive T cells.

Photorepair and excision repair removal of UV-induced pyrimidine dimers and (6-4) photoproducts in the tail fin of the medaka, Oryzias latipes.

Induction and repair of UV-B induced DNa damage in the tail fin of the Medaka, were examined immunohistochemicaly and by the enzyme-linked immunosorbent assay (ELISA). UV-induced DNA damage was detected only in the outermost layer of epithelial cells and did not differ in fishes having different degree of melanization. Both pyrimidine dimers and (6-4) photoproducts in the fin cells were removed by excision repair in the dark, the excision of (6-4) photoproducts being about twice as efficient as that of pyrimidine dimers. The rate of excision repair of UV-induced lesions in fin tissue was three to four times that in cultured Medaka cells, OL32. In the fin cells, reductions in the numbers of pyrimidine dimers and (6-4) photoproducts were seen after treatment with fluorescent light, whereas less reductions of pyrimidine dimers and no reductions of (6-4) photoproducts were observed in OL32 cells.