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Biomedical subjects

M Nagano

Publications and source records attributed to M Nagano.

At least 37 records · Page 2Linked to original sources

Primate spermatogonial stem cells colonize mouse testes.

In mice, transplantation of spermatogonial stem cells from a fertile male to the seminiferous tubules of an infertile recipient male results in progeny with donor-derived haplotype. Attempts to extend this approach by transplanting human testis cells to mice have led to conflicting claims that no donor germ cells persisted or that human spermatozoa were produced in the recipient. To examine this issue we used the baboon, a primate in which testis cell populations of several ages could be obtained for transplantation, and demonstrate that donor spermatogonial stem cells readily establish germ cell colonies in recipient mice, which exist for periods of at least 6 mo. However, differentiation of germ cells toward the lumen of the tubule and production of spermatozoa did not occur. The presence of baboon spermatogonial stem cells and undifferentiated spermatogonia in mouse seminiferous tubules for long periods after transplantation indicates that antigens, growth factors, and signaling molecules that are necessary for interaction of these cells and the testis environment have been preserved for 100 million years of evolutionary separation. Because germ cell differentiation and spermatogenesis did not occur, the molecules necessary for this process appear to have undergone greater divergence between baboon and mouse.

Animals↗

Point mutation (-69 G-->A) in the promoter region of cholesteryl ester transfer protein gene in Japanese hyperalphalipoproteinemic subjects.

Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester (CE) from HDL to apolipoprotein (apo) B-containing lipoproteins and plays a crucial role in reverse cholesterol transport, which is a major protective system against atherosclerosis. Genetic CETP deficiency is the most common cause of a marked hyperalphalipoproteinemia (HALP) in the Japanese, and various mutations have been identified in the coding region as well as in the exon/intron boundaries in the CETP gene. In the present study, we identified a novel mutation in the promoter region of the CETP gene. This mutation was a G-to-A substitution at the -69 nucleotide of the promoter region (-69 G-->A), corresponding to the second nucleotide of the PEA3/ETS binding site (CGGAA) located upstream of the putative TATA box. Four (2.0%) of 196 unrelated subjects with a marked HALP (HDL cholesterol >/=2.59 mmol/L=100 mg/dL) were revealed to be heterozygous for the -69 G-->A mutation, and the allelic frequency of the mutant was 0.0102 in the subjects with a marked HALP. The subjects with the -69 G-->A mutation had low plasma CETP levels. Reporter gene assay showed that this mutation markedly reduced the transcriptional activities in HepG2 cells (8% of wild type). These results suggested that this mutation would be dominant negative. In conclusion, a novel -69 G-->A mutation in the CETP gene causes the decreased transcriptional activity leading to HALP.

Carrier Proteins↗

Males in rural Bangladeshi communities are more susceptible to chronic arsenic poisoning than females: analyses based on urinary arsenic.

Spot urine samples were collected from the inhabitants of two rural communities in northwestern Bangladesh. We compared arsenic levels in the urine samples ([As](u); n = 346) with those in water from tube wells ([As](tw); range < 1-535 microg/L; n = 86) on an individual basis. The small variation of [As](u) within subjects and highly positive correlation with [As](tw) indicate that [As](u) is a useful indicator of exposure. Analyses of [As](u) showed that creatinine correction was necessary, that [As](u) only reflected recent exposure, and that there were substantial interindividual differences for a given [As](tw) level. To evaluate the toxic effects of arsenic exposure, we constructed a system for rating skin manifestations, which revealed distinct sex-related differences. Comparison of males and females in the same households confirmed that skin manifestations were more severe in the males, and in the males of one community a dose-response relationship between [As](u) and the degree of skin manifestation was evident. The results of this study indicate that [As](u) in spot urine samples can be used as an exposure indicator for As. They suggest that there might be sex-related, and perhaps community-related, differences in the relationship between [As](u) and skin manifestations, although several confounding factors, including sunlight exposure and smoking habits, might contribute to the observed sex difference. The existence of such differences should be further confirmed and examined in other populations to identify the subpopulations sensitive to chronic arsenic toxicity.

Adult↗

In vitro growth of mouse ovarian preantral follicles and the capacity of their oocytes to develop to the blastocyst stage.

Two groups of mouse preantral follicles with diameters of 125-150 and 151-175 microm were cultured individually for 6 days in a medium supplemented with FSH and fetal calf serum to determine their in vitro growth characteristics. Their oocyte capacity for maturation and development to the blastocyst stage following in vitro fertilization was also assessed. Antral formation rate at the end of culture was higher in the follicles of 151-175 microm (89%) than 125-150 microm (76%). The timing of antrum formation was different between the two follicle categories: most 151-175 microm follicles formed antra earlier than 125-150 microm follicles (days 4 and 5 vs. 5 and 6). However, follicle diameters at the time of antrum formation were the same regardless of the initial size and the culture period. Maturation rates of the oocytes derived from both categories of in vitro grown follicles (70 and 62%) were not different from those of oocytes from in vivo grown follicles (74%). The in vitro derived oocytes, however, showed less cleavage (30 and 35%) than the in vivo derived oocytes (89%). Although the oocytes from both follicle categories developed to the morula stage after in vitro fertilization, blastocysts were only obtained from oocytes derived from the 151-175 microm category. These results demonstrate that an individual follicle culture system using a medium with FSH and fetal calf serum supports in vitro growth of mouse preantral follicles with diameters of 151-175 microm to the preovulatory stage, and that their oocytes have the capability to develop to the blastocyst stage.

Animals↗

Relationship among growth, steroid production and immunolocalization of transforming growth factor-beta 1 in the normally developing mouse follicles cultured in vitro.

This study examined the relationship among growth, steroid production and transforming growth factor-beta 1 (TGF-beta 1) immunolocalization in the mouse follicles cultured in vitro to evaluate the hypothesis that normally developing follicles should express TGF-beta 1 in the granulosa cells around the time of antrum formation. Preantral follicles with 151-175 microns (large category) and 125-150 microns (small category) of initial diameters were used as models for normal and retarded follicles, respectively. Growth rate and timing of antrum formation in both categories were comparable to those of in-vivo grown follicles. At the time of antrum formation, follicular diameters were similar between the two follicle categories; however, antral follicles from the large category showed larger number of granulosa cells, higher estradiol production and proportion of follicles with TGF-beta 1 positive granulosa cells. Two days after antrum formation, there were no differences in the number of granulosa cells and the proportions of follicles with TGF-beta 1 positive granulosa or theca cells between the two categories. Temporal association in large follicles between the increase in estradiol production and proportion of follicles with TGF-beta 1 positive granulosa cells at the time of antrum formation supports our hypothesis. Furthermore, this study demonstrated the usefulness of the follicle culture system in the investigations of follicular physiology.

Animals↗

[Small cell carcinoma of the prostate: a case report].

A 81-year-old man was admitted to our department with the chief complaints of pollakisuria and difficulty in voiding. He presented with increased serum PSA level (over 100 ng/ml). We performed biopsy of the prostate and found a moderately differentiated adenocarcinoma. Various urological examinations showed metastases to paraaortic lymph nodes and systemic bones. He was started-on hormonal therapy. Nine months from the start of hormonal therapy, this therapy was effective and the serum PSA level was decreased to 14 ng/ml. Thereafter, the serum PSA level and the tumor volume were increased and he died 29 months from the start of treatment. The autopsy revealed small cell carcinoma with adenocarcinoma of the prostate.

Aged↗

[Recurrence of vesicoureteral reflux detected 19 years after ureterocystoneostomy: a case report].

A 21-year-old male, who had been operated on for bilateral vesicoureteral refluxes (VURs) with bilateral ureterocystoneostomy (Politano-Leadbetter's method) 19 years before, was admitted to our hospital due to recurrent VUR. Since the former operation, he had undergone voiding cystography (VCUG) twice for two years, and no refluxes were found. Moreover, no evidence of upper urinary tract deterioration was found by either intravenous pyelography (IVP) or renal ultrasound scanning taken the year before this admission. Nineteen years after the operation, the dilation of the left lower ureter was found on IVP and, consequently, he suffered from pyelonephritis. The VCUG revealed the recurrence of left VUR. Because of his allergic reaction to collagen, we again performed left ureterocystoneostomy (Politano-Leadbetter's method). At three months postoperatively, there was no VUR found on VCUG.

Adult↗

Malaria infections in crews of Japanese ships.

Plasmodium falciparum malaria is the most dangerous infection for seafarers in West Africa. In December 1998, five cases of this infection occurred among Japanese seafarers in West Africa, two of them died, one on board ship, and another died five days after the admission to the hospital in Reunion island, East Africa. Six other cases of falciparum malaria infection occurred among Japanese seafarers on another ship in December 1999. Three infected persons were admitted to hospitals in Abidjan (Ivory Coast) and Point Noire (Congo). In Japan, over 100 cases of imported malaria were recorded each year during the period from 1990 to 1997, and about 40% of these cases were falciparum infections. It is not known how many of them occurred among seafarers. We estimate that at least 5% of all malaria cases in Japan are seafarers. Measures to protect crews of ships against malaria are discussed.

Adult↗

Retrovirus-mediated gene delivery into male germ line stem cells.

The male germ line stem cell is the only cell type in the adult that can contribute genes to the next generation and is characterized by postnatal proliferation. It has not been determined whether this cell population can be used to deliberately introduce genetic modification into the germ line to generate transgenic animals or whether human somatic cell gene therapy has the potential to accidentally introduce permanent genetic changes into a patient's germ line. Here we report that several techniques can be used to achieve both in vitro and in vivo gene transfer into mouse male germ line stem cells using a retroviral vector. Expression of a retrovirally delivered reporter lacZ transgene in male germ line stem cells and differentiated germ cells persisted in the testis for more than 6 months. At least one in 300 stem cells could be infected. The experiments demonstrate a system to introduce genes directly into the male germ line and also provide a method to address the potential of human somatic cell gene therapy DNA constructs to enter a patient's germ line.

Animals↗

Health surveillance in the population living in a methyl mercury-polluted area over a long period.

It is important to follow up on the health status of inhabitants living in the methyl mercury-polluted area surrounding Minamata City, paying particular attention to diseases not only of the central nervous system but also of other organs. We have been carrying out such concentric studies for more than 10 years. We have previously studied the cause-specific standard mortality ratios in Minamata disease patients and reported that the SMRs for liver disease and renal disease were significantly raised in male and female patients, respectively. It was also found that complications arising from diabetes could be due to the large number of old people among the autopsy cases. The next step was to clarify the actual prevalence and incidence of liver disease, renal disease, and diabetes mellitus epidemiologically among the population in this area. The aim of this study was to determine the actual prevalence of these diseases and complaints, and to investigate the contribution of various risk factors to these diseases in this area. The study was a population-based cross-sectional mass screening survey. A case-control study was designed to estimate the role of various risk factors including methyl mercury exposure for these diseases. A mass multiple health examination survey was performed in 1500 subjects aged 40 years and older in Tsunagi Town, neighboring Minamata City, every summer since 1984. Tsunagi Town is located in a methyl mercury-polluted area and there are 36.9 certified Minamata disease patients per 1000 population. Data concerning liver disease, renal disease, and diabetes mellitus were collected on the basis of urine, hematological, physical, and ultrasonographic examinations. Data on risk factors and subjective complaints were collected by interview and other measures. The prevalence of these diseases was not higher in this methyl mercury-polluted area compared with other areas in Japan, contrary to what was expected based on standard mortality ratios and pathological findings. There were no positive correlations between those diseases and methyl mercury exposure. On the other hand, the population in the polluted area had more and a greater variety of complaints than those in the nonpolluted area. It is possible that not only neurological subjective complaints but also nonspecific complaints of the population in the polluted area might be influenced by past methyl mercury exposure. This health surveillance in the population living in a methyl mercury-polluted area must be maintained in the future.

Aged↗

Short-chain fatty acids inhibit the release and content of growth hormone in anterior pituitary cells of the goat.

The effects of short-chain fatty acids (SCFA: acetate, propionate, and butyrate) on growth hormone (GH)-releasing hormone (GHRH)-induced GH secretion from pituitary somatotrophs were assessed on isolated anterior pituitary cells of goats. Cells were cultured in Dulbecco's modified Eagle's medium for 3 days, either in the presence (1, 3, or 10 mM) or in the absence of each SCFA, and then stimulated with GHRH (10(-12) to 10(-7) M) for 30 min, again in the presence of and at the concentration of SCFA used over the previous 3 days. In the cells cultured in the absence of SCFA, the addition of SCFA to the medium during the GHRH stimulation period did not significantly change GHRH-induced GH release. However, in cells cultured in the presence of either propionate (3 or 10 mM) or butyrate (1, 3, or 10 mM), the addition of SCFA to the medium during GHRH stimulation significantly reduced the GHRH-induced GH release. The inhibitory effects of SCFA were dependent on the concentrations of SCFA and were greater for butyrate than for propionate. In the cells cultured in the presence of butyrate, but not in the absence, the total GH production (the sum of the released GH and the remaining GH after stimulation) was also significantly reduced. The GHmRNA expression was reduced in the cells cultured with 10 mM butyrate, whereas it was enhanced by the stimulation with 10(-7) M GHRH. These findings suggest that propionate and butyrate may inhibit GHRH-induced GH release and GH production by caprine anterior pituitary cells.

Acetates↗

Maternal program of apoptosis activated shortly after midblastula transition by overexpression of S-adenosylmethionine decarboxylase in Xenopus early embryos.

When we studied polyamine metabolism in Xenopus embryos, we cloned the cDNA for Xenopus S-adenosylmethionine decarboxylase (SAMDC), which converts SAM (S-adenosylmethionine), the methyl donor, into decarboxylated SAM (dcSAM), the aminopropyl donor, and microinjected its in vitro transcribed mRNA into Xenopus fertilized eggs. We found here that the mRNA injection induces a SAM deficient state in early embryos due to over-function of the overexpressed SAMDC, which in turn induces inhibition of protein synthesis. Such embryos developed quite normally until blastula stage, but stopped development at the early gastrula stage, due to induction of massive cell dissociation and cell autolysis, irrespective of the dosage and stage of the mRNA injection. We found that the dissociated cells were TUNEL-positive, contained fragmented nuclei with ladder-forming DNA, and furthermore, rescued completely by coinjection of Bcl-2 mRNA. Thus, overexpression of SAMDC in Xenopus embryos appeared to switch on apoptotic program, probably via inhibition of protein synthesis. Here, we briefly review our results together with those reported from other laboratories. After discussing the general importance of this newly discovered apoptotic program, we propose that the maternal program of apoptosis serves as a surveillance mechanism to eliminate metabolically severely-damaged cells and functions as a 'fail-safe' mechanism for normal development in Xenopus embryos.

Adenosylmethionine Decarboxylase↗

Species specificity of barnacle settlement-inducing proteins.

We previously isolated a larval settlement-inducing protein complex (SIPC) from adult extracts of the barnacle, Balanus amphitrite using a nitrocellulose membrane settlement assay. In the present study, we found that the extracts of other adult barnacles, Megabalanus rosa and Balanus eburneus, also induced the settlement of B. amphitrite cyprids although the inductive activity was slightly lower than that of conspecific extracts. Furthermore, we examined reactivity to anti-SIPC antibody in adult extracts from six species of Japanese barnacles other than B. amphitrite, brine shrimp and eight marine sessile organisms besides barnacles. The results showed that all barnacles examined contained SIPC-like proteins with slightly different molecular weight, while the other animals did not react to the antibody by immunoblot analysis. These findings suggest that species specificity in settlement-inducing proteins of barnacles is not so strict, but these proteins are characteristic to barnacle species.

Animals↗

CDK inhibitors suppress apoptosis induced by chemicals and by excessive expression of a cell death gene, reaper, in Drosophila cells.

The present study was aimed to investigate whether or not cyclin-dependent kinases (CDKs) participate in different cascades leading to apoptosis. We examined the effects of two CDK inhibitors, olomoucine (OLM) and butyrolactone-I (BL-I), on apoptosis induced in two kinds of Drosophila cell lines. Increases of caspase activity induced by actinomycin D, cycloheximide, H-7 or A23187 in a Drosophila neuronal cell line, ML-DmBG2-c2, and induced by excessive expression of a Drosophila cell death gene, reaper, in Drosophila S2 cells were suppressed by 24-h pretreatment of each CDK inhibitor. Concomitant with the suppression of the caspase activity, fragmentations of cells and DNA, representatives of apoptosis, were also inhibited. These results suggest that CDK(s) participates in progression of apoptosis. However, these effects of the CDK inhibitors were also observed even at lower doses which did not affect cell proliferation. Therefore, it was shown that apoptosis is not always related to cell cycle in Drosophila cells. It was also suggested that the target(s) of the CDK inhibitors locates upstream of caspase in the cascade(s) of apoptosis.

4-Butyrolactone↗

Calmodulin-dependent and -independent apoptosis in cell of a Drosophila neuronal cell line.

This study was undertaken to reveal apoptotic pathways in neurons using a Drosophila neuronal cell line derived from larval central nervous system. We could induce apoptotic cell death in the cells by a Ca2+ ionophore (A23187), a protein kinase inhibitor (H-7), an RNA synthesis inhibitor (actinomycin D) and a protein synthesis inhibitor (cycloheximide). All the apoptosis induced by each chemical required Ca2+ ions, although the origin of Ca2+ ions were different: apoptosis induced by A23187 was dependent on extracellular Ca2+ ions whereas those by the other three chemicals utilized intracellular Ca2+ ions. Furthermore, different reactions to W-7, a calmodulin inhibitor, were found: W-7 prevented the cell death by each of the three chemicals but not by A23187. Based on the results, we proposed that the apoptotic pathways are classified into two types in individual cells. One pathway induced by H-7, actinomycin D or cycloheximide is calmodulin-dependent (pathway H), and another induced by A23187 is calmodulin-independent (pathway A).

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

The validity of bioelectrical impedance phase angle for nutritional assessment in children.

BACKGROUND/PURPOSE: Bioelectrical impedance analysis (BIA) is a quick and noninvasive method for estimating body composition. Many prediction equations have been reported recently using bioelectrical impedance to calculate fat free mass (FFM) and fat mass (FM). These equations are based on the assumption that the composition and density of FFM are stable. In children, the composition and density of FFM vary according to age and clinical state, so the use of these equations is limited. However, phase angle is directly determined from resistance (Rz) and reactance (Xc) without equations and reflects body cell mass. The authors, therefore, investigated the validity of phase angle for nutritional assessment in children. METHODS: Bioelectrical impedance analysis and anthropometric measurements were performed in 81 patients, including 71 well-nourished and 10 malnourished children. RESULTS: Phase angle correlated with body weight (R = 0.818) and arm muscle circumference (r = 0.901) in well-nourished children. Malnourished patients showed lower phase angle than that of well-nourished children. CONCLUSION: Bioelectrical impedance phase angle is a useful parameter for nutritional assessment in children.

Body Composition↗

Graft healing in the bone tunnel in anterior cruciate ligament reconstruction.

The histologic sequence in the bone tunnel after anterior cruciate ligament reconstruction using the bone-patellar tendon-bone graft was investigated in this study. Eighteen adult mongrel dogs were used. After excision of the anterior cruciate ligament, the graft was routed through the bone tunnels and fixed with interference fit screws. After the dogs were sacrificed at intervals, the bone blocks containing the bone tunnels were isolated and processed for histologic examination. At the bone-bone interface, incorporation of the bone plug at each end of the graft was completed at 12 weeks. The structure of the tendon insertion of the grafted patellar tendon, consisting of four distinct zones, was observed without apparent necrotic and degenerative change for as long as 12 weeks. Between the tendon and the bone tunnel, a layer of hypercellular fibrous tissue gradually became mature with time. Thus, it appeared the morphologic characteristics and location of the reestablished attachment of the bone-patellar tendon-bone graft were more similar to those of the native anterior cruciate ligament compared with the graft to bone healing in the hamstring tendon graft.

Animals↗

Beneficial effects of vitamin E treatment in acute myocardial infarction.

BACKGROUND: Vitamin E (Vit E), an antioxidant, is considered to prolong survival in patients and animals after myocardial infarction. Because myocardial infarction is associated with arrhythmia and heart dysfunction, this study tested the hypothesis that early treatment with Vit E reduces mortality because of its protective effects against arrhythmia and cardiac dysfunction induced by acute myocardial infarction. METHODS: Rats were randomly divided into 4 groups: sham control, myocardial infarcted, Vit E-treated sham control, and Vit E-treated infarcted animals. Myocardial infarction was induced by ligation of the left anterior descending coronary artery. Treated animals received Vit E (25 mg/kg/d) through a gastric tube beginning 1 hour after the coronary occlusion, whereas control rats received tap water. RESULTS: Electrocardiograms (lead II) at 1, 3, 7, and 21 days after coronary occlusion in the untreated animals showed ST-segment elevation, abnormal Q waves, premature ventricular complex (PVC), and QTc prolongation. Conversely, Vit E-treated rats showed attenuated ST-segment changes, fewer abnormal Q waves, and decreased incidence of PVC after coronary occlusion. Total mortality was reduced from 38% to 16%, whereas the infarct size was decreased from 44.2% to 22.3% in infarcted rats treated with Vit E. The depression in left ventricular function as well as elevation of malondialdehyde content and conjugated diene formation in the 21-day infarcted rat hearts were prevented by Vit E treatment. CONCLUSION: These results indicate that Vit E may exert beneficial effects on the heart by reducing oxidative stress in acute myocardial infarction.

Animals↗