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Biomedical subjects

M Sakai

Publications and source records attributed to M Sakai.

At least 37 records · Page 2Linked to original sources
PubMed2005

Loop-mediated isothermal amplification: an emerging technology for detection of fish and shellfish pathogens.

Fish and shellfish diseases are a constant threat to the sustainability and economic viability of aquaculture. Early diagnosis plays a vital role in management of fish and shellfish diseases. Traditionally, various biochemical and serological tests have been used for fish disease diagnosis. However, the time and expertise required for such diagnoses makes it difficult for aquaculturists to easily adopt them under production conditions. Polymerase chain reaction and probe-based nucleic acid detection have become increasingly popular in fish and shellfish diagnostics. Recently, a novel technique called loop-mediated isothermal amplification (LAMP) has been developed, which is highly sensitive and rapid. LAMP has been used for the detection of bacterial, viral, fungal and parasitic diseases in both animal and plants. In aquaculture, LAMP-based detection of pathogens like Edwardsiella tarda, E. ictaluri, Nocardia seriolae, Tetracapsuloides bryosalmonae, white spot syndrome virus and infectious haematopoietic necrosis virus have been reported. In this review, the application of LAMP for the detection of aquaculture-associated pathogens is discussed.

Animals↗
PubMed2005

Clock gene expression in the submandibular glands.

Clock genes, which mediate molecular circadian rhythms, are expressed in a circadian fashion in the suprachiasmatic nucleus and in various peripheral tissues. To establish a molecular basis for circadian regulation in the salivary glands, we examined expression profiles of clock-related genes and salivary gland-characteristic genes. Clock-related genes-including Per1, Per2, Cry1, Bmal1, Dec1, Dec2, Dbp, and Reverbalpha-showed robust circadian expression rhythms in the submandibular glands in 12:12-hour light-dark conditions. In addition, a robust circadian rhythm was observed in amylase 1 mRNA levels, whereas the expression of other salivary-gland-characteristic genes examined was not rhythmic. The Clock mutation resulted in increased or decreased mRNA levels of Per2, Bmal1, Dec1, Dec2, and Dbp, and in Cry1-/- background, Cry2 disruption also increased or decreased mRNA levels of these clock-related genes and the amylase 1 gene. These findings indicate that the Clock- and Cry-dependent molecular clock system is active in the salivary glands.

ARNTL Transcription Factors↗
PubMed2005

Identification of expressed genes in carp (Cyprinus carpio) head kidney cells after in vitro treatment with immunostimulants.

We analysed a total of 530 expressed genes from the head kidney cells of common carp (Cyprinus carpio) treated by immunostimulants. Sequences of the cDNA clones were compared with sequences in the GenBank database. Immune-related genes identified after in vitro stimulation of carp head kidney cells were: BPI/LBP, C-type lectins, fucolectins, CC-chemokine, CXC-chemokine, CD18, cyclophilin, FcgammaR, G-CSFR, HSP 70, Ig heavy and light chains, NITR, integrin beta2-alpha, Mx, interleukin-1beta, beta thymosin, lysozyme G & C, MHC class II associated invariant chain 1 and 2, granulin, CAAT binding protein and tumour necrosis factor-alpha induced protein. The expression of these immune-related genes may be important for estimating the efficacy of immunostimulants.

Adjuvants, Immunologic↗
PubMed2004

Decidual and peripheral blood CD4+CD25+ regulatory T cells in early pregnancy subjects and spontaneous abortion cases.

Human pregnancy represents a situation of semiallograft to maternal host. Therefore, it has been reported that tolerance to the fetal allograft represents a mechanism for maintaining a pregnancy. CD4(+)CD25(bright) regulatory T cells are known to play an important role in the development and maintenance of tolerance in peripheral tissues. However, the potential role of CD4(+)CD25(bright) T cells in maintaining human pregnancy has not been reported. In this study, we show that early human pregnancy decidua contains an abundance of CD4(+)CD25(bright) T cells, which express CD152(CTLA-4) at a high level. CD4(+)CD25(bright) T cells mediate potent inhibition of autologous T-cell proliferation by anti-CD3 stimulation. Furthermore, these cells inhibit the proliferation of autologous CD4(+)CD25(-) T cells in a dose-dependent fashion. This suppressive function of decidual CD4(+)CD25(+) T cells required cell-to-cell contact. The proportion of decidual CD4(+)CD25(bright) T cells was significantly lower in specimens from spontaneous abortion compared to those from specimens from induced abortions. These results suggest that decidual CD4(+)CD25(bright) T cells contribute to the mechanisms mediating maternal immune tolerance of conceptus antigens and therefore might contribute to the maintenance of pregnancy.

Abortion, Spontaneous↗
PubMed2004

Serum granulysin is a marker for Th1 type immunity in pre-eclampsia.

Recent studies suggest that pre-eclampsia is associated with a Th1 predominant state and may be considered a failure of tolerance. Granulysin is a cytotoxic granule protein of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). Recently, we developed an enzyme-linked immunosorbent assay (ELISA) system for detecting serum granulysin, and reported that serum granulysin is a useful marker to evaluate the cell-mediated immunity. In this study, we show that the serum levels of granulysin were significantly elevated in pre-eclamptic patients compared with those in normal pregnancy subjects. In addition, the serum granulysin levels in pre-eclamptic patients were well associated with mean blood pressure, percentage of peripheral blood Th1 cells and Th1/Th2 ratios. The present results suggest that the serum granulysin levels would be a useful and novel serum marker to evaluate the Th1/Th2 balance, especially Th1 type immunity in pre-eclampsia.

Adult↗
PubMed2004

Detection of koi herpesvirus in common carp, Cyprinus carpio L., by loop-mediated isothermal amplification.

Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. In this study, using the LAMP method, a protocol for koi herpes virus (KHV) detection in common carp was designed. A set of four primers, two inner and two outer, were designed based on the sequence of the thymidine kinase (tk) gene of KHV. Time and temperature conditions for detection of KHV were optimized for 60 min at 65 degrees C. The detection limit using LAMP was found to be similar to that by polymerase chain reaction. In this study, we have developed a highly sensitive and rapid diagnostic procedure for detection of KHV infection in common carp.

Animals↗
PubMed2004

Auto-spermatophore extrusion reveals that the reproductive timer functions in the separated terminal abdominal ganglion in the male cricket.

Auto-spermatophore extrusion is a kind of spermatophore extrusion without genital coupling in the male cricket. It rarely occurred in intact males paired with a female, while it frequently occurred in all the males with the connectives cut under restraint and dissection. The time interval (SPaSE) between spermatophore preparation and auto-spermatophore extrusion was found to be comparable to that (RS2) of the time-fixed sexually refractory stage measured by the calling song. According to extracellular spike recording, the dorsal pouch motoneuron (mDP), which singly innervates the dorsal pouch muscles and is responsible for spermatophore extrusion, showed a burst discharge in association with auto-spermatophore extrusion with an interval similar to RS2 in males with the connectives transected between the 6th abdominal ganglion and the terminal abdominal ganglion (TAG) after spermatophore preparation. These results strengthened our previous conclusion that the reproductive timer for RS2 is located in the TAG, and demonstrated that it functions normally even in the TAG separated from the rest of the central nervous system.

Abdomen↗
PubMed2004

Thee effect of 5-HTP on the reproductive timer in the male cricket.

The post-copulatory sexually refractory stage in the male cricket Gryllus bimaculatus consists of the two substages: the first refractory stage (RS1, time-variable) between copulation and spermatophore preparation, and the second refractory stage (RS2, time-constant) between spermatophore preparation and the recommencement of courtship. To understand the mechanism of the timer for RS2, subcuticular or intraganglionic injection of biogenic amines (10(-2) mol l(-1)) was performed immediately after spermatophore preparation. RS2 was shortened by octopamine, 5-HT, 5-HTP and NA-5-HT. Among these, 5-HTP was most potent. It shortened RS2 to maximally about 38% of the control. The shortening effect continued for 4.5 h after subcuticular injection even when the hemolymph was washed out with saline at 1 hour after injection. Simultaneous injection of 5-HTP with the inhibitor (NSD-1015) of 5-HT synthesis enzyme nullified the effect of 5-HTP, indicating that the shortening effect was caused by 5-HT synthesized from extrinsic 5-HTP. Injection of the inhibitor (CHX) of protein synthesis had no effect of on RS2. These results suggest that the reproductive timer in the TAG may be controlled by 5-HT or a second messenger mediated by 5-HT.

5-Hydroxytryptophan↗
PubMed2003

Hypermethylation of retrotransposons in the liverwort Marchantia paleacea var. diptera.

Three DNA fragments encoding part of a reverse transcriptase similar to that of Ty1- copia retrotransposons were isolated from suspension-cultured cells of the liverwort Marchantia paleacea var. diptera. Digestion of total DNA with methylation-sensitive and -insensitive isoschizomers revealed that, like those of the subtelomeric region, CpG sequences in retrotransposon genes were hypermethylated. The methylation status and expression of these retrotransposons was less affected by 5-azadeoxycytidine treatment compared to the subtelomeric region, which was considerably demethylated.

Amino Acid Sequence↗
PubMed2003

Scavenger receptors for oxidized and glycated proteins.

Our present knowledge on chemically modified proteins and their receptor systems is originated from a proposal by Goldstein and Brown in 1979 for the receptor for acetylated LDL which is involved in foam cell formation, one of critical steps in atherogenesis. Subsequent extensive studies using oxidized LDL (OxLDL) as a representative ligand disclosed at least 11 different scavenger receptors which are collectively categorized as "scavenger receptor family". Advanced glycation endproducts (AGE) and their receptor systems have been studied independently until recent findings that AGE-proteins are also recognized as active ligands by scavenger receptors including class A scavenger receptor (SR-A), class B scavenger receptors such as CD36 and SR-BI, type D scavenger receptor (LOX-1) and FEEL-1/FEEL-2. Three messages can be summarized from these experiments; (i) endocytic uptake of OxLDL and AGE-proteins by macrophages or macrophage-derived cells is mainly mediated by SR-A and CD36, which is an important step for foam cell formation in the early stage of atherosclerosis, (ii) selective uptake of cholesteryl esters of high density lipoprotein (HDL) mediated by SR-BI is inhibited by AGE-proteins, suggesting a potential pathological role of AGE in a HDL-mediated reverse cholesterol transport system, (iii) a novel scavenger receptor is involved in hepatic clearance of plasma OxLDL and AGE-proteins.

Animals↗
PubMed2003

A case of body stalk anomaly arising in the second baby of a triplet pregnancy after in-vitro fertilization and embryo transfer.

We report a case of body stalk anomaly arising in the second baby of a triplet pregnancy after in-vitro fertilization and embryo transfer (IVF-ET). Body stalk anomaly or limb-body wall complex is a rare congenital anomaly with a series of similar clinical manifestations and poor prognosis. IVF-ET is an effective treatment for various types of infertility. We summarize and discuss herein the relation with the sequence of genesis for such malformations and multiple pregnancies after IVF-ET.

Abdominal Wall↗
PubMed2003

Sonographic assessment of the cervix before, during and after a uterine contraction is effective in predicting the course of labor.

OBJECTIVE: To investigate whether the degree of change in cervical length during a uterine contraction is predictive of subsequent progression of labor. METHODS: The subjects were 73 uncomplicated parturient women at term. We observed the cervix before, during and after a uterine contraction by transvaginal ultrasound in the first stage of labor and determined the degree of cervical shortening during the contraction relative to the cervical length before contraction. We related the degree of cervical shortening to labor patterns at the time of the ultrasound examination, which were retrospectively determined by reviewing the partogram. RESULTS: The cervix was shortened in length by about 50% on average during a uterine contraction in the normal course of labor. The degree of cervical shortening was significantly greater in the normal latent and active phases than it was in the prolonged latent phase, protracted active phase and false labor, whereas there were no differences between the former two phases nor between the latter three phases. Nulliparous and parous women exhibited almost the same degree of shortening in the normal latent and active phases. CONCLUSIONS: Real-time ultrasound observation of the cervix during uterine contraction could help differentiate inefficient uterine contractions from normal ones and thus predict the subsequent course of labor.

Adolescent↗
PubMed2003

Relationship of regional adiposity to serum leptin level in nonobese Japanese type 2 diabetic male patients.

BACKGROUND: The aim of the present study was to investigate the relationships between serum leptin levels and regional adipose fat area, BMI, and the measures of variables including serum insulin in nonobese Japanese type 2 diabetic patients. METHODS: A total of 121 nonobese Japanese type 2 diabetic patients [aged 35 to 83 years, body mass index (BMI) (15.4 to 26.8 kg/m(2))] were studied. They all were male patients. In conjunction with serum leptin level, BMI, glycosylated hemoglobin (HbA(1c)), and fasting concentrations of plasma glucose and serum insulin and lipids (triglycerides, total and HDL cholesterol) were measured. RESULTS: Univariate regression analysis showed that serum leptin levels were positively correlated to subcutaneous (r=0.566, P<0.0001) and visceral (r=0.481, P<0.001) fat area in our diabetic patients. Furthermore, serum leptin levels were positively correlated to serum insulin (r=0.517, P<0.0001), BMI (r=0.428, P<0.0001), serum triglycerides (r=0.279, P<0.005), and age (r=0.225, P<0.05). There was, however, no relationship between serum leptin levels and measures of other variables including total and HDL cholesterol. Multiple regression analyses showed that serum leptin levels were predicted by subcutaneous fat area (F=5.92, P<0.0001) and serum insulin level (F=5.60, P<0.0001), which explained 29.0% of the variability of serum leptin concentrations in our nonobese Japanese type 2 diabetic male patients. Visceral fat area, BMI, serum triglycerides, and age, however, were not independently associated with serum leptin levels in our patients. CONCLUSIONS: These results indicate that serum leptin levels are reflective of subcutaneous fat area in nonobese Japanese type 2 diabetic male patients.

Adipose Tissue↗
PubMed2003

Survivin inhibits apoptosis in cytotrophoblasts.

Survivin, a protein that inhibits apoptosis, is expressed in a variety of tumour cells. We detected survivin-specific mRNA and protein in normal placental tissues, two human choriocarcinoma cell lines (JEG-3 and BeWo), and a trophoblastic cell line (tPA30-1) by reverse transcription-polymerase chain reaction (RT-PCR), Northern blotting, and Western blotting. Immunohistochemically, survivin was localized to normal villous cytotrophoblasts, normal extravillous trophoblasts, cytotrophoblasts in hydatidiform mole, and choriocarcinoma cells. Antisense oligonucleotides for survivin dose-dependently induced apoptosis in two choriocarcinoma cell lines (JEG-3 and BeWo) and a trophoblastic cell line (tPA30-1), while sense oligonucleotides showed little effect. These findings suggest that survivin antagonizes apoptosis in cytotrophoblasts, extravillous trophoblasts, and choriocarcinoma cells.

Adult↗
PubMed2003

Decrease of T-helper 2 and T-cytotoxic 2 cells at implantation sites occurs in unexplained recurrent spontaneous abortion with normal chromosomal content.

BACKGROUND: In normal pregnancy, predominant type 2 cytokines help maintain pregnancy, and a T-helper (Th)1 type response is associated with unexplained recurrent spontaneous abortion (RSA). However, Th2 and T-cytotoxic (Tc)2 cells have not been localized at the implantation site in RSA. METHODS: Twenty-one cases with RSA were classified into RSA with normal chromosomal content (RSA-N, n = 10) and RSA with abnormal chromosomal content (RSA-A, n = 11). As a control, we selected 15 gestational age-matched cases of induced abortion with no history of spontaneous abortion. We immunostained paraffin-embedded decidual sections for a specific Th2 and Tc2 cell marker termed 'chemo-attractant receptor-homologous molecule expressed on Th2 cells (CRTH2)' and T-cell markers CD3 and CD8. The numbers and percentages of Th2 (CRTH2(+)CD8(-)CD3(+)) and Tc2 (CRTH2(+)CD8(+)) cells were compared between the decidua basalis and decidua parietalis. RESULTS: Th2 and Tc2 cells accumulated in the decidua basalis in normal pregnancy. Accumulation of Tc2 cells and both Th2 and Tc2 cells decreased in the decidua basalis in RSA-A and RSA-N respectively. The number and percentage of Th2, and Tc2 cells in the decidua parietalis were similar in normal pregnancy, RSA-A and RSA-N. CONCLUSION: Decreased Th2 and Tc2 cells at the implantation site may contribute to RSA-N.

Abortion, Habitual↗
PubMed2002

Serotonin precursor (5-hydroxytryptophan) has a profound effect on the post-copulatory time-fixed sexually refractory stage in the male cricket, Gryllus bimaculatus DeGeer.

This study addresses a potentially general basis of measuring time in a biological timer. Here, we examined the effects of biogenic amines on the time-fixed post-copulatory sexually refractory stage (ca. 1 h) which is defined as the time interval between spermatophore protrusion and the onset of a calling or a mating response in the reproductive cycle of the male cricket. For subcuticular injection of amines (0.15 ml, 10(-2) mol l(-1)), the interval of the refractory stage was shortened by octopamine, serotonin, 5-hydoxytryptophan and N-acetyl-serotonin but was unchanged by tryptophan, melatonin or 5-hydroxyindol-3-acetic acid. The effect of 5-hydoxytryptophan was most potent (maximum shortening, 38%) and long lasting (ca. 4.5 h) while other amines effected only the injected cycle. Injection of 5-hydoxytryptophan (180 nl, 10(-2) mol l(-1)) into the terminal abdominal ganglion also decreased the interval to a similar extent. Simultaneous injection of 5-hydoxytryptophan with the inhibitor of the serotonin synthesis enzyme reduced the 5-hydoxytryptophan effect suggesting that this effect results from synthesis of serotonin from 5-hydoxytryptophan. The protein synthesis inhibitor cycloheximide had no effect on the interval. These results suggest that the reproductive timer is regulated by serotonergic neurons in the terminal abdominal ganglion without protein synthesis during the interval of the time-fixed sexually refractory stage.

5-Hydroxytryptophan↗
PubMed2002

Effect of short-term low-intensity exercise on insulin sensitivity, insulin secretion, and glucose and lipid metabolism in non-obese Japanese type 2 diabetic patients.

The aim of the present study was to investigate the effects of short-term physical exercise that did not change body mass on insulin sensitivity, insulin secretion, and glucose and lipid metabolism in 39 non-obese Japanese type 2 diabetic patients. Insulin sensitivity and insulin secretion were estimated with homeostasis model assessment insulin resistance (HOMA-IR) and HOMA-B-cell function proposed by Matthews et al., respectively. All patients were hospitalized and were engaged in low-intensity exercise that consisted of walking and dumbbell exercise for successive 7 days. There were no changes in hospital diet and the dose of any medications used throughout the study. Fasting glucose, insulin, and lipids were measured before and after exercise. After exercise, serum triglyceride levels significantly decreased, but no significant changes were observed in total and HDL cholesterol concentrations. Fasting glucose, insulin, and HOMA-IR levels significantly decreased after exercise, but HOMA-B-cell function did not change during the study. There was no significant difference between BMI levels before and after exercise. From these results, it can be concluded that short-term (7 days) low-intensity physical exercise combined with hospital diet reduces serum triglycerides, insulin resistance, and fasting glucose levels without affecting BMI in non-obese Japanese type 2 diabetic patients.

Adult↗