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N Amino

Publications and source records attributed to N Amino.

At least 19 recordsLinked to original sources
PubMed2004

High-throughput differential screening of mRNAs by serial analysis of gene expression: decreased expression of trefoil factor 3 mRNA in thyroid follicular carcinomas.

To find mRNAs whose expression differs between thyroid follicular adenomas and carcinomas, a high-throughput analysis of mRNAs in these two tumours was performed. This method, named high-throughput differential screening by serial analysis of gene expression (HDSS), combines a modified method of serial analysis of gene expression (SAGE) and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). A total of 40 candidate tag sequences that showed extremely different expression levels between a follicular carcinoma and a follicular adenoma in the SAGE analysis were analysed by real-time quantitative RT-PCR, using RNAs from an additional four typical follicular carcinomas and adenomas. One sequence tag that represents trefoil factor 3 (TFF3) mRNA showed a clear difference in its expression level between adenomas and carcinomas. The expression levels of TFF3 mRNA in 48 follicular adenomas and 29 follicular carcinomas were measured by real-time quantitative RT-PCR using a specific probe for TFF3. They were significantly decreased in follicular carcinomas, especially in widely invasive types and those with evident metastases. These results indicate that the decreased expression of TFF3 mRNA is a marker of follicular carcinomas, especially those with a high risk of invasion or metastasis.

Adenocarcinoma, Follicular↗
PubMed2004

PGP9.5 mRNA could contribute to the molecular-based diagnosis of medullary thyroid carcinoma.

The protein gene product 9.5 (PGP9.5) is a ubiquitin hydrolase that is widely expressed in neuronal tissues at all stages of neuronal differentiation and is a known neuroendocrine marker. Medullary thyroid carcinoma (MTC) arises from parafollicular cells and is reported to overexpress several mRNAs such as RET, calcitonin, and CEA. These markers are thought to be useful in determining a molecular-based diagnosis of MTC. We examined the expression levels of PGP9.5 mRNA in 80 thyroid tissues using real-time quantitative reverse transcription (RT-PCR) and found that PGP9.5 mRNA was overexpressed in all 11 MTCs examined, both hereditary and sporadic, but not in other histological tumour types. Furthermore, by RT-PCR, PGP9.5 mRNA was detected only in aspirates from three medullary carcinomas, and not in aspirates from other tumour types. These results demonstrate that, in addition to the expression of RET, calcitonin and CEA, PGP9.5 mRNA expression may contribute to the molecular-based diagnosis of MTCs.

Adolescent↗
PubMed2003

Novel autoantibodies to pituitary gland specific factor 1a in patients with rheumatoid arthritis.

OBJECTIVE: We recently identified a new protein, pituitary gland specific factor 1a (PGSF1a), that is specifically transcribed in the pituitary gland. In our investigation of anti-PGSF1a antibody for pituitary diseases, we examined it in patients with RA and other autoimmune diseases. We unexpectedly discovered the frequent existence of anti-PGSF1a antibody in patients with RA. We therefore examined the prevalence of this antibody to understand its clinical significance in RA. METHODS: Anti-PGSF1a antibody was detected by radioligand assay using recombinant (35)S-labelled PGSF1a protein. Antibody activity is expressed as an index that was obtained by comparison with normal pooled serum. RESULTS: RA patients had a significantly higher mean anti-PGSF1a antibody index (n=46, 1.28+/-0.38, P < 0.001) than healthy controls (n=36, 1.04+/-0.13). Indices greater than the cut-off value (mean+2 S.D. of healthy controls) were found in 43.5% (20/46) and 10.0% (2/20) of patients with RA and osteoarthritis, respectively. There was no correlation between the activities of anti-PGSF1a antibodies and titres of rheumatoid factor (RF) or serum C-reactive protein concentrations, but RA patients with more erosive disease had a higher mean anti-PGSF1a antibody index. Four of eight sera samples obtained from RF-negative RA patients were positive for anti-PGSF1a antibodies. CONCLUSION: Anti-PGSF1a antibody is a useful new marker for the diagnosis of RA, especially for RF-negative RA, and may relate to clinical manifestations of RA.

Amino Acid Sequence↗
PubMed2002

Development of liver dysfunction after delivery is possibly due to postpartum autoimmune hepatitis. A report of three cases.

Autoimmune diseases, especially autoimmune thyroid disease, frequently develop after delivery due to the immune rebound mechanism. Most cases involve transient dysfunction of affected organs. We examined three patients who developed liver dysfunction after delivery. They were all diagnosed with definite or probable autoimmune hepatitis using the scoring system of the International Autoimmune Hepatitis Group. Moreover, all of them had anti-CYP2D6 antibodies detected by a sensitive radioligand assay. Our findings strongly suggest that liver dysfunction is induced by postpartum autoimmune hepatitis, and clinicians should be aware of this disease.

Adult↗
PubMed2002

Expression profile of active genes in the human pituitary gland.

To characterize transcripts abundantly expressed in the human pituitary gland in general as well as to isolate novel transcripts expressed specifically in this gland, we generated an expression profile of the active genes transcribed in it. A total of 1015 randomly collected 3prime prime or minute expressed sequence tags (ESTs) (gene signatures, GSs) were grouped into 527GS species. The results showed the relative expression levels of genes in the pituitary gland. The genes comprising more than 1% of total mRNA were prolactin, growth hormone and chromogranin B genes. When known genes were categorized, the genes for pituitary hormones were the most actively transcribed, followed by the genes for ribosomal proteins, nuclear proteins and secretory granule proteins. Through comparison of this gene expression profile with the BodyMap database containing profiles generated from 63 other human tissues, we obtained 11 genes which appeared to be specifically expressed in the pituitary gland. In addition to the eight known genes, we identified three novel pituitary-specific transcripts which encode putative proteins: pituitary gland specific factor 1a (PGSF1a), PGSF1b and PGSF2. This expression profile method is a novel approach to the isolation of pituitary-specific genes that may have important functions.

Amino Acid Sequence↗
PubMed2001

Overexpression of kalpha1 tubulin mRNA in thyroid anaplastic carcinoma.

The expression levels of (k)alpha1 tubulin in 84 benign and malignant thyroid tissues were measured by means of real-time quantitative reverse transcription-polymerase chain reaction. An increased expression of (k)alpha1 tubulin mRNA was observed in all of five anaplastic carcinomas and some of the papillary carcinomas. Expression levels of (k)alpha1 tubulin relative to thyroglobulin mRNA were slightly increased in papillary carcinomas and greatly increased in anaplastic carcinomas. Chemotherapeutic agents which are targeted to microtubules may be considered as an alternative choice for the treatment of anaplastic carcinomas and some differentiated carcinomas in which increased expression of (k)alpha1 mRNA is observed.

Carcinoma↗
PubMed2001

Quantitative measurement of thyroglobulin mRNA in peripheral blood of patients after total thyroidectomy.

Previous studies have reported the clinical usefulness of reverse transcription-polymerase chain reaction (RT-PCR) detection of thyroglobulin (TG) mRNA in the peripheral blood of patients with differentiated thyroid carcinoma. To evaluate this usefulness, we measured TG mRNA in the peripheral blood of patients diagnosed with thyroid carcinoma after total thyroidectomy by real-time quantitative RT-PCR using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control. Surprisingly, we detected TG mRNA in all samples obtained after total thyroidectomy, including those from 4 medullary carcinomas. Further, there was no statistical difference in expression levels of TG mRNA in the patients with or without metastasis, and no significant correlation was found between serum TG concentrations and the expression levels of TG mRNA. These results give rise to a question regarding the clinical applications of not only RT-PCR detection but also quantitative measurement of TG mRNA in peripheral blood.

Adolescent↗
PubMed2001

Screening for primary hyperparathyroidism (PHPT) in clinic patients: differential diagnosis between PHPT and malignancy-associated hypercalcemia by routine blood tests.

Screening for primary hyperparathyroidism (PHPT) by measurement of the serum calcium concentration detects one patient per 500-1000 individuals in Western countries, and one patient per 2500-5000 subjects in Japan. Among clinic patients, however, the presence of many false-positive cases due to malignancy-associated hypercalcemia (MAH) reduces the benefit of such screening. We evaluated a new method of screening for PHPT based on the results of routine blood tests using the hospital information system (HIS) at our hospital. This new method could distinguish PHPT from MAH. This study included 25179 blood samples in which the serum calcium (Ca), albumin (Alb), chloride (Cl) and inorganic phosphate (IP) concentrations had been measured between March, 1994 and February, 1995 at Osaka University Medical Hospital. The HIS was programmed to pick blood samples that satisfied Formula 1 [Ca(mEq/ml) > 0.3 x Alb(g/dl) + 4.1] and Formula 2 ([Cl(mEq/ml)-84] x [10 x Alb-15]/[IP(mg/dl)/3.1] > 400). Of data from 25179 blood samples collected, those from 54 patients satisfied both Formulae 1 and 2. The patients from which these samples were derived from were subject to further analysis: medical records were studied and the intact-parathyroid hormone concentration was measured if necessary. Of these 54 cases, 19 patients (35.2%) were subsequently diagnosed with PHPT, including two, who were newly diagnosed with PHPT by this screening procedure. Although 35 (64.8%) of 54 patients were false-positive, many of them were treated with blood purification therapies in the Department of Pediatrics or the Intensive Care Unit (ICU). On the other hand, there were four false-positive cases (7.4%) caused by MAH. False-negative case in this study was only one patient (5%), whose diagnosis was normocalcemic PHPT. When omitting samples from pediatric patients and those in ICU, this screening procedure for PHPT has the advantage of being able to differentiate this diagnosis from MAH.

Diagnosis, Differential↗
PubMed2001

Increase of CD5(+) B cells during adolescence in female mice.

We examined physiological changes in CD5(+) B lymphocytes in mice associated with aging (from 1 or 5 to 50 weeks of age). The most predominant populations of lymphocytes among these mice were mainly CD5(-) B cells in the spleen and peritoneal cavity, and T cells in the thymus. However, in the spleen, CD5(+) B cells increased from 1 to 15 weeks, and then decreased with aging. In the thymus, CD5(+) B cells increased from 3 to 9 weeks of age, and subsequently became more predominant than CD5(-) B cells. In the peritoneal cavity, CD5(+) B cells increased from 5 to 9 weeks of age, became the most predominant population in lymphocytes at 7 to 9 weeks of age, and decreased with aging. The proportion of CD5(+) B cells in total B cells increased from 5 to 7 or 9 weeks of age, and then decreased with aging, with the highest proportion at 9 weeks of age in the spleen (15%), thymus (94%), and peritoneal cavity (54%). These findings indicate that CD5(+) B cells increase physiologically during mouse adolescence, and subsequently decrease with aging.

Aging↗
PubMed2001

Characteristics of experimental autoimmune hypophysitis in rats: major antigens are growth hormone, thyrotropin, and luteinizing hormone in this model.

We produced experimental autoimmune hypophysitis (EAH) in rats and investigated its characteristics. Female Lewis rats were immunized by two injections with homologous pituitary homogenate and complete Freund's adjuvant. Blood was collected serially from the rats, and serum antibodies to pituitary antigens were examined. The rats were sacrificed 2 or 4 weeks after the final immunization, and histological examinations of the endocrine organs were carried out. Histological examination revealed slight, focal infiltration of mononuclear cells in the pituitary gland only in the rats immunized with the pituitary homogenate. Infiltration of mononuclear cells was not observed in the thyroid gland, pancreas, adrenal gland, or ovary. In the serological examination, antibodies to both cytosolic antigens and cytoplasmic particle antigens from the pituitary gland were detected by enzyme-linked immunosorbent assay (ELISA), and these antibody levels increased with time. Western blotting using the serum antibodies identified an immunoreactive protein of approximately 21.5 kDa among these antigens, and we confirmed that this protein was rat growth hormone (GH). Furthermore, antibodies to GH, thyrotropin (TSH), and luteinizing hormone (LH) were detected by ELISA. Antibodies to follicule stimulating hormone, prolactin, or adrenocorticotropin were not detected. These data suggest that several antigens from the pituitary gland are involved in EAH in rats, and that GH, TSH, and LH are major antigens among the pituitary antigens in this model.

Animals↗
PubMed2001

Large-scale analysis of mutations in RET exon 16 in sporadic medullary thyroid carcinomas in Japan.

Germline mutations in the RET proto-oncogene are the cause of multiple endocrine neoplasia type 2 (MEN 2A and 2B) and familial medullary thyroid carcinoma (FMTC). Some cases of sporadic medullary thyroid carcinoma (MTC) have also been reported to have mutations in the RET gene. However, two previous reports have given discrepant results on the frequency of the mutations in RET in sporadic MTCs in Japan. To clarify this problem, we analyzed mutations in RET exon 16 in 72 sporadic MTCs by means of the two methods used in the previous studies, direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Mutations in exon 16 were detected in only 2 of 72 cases of sporadic MTC. These results suggest that when a MTC has a mutation in RET exon 16, it is more likely to be a hereditary MTC than a sporadic one in Japan.

Adult↗
PubMed2001

Influence of breast-feeding on the production of cytokines.

PROBLEM: Recently, we reported increases in the production of interferon-gamma (IFN-gamma), interleukin-2 (IL-2), and IL-4 during the postpartum period. The present study was undertaken to investigate whether these increases might be explained by increased prolactin while breast-feeding. METHOD: Whole blood from 41 women who were breast-feeding, 13 women not breast-feeding, and 31 healthy non-pregnant women was stimulated with phorbol 12-myristate 13-acetate and ionomycin, and the levels of cytokines in the supernatant were measured by enzyme-linked immunosorbent assay. Their serum levels of prolactin were measured by enzyme immunoassay. RESULTS: Increases in IFN-gamma, IL-2, IL-4, and IL-10 production were observed in women who were breast-feeding but not in women who were not breast-feeding. Serum levels of prolactin correlated with the levels of IFN-gamma in culture supernatant. CONCLUSIONS: These results suggest that breast-feeding induces production of cytokines and that IFN-gamma production is enhanced by physiological concentrations of prolactin.

Breast Feeding↗
PubMed2001

Analysis of the KAL1 gene in 19 Japanese patients with Kallmann syndrome.

Kallmann syndrome is defined by the association of hypogonadotropic hypogonadism and anosmia. The KAL1 gene is responsible for the X-linked form of Kallmann syndrome. We analyzed the KAL1 gene in 19 Japanese patients with Kallmann syndrome, including 3 patients reported previously, using PCR-direct sequencing method. All of 19 patients were sporadic, except for 2 monozygotic twins. In this study, there were 3 kinds of mutations in the KAL1 gene. One of them was a novel mutation consisting of a G to A substitution in the acceptor site at the junction of intron 6/exon 7. None of the mutations have been reported in countries other than Japan. In male sporadic patients with Kallmann syndrome, the incidence of mutations in Japanese patients (14%: 2 of 14 cases) was slightly higher than that in patients in USA (8%). Also, we found 2 polymorphisms, which were always found together in 6 patients. The severity of hypogonadism was not related to the presence of mutations. Unilateral renal aplasia and mirror movement, which are frequently found in patients with the KAL1 gene mutation, were not related to the sites of mutations.

Adolescent↗
PubMed2001

Serum concentration of androstenediol and androstenediol sulfate in patients with hyperthyroidism and hypothyroidism.

Androstenediol (5-androsten-3beta, 17beta-diol, ADIOL) and androstenediol 3-sulfate (ADIOLS) are active metabolites of dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEAS), respectively, and have estrogenic activity and immunoregulatory function. We examined serum concentrations of ADIOL, ADIOLS, DHEA, DHEAS and pregnenolone sulfate (5-pregnen-3beta-ol-20-one sulfate, PREGS) in patients with Graves' thyrotoxicosis (male/female 9/14), hypothyroidism (11/20) and in normal controls (14/29). In hypothyroidism serum levels of all these steroids were significantly decreased in both genders. In hyperthyroidism, in contrast, serum levels of ADIOLS (male 1.49 +/- 0.69, female 0.64 +/- 0.31 micromol/l), DHEAS (male 7.43 +/- 3.91, female 5.13 +/- 2.03 micromol/l), and PREGS (male 1.13 +/- 0.58, female 1.07 +/- 0.85 micromol/l) were markedly increased, but serum concentrations of ADIOL and DEHA were not significantly different from controls (ADIOLS male 0.36 +/- 0.33, female 0.14 +/- 0.09 micromol/l; DHEAS male 2.88 +/- 1.70, female 1.86 +/- l1.03pmol/l; PREGS male 0.18 +/- 0.12, female 0.11 +/- 0.08 micromol/l; ADIOL male 3.76 +/- 1.35, female 1.91 +/- 1.17 nmol/l; DHEA male 9.23 +/- 3.49, female 13.5 +/- 10.8nmol/l). Serum concentrations of all these steroids correlated with the serum concentration of the thyroid hormones in these patients. Serum albumin and sex hormone-binding globulin concentrations were not related to these changes in the concentrations of steroids. These findings indicate that serum concentrations of ADIOLS, ADIOL, DHEAS, DHEA and PREGS were decreased in hypothyroidism, whereas serum ADIOLS, DHEAS and PREGS concentrations were increased but ADIOL and DHEA were normal in hyperthyroidism. Thyroid hormone may stimulate the synthesis of these steroids and sulfotransferase is speculated to be increased in hyperthyroidism. Increased ADIOLS might contribute to menstrual disturbances and gynecomastia in hyperthyroidism.

Adult↗
PubMed2001

Blocking type anti-tSH receptor antibodies detected by radioreceptor assay in Graves' disease.

We previously developed a radioreceptor assay which is presumably specific for detection of blocking-type anti-TSH receptor (TSHR) antibodies using unsolubilized porcine TSHR. Employing this assay, we measured blocking TSH-binding inhibitory immunoglobulin (TBII) in the sera from 30 untreated Graves' patients and compared the results with those of a bioassay measuring thyroid stimulation blocking antibodies (TSBAb). Blocking TBII was positive in 9 of 30 sera (30%) and the blocking TBII activity was correlated with the total TBII value, which was measured by the radioreceptor assay using solubilized porcine TSHR. On the other hand, TSBAb determined with the conventional bioassay was positive in only 2 sera (6.7%), and no correlation was observed with the TSAb activity. In some cases in which the TSAb activity was rather high, TSBAb could not be detected by bioassay, whereas blocking TBII was positive. There was no correlation of blocking TBII activity with goiter size, thyroid hormone level, or proptosis. However, there was a tendency for anti-thyroid therapy to require a shorter time for FT4 normalization in blocking TBII subjects, suggesting that blocking-type anti-TSHR antibody plays some role in the pathophysiology of Graves' disease. In conclusion, blocking type anti-TSHR antibodies are often found in the sera of Graves' patients when the blocking-specific radioreceptor assay is applied.

Adolescent↗
PubMed2001

[Aspiration biopsy-RT-PCR(ABRP): lesson from its success].

Molecular-based diagnosis of thyroid carcinomas can be more easily established by utilizing specific mRNAs that are expressed only in cancer tissues. In a previous study, we introduced a new method of preoperatively diagnosing thyroid carcinomas. This technique, aspiration biopsy-RT-PCR(ABRP), facilitated simultaneous cytological and molecular-based diagnoses by extracting RNA from cells remaining within the needle used for fine needle aspiration biopsies(FNABs). ABRP provides both RNA information and a cytological diagnosis without further invasion to the patient. We proved that by ABRP detection of oncofetal fibronectin(onfFN) mRNA in FNABs, papillary and anaplastic carcinomas are accurately diagnosed preoperatively. Further, by real-time monitoring RT-PCR measuring onfFN mRNA, a fully automated system was established. It is not clarified, however, why cancer-specific mRNAs, especially those overexpressed in fetal tissues, can clearly distinguish benign tissues from carcinomas, while genomic alternation such as mutations in RAS or P53 gene cannot. Further, a widely accepted hypothesis, multi-step carcinogenesis, does not explain some of the clinical and experimental evidence from thyroid carcinomas. Considering these facts, we propose a new concept of thyroid carcinogenesis called "germ-cell carcinogenesis", in which cancer cells are derived from the remnant of fetal thyroid germ cells(thyroblasts) instead of normal thyroid follicular cells.

Biomarkers, Tumor↗
PubMed2000

Loop-mediated isothermal amplification of DNA.

We have developed a novel method, termed loop-mediated isothermal amplification (LAMP), that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions. This method employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. An inner primer containing sequences of the sense and antisense strands of the target DNA initiates LAMP. The following strand displacement DNA synthesis primed by an outer primer releases a single-stranded DNA. This serves as template for DNA synthesis primed by the second inner and outer primers that hybridize to the other end of the target, which produces a stem-loop DNA structure. In subsequent LAMP cycling one inner primer hybridizes to the loop on the product and initiates displacement DNA synthesis, yielding the original stem-loop DNA and a new stem-loop DNA with a stem twice as long. The cycling reaction continues with accumulation of 10(9) copies of target in less than an hour. The final products are stem-loop DNAs with several inverted repeats of the target and cauliflower-like structures with multiple loops formed by annealing between alternately inverted repeats of the target in the same strand. Because LAMP recognizes the target by six distinct sequences initially and by four distinct sequences afterwards, it is expected to amplify the target sequence with high selectivity.

Base Sequence↗