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Eitetsu Koh

Publications and source records attributed to Eitetsu Koh.

At least 19 recordsLinked to original sources
PubMed2006

Liquid chromatography-mass spectrometric assay of androstenediol in prostatic tissue: influence of androgen deprivation therapy on its level.

Androstenediol (Adiol, androst-5-ene-3beta,17beta-diol) is suspected of being an endogenous proliferation agent of prostate cancer (PCa) even after androgen deprivation therapy (ADT). A liquid chromatography-electron capture atmospheric pressure chemical ionization-mass spectrometric (LC-ECAPCI-MS) method for the determination of Adiol in prostatic tissue was developed and validated for evaluating the influence of ADT on the prostatic Adiol level. After derivatization of Adiol with 4-nitrobenzoyl chloride, the detection response of the derivative was increased 150 times more than that of intact Adiol. The LC-MS method was specific and reliable for the measurement of a trace amount of Adiol in 30 mg of tissue. The clinical study using the developed method showed that the prostatic Adiol level was not changed by ADT. That is, the prostatic Adiol levels of PCa patients with ADT (n = 12), benign prostate hypertrophy patients without ADT (n = 8) and bladder cancer patients (without prostatic disease) (n = 6) were 0.83 +/- 0.28, 0.62 +/- 0.31 and 0.71 +/- 0.28 ng g(-1)tissue, respectively, and there was no significant difference between these groups.

Androstenediol↗
PubMed2006

Is a genetic defect in Fkbp6 a common cause of azoospermia in humans?

FK506-binding protein 6 (Fkbp6) is a member of a gene family containing a prolyl isomerase/FK506-binding domain and tetratricopeptide protein-protein interaction domains. Recently, the targeted inactivation of Fkbp6 in mice has been observed to result in aspermic males and the absence of normal pachytene spermatocytes. The loss of Fkbp6 results in abnormal pairing and a misalignment of the homologous chromosomes, and in non-homologous partner switches and autosynapsis of the X chromosome cores in meiotic spermatocytes. In this study, we analyzed whether human FKBP6 gene defects might be associated with human azoospermia. We performed a mutation analysis in all the coding regions of the human FKBP6 gene in 19 patients with azoospermia resulting from meiotic arrest. The expression of the human FKBP6 gene was specific to the testis, and a novel polymorphism site, 245C --> G (Y60X) could be found in exon 3. Our findings suggest that the human FKBP6 gene might be imprinted in the testis based on an analysis using two polymorphism sites.

Amino Acid Sequence↗
PubMed2006

[Disorder of AZF].

Explore the source record for details and available documents.

Base Sequence↗
PubMed2006

Reevaluation of azoospermic factor c microdeletions using sequence-tagged site markers with confirmed physical positions from the GenBank database.

OBJECTIVE: To investigate microdeletions and palindrome complexes in the azoospermic factor (AZF) c regions of the Y chromosome, by polymerase chain reaction (PCR) with sequence-tagged site (STS) markers with known physical positions, after verification of the exact physical locations of candidate STSs and exclusion of those that would give ambiguous results on PCR. DESIGN: Retrospective STS deletion study in infertile Japanese men. SETTING: University hospital and reproductive clinic. PATIENT(S): A total of 410 men with nonobstructive azoospermia and severe oligospermia (sperm concentrations of <5 x 10(6)/mL). INTERVENTION(S): Polymerase chain reaction was performed for all STS markers confirmed to be physically located in the partial AZFb and AZFc regions of the Y chromosome. All STSs were retrieved from the University of California at Santa Cruz database, and their location and specificity were verified. MAIN OUTCOME MEASURE(S): Presence or absence of appropriately sized PCR products. RESULT(S): Sixty-nine markers were retrieved, 32 of which were not specific to the long arm of the Y chromosome (Yq). The markers retained for test use were contiguous deletions classified as P1+P2 (AZFc) or P3 proximal/P1 (AZFb+c). The prevalence of accurately mapped microdeletions was 5.1% (21 of 410) in this patient population. CONCLUSION(S): Although noncontiguous deletions were observed, considerable confusion remained until the genome sequence was finally determined because many of the STSs were either repetitive sequences or polymorphic between individuals or races.

Animals↗
PubMed2006

Inter/intra investigator variation in orchidometric measurements of testicular volume by ten investigators from five institutions.

AIM: To perform quality control studies on testicular volume measurements for a multi-center epidemiological study of male reproductive function. METHODS: We constructed a data matrix with a balanced assignment for 2 consecutive days by ten investigators (andrological career: 4-21 years) from five institutions and 12 male volunteers aged 20-26 years. Testicular volume was measured by Prader's orchidometer. A skilled technician also performed an ultrasound estimate of testicular volume. RESULTS: A statistically significant inter-investigator variation was found for both testes (P < 0.05). In addition, there was a statistically significant investigator-by-volunteer interaction in testicular volume measurement (P < 0.01). However, there was no statistically significant difference in the two measurements performed on consecutive days for either testis. The testicular volumes for both the right and left testes as estimated by ultrasonography were smaller than results using the orchidometer. However, there was no statistical significance (P > 0.05). The difference in experiences of the investigators did not significantly correlate with accuracy of measurements in either testis. CONCLUSION: The present study revealed significant differences in the results of estimation of testicular volume among the ten investigators, but intra-investigator variation was not considerable. Improved training and proper standardization of the measurement will be necessary before starting a multi-center study based on an andrological examination.

Adult↗
PubMed2005

Endobronchial ultrasound guided transbronchial needle aspiration for staging of lung cancer.

During the staging process of lung cancer, accurate mediastinal lymph node staging is one of the important factors which affect patient management. The purpose of the current study was to evaluate the usefulness of direct real-time endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) for staging and diagnosis of lung cancer in patients with mediastinal lymph nodes suspected of malignancy and to assess the impact of this method in patient management. One hundred and eight patients with mediastinal lymph nodes with known or suspected lung cancer were included. The convex probe EBUS integrated with a convex scanning probe on its tip was used in all cases. Final diagnosis was based on cytology, surgical results, and/or clinical follow-up. In 105 patients, EBUS-TBNA was successfully performed to obtain samples from 163 lymph nodes. With respect to the correct prediction of lymph node stage, EBUS-TBNA had a sensitivity of 94.6%, specificity of 100%, positive predictive value of 100%, negative predictive value of 89.5%, and diagnostic accuracy rate of 96.3%. In the 20 suspected lung cancer cases, mediastinal lymph node was used for tissue diagnosis of malignancy as well as staging. As a result of EBUS-TBNA, 29 mediastinoscopies, 8 thoracotomies, 4 thoracoscopies, and 9 CT-guided PCNB were avoided. The procedure was uneventful without complications. EBUS-TBNA is a safe and sensitive method for lymph node staging in patients with lung cancer. It spares invasive staging procedures which has a major impact on patient management.

Adenocarcinoma↗
PubMed2005

Determination of prostatic androgens in 10 mg of tissue using liquid chromatography-tandem mass spectrometry with charged derivatization.

A practical liquid chromatography-tandem mass spectrometric (LC-MS-MS) method for the determination of prostatic 5alpha-dihydrotestosterone (DHT) and testosterone (T) has been developed. The prostatic androgens were extracted with MeOH-H2O (3:7, v/v), purified with an Oasis HLB cartridge, derivatized with the permanently charged reagent 2-hydrazino-1-methylpyridine (HMP), and subjected to LC-MS-MS analysis using electrospray ionization (ESI) operated in the positive ion mode. The derivatization with HMP was very effective at increasing the detectability using the positive-ESI-MS. The method allowed the reproducible and accurate quantification of ng g(-1) tissue levels of prostatic androgens in 10 mg of tissue. That is, the intra- and inter-assay coefficients of variation were below 8.1 and 9.3%, respectively, and the analytical recoveries of the androgens were quantitative. The limits of quantitation for DHT and T were both 1.0 ng g(-1) tissue. The developed method was used to determine DHT and T in the prostates of patients with benign prostatic hyperplasia and prostate cancer, and satisfactory results were obtained.

Chromatography, Liquid↗
PubMed2005

Determination of sulfates of androsterone and epiandrosterone in human serum using isotope diluted liquid chromatography-electrospray ionization-mass spectrometry.

A promising liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) for analysis of the sulfates of 5alpha-androgen, androsterone and epiandrosterone (A-S and EpiA-S) in human serum was developed. The method was used to assess one of the markers of 5alpha-reductase activity of males including patients with prostate cancer (PC). After deproteinization with acetonitrile, the androgen sulfates and the internal standard, [7,7,16,16-2H4]dehydroepiandrosterone-S, were extracted from human serum using a solid-phase extraction cartridge and washed with hexane. The extract was reconstituted and applied to the LC/ESI-MS system operated in the selected ion monitoring mode. The method was validated over the range 0.02-5 microg/mL (A-S) and 0.005-1.5 microg/mL (EpiA-S) using 10 microL of human serum. The method was a concise procedure without chemical or enzymatic hydrolysis of the conjugates, purification by high-performance liquid chromatography and/or derivatization, and proved to be satisfactory in its reproducibility and accuracy. The levels of these androgen sulfates tended to decrease during aging, and the A-S levels in the sera obtained from both healthy males and patients with PC were correlated with their EpiA-S levels.

Adult↗
PubMed2004

Correlation between interleukin 6 production and tumor proliferation in non-small cell lung cancer.

UNLABELLED: Interleukin 6 (IL-6) facilitates the differentiation of B cells to immunoglobulin-secreting cells and is reported to be a proliferative factor in some tumors. In this study, we examined IL-6 production in non-small cell lung carcinoma (NSCLC) and the proliferation of tumor cells following IL-6 treatment in vitro and in vivo. We analyzed the expression of IL-6 mRNA and protein in a series of 15 human lung cancer cell lines (four adenocarcinomas, five squamous cell carcinomas, two large cell carcinomas, and four small cell carcinomas) by reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). We established an IL-6-producing cell line (ABC-1#IL-6) by transfecting a human IL-6 cDNA into a human non-IL-6-producing NSCLC cell line (ABC-1). These two cell lines were used to determine tumor cell proliferation both in vivo and in vitro in order to clarify the effect of IL-6 on tumor growth and metastasis. Athymic nude mice, SCID mice, and BALB/c mice were subcutaneously inoculated with these two cell lines, and body weight, tumor growth, and tumor doubling time were measured. The presence of IL-6 and tumor-infiltrating lymphocytes (TILs) within tumor tissues was examined by immunohistochemical staining. RESULTS: Eight of 15 (53%) lung cancer cell lines expressed both IL-6 mRNA and protein. Tumor lesions of both cell lines developed in nude and SCID mice, although no such lesions of either cell lines developed in BALB/c mice. The tumor doubling time in nude and SCID mice was 2.97+/-1.22 days and 2.45+/-1.32 days, respectively, in mice inoculated with the cell line ABC-1#IL-6. These doubling times were statistically significantly shorter than those evident in mice inoculated with the control original ABC-1 cell line (nude, p=0.0337; SCID, p=0.0119; unpaired t-test). The rates of cell proliferation in vitro of the ABC-1#IL-6 and original ABC-1 cells lines were comparable ( p=0.1441, unpaired t-test). Immunohistochemical staining revealed strong IL-6 expression in tumors derived from the IL-6-producing cell line but not in tumors derived from the original ABC-1 cell line (both in nude and SCID mice). CONCLUSION: 53% of lung cancer cell lines produce IL-6 mRNA and protein. Although IL-6 itself does not influence tumor cell proliferation in vitro, an association between IL-6 expression and tumor proliferation was found in vivo in nude and SCID mice. An anti-IL-6 reagent could provide a novel therapeutic strategy in patients with IL-6-producing lung tumors.

Adenocarcinoma↗
PubMed2004

The adrenal androgen androstenediol is present in prostate cancer tissue after androgen deprivation therapy and activates mutated androgen receptor.

Despite an initial response to androgen deprivation therapy, prostate cancer (PCa) progresses eventually from an androgen-dependent to an androgen-independent phenotype. One of the mechanisms of relapse is antiandrogen withdrawal phenomenon caused by mutation of 877th amino acid of androgen receptor (AR). In the present study, we established a method to measure the concentration of androstenediol (adiol) in prostate tissue. We found that adiol maintains a high concentration in PCa tissue even after androgen deprivation therapy. Furthermore, adiol is a stronger activator of mutant AR in LNCaP PCa cells and induces more cell proliferation, prostate-specific antigen (PSA) mRNA expression, and PSA promoter than dihydrotestosterone (DHT). Because antiandrogen, bicalutamide, blocked adiol activity in LNCaP cells, it was suggested that adiol effect was mediated through AR. However, high concentration of bicalutamide was necessary to block completely adiol activity. These effects were specific to LNCaP cells because adiol had less effect in PC-3 PCa cells transfected with wild-type AR than DHT and had similar effect in PC-3 cells transfected with mutant AR. The mechanism that adiol activates mutant AR in LNCaP cells did not result from the increased affinity to mutant AR or from AR's association with coactivator ARA70. However, low concentration of adiol induced more AR nuclear translocation than DHT in LNCaP cells and not PC-3 cells transfected with AR. These results indicate that adiol may cause the progression of PCa even after hormone therapy.

Androgen Antagonists↗
PubMed2004

[Reference ranges of total serum and free testosterone in Japanese male adults].

PURPOSE: To establish reference range of serum Total Testosterone (T-T) and Free Testosterone (F-T) in Japanese male adults. SUBJECTS AND METHOD: Among 1,172 male adults, who daily lived their healthy life, 1,143 subjects in the year range from 20 to 77 years old, who had serum LH concentration within its reference range (For 20-70 years old: 1.1-25.9 mIU/mL), were selected. As diurnal rhythm of both T-T and F-T was observed, blood samples were collected in the morning when T-T and F-T concentration were relatively stable at their high concentration levels. The collected samples were stored at -20 degree C until they were used for assays. RESULTS: Reference range for T-T has decided to express by the mean +/- 2SD calculated from the entire test results, because influence of aging on the results was negligible, Reference range of T-T has established as 2.01-7.50 ng/mL. The other hand, reference ranges of F-T classified for every decade have decided to express by the mean +/- 2SD of each decade subgroup, because great influence of aging on F-T was observed. For each decade from 20 years on and > 70 years, reference ranges of F-T have established as: 8.5-27.9 pg/mL, 7.6-23.1 pg/mL, 7.7-21.6 pg/mL, 6.9-18.4 pg/mL, 5.4-16.7 pg/mL, and 4.5-13.8 pg/ mL, respectively. CONCLUSION: Reference ranges of serum T-T and F-T in Japanese male adults have established. And the value of Young Adult Mean (YAM) of F-T calculated for a group of the ages between 20 and 39 years have been proposed as a guideline of requirement for the Androgen Replacement Therapy (HRT). The value of 80% and 70% of YAM were 12.4 pg/mL and 10.9 pg/mL, respectively.

Adult↗
PubMed2004

[Impaired spermatogenesis and genes].

The crucial genes for spermatogenesis have not yet been identified, but the AZF regions in the Y chromosome are now thought to show a major correlation with spermatogenesis. Recently, a complex of palindromes exists in the Yq, and is conserved in a highly repetitive sequence. Therefore, the PCR based sequence tagged site (STS) could not be accurately decided their unique position. This review demonstrated that there was not a high frequency of deletion of AZF-specific genes than expected using usual STS-PCR. However, the frequency of expression in AZF genes is relatively low. This discrepancy is probably due to repetitive sequence of complex palindromes in the Yq.

Base Sequence↗
PubMed2003

Simultaneous determination of androstenediol 3-sulfate and dehydroepiandrosterone sulfate in human serum using isotope diluted liquid chromatography-electrospray ionization-mass spectrometry.

A simple method for simultaneous determination of androstenediol 3-sulfate (Adiol-3S) and dehydroepiandrosterone sulfate (DHEA-S) in human serum using isotope diluted liquid chromatography-electrospray ionization-ion trap-mass spectrometry (LC-ESI-ion trap-MS) was developed. After addition of deuterated internal standards ([2H5]Adiol-3S and [2H4]DHEA-S), human serum (100 microl) was deproteinized with acetonitrile and then applied to a solid-phase extraction cartridge, Oasis HLB. The obtained steroid sulfates fraction was washed with hexane and then analyzed by LC-ESI-MS operated in the negative ion mode. The quantification ranges of Adiol-3S and DHEA-S were 10-400 ng/ml and 0.05-8 microg/ml, respectively. The method does not require the chemical or enzymatic hydrolysis of the conjugates and purification with high-performance liquid chromatography, and shows satisfactory reproducibility and accuracy. The concentrations of these sulfates in the sera of healthy male volunteers (n=14) were 19.2-245.3 mg/ml (Adiol-3S) and 0.175-5.16 microg/ml (DHEA-S), and those of patients with prostate cancer (n=19) were 15.3-182.7 ng/ml (Adiol-3S; four samples, not detectable) and 0.110-2.421 microg/ml (DHEA-S).

Aged↗
PubMed2003

Androgen-independent growth in LNCaP cell lines and steroid uridine diphosphate-glucuronosyltransferase expression.

AIM: To investigate the mechanism of androgen-independent growth of prostate cancer after androgen ablation in LNCaP cells and the effect of glucuronidation activity. METHODS: To establish androgen-independent growth in prostate cancer LNCaP-SF, continuous passage was performed in androgen-stripped medium and the cells were evaluated for glucuronidation activity. The expression vector of antisense uridine diphosphate glucuronosyl-transferase (UGT) 2B15 cDNA was also constructed and evaluated. RESULTS: LNCaP-SF lead to a higher expression in UGT2B15 and their glucuronidation activity is 2.5 times higher than that of LNCaP cells. Significantly fewer LNCaP and LNCaP-SF than control were transfected with the antisense UGT2B15 cDNA, suggesting that UGT2B15 plays an important part in the glucuronidation activity of androgens in both cells. CONCLUSION: The alteration of UGT2B15 expression in LNCaP-SF cells is proposed as a biological characteristic involved in the growth of hormone-refractory prostate cancer.

Androgens↗
PubMed2002

Differential expression of 17beta-hydroxysteroid dehydrogenase isozyme genes in prostate cancer and noncancer tissues.

BACKGROUND: The adrenal steroids dehydroepiandrosterone and androstenediones are converted into active androgen testosterone in prostatic tissues. Different 17beta-hydroxysteroid dehydrogenase (17betaHSD) isozymes are characterized by either oxidation or reduction reactions. These redox reactions represent an important step in both biosynthesis and metabolism of androgens. This study presents the differential expression of 17betaHSD isozyme genes in cancerous and noncancerous prostate tissues of in vivo samples. METHODS: Thirty-four fresh specimens of transrectal prostatic needle biopsy were obtained; 11 were pathologically diagnosed as adenocarcinoma and 23 as without malignancy. The gene expression levels of five isozymes (type 1-5) of 17betaHSD were evaluated. The quantification of gene expression was assessed by means of the real-time polymerase chain reaction. RESULTS: The expression levels of the type 3 17betaHSD gene with malignancy were significantly higher than those in prostatic tissues without malignancy, and those of type 2 17betaHSD with malignancy were significantly lower than those in nonmalignant tissues. There were no significant differences in 17betaHSD type 1, type 4, and type 5 gene expression in cancerous and noncancerous tissues. CONCLUSION: Our results suggest that 17betaHSD type 2 and type 3 play an important role in the conversion of adrenal steroids into potential androgens in prostate cancer tissue.

17-Hydroxysteroid Dehydrogenases↗