bluuebunny/crossref_metadata_2025 huggingface dataset:
Population‐wide Infant Screening for HLA‐Based Type 1 Diabetes Risk via Dried Blood Spots from the Public Health Infrastructure
Authors: WION EMILY , BRANTLEY MICHAEL , STEVENS JEFF , GALLINGER SUSAN , PENG HUI , GLASS MICHAEL , HAGOPIAN WILLIAM
Published: January 2004

Abstract: The frequency of type 1 diabetes mellitus (T1DM)‐associated HLA DQ alleles in the U.S. Pacific Northwest is as high as in Scandinavia, which has the highest T1DM incidence in the world. The high regional rate of islet autoimmunity observed among DPT‐1 relatives supports this notion. Fortunately, Washington State archives dried blood spots after legislature‐mandated newborn screening. The Diabetes Evaluation in Washington (DEW‐IT) study aims to show that population‐based prospective prediction of T1DM by HLA genotype screening followed by autoantibody surveillance can be performed within the public health infrastructure.
Vaccinations May Induce Diabetes‐Related Autoantibodies in One‐Year‐Old Children
Authors: WAHLBERG J. , FREDRIKSSON J. , VAARALA O. , LUDVIGSSON J. ,   ABIS STUDY GROUP
Published: January 2004

Abstract: Vaccinations have been discussed as one among many environmental candidates contributing to the immune process that later may lead to type 1 diabetes. ABIS (All Babies in Southeast Sweden) is a prospective cohort study following a nonselected birth cohort of general population. In a randomly selected sample collection from 4400 children, GADA and IA‐2A have been determined at the age of 1 year. The information on vaccinations was collected from questionnaires answered by the parents and was related to β cell autoantibodies. When studying the induction of autoantibodies using the autoantibody level of 90th percentile as cutoff level, hemophilus influenza B (HIB) vaccination appeared to be a risk factor for IA‐2A [OR 5.9 (CI 1.4‐24.4; p= 0.01)] and for GADA [OR 3.4 (CI 1.1‐10.8; p= 0.04)] in logistic regression analyses. Furthermore, the titers of IA‐2A were significantly higher (p < 0.01 in Mann‐Whitney test) in those children who had got HIB vaccination. When 99th percentile was used as cutoff to identify the children at risk of type 1 diabetes, BCG vaccination was associated with increased prevalence of IA‐2A (p < 0.01). We conclude that HIB vaccination may have an unspecific stimulatory polyclonal effect increasing the production of GADA and IA‐2A. This might be of importance under circumstances when the β cell‐related immune response is activated by other mechanisms.
Inhibition of STAT4 Activation by Lisofylline Is Associated with the Protection of Autoimmune Diabetes
Authors: YANG ZANDONG , CHEN MENG , FIALKOW LAWRENCE B. , ELLETT JUSTIN D. , WU RUNPEI , NADLER JERRY L.
Published: January 2004

Abstract: We investigated the signal transduction pathway of IL‐12 and showed that lisofylline (LSF) inhibited the signal transducer and activator of transcription factor‐4 (STAT4) activation. Interruption of IL‐12‐mediated STAT4 activation prevented autoimmune diabetes in NOD mice.
Reduced Thymic Expression of Islet Antigen Contributes to Loss of Self‐Tolerance
Authors: MATHEWS C. E. , PIETROPAOLO S. L. , PIETROPAOLO M.
Published: January 2004

Abstract: Type 1 diabetes (T1DM) results from a failure of central and peripheral tolerance to islet cell antigens. ICA69 belongs to a group of molecules expressed predominantly in neuroendocrine tissues (including pancreatic islets), which are targets of autoimmune responses in T1DM. These molecules are also expressed in the thymus and peripheral lymphoid organs by dendritic cells. The aim of the present study was to evaluate possible variation in thymic ICA69 expression, comparing diabetes‐resistant controls to T1DM‐prone NOD mice. Thymic tissue was retrieved from 3‐ to 6‐week‐old female B6, NOD‐H2b, and NOD mice. Paraffin‐embedded sections were stained with an ICA69‐specific antibody in an immunoperoxidase assay. ICA69 staining of thymic sections from B6 and NOD.H2b showed strong and continual staining, yet the sections from the NOD mice showed significantly reduced staining for ICA69. Corroboration of the reduced level of ICA69 in the thymus of NOD mice has been obtained via analysis for the expression of ICA69 versus other candidate autoantigens (glutamic acid decarboxylase 65, glutamic acid decarboxylase 67, and insulin 2) in the thymus. Real‐time PCR analysis, using cDNA generated from the thymus, displayed that the expression of GAD65, GAD67, and INS2 were equivalent when comparing NOD at any age to B6, BALB/cJ, and ALR/LtJ. In marked contrast, the level of ICA69 in the thymus of the NOD mice examined was significantly reduced when compared to the controls. In fact, the real‐time PCR analysis strongly suggested that ICA69 was not expressed in the thymus of NOD mice. These findings support the hypothesis that the level of thymic ICA69 expression may be of importance in regulating self‐tolerance in T1DM.
Dietary Microbial Toxins and Type 1 Diabetes
Authors: MYERS M. A. , HETTIARACHCHI K. D. , LUDEMAN J. P. , WILSON A. J. , WILSON C. R. , ZIMMET P. Z.
Published: January 2004

Abstract: Toxins may promote type 1 diabetes by modifying or damaging the β cell causing release of autoantigens. Streptomyces is a common soil bacterium that produces many toxic compounds. Some Streptomyces can infect vegetables, raising the possibility of dietary exposure to toxins. We aimed to identify toxins that erode cellular proton gradients in extracts of Streptomyces and infested vegetables and to establish the effect of low doses of these toxins on pancreatic islets in mice. The vacuolar ATPase inhibitors, bafilomycin and concanamycin, and the ionophore, nigericin, were identified in extracts from 4 of 13 Streptomyces isolated from infested potatoes and in potatoes themselves. Injection of bafilomycin A1 into mice impaired glucose tolerance, reduced islet size, and decreased relative β cell mass. Thus, exposure to small quantities of bafilomycin in the diet may contribute to the cause of type 1 diabetes.
Prevention of Autoimmune Diabetes through Immunostimulation with Q Fever Complement‐Fixing Antigen
Authors: SILVA D. G. , CHARLTON B. , COWDEN W. , PETROVSKY N.
Published: January 2004

Abstract: The most promising strategies for prevention of type 1 diabetes seem to be in the categories of immunomodulation (e.g., nondepleting anti‐CD3, Diapep, linomide) and/or immunostimulation (e.g., QFA, BCG). We are currently undertaking a research program directed toward better understanding of immunostimulants to help maximize the likelihood of success of future human clinical trials for diabetes prevention. This program is focused on the key areas of optimization of vaccine dose and route of administration, development of surrogate immune markers, and elucidation of the mechanism of protection. The mechanism whereby QFA protects against diabetes currently is not known. The elucidation of the mechanism should help identify the optimal way in which to administer QFA to provide diabetes protection. It may also assist the development of even more potent immunostimulatory vaccines.
AIRE‐1 (Autoimmune Regulator Type 1) as a Regulator of the Thymic Induction of Negative Selection
Authors: PARK YONGSOO , MOON YOOMI , CHUNG HEE‐YONG
Published: January 2004

Abstract: The monogenic autoimmune syndrome, APS‐1 (autoimmune polyglandular syndrome type 1), is characterized by the loss of self‐tolerance to multiple organs. Although mutations in the AIRE (autoimmune regulator) gene are responsible for the APS‐1, the function of AIRE is not known. AIRE may determine thymic induction of tolerance to self‐antigens in multiple organs. To study the function of AIRE in induction of self‐tolerance, an in vitro negative selection system was made using 106 DO11.10 TCR transgenic thymocytes, 105 antigen‐presenting cells (APC), and the different constructs of ovalbumin (OVA). In this system, the addition of the immunodominant epitopes of OVA peptide, the antigenic ligand for the DO11.10, made the thymocytes apoptotic and negatively selected. Overexpression of the AIRE gene in APC using retroviral transduction did not cause more thymocytes to become apoptotic. However, the suppression of the expression of AIRE in APC using the dominant‐negative gene made the recovery rates of the thymocytes higher than those with the expression of LacZ as a control, and consequently inducing loss of self‐tolerance. From these studies, it might be possible to suggest that the AIRE gene might regulate thymic induction of the negative selection process. The target genes for transcriptional regulation by AIRE have been investigated to study the influence of AIRE expression on other proteins in antigen presentation. The expression level of B7.1 was higher in APC expressing the dominant‐negative form of AIRE. The target gene regulated by AIRE in transcription will be screened using cDNA microarray.
Association of Interleukin‐18 Gene Promoter Polymorphisms in Type 1 Diabetes and Autoimmune Thyroid Disease
Authors: IDE AKANE , KAWASAKI EIJI , ABIRU NORIO , SUN FUYAN , FUKUSHIMA TETSUYA , ISHII REIKO , TAKAHASHI RYOKO , KUWAHARA HIRONAGA , FUJITA NARUHIRO , KITA ATSUSHI , IMAIZUMI MISA , OSHIMA KATSUYA , USA TOSHIRO , UOTANI SHIGEO , EJIMA ERI , YAMASAKI HIRONORI , ASHIZAWA KIYOTO , YAMAGUCHI YOSHIHIKO , EGUCHI KATSUMI
Published: January 2004

Abstract: Type 1 diabetes is a heterogeneous autoimmune disease and is often associated with other organ‐specific autoimmune diseases, including autoimmune thyroid disease (AITD). IL‐18 is a potent proinflammatory cytokine capable of inducing IFN‐γ production that is associated with the development of type 1 diabetes and AITD. The gene for IL‐18 is located near Idd2 and has been reported to be associated with a susceptibility to type 1 diabetes. To test the putative involvement of IL‐18 gene polymorphism in predisposition to type 1 diabetes and AITD, we conducted a case‐control study in Japanese population. The SNPs at position −607 (C/A) and −137 (G/C) in the promoter region of the IL‐18 gene were analyzed by sequence‐specific PCR in 74 nondiabetic patients with AITD, 47 type 1 diabetic patients with AITD, and 114 normal controls. There was no significant increase in the genotype and allele frequencies not only in nondiabetic patients with AITD compared with normal controls, but also in type 1 diabetic patients with AITD compared with normal controls. The distribution of IL‐18 gene haplotypes was also similar between both patient groups and normal controls. These results suggest that polymorphisms of the IL‐18 gene are not associated with a susceptibility to AITD and type 1 diabetes coexistent with AITD in Japanese population.
Epitope Analysis of GAD65 Autoantibodies in Japanese Patients with Autoimmune Diabetes
Authors: KAWASAKI EIJI , ABIRU NORIO , IDE AKANE , SUN FUYAN , FUKUSHIMA TETSUYA , TAKAHASHI RYOKO , KUWAHARA HIRONAGA , FUJITA NARUHIRO , KITA ATSUSHI , OSHIMA KATSUYA , UOTANI SHIGEO , YAMASAKI HIRONORI , YAMAGUCHI YOSHIHIKO , EGUCHI KATSUMI
Published: January 2004

Abstract: Type 1 diabetes is an organ‐specific autoimmune disease characterized by T cell‐mediated destruction of pancreatic β cells. In Japanese population, the incidence of type 1 diabetes in children is very low compared to European countries. However, there are more patients with type 1 diabetes in adults, including latent autoimmune diabetes in adults (LADA). The circulating autoantibodies to multiple islet autoantigens including GAD, insulin, and IA‐2 are the important immunological features of type 1 diabetes. The prevalences of anti‐islet autoantibodies in patients with Japanese type 1 diabetes are 60‐70% for GAD autoantibodies, 45‐50% for insulin autoantibodies (IAA), and 60‐65% for IA‐2 autoantibodies at disease onset, which are similar to those reported in Caucasian patients. With combinatorial analysis of these autoantibodies, 90% of patients express at least one of these autoantibodies and are classified as type 1A diabetes. Although the majority of patients with type 1 diabetes are young, lean, and ketosis‐prone, there are a number of patients with type 1 diabetes initially diagnosed as having type 2 diabetes at disease onset called LADA. These patients with LADA often progress toward an insulin‐deficient state within several years after diagnosis. High levels of GAD autoantibodies have a high predictive value for future insulin deficiency in LADA. Further, epitope analysis of GAD65 autoantibodies may be helpful to predict future insulin dependency in LADA patients. In conclusion, Japanese patients with type 1 diabetes are clinically heterogeneous and the determination of immunological features are helpful to clarify the characteristics of the Japanese type 1 diabetic syndrome.
The Attitudes of Cancer Patients and Their Families Toward the Disclosure of Terminal Illness
Authors: Yun Young Ho , Lee Chang Geol , Kim Si-young , Lee Sang-wook , Heo Dae Seog , Kim Jun Suk , Lee Keun Seok , Hong Young Seon , Lee Jung Suk , You Chang Hoon
Published: January 2004

Purpose

To ascertain the attitude of cancer patients and their families toward disclosure of terminal illness to the patient.

Patients and Methods

We constructed a questionnaire that included demographic and clinical information and delivered it to 758 consecutive individuals (433 cancer patients and 325 families that have a relative with cancer) at seven university hospitals and one national cancer center in Korea.

Results

380 cancer patients and one member from each of 281 families that have a relative with cancer completed the questionnaire. Cancer patients were more likely than family members to believe that patients should be informed of the terminal illness (96.1% v 76.9%; P < .001). Fifty percent of the family members and 78.3% of the patients thought that the doctor in charge should be the one who informs the patient. Additionally, 71.7% of the patients and 43.6% of the family members thought that patients should be informed immediately after the diagnosis. Stepwise multiple logistic regression indicated that the patient group was more likely than the family group to want the patient to be informed of the terminal illness (odds ratio [OR], 9.76; 95% CI, 4.31 to 22.14), by the doctor (OR, 4.00; 95% CI, 2.61 to 6.11), and immediately after the diagnosis (OR, 3.64; 95% CI, 2.45 to 5.41).

Conclusion

Our findings indicated that most cancer patients want to be informed if their illness is terminal, and physicians should realize that the patient and the family unit may differ in their attitude toward such a disclosure. Our results also reflect the importance of how information is given to the patient.
Outcome of Elderly Patients With Recurrent or Metastatic Head and Neck Cancer Treated With Cisplatin-Based Chemotherapy
Authors: Argiris Athanassios , Li Yi , Murphy Barbara A. , Langer Corey J. , Forastiere Arlene A.
Published: January 2004

Purpose

To evaluate the outcome of elderly patients with head and neck cancer undergoing palliative chemotherapy.

Patients and Methods

We analyzed combined data from two mature phase III randomized trials conducted by the Eastern Cooperative Oncology Group (ECOG; trial E1393, which compared cisplatin plus paclitaxel at two dose levels, and trial E1395, which compared cisplatin plus fluorouracil to cisplatin plus paclitaxel) to evaluate the toxicity, objective response rates, and survival of patients 70 years or older versus their younger counterparts. All patients had previously untreated recurrent or metastatic squamous cell carcinoma of the head and neck and ECOG performance status 0 or 1.

Results

Fifty-three elderly patients were enrolled from a total of 399 eligible participants (13%). Elderly patients had similar objective response rates (28% v 33%) and median time to progression (5.25 v 4.8 months) compared with younger patients. The median survival was 5.3 v 8 months (Wilcoxon P = .06; log-rank P = .17) and the 1-year survival 26% v 33% for elderly and younger patients, respectively. Elderly patients had a significantly higher incidence of severe nephrotoxicity, diarrhea, and thrombocytopenia. A higher rate of toxic deaths was noted in the elderly but did not reach statistical significance (13% v 8%; P = .29).

Conclusion

Elderly patients were underrepresented in these studies. Fit elderly patients with recurrent or metastatic head and neck cancer sustained increased toxicities with cisplatin-based doublets but had comparable survival outcomes compared with younger patients. Strategies to ameliorate toxicities should be pursued in the elderly.
In Boys with Abnormal Developmental Tempo, Maturation of the Skeleton and the Hypothalamic-Pituitary-Gonadal Axis Remains Synchronous
Authors: Flor-Cisneros Armando , Leschek Ellen W. , Merke Deborah P. , Barnes Kevin M. , Coco Marilena , Cutler Gordon B. , Baron Jeffrey
Published: January 2004

The primary mechanism that initiates puberty is unknown. One possible clue is that pubertal maturation often parallels skeletal maturation. Conditions that delay skeletal maturation also tend to delay the onset of puberty, whereas conditions that accelerate skeletal maturation tend to hasten the onset of puberty. To examine this relationship, we studied boys with congenital adrenal hyperplasia (n = 13) and familial male-limited precocious puberty (n = 22), two conditions that accelerate maturational tempo, and boys with idiopathic short stature (n = 18) in which maturational tempo is sometimes delayed.

In all three conditions, the onset of central puberty generally occurred at an abnormal chronological age but a normal bone age. Boys with the greatest skeletal advancement began central puberty at the earliest age, whereas boys with the greatest skeletal delay began puberty at the latest age. Furthermore, the magnitude of the skeletal advancement or delay matched the magnitude of the pubertal advancement or delay. This synchrony between skeletal maturation and hypothalamic-pituitary-gonadal axis maturation was observed among patients within each condition and also between conditions. In contrast, the maturation of the hypothalamic-pituitary-gonadal axis did not remain synchronous with other maturational processes including weight, height, or body mass index.

We conclude that in boys with abnormal developmental tempo, maturation of the skeleton and the hypothalamic-pituitary-gonadal axis remains synchronous. This synchrony is consistent with the hypothesis that in boys, skeletal maturation influences hypothalamic-pituitary-gonadal axis maturation.
Dysmetabolic Syndrome in a Man with a Novel Mutation of the Aromatase Gene: Effects of Testosterone, Alendronate, and Estradiol Treatment
Authors: Maffei Laura , Murata Yoko , Rochira Vincenzo , Tubert Gloria , Aranda Claudio , Vazquez Marcela , Clyne Colin D. , Davis Susan , Simpson Evan R. , Carani Cesare
Published: January 2004

We present the fourth case of an adult man (29 yr old) affected by aromatase deficiency resulting from a novel homozygous inactivating mutation of the CYP19 (P450arom) gene. At first observation, continuing linear growth, eunuchoid body proportions, diffuse bone pain, and bilateral cryptorchidism were observed. The patient presented also a complex dysmetabolic syndrome characterized by insulin resistance, diabetes mellitus type 2, acanthosis nigricans, liver steatohepatitis, and signs of precocious atherogenesis. The analysis of the effects induced by the successive treatment with high doses of testosterone, alendronate, and estradiol allows further insight into the roles of androgens and estrogens on several metabolic functions. High doses of testosterone treatment resulted in a severe imbalance in the estradiol to testosterone ratio together with the occurrence of insulin resistance and diabetes mellitus type 2. Estrogen treatment resulted in an improvement of acanthosis nigricans, insulin resistance, and liver steatohepatitis, coupled with a better glycemic control and the disappearance of two carotid plaques. Furthermore, the study confirms previous data concerning the key role of estrogens on male bone maturation, at least in part, and regulation of gonadotropin secretion. The biopsy of the testis showed a pattern of total germ cell depletion that might be due to the concomitant presence of bilateral cryptorchidism. Thus, a possible role of estrogen in male reproductive function is suggested but without revealing a direct cause-effect relationship.

Data from this case provide new insights into the role of estrogens in glucose, lipid, and liver metabolism in men. This new case of aromatase deficiency confirms previous data on bone maturation and mineralization, and it reveals a high risk for the precocious development of cardiovascular disease in young aromatase-deficient men.
Mutations in the Seipin and AGPAT2 Genes Clustering in Consanguineous Families with Berardinelli-Seip Congenital Lipodystrophy from Two Separate Geographical Regions of Brazil
Authors: Gomes Karina Braga , Fernandes Ana Paula , Ferreira Alessandro Clayton Souza , Pardini Hermes , Garg Abhimanyu , Magré Jocelyne , Pardini Victor Cavalcanti
Published: January 2004

Berardinelli-Seip congenital lipodystrophy (BSCL) is characterized by a near total congenital absence of fat and predisposition to develop diabetes mellitus. In this study, we investigated the presence of mutations in the Seipin and 1-acylglycerol phosphate acyltransferase 2 (AGPAT2) genes in 32 affected subjects with BSCL from 17 consanguineous pedigrees living in two separate geographical regions, the northeastern and southeastern regions, of Brazil. All, except one, of the 22 BSCL subjects from 15 families living in the northeastern region were found to have a homozygous 669insA mutation in the Seipin gene. In contrast, all 10 BSCL subjects from two families living in the southeastern region were found to a have a homozygous 1036-bp deletion including exons 3 and 4 of AGPAT2. These results support genetic heterogeneity among BSCL patients in Brazil. Our finding of a single mutation in the Seipin and AGPAT2 genes in the pedigrees from the northeastern and southeastern regions, respectively, will be useful in genetic counseling of subjects from these large pedigrees from Brazil.
Do Height-Related Variations in Insulin-Like Growth Factors Underlie the Associations of Stature with Adult Chronic Disease?
Authors: Gunnell D. , Oliver S. E. , Donovan J. L. , Peters T. J. , Gillatt D. , Persad R. , Hamdy F. C. , Neal D. E. , Holly J. M. P.
Published: January 2004

Tall people, particularly those with long legs, have an increased risk of developing cancer but a reduced risk of cardiovascular disease and type II diabetes. We examined associations of stature and body mass index with IGF-I, IGF-II, and IGF binding protein (IGFBP)-2 and IGFBP-3 in 274 men aged 50–70 yr to investigate whether variations in growth factor levels underlie associations of anthropometry with a number of adult diseases. Height and leg and trunk length were not strongly associated with circulating levels of IGF-I, IGF-II, or IGFBP-3. The molar ratio of IGF-I/IGFBP-3 increased with increases in the leg/trunk length ratio (P = 0.06). IGFBP-2 was positively associated with leg length and inversely associated with trunk length. Mean levels of IGFBP-2 (in nanograms per milliliter) across quartiles of increasing leg length were 504.4 493.6, 528.7, and 578.8 (Ptrend = 0.06), and for trunk length were 615.2, 507.2, 498.6, 488.5 (Ptrend < 0.01), suggesting that variations in IGFBP-2, or a factor influencing its levels in the circulation, may contribute to biological mechanisms underlying height-disease associations. We conclude that whereas growth-influencing exposures during childhood, which may operate through effects on IGF-I levels, have long-term influences on disease risk, they do not necessarily program IGF-I levels throughout life. The associations of anthropometry with IGFBP-2 merit additional investigation.
Vitamin D Status as a Determinant of Peak Bone Mass in Young Finnish Men
Authors: Välimäki Ville-Valtteri , Alfthan Henrik , Lehmuskallio Eero , Löyttyniemi Eliisa , Sahi Timo , Stenman Ulf-Håkan , Suominen Harri , Välimäki Matti J.
Published: January 2004

Severe vitamin D deficiency causes rickets, but scarce data are available about the extent to which vitamin D status determines the development of the peak bone mass in young adults. Our aim was to evaluate the prevalence of vitamin D deficiency [serum 25-hydroxyvitamin D (25-OHD) less than the lower limit of the reference range of 20–105 nmol/liter] and the relationship between vitamin D status and peak bone mass among young Finnish men. A cross-sectional study of determinants of peak bone mass with data on lifestyle factors collected retrospectively was performed in 220 young men, aged 18.3–20.6 yr. One hundred and seventy men were recruits of the Finnish Army, and 50 were men of similar age who had postponed their military service for reasons not related to health. Bone mineral content, bone mineral density, and scan area were measured in lumbar spine and upper femur by dual energy x-ray absorptiometry. Serum 25-OHD concentrations were followed prospectively for 1 yr. In July 2000, only 0.9% of the men had vitamin D deficiency, but 6 months later, in the winter, the respective percentage was 38.9%. After adjusting for age, height, weight, exercise, smoking, calcium, and alcohol intake, there existed a positive correlation between serum 25-OHD and bone mineral content at lumbar spine (P = 0.057), femoral neck (P = 0.041), trochanter (P = 0.010), and total hip (P = 0.025). The correlation coefficients for the bone mineral densities at the four measurement sites were 0.035, 0.061, 0.056, and 0.068, respectively. No correlation was found to scan area. We conclude that vitamin D deficiency is very common in Finnish young men in the winter, and it may have detrimental effects on the acquisition of maximal peak bone mass. As in Finland vitamin D supplementation to infants is now stopped at the age of 3 yr, it can be asked whether at our latitude it should be continued from that age onward, not for the prevention of rickets, but as prophylaxis for osteoporosis.
A Coated Tube Assay for the Detection of Blocking Thyrotropin Receptor Autoantibodies
Authors: Minich Waldemar B. , Lenzner Cornelia , Bergmann Andreas , Morgenthaler Nils G.
Published: January 2004

We developed a coated tube assay to discriminate TSH-receptor-stimulating autoantibodies [thyroid-stimulating antibodies (TSAb)] from those autoantibodies blocking TSH binding without intrinsic activation [thyroid-blocking antibodies (TBAb)]. The wild-type TSH receptor in the TSH binding-inhibitory assay was exchanged for a chimeric receptor where a TSAb epitope (amino acids 8–165) was replaced by comparable LH-R residues. Binding of 125I-labeled TSH to this chimera could be inhibited by sera containing TBAb up to 95%. Sera from 316 patients with Graves’ disease and 17 with autoimmune thyroid disease were grouped according to their bioassay activity. At the decision threshold, the chimera A assay had a sensitivity of 78.0% for TBAb with a specificity of 90.2%. In detail, 19 of 22 (86.4%) TBAb sera and 15 of 23 (65.2%) TSAb/TBAb sera were positive but only 32 of 216 (14.0%) TSAb sera and 5 of 72 (6.9%) bioassay negative sera. There was a weak but significant positive correlation (r = 0.46) between the chimera assay and the bioassay for TBAb. This is the first report of a coated tube assay for the determination of TBAb employing an adaptation of the TSH binding-inhibitory format, which could be a useful alternative to the bioassay.
Effects of the Phytoestrogen Genistein on the Circulating Soluble Receptor Activator of Nuclear Factor κB Ligand-Osteoprotegerin System in Early Postmenopausal Women
Authors: Crisafulli Alessandra , Altavilla Domenica , Squadrito Giovanni , Romeo Adolfo , Adamo Elena B. , Marini Rolando , Inferrera Maria A. , Marini Herbert , Bitto Alessandra , D’anna Rosario , Corrado Francesco , Bartolone Sebastiano , Frisina Nicola , Squadrito Francesco
Published: January 2004

We investigated the serum levels of both receptor activator of nuclear factor κB ligand (RANKL) and its decoy receptor osteoprotegerin (OPG) in postmenopausal healthy women after a 1-yr therapy with genistein, (n = 30; 54 mg/d), hormone replacement therapy (n = 30; 1 mg/d 17β-estradiol combined with norethisterone acetate) and placebo (n = 30). By comparison with placebo, the soluble RANKL (sRANKL)/OPG ratio was lower in the genistein group (−69 ± 7%; P < 0.01 vs. placebo 81 ± 24%) and in hormone replacement therapy-treated women (−11 ± 2%; P < 0.01 vs. placebo). A positive correlation (r = 0.63; P < 0.01) was found between 1-yr percentage change in sRANKL/OPG ratio and 1-yr change in urinary deoxypyridinoline, a bone resorption marker. A negative correlation was observed between 1-yr percentage change in sRANKL/OPG ratio and 1-yr change in femoral neck bone mineral density (r = −0.7; P < 0.01). Our findings suggest that the sRANKL-OPG system may mediate the beneficial effects of genistein on bone remodeling in postmenopausal women.
Anti-Müllerian Hormone Serum Concentrations in Normoovulatory and Anovulatory Women of Reproductive Age
Authors: Laven Joop S. E. , Mulders Annemarie G. M. G. J. , Visser Jenny A. , Themmen Axel P. , de Jong Frank H. , Fauser Bart C. J. M.
Published: January 2004

Anti-Müllerian hormone (AMH) concentrations correlate with the number of antral follicles as well as age and constitute an endocrine marker for ovarian aging. In normogonadotropic anovulatory infertile women [World Health Organization (WHO) class 2], the number of early antral follicles is usually increased. To investigate whether AMH concentrations are increased, serum levels in 128 WHO 2 women were compared with those in 41 normoovulatory premenopausal women of similar age.

Serum AMH concentrations are significantly (P < 0.001) elevated in WHO 2 patients [median, 7.6 μg/liter (range, 0.1–40.0)], compared with controls [median, 2.1 μg/liter (0.1–7.4)]. In 106 patients presenting with polycystic ovaries (PCOs) (≥12 follicles/ovary measuring 2–9 mm and/or an ovarian volume > 10 ml), AMH levels were elevated [9.3 μg/liter (1.8–40.0)], compared with 22 patients without PCOs [6.4 μg/liter (0.1–22.1)] (P < 0.0001). In WHO 2 patients, AMH concentrations correlated with features characteristic for polycystic ovary syndrome such as LH concentrations (r = 0.331; P = 0.0001), testosterone levels (r = 0.477, P = 0.0001), mean ovarian volume (r = 0.421; P = 0.0001), and the number of ovarian follicles (r = 0.308; P = 0.0001). AMH levels correlated well with age in WHO 2 patients (r = −0.248; P = 0.002) as well as in controls (r = −0.465; P = 0.005). However, the relative decline in AMH with age is less pronounced in WHO 2 patients. In a subset of patients no significant correlation was found between AMH serum concentrations and the FSH response dose, the duration of stimulation, and the total number of ampoules of FSH used.

In conclusion, serum AMH concentrations are elevated in WHO 2 women, especially in those patients exhibiting PCOs. Because AMH concentrations correlated well with other clinical, endocrine, and ultrasound markers associated with polycystic ovary syndrome, AMH may be used as a marker for the extent of the disease. A less pronounced AMH decrease over time in these women may suggest retarded ovarian aging. The latter hypothesis, however, should be confirmed by longitudinal studies.
The Insulin Secretagogues Glibenclamide and Repaglinide Do Not Influence Growth Hormone Secretion in Humans but Stimulate Glucagon Secretion during Profound Insulin Deficiency
Authors: Østergård Torben , Degn Kristine B. , Gall Mari-Anne , Carr Richard D. , Veldhuis Johannes D. , Thomsen Mads K. , Rizza Robert A. , Schmitz Ole
Published: January 2004

In vitro data have recently suggested that sulfonylureas (SUs) enhance GH secretion by modulating the effects of GHRH and somatostatin in pituitary cells. The present study was undertaken to explore in more detail a possible influence of a single dose of SU (glibenclamide) and a non-SU (repaglinide) insulin secretagogue on circulating GH dynamics. Ten C-peptide-negative type 1 diabetic individuals were examined on three occasions in random order. Either glibenclamide (10.5 mg), repaglinide (8 mg), or placebo was administered after overnight normalization of plasma glucose by iv insulin infusion. Subsequently, GH concentrations were measured regularly after stimulation with GHRH (bolus 0.1 μg/kg) alone and during concomitant infusion with somatostatin (7 ng·kg–1·min–1). Insulin was replaced at baseline levels (0.25 mU·kg–1·min–1) and plasma glucose clamped at 5–6 mmol/liter. Overall, there were no significant statistical differences in GH responses determined as either GH peak concentrations, integrated levels of GH, or secretory burst mass of GH during the experimental protocol. In contrast, plasma glucagon concentrations were significantly increased during glibenclamide and repaglinide exposure. The present experimental design does not support the hypothesis that acute administration of pharmacological doses of the oral antihyperglycemic agents glibenclamide and repaglinide per se enhance GH release in humans. Additionally, this study shows that these potassium channel inhibitors seem to stimulate glucagon secretion in people who have severe intraislet insulin deficiency (e.g. type 1 diabetes). However, extrapolation of our findings to type 2 diabetic individuals should be done with some caution.
Recombinant Human Thyrotropin Stimulation of Fluoro-d-Glucose Positron Emission Tomography Uptake in Well-Differentiated Thyroid Carcinoma
Authors: Chin Bennett B. , Patel Pavni , Cohade Christian , Ewertz Marge , Wahl Richard , Ladenson Paul
Published: January 2004

TSH stimulates thyrocyte metabolism, glucose transport, and glycolysis. 2-Deoxy-2-[18F]fluoro-d-glucose (FDG) is a glucose analog used in positron emission tomography (PET) to detect occult well-differentiated thyroid carcinoma. The objective of this study was to examine the effects of recombinant human TSH (rTSH) on FDG PET uptake in patients with residual or recurrent disease. Seven patients with well-differentiated thyroid carcinoma, negative 131-I scintigraphy, and biochemical evidence of residual disease were randomized and prospectively studied with FDG PET both on thyroid hormone suppression and rTSH stimulation within 1 wk. All lesions seen on the TSH suppression scans were seen on the rTSH stimulation studies. rTSH stimulation studies identified four additional lesions not seen on TSH suppression. One patient was positive on rTSH stimulation alone. The mean (2.54 ± 0.72 vs. 1.79 ± 0.88) and maximum (2.49 ± 0.95 vs. 1.74 ± 0.81) lesion to background ratios were significantly higher with rTSH stimulation, compared with TSH suppression (P = 0.02 for both). rTSH stimulation improves the detectability of occult thyroid metastases with FDG PET, compared with scans performed on TSH suppression.
Adiponectin, an Adipocyte-Derived Protein, Predicts Future Insulin Resistance: Two-Year Follow-Up Study in Japanese Population
Authors: Yamamoto Yukihiro , Hirose Hiroshi , Saito Ikuo , Nishikai Kanako , Saruta Takao
Published: January 2004

It has been reported that the serum adiponectin level was negatively correlated with body mass index (BMI), insulin resistance index, and triglycerides and was positively correlated with high-density lipoprotein cholesterol in several cross-sectional studies. However, the causal relationship has not been elucidated. We investigated whether the baseline adiponectin level could predict subsequent changes in insulin resistance, lipid profile, or body weight in a 2-yr longitudinal study. This study included 590 male Japanese subjects, aged 30–65 yr, who received annual health checkups in both 2000 and 2002. Blood pressure, heart rate, and anthropometric and metabolic parameters, including serum insulin and adiponectin levels, were determined. The insulin resistance index was calculated based on homeostasis model assessment. Baseline adiponectin level was not correlated with the subsequent change in lipid profile or BMI in 2 yr after adjustment for each baseline value. However, the baseline adiponectin level was negatively correlated with subsequent changes in insulin and insulin resistance index based on homeostasis model assessment, even after adjustment for change in BMI (r = −0.162 and r = −0.140, respectively). These findings suggest that the serum adiponectin concentration predicts subsequent changes in insulin resistance, but not in lipid profile or body weight.
The Metabolic Syndrome in Overweight Hispanic Youth and the Role of Insulin Sensitivity
Authors: Cruz Martha L. , Weigensberg Marc J. , Huang Terry T.-K. , Ball Geoff , Shaibi Gabriel Q. , Goran Michael I.
Published: January 2004

The prevalence of the metabolic syndrome is highest among Hispanic adults. However, studies exploring the metabolic syndrome in overweight Hispanic youth are lacking. Subjects were 126 overweight children (8–13 yr of age) with a family history for type 2 diabetes. The metabolic syndrome was defined as having at least three of the following: abdominal obesity, low high-density lipoprotein (HDL) cholesterol, hypertriglyceridemia, hypertension, and/or impaired glucose tolerance. Insulin sensitivity was determined by the frequently sampled iv glucose tolerance test and minimal modeling. The prevalence of abdominal obesity, low HDL cholesterol, hypertriglyceridemia, systolic and diastolic hypertension, and impaired glucose tolerance was 62, 67, 26, 22, 4, and 27%, respectively. The presence of zero, one, two, or three or more features of the metabolic syndrome was 9, 22, 38, and 30%, respectively. After controlling for body composition, insulin sensitivity was positively related to HDL cholesterol (P < 0.01) and negatively related to triglycerides (P < 0.001) and systolic (P < 0.01) and diastolic blood pressure (P < 0.05). Insulin sensitivity significantly decreased (P < 0.001) as the number of features of the metabolic syndrome increased. In conclusion, overweight Hispanic youth with a family history for type 2 diabetes are at increased risk for cardiovascular disease and type 2 diabetes, and this appears to be due to decreased insulin sensitivity. Improving insulin resistance may be crucial for the prevention of chronic disease in this at-risk population.
Effect of ketoprofen and its enantiomers on the renal disposition of methotrexate in the isolated perfused rat kidney
Authors: Karpf D M , Kirkegaard A L , Evans A M , Nation R L , Hayball P J , Milne R W
Published: January 2004

Non-steroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit the renal tubular secretion of methotrexate. However, the relative contribution of the active S- and inactive R-enantiomers is unknown. This study examined the effect of racemic ketoprofen and its enantiomers on the renal disposition of methotrexate in the isolated perfused rat kidney (IPK). Nineteen kidneys were divided between a control and three treatment groups. Controls were perfused with methotrexate alone (25 μg mL−1, n = 5) over three 30-min periods. Treatment groups were perfused with methotrexate (25 μg mL−1) for the first period, followed by a second period of methotrexate (25 μg mL−1) plus R- (n = 5), S- (n = 5) or RS-ketoprofen (n = 4) at 25 μg mL−1, and a third period of methotrexate (25 μg mL−1) plus R-, S- or RS-ketoprofen (50 μg mL−1). Perfusate and urine were collected over 10-min intervals. Methotrexate was measured by HPLC and its binding in perfusate by ultrafiltration. The clearance ratio (CR) for methotrexate was obtained by dividing the renal clearance by the product of its fraction unbound and the glomerular filtration rate. During control experiments, there was no significant change in the CR over 90 min. R-, S- and RS-ketoprofen at 50 μg mL−1 reduced the CR of methotrexate significantly, but there was no difference between the three groups. While the enantiomers of ketoprofen reduced the renal excretion of methotrexate, the interaction was not enantioselective.
A bZIP/bRLZ transcription factor required for DIF signaling inDictyostelium
Authors: Thompson Christopher R. L. , Fu Qing , Buhay Caroline , Kay Robert R. , Shaulsky Gad
Published: January 2004

The intermingled differentiation and sorting out of Dictyosteliumprestalk-O and prespore cells requires the diffusible signaling molecule DIF-1, and provides an example of a spatial information-independent patterning mechanism. To further understand this patterning process, we used genetic selection to isolate mutants in the DIF-1 response pathway. The disrupted gene in one such mutant, dimA–, encodes a bZIP/bRLZ transcription factor, which is required for every DIF-1 response investigated. Furthermore, the dimA– mutant shows strikingly similar developmental defects to the dmtA– mutant,which is specifically defective in DIF-1 synthesis. However, key differences exist: (1) the dmtA– mutant responds to DIF-1 but does not produce DIF-1; (2) the dimA– mutant produces DIF-1 but does not respond to DIF-1; and (3) the dimA– mutant exhibits cell autonomous defects in cell type differentiation. These results suggest that dimA encodes the key transcriptional regulator required to integrate DIF-1 signaling and subsequent patterning in Dictyostelium.
Drosophilaventral furrow morphogenesis: a proteomic analysis
Authors: Gong Lei , Puri Mamta , Ünlü Mustafa , Young Margaret , Robertson Katherine , Viswanathan Surya , Krishnaswamy Arun , Dowd Susan R. , Minden Jonathan S.
Published: January 2004

Ventral furrow formation is a key morphogenetic event during Drosophila gastrulation that leads to the internalization of mesodermal precursors. While genetic analysis has revealed the genes involved in the specification of ventral furrow cells, few of the structural proteins that act as mediators of ventral cell behavior have been identified. A comparative proteomics approach employing difference gel electrophoresis was used to identify more than fifty proteins with altered abundance levels or isoform changes in ventralized versus lateralized embryos. Curiously, the majority of protein differences between these embryos appeared well before gastrulation, only a few protein changes coincided with gastrulation,suggesting that the ventral cells are primed for cell shape change. Three proteasome subunits were found to differ between ventralized and lateralized embryos. RNAi knockdown of these proteasome subunits and time-dependent difference-proteins caused ventral furrow defects, validating the role of these proteins in ventral furrow morphogenesis.
Chaperone-mediated folding and assembly of myosin in striated muscle
Authors: Srikakulam Rajani , Winkelmann Donald A.
Published: January 2004

De novo folding and assembly of striated muscle myosin was analyzed by expressing a GFP-tagged embryonic myosin heavy chain (GFP-myosin) in post-mitotic C2C12 myocytes using replication defective adenoviruses. In the early stages of muscle differentiation, the GFP-myosin accumulates in bright globular foci and short filamentous structures that are later replaced by brightly fluorescent myofibrils. Time-lapse microscopy shows that the intermediates are dynamic and are present in elongating and fusing myocytes and in multinucleated myotubes. Immunostaining reveals the co-localization of the molecular chaperones Hsc70 and Hsp90 with the GFP-myosin in the intermediates, but not in the mature myofibrils. Uninfected cells have similar intermediates suggesting a common pathway for myosin maturation. Two conformation-sensitive antibodies that bind the unfolded motor domain and the coiled-coil conformation of the rod demonstrate that in the intermediates, the myosin rod is folded but the motor domain is not folded. Electron microscopy reveals that the intermediates contain loose filament bundles surrounded by a protein rich matrix. Geldanamycin, a specific inhibitor of Hsp90, reversibly blocks myofibril assembly and triggers accumulation of myosin folding intermediates. We conclude that multimeric complexes of nascent myosin filaments associated with Hsc70 and Hsp90 are intermediates in the folding and assembly pathway of muscle myosin.
A domain of Rad9 specifically required for activation of Chk1 in budding yeast
Authors: Blankley Richard T. , Lydall David
Published: January 2004

The Rad9 protein is a key adaptor protein in Saccharomyces cerevisiae DNA damage checkpoint pathways. Its adaptor function is to link the activity of the Mec1 kinase to the activation of two parallel signalling pathways dependent on the Rad53 and Chk1 kinases. The mechanisms by which Rad9 interacts with, and activates, Rad53 are well understood. However, little was known about how Rad9 facilitates the activation of Chk1. We show here that the N-terminus of Rad9 is specifically important for phosphorylation and activation of the Chk1 kinase but not for the phosphorylation and activation of the Rad53 kinase. The Chk1 activation domain (CAD) of Rad9 is specifically important for signalling cell-cycle arrest after cdc13-1- and yku70Δ-induced telomere damage but not for tolerating ultraviolet-induced damage or inhibiting nuclease activity at telomeres. This work extends data showing that separable domains within the Rad9 adaptor protein allow it to activate two distinct kinase signalling pathways independently of each other.
Electrogenic proton-regulated oxalate/chloride exchange by lobster hepatopancreatic brush-border membrane vesicles
Authors: Gerencser George A. , Robbins Frank , Zhang Jianliang , Ahearn Gregory A.
Published: January 2004

The transport of [14C]oxalate (Ox2–) by epithelial brush-border membrane vesicles (BBMV) of lobster (Homarus americanus) hepatopancreas, formed by a magnesium precipitation technique, was stimulated by an outward Cl– gradient (in >out). By contrast, Ox2– uptake was not enhanced by an inward Na+ or K+ transmembrane gradient. Generation of an inside-positive membrane potential by K+ in the presence of valinomycin stimulated Ox2–/Cl– exchange,while an inside-negative membrane potential generated by K+ efflux in the presence of valinomycin inhibited this process. Neither Ox2–/Ox2– nor Ox2–/SO42– transport exchange were affected by alterations of transmembrane potential. An inwardly directed proton gradient, or the presence of low bilateral pH, enhanced Ox2–/Cl– exchange, yet the H+gradient alone could not stimulate Ox2– uptake in Cl–-equilibrated BBMV or in vesicles lacking internal Cl–. The stilbenes 4-acetamido-4′-isothiocyanotostilbene-2,2′-disulfonic acid (SITS)and 4,4′-diisothiocyano-2,2′-disulfonic stilbene (DIDS) strongly inhibited Ox2–/Cl– exchange. Oxalate influx occurred by a combination of carrier-mediated transfer, exhibiting Michaelis–Menten kinetics, and nonsaturable `apparent diffusion'. Apparent kinetic constants for Ox2–/Cl–exchange were Kt=0.20 mmol l–1 and Jmax=1.03 nmol l–1 mg–1protein 7 s–1. 36Cl– influx into oxalate-loaded BBMV was stimulated by an inside-negative transmembrane potential compared with short-circuited vesicles. These results suggest that Ox2–/Cl– exchange in crustacean hepatopancreatic BBMV occurred by an electrogenic carrier mechanism exhibiting a 1:1 flux ratio that was modulated by an external proton-sensitive regulatory site.
Water calcium concentration modifies whole-body calcium uptake in sea bream larvae during short-term adaptation to altered salinities
Authors: Guerreiro Pedro M. , Fuentes Juan , Flik Gert , Rotllant Josep , Power Deborah M. , Canario Adelino V. M.
Published: January 2004

Whole-body calcium uptake was studied in gilthead sea bream larvae(9–83 mg) in response to changing environmental salinity and[Ca2+]. Calcium uptake increased with increased fish size and salinity. Fish exposed to calcium-enriched, diluted seawater showed increased calcium uptake compared with fish in diluted seawater alone. Calcium uptake was unchanged in Na+-enriched, diluted seawater. Overall,[Ca2+], and not salinity/osmolarity per se, appears to be the main factor contributing to calcium uptake. By contrast, drinking was reduced by a decrease in salinity/osmolarity but was little affected by external [Ca2+]. Calculations of the maximum contribution from drinking-associated calcium uptake showed that it became almost insignificant(less than 10%) through a strong decrease in drinking rate at low salinities(0–8‰). Diluted seawater enriched in calcium to the concentration present in full-strength seawater (i.e. constant calcium, decreasing salinity)restored intestinal calcium uptake to normal. Extra-intestinal calcium uptake also benefited from calcium addition but to a lesser extent.