388 Oxidative Stress in Overweight Children and Adolescents

v o l u m e 5 5 su p p l e m e n t 1 N J a n u a r y 2 0 0 7 PCOS is less clear. Purpose of Study: The long-term goal of our study is to determine whether raised peripubertal T levels in female macaques lead to the hypothalamic-pituitary-ovarian sequelae of PCOS. During the first phase of the study presented here our goals are threefold: to obtain normative data for peripubertal T levels in female macaques, to choose a raised T target range, and to devise a method for raising the T levels. Methods Used: Sera obtained from five female macaques starting at prepuberty (age 12 months) and through menarche (average age 29 months) were used to determine normative T data. The commercially available RIA T assay DSL-4100 was used. We used surgically placed Silastic tube implants to raise T levels to target range in eight experimental animals. Six control animals were implanted with cholesterol-containing Silastic implants. Summary of Results: The mean peripubertal T in female macaques was 0.36 6 0.16 ng/mL. Since obese peripubertal adolescent girls have T levels 3 to 4 times the normal-weight controls, we chose a target T range of 0.12 to 0.16 ng/mL. Subcutaneously placed Silastic implants containing 5 mm of T:cholesterol (1:12 or 1:15) are used to achieve the target T range. Monkeys are phlebotomized weekly and the frequency of implant change (on average every 6 weeks) is determined by the serum T levels. Conclusions: Peripubertal female macaques have a low circulating level of T, which can be raised reliably to a narrow target range with surgically placed Silastic tubes and close monitoring of resultant circulating T levels. We will maintain the raised T levels throughout puberty as we compare the T-treated animals’ ovarian changes (menstrual/ovarian follicles) and hypothalamic-pituitary parameters (LH pulse and response to GnRH/progesterone) with those in control animals. This work is supported by the following NIH grants: HD18185, DK007674-13, and RR00163. 388 385 POTENTIAL MALE CONTRACEPTIVE TARGET: GERM CELL–SPECIFIC VASA PROTEIN LOCALIZATION IN MONKEY AND HUMAN TESTES. K. Ma,1 F. Berglund,1 M. Crespo,1 Y.H. Lue,1 R.S. Swerdloff,1 A.P. Sinha Hikim,1 Y.X. Liu,2 Y.G. Cui,3 X.H. Wang,4 J.H. Sha,3 Z.M. Zhou,3 C. Wang,1 1LABioMed at Harbor-UCLA Medical Center, Torrance, CA; 2Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Science, Beijing, China; 3First Affiliated Hospital and Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China; 4Jiangsu Family Planning Research Institute, Nanjing, China. We previously demonstrated that pachytene and diplotene spermatocytes and round spermatids are the most susceptible cells to undergo apoptosis in response to either a withdrawal of intratesticular testosterone, increased testicular temperature, or a combination of both in rats and monkeys. Our study focuses on vasa (DDX4 or MvH), a specific meiotic and/or postmeiotic molecule essential for germ cell survival and development. Vasa is a member of the DEAD (Asp-Glu-Ala-Asp) box family of ATP-dependent RNA helicases and participates in RNA unwinding, translation initiation, and RNA turnover. Previously, the vasa gene was found to be conserved across species in Caenorhabditis elegans, Xenopus, zebra fish, mice, and rats. In mice, vasa protein has been localized in spermatocytes and round spermatids and vasa knockout mice were infertile due to an arrest of spermatogenesis at zygotene spermatocytes. The objective of this study is to compare vasa protein expression and localization in rat, monkey, and human testes. Testicular samples were obtained from groups of four adult rats, four adult monkeys, and four fertile men. Bouin’s fixed, paraffin-embedded testicular sections were used for vasa immunohistochemistry. Vasa proteins were predominantly localized in the cytoplasm of pachytene and diplotene spermatocytes and round spermatids across all three species. The vasa proteins were condensed in the chromatoid bodies of late spermatocytes and round spermatids. Our studies also confirm vasa as an evolutionary conserved germ cell–specific molecule since vasa proteins were present in rat, monkey, and human testes. The function of vasa will be determined in the monkey and human testes after interventions to regulate spermatogenesis. Further investigation of vasa in the human testes may lead to a potential target for male contraception. 386 LARGE INDIVIDUAL DIFFERENCES IN THE LEVEL OF PHYSICAL ACTIVITY ARE APPARENT THROUGH THE PUBERTAL TRANSITION. M.O. Cardenas,1 J.L. Cameron,1,2 1Oregon Health & Science University, Portland, OR; 2Oregon National Primate Research Center, Portland, OR. Background: A number of epidemiologic studies have shown an association between low levels of physical activity and elevated weight gain, but most reports of physical activity have been made by selfreport. Recently, we directly measured physical activity using three-way accelerometers in adult female rhesus monkeys and showed that sedentary monkeys gained significantly more weight than active monkeys (Sullivan et al, Am J Physiol Regul Integ Comp Physiol 2006;291:R633–42). An eightfold variation in daily physical activity was found between the least and most active monkeys, and the physical activity level was very stable over time. Purpose: Over adolescent development, human studies have shown a dramatic decrease in average physical activity level. In this study, we sought to determine if individual differences in physical activity already exist prepubertally and whether an individual’s prepubertal level of physical activity predicts their postpubertal level of physical activity. Methods: Forty-three rhesus monkeys were studied over a 1-year period between 2005 and 2006. Monkeys were living in seminatural conditions in large outdoor troops at the Oregon National Primate Research Center. Each year the monkeys were brought indoors for a 10-day period in the summer and each monkey had physical activity monitored one summer by a three-way accelerometer (set to record activity counts each minute) that was attached to a loose-fitting metal collar. During the 10-day period of study, monkeys were housed individually in single cages. Results: There was a 10.5-fold difference in physical activity level among monkeys. Over the 1-year study period there was a 2.6-fold decrease in physical activity. There was a significant correlation (r 5 .519, , .001) in activity levels measured when monkeys were 2 years old (ie, prepubertal) and 3 years old (ie, postpubertal). Conclusion: We conclude that like humans, monkeys experience a decrease in physical activity as they transition through puberty; however, prepubertal physical activity level is a strong predictor of postpubertal physical activity level. These findings suggest that sedentary adolescents will remain sedentary in adulthood and that these individuals will be at risk of obesity. 387 RAISING PERIPUBERTAL TESTOSTERONE LEVELS IN FEMALE MACAQUES: FIRST PHASE OF A PROSPECTIVE STUDY TO EVALUATE THE ROLE OF PERIPUBERTAL HYPERANDROGENEMIA IN THE PATHOGENESIS OF POLYCYSTIC OVARY SYNDROME. A. Bahar, O. Slayden, D. Hess, J. Cameron, R.L. Stouffer, Oregon National Primate Research Center, Oregon Health & Science University, Portland, OR. Background: Polycystic ovary syndrome (PCOS), a syndrome of hyperandrogenism, polycystic ovaries, and oligo-anovulation, is a major cause of infertility in American women. Observational studies suggest that women first develop PCOS in adolescence. Recent studies show that testosterone (T) levels are elevated in prepubertal obese girls in comparison with normal weight age and puberty-matched controls and this trend continues throughout puberty. Although insulin resistance is a common finding in PCOS and is implicated in the pathogenesis of PCOS, the causal role of T in the development of OXIDATIVE STRESS IN OVERWEIGHT CHILDREN AND ADOLESCENTS. W.N. Evans, G.A. Mayman, R.J. Acherman, K.A. Cass, C.F. Luna, A.J.C. Collazos, H. Restrepo, Children’s Heart Center and University of Nevada, School of Medicine, Las Vegas, NV. Background: In adult studies, elevated concentrations of oxidative stress markers have been associated with obesity and other risk factors for cardiovascular disease. Objective: To compare concentrations of urinary oxidative stress markers between overweight and normal-weight children and adolescents. Methods and Patients: This pilot study compared data from 23 children and adolescents with a body mass index (BMI) $ 95th percentile, in good health conditions except for their obesity, with a group of 17 normal BMI percentile peers. Oxidative stress test, which includes 8-epi-PGF2£ and 8-epi-PGF2£/ creatinine ratio, were measured in a spot urine morning sample. Spearman correlation and nonparametric tests were used for statistical analysis. Results: The group of overweight children was composed of 14 boys and 9 females, with a mean age of 12.3 6 2.0 years and a mean BMI Z-score of 2.1 6 0.4; the group of normal-weight children had 12 boys and 5 girls, with a mean age of 13.7 6 2.2 and a mean BMI Z-score of 20.5 6 1.3. There was no significant difference between groups in 8-epi-PG F2£ (2,561.8 6 1,578.2 vs 2,601.5 6 1,337.2 pg/mL, p 5 .7) or 8-epi-PGF2£/creatinine ratio (17.2 6 5.2 vs 17.9 6 5.9, p 5 .8). 8-epi-PG F2£ was not correlated with BMI or BMI Z-score. Conclusion: In this group of overweight children and adolescents, oxidative stress markers were not significantly elevated or correlated with BMI. 389 PARATHYROID CARCINOMA: A CASE REPORT. P. Sahasranam, M.T. Tran, T.C. Friedman, Charles R. Drew University of Medicine and Science, Los Angeles, CA. A 53-year-old man with no past medical history was admitted with complaints of hematuria and flank and abdominal pain, for a duration of 1 week. He also complained of bone pain, fatigue, nausea, weight loss, and an enlarging new neck mass 1 month prior to the presentation. He admitted to a history of smoking in the past and no significant family history. On admission, his vital signs were stable. Initial laboratory results showed serum calcium 17.3 mg/dL (normal 8.5–10.5 mg/dL), serum albumin 2.9 g/ dL (normal 3.0–5.0 g/dL), blood urea nitrogen 48 mg/dL (normal 7–18 mg/dL), serum creatinine 3.4 mg/dL (normal 0.5–1.2 mg/dL), serum phosphorus 4.3 mg/dL (normal 2.5–4.6 mg/dL), serum magnesium 2.2 mg/dL (normal 1.8–2.8 mg/dL), urine analysis positive for moderate amount of blood, alkaline phosphatase 121 IU/L (normal 44–147 IU/L), hemoglobin 8.4 g/dL (normal 13–18 g/dL), hematocrit 24.5 (normal 37–49%), and intact PTH 1,491 pg/mL (normal 10–65 pg/dL). A sestamibi scan done subsequently was negative. A neck sonogram showed a complicated left neck mass measuring 3.4 cm 3 2.2 cm 3 1.7 cm with a thick septum and a cystic component. A renal sonogram showed bilateral renal cysts and two stones each measuring 10 mm on the left kidney. The chest radiograph was normal. He was hydrated for a week with improvement in his laboratory tests showing serum calcium decreased to 9.3 mg/dL and a serum creatinine of 1.8 mg/dL. He underwent a total thyroidectomy and parathyroidectomy. The pathology showed bilateral parathyroid carcinoma infiltrating the capsule, invading the capsule, and adhering to the thyroid, with evidence of lymphovascular invasion. The immunohistochemical staining was negative for p53 and Ki67. After surgery, his symptoms significantly improved and laboratory tests done 2 weeks later showed the following: calcium 7.8 mg/dL, intact PTH 7 pg/mL, TSH 15 mIU/L, and FT4 0.6 mg/dL. A DXA scan showed osteoporosis. Postoperative management included treatment of hypothyroidism, hypoparathyroidism, osteoporosis, and chronic renal insufficiency. Two years later, he continues to be asymptomatic on medications with normal biochemical parameters. A high degree of suspicion is required to diagnose parathyroid carcinoma. A better knowledge and understanding of this condition would aid in early diagnosis and possibly increase in survival rate. 390 A RARE CAUSE OF POLYURIA. F. Wolf, D. Cook, Oregon Health & Sciences University, Portland, OR. Case History: A 36-year-old female presents with a lifelong history of polydipsia and polyuria of 4 to 8 L/day. She is on no medications. Her father and two paternal uncles have the same symptoms. Evaluation: She has no electrolyte or glucose abnormalities; her serum sodium is 141. Maximum urine osmolality after water deprivation is 36 mOsm/kg and does not respond to exogenous vasopressin (DDAVP). Antidiuretic hormone (ADH) level during the test is 10.2 pg/mL (normal , 4.8). Because her family history is suggestive of an autosomal dominant (AD) disorder, genetic studies are done and reveal an E258K mutation of the aquaporin 2 (AQP2) gene. Discussion: Less than 1% of patients (only seven families) diagnosed with familial nephrogenic diabetes insipidus (NDI) have mutations in the AQP2 gene on chromosome 12q13 that are inherited in an AD fashion. AQPs are expressed as homotetramers; however, monomers are functional. When expressed in oocytes, AQP2 mutants form heterotetramers with wt-AQP2, which are missorted and retained in the Golgi apparatus and other organelles. In comparison, autosomal recessive (AR) mutations of AQP2, which comprise 9% of all NDI, lead to misfolding and trapping of the mutants in the endoplasmic reticulum and a more severe phenotype. This includes an earlier presentation of symptoms and a complete lack of response to DDAVP with urine concentrating ability less than 200 mosmol/kgH2O when compared to AD AQP2 mutations. Mutations of the vasopressin type 2 receptor (V2R) are the most common cause of familial NDI, responsible for over 90% of cases. These loss-of-function mutations are inherited in an X-linked J Investig Med: first published as on 12 January 2016. Downloaded from file:/ on November 23, 2023 by guest. Protected by copyright. administration of exogenous testosterone. The JNK (c-Jun NH2-terminal kinase) signaling pathway has been implicated in the activation of apoptosis in various cell systems by stimulating the intrinsic pathway, but its role in testicular germ cell death is unclear. The goal of this study was to define the role of JNK in male germ cell apoptosis in monkeys after mild testicular hyperthermia or deprivation of intratesticular T or the combination of both interventions. Study Design: Groups of eight adult cynomolgus monkeys received one of the following treatments: (1) two empty Silastic implants (C); (2) two 5.5 cm T-implants (T); (3) daily exposure of testes to heat (43uC for 30 minutes) for 2 consecutive days (H); and (4) two T-implants plus exposure of the testes to heat for 2 consecutive days (T + H). Testicular biopsies were performed before and at 3, 8, and 28 days during treatment. Results: Activation of JNK, as evidenced by increase in phospho-c-Jun in testis lysates, was detected in all treatment groups on day 3. Compared with controls, where no staining was detected, a strong phospho-c-Jun staining was detected in the nuclei of apoptotic germ cells in all treatment groups and in the Sertoli cell nuclei at day 8 in H and H + T groups. To further define the role of JNK in apoptotic signal transduction, we examined the expression of JNK1, JNK2, and JNK3 in testes after these interventions. In the control testes, the expression of JNKs was localized in the Sertoli cell cytoplasm. Costaining for JNK2 and -3 and for TUNEL shows expression of both of these isoforms only in those germ cells undergoing apoptosis when compared with controls where these proteins were detected in cytoplasm. In contrast, JNK1 was detected in the Sertoli cell nuclei at day 8 in H and H + T groups. Conclusion: Our results indicate that (1) the JNK pathway may play a role in male germ cell apoptosis in monkeys; (2) JNK isoforms could have preferential effects on testis function; and (3) Sertoli cells participate in germ cell apoptosis triggered by heat stress via JNK signaling. S 139