Effects of hyperandrogenemia and increased adiposity on reproductive and metabolic parameters in young adult female monkeys.

Many patients with hyperandrogenemia are overweight or obese, which exacerbates morbidities associated with polycystic ovary syndrome (PCOS). To examine the ability of testosterone (T) to generate PCOS-like symptoms, monkeys received T or cholesterol (control) implants (n = 6/group) beginning prepubertally. As previously reported, T-treated animals had increased neuroendocrine drive to the reproductive axis [increased luteinizing hormone (LH) pulse frequency] at 5 yr, without remarkable changes in ovarian or metabolic features. To examine the combined effects of T and obesity, at 5.5 yr (human equivalent age: 17 yr), monkeys were placed on a high-calorie, high-fat diet typical of Western cultures [Western style diet (WSD)], which increased body fat from <2% (pre-WSD) to 15-19% (14 mo WSD). By 6 mo on WSD, LH pulse frequency in the controls increased to that of T-treated animals, whereas LH pulse amplitude decreased in both groups and remained low. The numbers of antral follicles present during the early follicular phase increased in both groups on the WSD, but maximal follicular size decreased by 50%. During the late follicular phase, T-treated females had greater numbers of small antral follicles than controls. T-treated monkeys also had lower progesterone during the luteal phase of the menstrual cycle. Although fasting insulin did not vary between groups, T-treated animals had decreased insulin sensitivity after 1 yr on WSD. Thus, while WSD consumption alone led to some features characteristic of PCOS, T + WSD caused a more severe phenotype with regard to insulin insensitivity, increased numbers of antral follicles at midcycle, and decreased circulating luteal phase progesterone levels.

The RHOX homeobox gene cluster is selectively expressed in human oocytes and male germ cells.

STUDY QUESTION: What human tissues and cell types express the X-linked reproductive homeobox (RHOX) gene cluster? SUMMARY ANSWER: The RHOX homeobox genes and proteins are selectively expressed in germ cells in both the ovary and testis. WHAT IS KNOWN ALREADY: The RHOX homeobox transcription factors are encoded by an X-linked gene cluster whose members are selectively expressed in the male and female reproductive tract of mice and rats. The Rhox genes have undergone strong selection pressure to rapidly evolve, making it uncertain whether they maintain their reproductive tissue-centric expression pattern in humans, an issue we address in this report. STUDY DESIGN, SIZE, DURATION: We examined the expression of all members of the human RHOX gene cluster in 11 fetal and 8 adult tissues. The focus of our analysis was on fetal testes, where we evaluated 16 different samples from 8 to 20 weeks gestation. We also analyzed fixed sections from fetal testes, adult testes and adult ovaries to determine the cell type-specific expression pattern of the proteins encoded by RHOX genes. PARTICIPANTS/MATERIALS, SETTING, METHODS: We used quantitative reverse transcription-polymerase chain reaction analysis to assay human RHOX gene expression. We generated antisera against RHOX proteins and used them for western blotting, immunohistochemical and immunofluorescence analyses of RHOXF1 and RHOXF2/2B protein expression. MAIN RESULTS AND THE ROLE OF CHANCE: We found that the RHOXF1 and RHOXF2/2B genes are highly expressed in the testis and exhibit low or undetectable expression in most other organs. Using RHOXF1- and RHOXF2/2B-specific antiserum, we found that both RHOXF1 and RHOXF2/2B are primarily expressed in germ cells in the adult testis. Early stage germ cells (spermatogonia and early spermatocytes) express RHOXF2/2B, while later stage germ cells (pachytene spermatocytes and round spermatids) express RHOXF1. Both RHOXF1 and RHOXF2/2B are expressed in prespermatogonia in human fetal testes. Consistent with this, RHOXF1 and RHOXF2/2B mRNA expression increases in the second trimester during fetal testes development when gonocytes differentiate into prespermatogonia. In the human adult ovary, we found that RHOXF1 and RHOXF2/2B are primarily expressed in oocytes. LIMITATIONS, REASONS FOR CAUTION: While the average level of expression of RHOX genes was low or undetectable in all 19 human tissues other than testes, it is still possible that RHOX genes are highly expressed in a small subset of cells in some of these non-testicular tissues. As a case in point, we found that RHOX proteins are highly expressed in oocytes within the human ovary, despite low levels of RHOX mRNA in the whole ovary. WIDER IMPLICATIONS OF THE FINDINGS: The cell type-specific and developmentally regulated expression pattern of the RHOX transcription factors suggests that they perform regulatory functions during human fetal germ cell development, spermatogenesis and oogenesis. Our results also raise the possibility that modulation of RHOX gene levels could correct some cases of human infertility and that their encoded proteins are candidate targets for contraceptive drug design.

Elevated androgens during puberty in female rhesus monkeys lead to increased neuronal drive to the reproductive axis: a possible component of polycystic ovary syndrome.

BACKGROUND: Hyperandrogenemia is associated with several clinical disorders in which both reproductive dysfunction and metabolic changes may coexist [i.e. polycystic ovary syndrome (PCOS), obesity and congenital adrenal hyperplasia]. Moreover, there is growing evidence that the elevated levels of circulating androgens in obese girls may lead to an increased neuroendocrine drive to the reproductive axis, similar to that associated with PCOS. METHODS: To test whether androgen exposure in the childhood and adolescent period could lead to pubertal alterations in LH secretory patterns, female rhesus monkeys received subcutaneous testosterone implants prepubertally beginning at 1 year of age, maintaining a 3.7-fold increase (P = 0.001) in circulating testosterone levels over cholesterol-implant controls (n = 6/group) into the post-pubertal period. In early adulthood, pulsatile secretion of LH was measured over 12 h during the early follicular phase of a menstrual cycle, and responsiveness of the pituitary to gonadotrophin-releasing hormone was determined. In addition, ultrasounds were performed to assess ovarian morphology and glucose tolerance testing was performed to assess insulin sensitivity. RESULTS: The timing of menarche was similar between groups. Testosterone-treated animals had a significantly greater LH pulse frequency during the early follicular phase compared with controls (P = 0.039) when measured at 5 years of age. There was a larger LH response to GnRH when testosterone-treated animals were 4 years of age (P = 0.042), but not when the animals were 5 years old (P = 0.57). No differences were seen in insulin sensitivity or ovarian morphology, and the groups showed similar rates of ovulation in early adulthood. CONCLUSIONS: Exposure to increased levels of androgens over the course of pubertal development appears to trigger physiological changes in the neural drive to the reproductive axis that resemble those of obese hyperandrogenemic girls in early adulthood and are characteristic of PCOS.

Reproductive endocrinology of adolescent polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is the most common endocrinopathy among reproductive-aged women, and it typically presents during adolescence. The objective of this review is to describe the clinical manifestations of PCOS in adolescent girls and the underlying basis for the altered reproductive physiology. Recognising adolescents at risk for PCOS and taking the appropriate steps to reduce circulating androgen levels is critical in reducing the clinical symptomatology of this disorder, and the development of adulthood infertility, diabetes, and metabolic syndrome in patients with PCOS.

New Cs sputter ion source with polyatomic ion beams for secondary ion mass spectrometry applications.

A simple design for a cesium sputter ion source compatible with vacuum and ion-optical systems as well as with electronics of the commercially available Cameca IMS-4f instrument is reported. This ion source has been tested with the cluster primary ions of Si(n)(-) and Cu(n)(-). Our experiments with surface characterization and depth profiling conducted to date demonstrate improvements of the analytical capabilities of the secondary ion mass spectrometry instrument due to the nonadditive enhancement of secondary ion emission and shorter ion ranges of polyatomic projectiles compared to atomic ones with the same impact energy.

[Polycystic ovaries in 2001: physiology and treatment]. / Ovaires polykystiques en 2001: physiologie et thérapeutique.

Clinical characteristics of PCOS Syndrome Two fundamental characteristics: hyperandrogenism and anovulation which lead to hirsutism and oligo-or amenorrhea. Other features include obesity, acanthosis nigricans, and metabolic disruption (insulin resistance with hyperinsulinemia, glucose intolerance, or type II diabetes mellitus). Complementary tests Serum testosterone and DHEA-S levels: to exclude androgen-producing tumors. Serum 17-hydroxyprogesterone level: to exclude congenital adrenal hyperplasia, 21-hydroxylase deficiency. Ultrasound: increased size of the ovaries and central stroma with presence of peripheral follicular cysts (8-10) measuring about 8 mm in diameter. Pathophysiology Therapeutic approaches Therapeutic approaches

Pancreatitis secondary to hyperparathyroidism during pregnancy.

BACKGROUND: It has been suggested that magnesium can be used to reduce serum calcium levels seen with hyperparathyroidism during pregnancy, thus reducing maternal and fetal risk. CASE: A young woman presented at 32 weeks' gestation with abdominal pain from pancreatitis caused by hyperparathyroidism from a parathyroid adenoma. She was started on magnesium sulfate tocolysis for preterm labor. During treatment, serum parathyroid hormone was undetectable, but serum calcium and vitamin D-1,25 were elevated. When magnesium was discontinued, her vitamin D-1,25 was suppressed and the parathyroid hormone was elevated. CONCLUSION: For some patients, because of persistent hypercalcemia, magnesium sulfate might not be a viable treatment option for hyperparathyroidism during pregnancy.

A model for initial care of the infertile couple.

Infertility affects an estimated 6.1 million women in the United States. Although more than 1.2 million women seek medical care for infertility each year, the majority drop out of the treatment process before achieving a successful pregnancy. An expert panel of five reproductive endocrinologists developed the Individualized Infertility Care Plan model to aid obstetrician-gynecologists and primary care physicians in streamlining the critical period encompassing diagnosis and initial treatment of the patient with uncomplicated infertility. The model emphasizes the importance of the physician's establishing a strong, lasting relationship with the infertile patient.

Pregnancy-associated plasma protein-A gene expression in human ovaries is restricted to healthy follicles and corpora lutea.

Recently, Pregnancy-Associated Plasma Protein-A (PAPP-A) in human follicular fluid was identified as an insulin-like growth factor binding protein-4 protease (IGFBP-4ase). The ability of IGFBP-4ase to inactivate the FSH antagonist, IGFBP-4, has suggested a possible role for PAPP-A in regulating FSH action. Despite growing interest in this protease, the question of whether the PAPP-A gene is expressed in ovaries of normal cycling women is unknown. To fill this basic gap in our knowledge, we have identified the cellular sites of PAPP-A gene expression in normal human ovaries by in situ hybridization. PAPP-A mRNA was low or undetectable in preantral follicles, small (1-2 mm) healthy and atretic antral follicles, larger atretic antral follicles, surface epithelium, tunica albuginea and connective tissue cells. In contrast, an intense PAPP-A hybridization signal was evident in the healthy antral follicles examined from 5 mm to the preovulatory stage. In these follicles, the signal was restricted to the granulosa cells (GC). An intense signal for PAPP-A mRNA was also present in healthy corpora lutea (CL), being localized to a subset of large luteal cells. Collectively, these results provide the first evidence that the gene encoding PAPP-A is expressed in ovaries of normal cycling women and show that the gene is expressed almost exclusively in healthy GC and CL cells. The restricted pattern of PAPP-A expression in normal human ovaries suggests that PAPP-A may be a functional marker of the dominant follicle and its product, the CL. Although the physiological function of ovarian PAPP-A remains to be identified, we hypothesize it might play a role in controlling survival, growth, and/or differentiation of the dominant follicle and CL by inactivating the gonadotropin antagonist, IGFBP-4.

Reversed-phase high-performance liquid chromatographic assay for the adenovirus type 5 proteome.

An RP-HPLC assay was developed for a recombinant adenovirus type 5. During chromatography, intact adenovirus dissociated into its structural components (DNA and proteins) and the viral proteome was separated yielding a characteristic fingerprint. The individual components were identified by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy, N-terminal sequencing and amino acid composition. The assay was utilized to measure adenovirus particle concentration through quantification of structural proteins. Each structural protein provided independent measurement of virus concentration allowing verification of accuracy. The assay sensitivity is at or below 2 x 10(8) particles. Contrary to the benchmark spectrophotometric assay, the RP-HPLC assay was shown to be insensitive to contaminants common for partially purified adenovirus preparations.

Diagnosis of polycystic ovary syndrome.

The clinical features of polycystic ovary syndrome (PCOS) include hirsutism and irregular menses, which are the results of ovarian hyperandrogenism and chronic, unopposed estrogen secretion. The discovery that most women with PCOS are insulin-resistant and have compensatory hyperinsulinemia, with increased risk for type 2 diabetes mellitus, designates this condition as a reproductive-metabolic disorder. That the symptoms of PCOS may be mimicked by other endocrine disorders of the ovary and adrenal glands warrants careful evaluation to exclude these associated conditions.

A functional bone morphogenetic protein system in the ovary.

Bone morphogenetic proteins (BMPs) comprise a large group of polypeptides in the transforming growth factor beta superfamily with essential physiological functions in morphogenesis and organogenesis in both vertebrates and invertebrates. At present, the role of BMPs in the reproductive system of any species is poorly understood. Here, we have established the existence of a functional BMP system in the ovary, replete with ligand, receptor, and novel cellular functions. In situ hybridization histochemistry identified strong mRNA labeling for BMP-4 and -7 in the theca cells and BMP receptor types IA, IB, and II in the granulosa cells and oocytes of most follicles in ovaries of normal cycling rats. To explore the paracrine function of this BMP system, we examined the effects of recombinant BMP-4 and -7 on FSH (follicle-stimulating hormone)-induced rat granulosa cytodifferentiation in serum-free medium. Both BMP-4 and -7 regulated FSH action in positive and negative ways. Specifically, physiological concentrations of the BMPs enhanced and attenuated the stimulatory action of FSH on estradiol and progesterone production, respectively. These effects were dose- and time-dependent. Furthermore, the BMPs increased granulosa cell sensitivity to FSH. Thus, BMPs have now been identified as molecules that differentially regulate FSH-dependent estradiol and progesterone production in a way that reflects steroidogenesis during the normal estrous cycle. As such, it can be hypothesized that BMPs might be the long-sought "luteinization inhibitor" in Graafian follicles during their growth and development.

How does warfarin affect the activated coagulation time?

BACKGROUND: The activated coagulation time (ACT) is a rapid measurement of a patient's level of heparin anticoagulation during cardiac catheterization. Patients receiving warfarin therapy occasionally are seen at the catheterization laboratory for emergent procedures. The effects of warfarin on ACT activity have not been previously described. We compared the ACT and the international normalized ratios (INR) in 77 patients receiving warfarin and 57 patients who were not receiving any anticoagulation (controls). RESULTS: Both the mean ACT (131+/-17.0 seconds) and INR (2.5+/-0.90 seconds) of the anticoagulated patients differed from the controls (ACT=115+/-14.5 seconds, INR=1.0+/-0.10 seconds; P< 0.05). The ACT increased linearly with INR in the warfarin group (r=0.70, P< .001). There was no relation between ACT and INR in the control group. CONCLUSION: Patients receiving warfarin therapy will have a linear increase in ACT develop similar to patients receiving heparin therapy.

Evidence for a neutrophil-interleukin-8 system in human folliculogenesis.

OBJECTIVE: This study was conducted to determine whether polymorphonuclear leukocytes (neutrophils) and the potent chemoattractant interleukin-8 are associated with follicle development in the normal human ovary. STUDY DESIGN: We performed a morphometric analysis of neutrophils in 268 human ovarian follicles, of which 199 were preantral and 69 were antral. In each antral follicle the numbers of mitotic, apoptotic, and total granulosa cells were counted to determine healthy and atretic follicles. Interleukin-8 protein and messenger ribonucleic acid were detected by immunohistochemistry and in situ hybridization, respectively. RESULTS: Antral follicles contained relatively large numbers of neutrophils within the theca vasculature. The density of neutrophil was twofold greater (p < 0.05) in atretic versus healthy follicles. The neutrophil index (neutrophils/granulosa cells x 1000) was inversely correlated to the number of granulosa cells per follicle. Immunoreactive interleukin-8 was detected in the theca and granulosa cells of most all antral follicles examined. Interleukin-8 messenger ribonucleic acid was demonstrated in theca and granulosa cells of some but not all follicles examined. CONCLUSIONS: Neutrophils are present in the theca of developing antral follicles, increase in number during atresia, and are associated with expression of interleukin-8 in the follicle wall.

Identification of a linked set of genes in Serpulina hyodysenteriae (B204) predicted to encode closely related 39-kilodalton extracytoplasmic proteins.

A tandem pair of nearly identical genes from Serpulina hyodysenteriae (B204) were cloned and sequenced. The full open reading frame of one gene and the partial open reading frame of the neighboring gene appear to encode secreted proteins which are homologous to, yet distinct from, the 39-kDa extracytoplasmic protein purified from the membrane fraction of S. hyodysenteriae. We have designated these newly identified genes vspA and vspB (for variable surface protein).

Ophthalmic disorders among the homeless and nonhomeless in Los Angeles.

BACKGROUND: Few studies have addressed ocular disorders among the homeless and nonhomeless poor. METHODS: To better understand the health problems of the homeless, ophthalmic disorders were analyzed among 512 homeless and 413 nonhomeless poor individuals receiving vision-screening examinations in Los Angeles. RESULTS: Significantly, more 4- to 17-year-old nonhomeless poor were diagnosed with uncorrected visual acuity worse than or equal to 20/50 without correction (p = 0.001), total refractive errors (p < 0.0005), astigmatism (p = 0.001), and myopia (p < 0.0005) than were a control group of 4- to 17-year-old homeless individuals. More homeless individuals had extraocular muscle imbalance (p < 0.040), but fewer had external eye diseases (p2 = 0.016) than the nonhomeless poor, when age adjusted. In addition, higher rates of glaucoma and cataracts were observed in both homeless and poor nonhomeless populations than in the general population. CONCLUSIONS: Health care professionals should provide vision screenings intended to detect these ocular disorders. Screening and correction of myopia and glaucoma, in particular, can greatly improve the quality of life for those treated.

Multisite electromyographic analysis of therapeutic touch and qigong therapy.

The influence of complementary healing treatment on paraspinal electromagnetic activity at specific neuromuscular sites was examined in an exploratory pilot study that used a multisite surface electromyographic (sEMG) assessment procedure. The study was a replication and extension of previous research that indicated that complementary healing had a significant effect in normalizing the activity of the "end organ" for the central nervous system (CNS). Multisite sEMG electrodes were placed on the frontalis, cervical (C4), thoracic (T6), and lumbosacral (L3) paraspinals of 44 subjects who were divided into three groups: (1) students/patients of a Qigong practitioner (n = 16); (2) students/patients of a therapeutic touch (TT) practitioner (n = 14); and (3) nonbelievers in complementary healing (n = 14). A traditional ABAC experimental design was used with each subject evaluated for one 20-minute session that included four 5-minute segments. The purpose of this study was to measure the variable energizing effect of Qigong therapy along with the anecdotally and experimentally established relaxation effect of TT therapy relative to patient belief and expectancy. Treatment sessions consisted of Qigong and a modified form of TT intervention for all three groups. Due to the double-blind nature of the study, however, group 1 subjects were aware of only the Qigong intervention; group 2 subjects were aware of only the TT intervention, and group 3 subjects were informed that the study was designed to assess the neuromuscular activity of individuals in a seated position. The results indicated a statistically significant rise in electromagnetic activity for group 1 during the Qigong intervention segment (p < .024). Group 2 demonstrated a modest although overall nonsignificant decrease in multisite sEMG levels for both treatment protocols, whereas group 3 exhibited relatively consistent neuromuscular activity for both control and treatment segments. The results of this study are considered preliminary in nature, however, due to the potential influence of several confounds including psychophysiological factors, established behavior patterns, and the possibility for information transfer due to sensory cues.

Pulsatile gonadotrophin secretion in women with polycystic ovary syndrome after gonadotrophin-releasing hormone agonist treatment.

In polycystic ovary syndrome (PCOS), increased luteinizing hormone (LH) pulse frequency has been attributed to either the hypothalamic gonadotrophin-releasing hormone (GnRH) pulse generator or ovarian oestrogen feedback. To address this issue, a detailed examination of pulsatile LH secretion was undertaken during recovery from GnRH agonist (GnRHa) suppression. Each of six women with PCOS and six normal ovulatory women received daily GnRHa treatment for 14 weeks. Frequent blood samples were collected and assayed for gonadotrophins, androgens and oestrogens before, during and up to 4 weeks after treatment. Women with PCOS had higher basal LH pulse frequency and amplitude and increased serum concentrations of LH, androstenedione, testosterone and oestrone than controls. After 3 months of GnRHa treatment, all these parameters were suppressed with no differences observed between the two groups. One week after cessation of GnRHa, LH pulse frequency promptly returned to pre-treatment range with no between-group differences noted, whereas LH pulse amplitude, serum gonadotrophins and ovarian steroids remained maximally suppressed and equivalent in the two groups. Subsequent LH pulse frequency remained constant while LH pulse amplitude and circulating concentrations gradually increased in parallel with a return of serum oestrogen to pre-treatment values. Despite comparable resumption of LH secretion in the two groups, corresponding androgen concentrations in women with PCOS were greater than those of normal ovulatory women. Thus, the immediate restoration of LH pulse frequency after discontinuing GnRHa treatment is largely independent of ovarian oestrogen production and reflects primacy of the GnRH pulse generator in determining basal LH pulse frequency. Equivalent LH pulse frequency rates in the two groups during the recovery period do not suggest an intrinsic hypothalamic-pituitary hyperactivity in PCOS.