Identification of a unique anti-Ro60 subset with restricted serological and molecular profiles.

Anti-Ro60 is one of the most common and clinically important serum autoantibodies that has a number of diagnostic and predictive capabilities. Most diagnostic laboratories report this simply as a qualitative positive/negative result. The objective of this study was to examine the clinical and serological relevance of a novel subset of anti-Ro60 in patients who display low levels of anti-Ro60 (anti-Ro60low ). We retrospectively identified anti-Ro60 sera during a 12-month period at a major immunopathology diagnostic laboratory in Australia. These all were anti-Ro60-precipitin-positive on the diagnostic gold standard counter-immuno-electrophoresis (CIEP). Lineblot immunoassay was used to stratify patients into either anti-Ro60low or anti-Ro60high subsets. We compared the medical and laboratory parameters associated with each group. Enzyme-linked immunosorbent assay (ELISA) and mass spectrometry techniques were used to analyse the serological and molecular basis behind the two subsets. Anti-Ro60low patients displayed less serological activity than anti-Ro60high patients with less intermolecular spreading, hypergammaglobulinaemia and less tendency to undergo anti-Ro60 isotype-switching than anti-Ro60high patients. Mass spectrometric typing of the anti-Ro60low subset showed restricted variable heavy chain subfamily usage and amino acid point mutations. This subset also displayed clinical relevance, being present in a number of patients with systemic autoimmune rheumatic diseases (SARD). We identify a novel anti-Ro60low patient subset that is distinct from anti-Ro60high patients serologically and molecularly. It is not clear whether they arise from common or separate origins; however, they probably have different developmental pathways to account for the stark difference in immunological maturity. We hence demonstrate significance to anti-Ro60low and justify accurate detection in the diagnostic laboratory.

Precipitating anti-dsDNA peptide repertoires in lupus.

Anti-double-stranded (ds)DNA autoantibodies are prototypical serological markers of systemic lupus erythematosus (SLE), but little is known about their immunoglobulin variable (IgV) region composition at the level of the secreted (serum) proteome. Here, we use a novel proteomic workflow based on de novo mass spectrometric sequencing of anti-dsDNA precipitins to analyse IgV subfamily expression and mutational signatures of high-affinity, precipitating anti-dsDNA responses. Serum anti-dsDNA proteomes were oligoclonal with shared (public) expression of immunoglobulin (Ig)G heavy chain variable region (IGHV) and kappa chain variable region (IGKV) subfamilies. IgV peptide maps from eight subjects showed extensive public and random (private) amino acid replacement mutations with prominent arginine substitutions across heavy (H)- and light (L)-chains. Shared sets of L-chain complementarity determining region 3 (CDR3) peptides specified by arginine substitutions were sequenced from the dominantly expressed IGKV3-20 subfamily, with changes in expression levels of a clonal L-chain CDR3 peptide by quantitative multiple reaction monitoring (MRM) paralleling the rise and fall of anti-dsDNA levels by Farr radioimmunoassays (RIA). The heavily mutated IgV peptide signatures of precipitating anti-dsDNA autoantibody proteomes reflect the strong selective forces that shape humoral anti-dsDNA responses in germinal centres. Direct sequencing of agarose gel precipitins using microlitre volumes of stored sera streamlines the antibody sequencing workflow and is generalizable to other precipitating serum antibodies.

Lupus anti-ribosomal P autoantibody proteomes express convergent biclonal signatures.

Lupus-specific anti-ribosomal P (anti-Rib-P) autoantibodies have been implicated in the pathogenesis of neurological complications in systemic lupus erythematosus (SLE). The aim of the present study was to determine variable (V)-region signatures of secreted autoantibody proteomes specific for the Rib-P heterocomplex and investigate the molecular basis of the reported cross-reactivity with Sm autoantigen. Anti-Rib-P immunoglobulins (IgGs) were purified from six anti-Rib-P-positive sera by elution from enzyme-linked immunosorbent assay (ELISA) plates coated with either native Rib-P proteins or an 11-amino acid peptide (11-C peptide) representing the conserved COOH-terminal P epitope. Rib-P- and 11-C peptide-specific IgGs were analysed for heavy (H) and light (L) chain clonality and V-region expression using an electrophoretic and de-novo and database-driven mass spectrometric sequencing workflow. Purified anti-Rib-P and anti-SmD IgGs were tested for cross-reactivity on ELISA and their proteome data sets analysed for shared clonotypes. Anti-Rib-P autoantibody proteomes were IgG1 kappa-restricted and comprised two public clonotypes defined by unique H/L chain pairings. The major clonotypic population was specific for the common COOH-terminal epitope, while the second shared the same pairing signature as a recently reported anti-SmD clonotype, accounting for two-way immunoassay cross-reactivity between these lupus autoantibodies. Sequence convergence of anti-Rib-P proteomes suggests common molecular pathways of autoantibody production and identifies stereotyped clonal populations that are thought to play a pathogenic role in neuropsychiatric lupus. Shared clonotypic structures for anti-Rib-P and anti-Sm responses suggest a common B cell clonal origin for subsets of these lupus-specific autoantibodies.

An immunodominant La/SSB autoantibody proteome derives from public clonotypes.

The La/SSB autoantigen is a major target of long-term humoral autoimmunity in primary Sjögren's Syndrome (SS) and systemic lupus erythematosus. A majority of patients with linked anti-Ro60/Ro52/La responses target an NH2-terminal epitope designated LaA that is expressed on Ro/La ribonucleoprotein complexes and the surface membrane of apoptotic cells. In this study, we used high-resolution Orbitrap mass spectrometry to determine the clonality, isotype and V-region sequences of LaA-specific autoantibodies in seven patients with primary SS. Anti-LaA immunoglobulin (Ig)Gs purified from polyclonal sera by epitope-specific affinity chromatography were analysed by combined database and de-novo mass spectrometric sequencing. Autoantibody responses comprised two heavily mutated IgG1 kappa-restricted monoclonal species that were shared (public) across unrelated patients; one clonotype was specified by an IGHV3-30 heavy chain paired with IGKV3-15 light chain and the second by an IGHV3-43/IGKV3-20 pairing. Shared amino acid replacement mutations were also seen within heavy and light chain complementarity-determining regions, consistent with a common breach of B cell tolerance followed by antigen-driven clonal selection. The discovery of public clonotypic autoantibodies directed against an immunodominant epitope on La, taken together with recent findings for the linked Ro52 and Ro60 autoantigens, supports a model of systemic autoimmunity in which humoral responses against protein-RNA complexes are mediated by public sets of autoreactive B cell clonotypes.

Hierarchical self-tolerance to T cell determinants within the ubiquitous nuclear self-antigen La (SS-B) permits induction of systemic autoimmunity in normal mice.

Systemic autoimmune diseases are frequently associated with clustering of high titer autoantibody responses towards nuclear self-antigens. Little is known, however, about the extent of immune tolerance to the target nuclear antigens or the events leading to the complex autoantibody responses that are characteristic of systemic autoimmunity. To address these issues, we have examined the mouse immune response to La autoantigen (mLa) and the homologous human La antigen (hLa), which are components of the La(SS-B)/Ro(SS-A) ribonucleoprotein (RNP) complex targeted in systemic lupus erythematosus and primary Sjögren's syndrome. The findings reveal the presence of hierarchical T cell tolerance involving multiple autodeterminants within the La autoantigen expressed by normal H-2k and H-2a mice. At one end of this spectrum, there was no detectable T or B cell autoimmunity observed in mice that were immunized with the immunodominant mLa287-301 determinant, which differed by a single residue in its core sequence from the homologous but highly immunogenic human La288-302 determinant. Interestingly, the mLa287-301 peptide acted as an altered peptide ligand that specifically antagonized the activation of an hLa288-302-specific T cell hybridoma. In contrast to the tolerogenic mLa287-301 determinant, a range of autoimmune potential was identified among poorly tolerizing, subdominant self-peptides present within mouse La autoantigen. Notably, immunization of normal mice with the autologous subdominant La25-44 and La106-129 determinants resulted in limited or no detectable autoantibody response. In contrast, immunization with the subdominant mouse La13-30 determinant induced a proliferative T cell response associated with the appearance of specific autoantibodies recognizing multiple intrastructural (La) and intermolecular components (Ro) of the murine La/Ro RNP. The findings suggest how diversified autoimmunity might follow initiation of immunity to simple peptide mimics of poorly tolerogenic determinants that are present within ubiquitous self-antigens.

Plasma testosterone levels in the male rhesus: influences of sexual and social stimuli.

Four adult male rhesus monkeys were provided access individually to a group of receptive females. Each male assumed dominance and engaged in frequent copulations. Plasma testosterone levels increased two- to threefold during this period. Next, each male was subjected to sudden and decisive defeat by a large all-male group, and plasma testosterone fell following this experience. Two males were later reintroduced to the females, and plasma testosterone rose rapidly to the previous elevated levels.

Primates.

Nonhuman primates demonstrate marked similarities to humans in almost all aspects of their anatomy, endocrinology, and physiology. These similarities underlie the value of these animals for appropriate studies in neurobiology, immunology, pathology, reproductive biology, teratology, neonatology, endocrinology, cardiology, and psychology. Investigations with nonhuman primates has made, and continues to make, significant contributions to biomedical and behavioral research. This review provides an overview of basic and applied studies for which primates are appropriate subjects and a summary of the advantages and problems of using nonhuman primates in research.

Prevalence of sicca symptoms in a South Australian cohort with systemic sclerosis.

BACKGROUND: The presence of sicca symptoms is a frequent finding in patients with systemic sclerosis (SSc). The aim of this study was to examine the prevalence of sicca symptoms in a South Australian cohort of SSc patients and correlate this to a number of parameters, including autoantibody status, use of anticholinergic medication, age and the presence of functional anti-muscarinic-3 receptor (M3R)-blocking antibodies. METHODS: A screening questionnaire was sent out to all patients on the South Australian Scleroderma Register from the years 1998-2006 to determine the prevalence of sicca symptoms. A subset of patients on the register had ocular sicca symptoms tested by use of Schirmer's strips to validate the accuracy of the questionnaire. Eight patients were tested for anti-M3R-blocking antibodies using a functional physiological assay. RESULTS: One hundred and ninety-three SSc patients took part in this study. Sicca symptoms were present in 59% of patients with the limited form of SSc, compared with 49% of patients with the diffuse form and 40% of patients with the overlap syndrome. The use of anticholinergic medication or thyroxine was associated with higher sicca scores in SSc patients. SS-A and SS-B autoantibodies (seen in Sjögren's syndrome) were detected in eight patients in this study. The detection of anti-M3R-blocking antibodies correlated well to presence of sicca. CONCLUSION: This study confirmed that sicca symptoms are found in a high proportion of patients with SSc, especially those with the limited variant. Further testing of larger numbers of SSc patients with sicca for anti-M3R-blocking antibodies will be needed before more definitive conclusions can be drawn. Physicians should be made aware that sicca symptoms are a frequent cause of morbidity for SSc patients*.

Prevalence of natural infection with simian immunodeficiency virus and simian T-cell leukemia virus type I in a breeding colony of sooty mangabey monkeys.

The seroprevalence of antibodies to simian immunodeficiency virus (SIVsmm) and simian T-cell leukemia virus type I (STLV-I) in a captive breeding colony of sooty mangabey monkeys was determined, and infection by SIVsmm was confirmed in all cases by virus isolation. Among 138 animals tested, 57 and 33% were infected with SIVsmm and STLV-I, respectively. While the proportion of female mangabeys (66%) differed significantly (P less than 0.01) from the proportion of male mangabeys (42%) infected with SIVsmm, the proportions of males and females infected with STLV-I were similar, suggesting independent transmission of the two viruses. Among mangabeys less than 1 year old, none were infected with STLV-I and only five of 27 mangabeys, all of which were at least 6 months old when first tested, were infected with SIVsmm. The data document that natural infection of sooty mangabey monkeys with SIVsmm or in association with STLV-I infection does not result in increased disease or mortality, and that transmission of both SIVsmm and STLV-I appears to occur primarily through sexual activity.

Ontogeny of luteinizing hormone secretion and first ovulation in seasonal breeding rhesus monkeys.

A longitudinal analysis assessed the role of nongonadal control and estradiol (E2) inhibition of LH secretion during puberty in female rhesus monkeys and examined how maturational changes in LH release were related to GH secretion, ponderal growth, and seasonal changes in the environment. Three groups of spring-born monkeys housed outdoors were studied from 12 months of age in May 1982 through 43 months of age in December 1984: gonadally intact (INT; n = 8), ovariectomized (OVX; n = 5), and OVX E2 treated (E2OVX; n = 5; E2, approximately 45 pg/ml). Suppression of bioactive levels of LH by E2 was functional once changes in nongonadal control of LH release occurred, as basal LH levels increased significantly earlier in OVX (23.7 +/- 2.1 months) than either E2OVX (29.6 +/- 0.4 months) or INT (29.6 +/- 1.0 months). Although these increases in LH were not seasonally limited, higher levels were achieved if these initial increases occurred during the fall. Menarche in INT females occurred after the initial increases in LH (30.9 +/- 0.1 months). First ovulation for INT females was seasonally restricted, occurring either at 31.5 +/- 0.4 months in December/January (n = 3) or 11 months later at 42.5 +/- 0.4 months in October/November (n = 5). No ovulations occurred during the intervening spring and summer period, as serum levels of LH were significantly lower than in the preceding perimenarchial fall months. After the initial maturational increases, LH levels declined significantly in OVX and E2OVX females in late winter at 33 months before rising again the next fall. The timing and extent of the maturational decrease in E2 inhibition of LH secretion were related to developmental increases in serum GH and not to body weight. Furthermore, females who had a normal first ovulation shortly after menarche at 31 months had significantly higher serum GH levels yet similar growth rates and body weights compared to their age-mates. First ovulation at 43 months in the remaining females occurred after sustained elevations in serum GH, similar to those observed in the early ovulating females. These data indicate that both nongonadal control and steroid inhibition of LH secretion operate during puberty in female rhesus monkeys and support the hypothesis that the final stages of puberty are characterized by a maturational decrease in E2 negative feedback of LH secretion.(ABSTRACT TRUNCATED AT 400 WORDS)

Endocrine control of the seasonal occurrence of ovulation in rhesus monkeys housed outdoors.

In female rhesus monkeys (n = 12) housed in a seminatural environment, serum gonadotropin and steroid levels fluctuated annually in a pattern indicative of a seasonally restricted period of ovulatory cycles in the fall and winter and anovulatory cycles in the spring and summer. This seasonal endocrine rhythm occurred independent of pregnancy and lactation, although the presence of a suckling infant delayed the onset of ovulation in the fall by 81 +/- 3.7 days (Dec. 4 vs. Sept 14). Except for serum PRL, levels of gonadotropin and ovarian hormones were similar in lactating and nonlactating females during the spring and summer anovulatory months. Serum levels of LH (less than 10 ng/ml), FSH (less than 4 micrograms/ml), and 17 beta-estradiol (E2; less than 30 pg/ml) were low throughout the summer anovulatory period, exhibiting a significant rise approximately 20 days before first ovulation. Serum progesterone levels were low throughout the 100 days before ovulation (less than 0.5 ng/ml) and did not rise until ovulation occurred. PRL levels remained elevated (greater than 20 ng/ml) in lactating females until 50 days before the first ovulation of the breeding season, but were low throughout the ovulatory, anovulatory, and ensuing ovulatory periods (less than 10 ng/ml). During the breeding period, females exhibited from two to six ovulations. Although the first ovulation of the breeding season occurred within a 40-day period for all females, a subset (n = 6) ceased ovulations significantly earlier than the remaining six females (Jan. 26 vs. March 3). The early cessation of ovulation for these females was associated with significantly lower body weight. After the last ovulation, FSH and E2 fell and remained low, at levels indistinguishable from those of the ensuing spring-summer anovulatory period. In contrast, in females who ceased ovulations later in the breeding season, the period following the luteal phase of the last ovulation was characterized by E2 and gonadotropin levels that were intermediate between those of the anovulatory months and normal follicular phase values. Serum progesterone levels were slightly but significantly elevated following the last ovulation for both groups of females. These data indicate that low basal levels of gonadotropin secretion during the seasonal anovulatory period may result from diminished GnRH secretion or from an alteration in pituitary sensitivity to GnRH stimulation. These data further suggest that the timing of ovulations and associated changes in the neuroendocrine system controlling gonadotropin secretion may be mediated by an environmental variable.

Effects of a natural versus artificial environment on the tempo of maturation in female rhesus monkeys.

To identify factors that regulate the tempo of growth and puberty, the present study examined how the environment influenced the timing of menarche and first ovulation in rhesus monkeys and how these events were related to differential rates of growth. Spring-born females were raised from 12 months of age under natural outdoor conditions (OH; n = 6) or indoors (IH; n = 9) under a controlled photoperiod (12h of light, 12h of darkness) and temperature (20-23 C). Ages at the initial increases in serum bioactive LH levels (27.7 +/- 0.7 vs. 31.4 +/- 1.0 months), menarche (26.0 +/- 0.7 vs. 32.5 +/- 0.9 months), and first ovulation (33.9 +/- 1.4 vs. 43.5 +/- 0.3 months) were significantly advanced in IH compared to OH females. First ovulation for the OH females occurred exclusively in October and November of the fourth year, whereas the distribution of first ovulation of IH females was biomodal, with seven of nine occurring in November or December at 31.8 +/- 0.5 months, and two of nine ovulating in September or October at 41.2 +/- 0.5 months. Serum levels of PRL varied seasonally in OH females throughout development, with peaks in July and nadirs in October. A similar rhythm was observed for IH females during the first 12 months of indoor housing, after which point the period decreased from 11.9 +/- 0.5 to 9.3 +/- 0.6 months. Overall increments in body weight did not differ between groups. An acceleration of growth in both crown-rump and tibial lengths occurred just before menarche in both groups, and this occurred at about 26 months for IH and about 32 months for OH females. Skeletal maturity was significantly advanced at 27 months in IH females and at every chronological age thereafter. Serum concentrations of somatomedin-C and GH paralleled group differences in bone maturation. Both hormones were significantly elevated by 16-18 months of age in IH animals compared to OH females and remained so until 34-36 months of age. Although a distinct seasonal rhythm in both GH and somatomedin-C was evident in OH animals, no such pattern was observed in IH females. These data suggest that exposure to an outdoor environment moderates the tempo of both sexual and skeletal maturation. The acceleration in reproductive development in animals exposed to a constant environment was associated with an acceleration in bone maturation, suggesting that common factors may be responsible for the initiation of both events.

Gonadal status influences developmental patterns of serum prolactin in female rhesus monkeys housed outdoors.

Although estradiol (E2) is thought to facilitate PRL secretion under certain conditions in female primates, the role of E2 or other ovarian products in the control of PRL secretion during puberty is unknown. The present study examined the influence of gonadal status on serum PRL levels in prepubertal monkeys housed in an outdoor environment. Basal levels of PRL were examined in three groups of spring-born females that were studied from 12 to 28 months of age (May 1982 to September 1983): gonadally intact (INT; n = 8), ovariectomized (OVX; n = 5), and ovariectomized and treated chronically with E2 (E2OVX; n = 5). All groups exhibited a significant annual rhythm in PRL levels with peaks (10-20 ng/ml) at 14 and 26 months (June to July) and a nadir (less than 2 ng/ml) at 19 months (November to December). Basal PRL levels were significantly higher from 12 to 15 months (May to August) in E2OVX subjects, with OVX having significantly greater concentrations than INT females. Group differences were not evident during the period of minimal secretion from 16 to 20 months (September to January). Finally, levels were again significantly higher in both E2OVX and OVX subjects during the subsequent period from 21 to 27 months (February to August). Although serum levels of E2 were lower in INT (30.9 +/- 2.3 pg/ml) than E2OVX females (51.4 +/- 4.0 pg/ml), group differences in PRL could not be attributed to E2 since OVX females, with no measurable levels of E2 (less than 15 pg/ml), had intermediate levels of PRL. These data suggest that during primate maturation serum PRL levels are dampened by some product of the ovary. Furthermore, whether age specific or environmentally mediated, this rhythm indicates an annual alteration in PRL release, with absolute levels enhanced by E2 replacement after ovariectomy, for prepubertal monkeys housed outdoors. In addition, a significant maturational increase in PRL levels was observed only in INT females when serum levels of PRL were compared for the ages 12-16 months to 24-28 months corresponding to the period between May and September for 2 successive years. These changes in PRL were not related to age-dependent changes in serum E2. Acute treatment of both E2OVX and OVX females with a single injection of E2-benzoate at three different ages did not induce any changes in serum PRL.(ABSTRACT TRUNCATED AT 400 WORDS)

Activity of the hypothalamic-pituitary-adrenal axis is altered by aging and exposure to social stress in female rhesus monkeys.

Age-related changes in glucocorticoid negative feedback inhibition of hypothalamic CRF and pituitary ACTH are observed in rodents. Attempts to study similar effects in humans have produced mixed results due in part to the difficulty in matching older subjects on social and lifestyle variables. The present study used female rhesus monkeys as a model for women by comparing young adult (n = 20) to old (n = 20) females to test the hypotheses that the hypothalamic-pituitary-adrenal axis is altered in older animals and that this difference is exacerbated by exposure to social stress. The effects of age on the response to two doses of dexamethasone and two doses of CRF were assessed in females living in a stable social environment (control) and in socially stressed females removed from their group and housed temporarily in a remote, nonsocial environment (separated). The suppression of serum cortisol was not different between the two doses of dexamethasone. Before dexamethasone administration (2100 h), serum cortisol was significantly higher in old control females than in either young or old separated females, who were not different from one another. The young control females had baseline cortisol concentrations significantly lower than all other groups. Serum cortisol was suppressed approximately 75% below baseline values in all groups by 10 h after dexamethasone administration. Age significantly affected serum cortisol after dexamethasone, as the old control group showed a release from suppression 19 h posttreatment compared to the young control group and compared to the separated groups. Social condition had a significant effect on the responses of serum cortisol and plasma ACTH to CRF administration. At baseline (0930 h), serum cortisol was significantly higher in young controls compared with older controls, with both separated groups having intermediate values. Similarly, plasma ACTH at baseline was significantly higher in young controls compared to all other groups. Social separation significantly diminished the elevation of both serum cortisol and ACTH after stimulation with either dose of CRF. Control females showed a prolonged increase in plasma ACTH through 60 min and an increase in serum cortisol through 120 min after CRF. In contrast, these hormones either declined by 60 min or did not increase in socially separated females after CRF administration. These data suggest that the circadian rhythm in serum cortisol may be affected by aging, as levels were higher in the evening and lower in the morning in old control compared to young control females. The effect of age on the response to dexamethasone treatment among the control groups lends support to the hypothesis that the sensitivity of glucocorticoid negative feedback diminishes with aging. Although age did not affect the response to CRF, social separation diminished the elevation in both serum cortisol and plasma ACTH. Whether this effect was due to stress-induced down-regulation of pituitary CRF receptors remains to be determined.

Effects of growth hormone on neonatal growth in nursing rhesus monkeys.

The effects of recombinant human GH treatment of either nursing mothers or their infants on neonatal growth in rhesus monkeys was determined. Growth rates of infants treated daily from birth with GH (INFGH; n = 9; 100 micrograms/kg, sc) were compared to those of infants given saline (INFc; n = 10), infants whose mothers received saline from the second trimester of pregnancy through 7 weeks postpartum (CON; n = 9), infants of mothers who received GH during pregnancy only from the second trimester to parturition (PRG; n = 8), infants of mothers who received GH during lactation only from parturition through 7 weeks postpartum (LAC; n = 9), and infants of mothers who received GH during the second trimester of pregnancy through 7 weeks postpartum (PRG/LAC; n = 8). Mothers receiving GH were given 250 micrograms/kg, sc, Monday, Wednesday, and Friday. Infants were allowed to nurse ad libitum. Although infant birth weights were similar among the six groups, body weights at 7 weeks of age were significantly greater in PRG/LAC infants (0.77 +/- 0.03 kg) compared to those in CON (0.66 +/- 0.02 kg), INFc (0.62 +/- 0.03 kg), LAC (0.62 +/- 0.04 kg), and INFGH infants (0.62 +/- 0.01 kg), with infants of PRG mothers intermediate (0.71 +/- 0.02 kg) between them. By 35 weeks of age, after infants had been weaned by their mothers, body weights were similar among all groups. Serum concentrations of insulin-like growth factor-I (IGF-I) rose significantly in all infants during the study period. Although IGF-I levels did not vary significantly among the treatment groups, average concentrations of IGF-I were significantly related to weight gains. Analyses of milk composition revealed that total protein, lactose, and IGF-I levels were similar among groups, whereas the percentage of fat in the milk was significantly higher in PRG/LAC mothers. Milk protein content was significantly related to weight gain. These data suggest that neonatal body weight gain can be accelerated in nursing infants whose mothers have received GH from at least the second trimester of pregnancy through the lactational interval. Since infants of mothers receiving GH during lactation only were not different from controls, the effect of GH in this treatment paradigm may be mammogenic rather than galactopoietic per se.

Effects of growth hormone on the tempo of sexual maturation in female rhesus monkeys.

The signal that initiates and maintains the developmental change in LHRH and, consequently, LH secretion in primates, thus regulating the tempo of puberty, is not known. Given the close association between reproductive development and bone maturation, we examined the hypothesis that GH was involved in developmental increases in LH release, specifically by augmenting the decrease in estradiol (E2) negative feedback inhibition of LH that characterizes late puberty in primates. Recombinant human GH (rhGH; 250 micrograms/kg) was given (sc) three times weekly to immature female rhesus monkeys to determine if developmental increases in basal serum LH would occur at an earlier age, and if menarche and first ovulation also would be advanced. The study groups included intact females receiving rhGH (INT + GH; n = 5), intact control animals (INT; n = 6), ovariectomized females receiving E2 plus rhGH (E2OVX + GH; n = 5), and E2-treated ovariectomized control monkeys (E2OVX; n = 4). The females were studied from 20 months of age until their serum LH levels increased (E2OVX groups) or until the occurrence of first ovulation (intact groups). After 12 months of rhGH treatment, the crown-rump lengths were significantly increased, regardless of ovarian status, an effect maintained in the intact females through 21 months of treatment. The mean age at the time of the initial rise in serum LH was advanced by rhGH treatment in intact females (29.6 +/- 0.4 vs. 31.3 +/- 0.3 months), but not E2OVX females. Subsequent maturational elevations in LH secretion were similar in the E2OVS + GH and E2OVX animals even after an incremental increase in E2. Ages at menarche were similar in the INT + GH and INT groups, whereas first ovulation was significantly advanced in three of five INT + GH females (31.5 +/- 0.7 months) compared to that in INT females (43.5 +/- 0.3 months). The remaining INT + GH females ovulated at an age (42.4 +/- 0.4 months) similar to that of INT females. Those females that ovulated by 32 months had higher skeletal maturity scores than the later ovulating INT females, with the other INT + GH females being intermediate. Furthermore, rhGH resulted in a significant increase in serum E2 levels within 12 h of injection, which remained elevated through 24 h. This effect of rhGH on ovarian E2 secretion did not occur until females had shown elevations in basal serum LH.(ABSTRACT TRUNCATED AT 400 WORDS)

An enhanced chemiluminescence detection system combined with a modified immunoblot technique for the detection of low molecular weight IgM in sera from healthy adults and neonates.

An enhanced chemiluminescence detection system combined with a modified immunoblot technique is described for the detection of low molecular weight IgM (LMW IgM) in human sera and cell culture supernatants at levels down to 2 pg/ml. This detection system is reliable, specific and more sensitive than the previously described chromogenic detection system. Importantly, this method has, for the first time, revealed LMW IgM in the sera from all 34 healthy adults and 26 neonatal cord bloods tested. A significant linear correlation was observed between the LMW IgM and the total circulating IgM in both the healthy adult sera (r = 0.88, p less than 0.001) and cord blood sera (r = 0.98, p less than 0.001).

Nocturnal changes in serum melatonin during female puberty in rhesus monkeys: a longitudinal study.

Diurnal concentrations of serum melatonin were determined longitudinally in female rhesus monkeys throughout sexual maturation to ascertain how levels varied with advancing age and reproductive onset. Females were housed either in outdoor enclosures (n = 8) exposed to ambient environmental conditions, or indoors (n = 4) under a photoperiod of 12 h light: 12 h darkness and fixed temperature of 20-23 degrees C. Animals were studied from immaturity (15 months) through first ovulation and were additionally compared with fully adult female rhesus monkeys (n = 5) studied during the annual breeding season. The diurnal melatonin pattern was described for the developing females in the summer, autumn and winter in 3 successive years from samples collected at 10.00, 18.00, 22.00, 02.00, 06.00 and 10.00 h. Nocturnal levels of melatonin declined significantly during development in both indoor- and outdoor-housed females with a progressive decrease up to 33 months of age. Daytime values were consistently low but exhibited a slight decline also with age. Nocturnal values in all months sampled fell significantly with greater decreases occurring at the earliest ages. Furthermore, superimposed upon this developmental change, animals housed outdoors responded to seasonal changes in photoperiod with diurnal increases in melatonin occurring after sunset. The females in the present study exhibited first ovulation at two distinct ages: 32-37 months ('early', n = 6) and 41-45 months ('later', n = 5). One female did not ovulate within the study period. Although nocturnal levels of serum melatonin were similar between the two groups up to 29 months of age, a post-hoc analysis revealed that concentrations were significantly lower by 34 months of age for the early group, a time coincident with first ovulation. Nocturnal levels of melatonin remained high, relative to the early group, in the later ovulating females until 43 months of age, coincident with first ovulation for these animals. The diurnal pattern of serum melatonin at first ovulation, regardless of chronological age, was similar to that observed during the ovulatory season for adult female rhesus monkeys. These data suggest that nocturnal melatonin concentrations decline with advancing chronological age in prepubertal female rhesus monkeys.(ABSTRACT TRUNCATED AT 400 WORDS)