Antide-induced suppression of pituitary gonadotropin and ovarian steroid secretion in cynomolgus monkeys: premature luteolysis and prolonged inhibition of folliculogenesis following single treatment

Administration of high-dose Antide to ovariectomized monkeys results in rapid, prolonged, and reversible inhibition of gonadotropin secretion. The present study examined whether similar long-term control would be manifested in the menstrual cycle of intact primates. Antide administration at a dose of either 3.0 or 18.0 mg/kg induced rapid suppression of bioassayable LH concentrations, precipitating a concurrent fall in serum progesterone concentrations from 7.9 +/- 3.6 and 5.8 +/- 1.0 ng/ml (mean +/- SEM) on the day of injection to 0.6 +/- 0.2 and 0.5 +/- 0.1 ng/ml by 2 days post-treatment, respectively. This Antide-induced luteolysis was accompanied by the premature onset of menses within 3 days. The next menses following Antide administration was delayed. Ultimately, folliculogenesis culminating in normal follicular-phase estradiol production, ovulation, and subsequent normal luteal-phase progesterone production did occur in all treated monkeys. Menses resumed 54 +/- 9 and 75 +/- 13 days after treatment with 3.0 and 18.0 mg/kg Antide, respectively. No allergic cutaneous or peripheral reactions were seen, even at the highest dose of Antide. Thus, the long duration of action of high-dose Antide reported earlier in ovariectomized monkeys is also demonstrated in intact primates. These findings, along with the apparent absence of histamine-release effects even at high doses, suggest that Antide is a GnRH antagonist deserving clinical evaluation.     

Arachidonic acid metabolism and prostaglandin production by primate preimplantation blastocysts.

Preimplantation embryos of many species are known to synthesize prostaglandins. These tissue hormones are believed to influence embryonic metabolism, as well as embryo-maternal interaction during implantation although their putative role(s) remains obscure. Here, prostaglandin production by blastocysts from cynomolgus monkeys (Macaca fascicularis) was examined qualitatively during in vitro culture. Tritium labelled arachidonic acid was metabolized to 6 keto-prostaglandin F1 alpha, 2,3-dinor-prostaglandin F1 alpha and thromboxane B2, as characterized by HPLC separation. Also, 6-keto-prostaglandin F1 alpha, and thromboxane B2 as characterized by HPLC separation. Also, 6-keto-prostaglandin F1 alpha and thromboxane B2 were identified by specific RIA's. Our data suggest that the main arachidonic acid metabolites produced by blastocysts of cynomolgus monkeys are prostacyclin and thromboxane.     

Pregnancy diagnosis in squirrel monkeys: hemagglutination test, radioimmunoassay, and bioassay of chorionic gonadotropin.

The Nonhuman Primate Pregnancy Test may be useful for diagnosis of pregnancy in squirrel monkeys between 40 and 60 days in pregnancy. However, single determinations have an inherent 10% risk of false negative responses caused by low chorionic gonadotropin levels; thus, initial negative test responses should be followed within one week by an independent confirmatory test. Preliminary results with this hemagglutination inhibition test compare favorably with bioassay and may be useful, in conjunction with conventional uterine palpation, for diagnosis of pregnancy in squirrel monkeys.     

Pregnancy diagnosis in owl monkeys (Aotus trivirgatus): evaluation of the hemagglutination inhibition test for urinary chorionic gonadotropin.

The Subhuman Primate Pregnancy Test was evaluated as a means of detecting urinary chorionic gonadotropin to aid in pregnancy diagnosis in owl monkeys. Using radioimmunoassay, the excretion pattern of chorionic gonadotropin from pregnant owl monekys was delineated, the hormone being detected from 16 weeks prepartum until birth. By comparison, the pregnancy test kit detected chorionic gonadotropin between the fourteenth week prepartum and the last week of gestation with 94% accuracy. In a 2-year study using a simplified urine collection technique, the Subhuman Primate Pregnancy Test was shown to be a valuable procedure for diagnosing pregnancy and detecting spontaneous abortions in owl monkeys.     

Antigenic similarities to HCG subunits among chorionic gonadotropins of nonhuman primates.

Comparison of antigenic similarity between human chorionic gonadotropin (HCG) subunits and the chorionic gonadotropins of six species of nonhuman primates indicates marked similarity of antigenic determinants between both subunits of HCG and the chorionic gonadotropins of chimpanzees, gorillas, and orangutans. Antisera to HCG subunits (alpha or beta) did not cross-react with the chorionic gonadotropins of baboons, macaques, or marmosets. Because of the relative availability of chimpanzees for laboratory studies, we suggest that chimpanzees may be the optimal nonhuman primate model for determining the advisability of vaccinations in man using conjugates of HCG fragments to achieve fertility control or for suppression of HCG-producing neoplasms.     

Association of 25-Hydroxyvitamin D status and genetic variation in the vitamin D metabolic pathway with FEV1 in the Framingham Heart Study

Background

Vitamin D is associated with lung function in cross-sectional studies, and vitamin D inadequacy is hypothesized to play a role in the pathogenesis of chronic obstructive pulmonary disease. Further data are needed to clarify the relation between vitamin D status, genetic variation in vitamin D metabolic genes, and cross-sectional and longitudinal changes in lung function in healthy adults.

Methods

We estimated the association between serum 25-hydroxyvitamin D [25(OH)D] and cross-sectional forced expiratory volume in the first second (FEV₁) in Framingham Heart Study (FHS) Offspring and Third Generation participants and the association between serum 25(OH)D and longitudinal change in FEV₁ in Third Generation participants using linear mixed-effects models. Using a gene-based approach, we investigated the association between 241 SNPs in 6 select vitamin D metabolic genes in relation to longitudinal change in FEV₁ in Offspring participants and pursued replication of these findings in a meta-analyzed set of 4 independent cohorts.

Results

We found a positive cross-sectional association between 25(OH)D and FEV₁ in FHS Offspring and Third Generation participants (P = 0.004). There was little or no association between 25(OH)D and longitudinal change in FEV₁ in Third Generation participants (P = 0.97). In Offspring participants, the CYP2R1 gene, hypothesized to influence usual serum 25(OH)D status, was associated with longitudinal change in FEV₁ (gene-based P < 0.05). The most significantly associated SNP from CYP2R1 had a consistent direction of association with FEV₁ in the meta-analyzed set of replication cohorts, but the association did not reach statistical significance thresholds (P = 0.09).

Conclusions

Serum 25(OH)D status was associated with cross-sectional FEV₁, but not longitudinal change in FEV₁. The inconsistent associations may be driven by differences in the groups studied. CYP2R1 demonstrated a gene-based association with longitudinal change in FEV₁ and is a promising candidate gene for further studies.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-015-0238-y) contains supplementary material, which is available to authorized users.     

Neutrophils exhibit distinct phenotypes toward chitosans with different degrees of deacetylation: implications for cartilage repair

Introduction  

Osteoarthritis is characterized by the progressive destruction of cartilage in the articular joints. Novel therapies that promote resurfacing of exposed bone in focal areas are of interest in osteoarthritis because they may delay the progression of this disabling disease in patients who develop focal lesions. Recently, the addition of 80% deacetylated chitosan to cartilage microfractures was shown to promote the regeneration of hyaline cartilage. The molecular mechanisms by which chitosan promotes cartilage regeneration remain unknown. Because neutrophils are transiently recruited to the microfracture site, the effect of 80% deacetylated chitosan on the function of neutrophils was investigated. Most studies on neutrophils use preparations of chitosan with an uncertain degree of deacetylation. For therapeutic purposes, it is of interest to determine whether the degree of deacetylation influences the response of neutrophils to chitosan. The effect of 95% deacetylated chitosan on the function of neutrophils was therefore also investigated and compared with that of 80% deacetylated chitosan.     

Purification and characterization of Dolichos lablab lectin

The mannose/glucose-binding Dolichos lablab lectin (designated DLL) has been purified from seeds of Dolichos lablab (hyacinth bean) to electrophoretic homogeneity by affinity chromatography on an ovalbumin- Sepharose 4B column. The purified lectin gave a single symmetric protein peak with an apparent molecular mass of 67 kDa on gel filtration chromatography, and five bands ranging from 10 kDa to 22 kDa upon SDS-PAGE. N-Terminal sequence analysis of these bands revealed subunit heterogeneity due to posttranslational proteolytic truncation at different sites mostly at the carboxyl terminus. The carbohydrate binding properties of the purified lectin were investigated by three different approaches: hemagglutination inhibition assay, quantitative precipitation inhibition assay, and ELISA. On the basis of these studies, it is concluded that the Dolichos lablab lectin has neither an extended carbohydrate combining site, nor a hydrophobic binding site adjacent to it. The carbohydrate combining site of DLL appears to most effectively accommodate a nonreducing terminal alpha-d-mannosyl unit, and to be complementary to the C-3, C-4, and C-6 equatorial hydroxyl groups of alpha-d-mannopyranosyl and alpha-d-glucopyranosyl residues. DLL strongly precipitates murine IgM but not IgG, and the recent finding that this lectin interacts specifically with NIH 3T3 fibroblasts transfected with the Flt3 tyrosine kinase receptor and preserves human cord blood stem cells and progenitors in a quiescent state for prolonged periods in culture, make this lectin a valuable tool in biomedical research.      

The structure, catalytic mechanism and regulation of adenylyl cyclase

The recent structure determinations of the mammalian effector enzyme adenylyl cyclase reveal the structure of its catalytic core, provide new insights into its catalytic mechanism and suggest how diverse signaling molecules regulate its activity.