RESUMO
By means of two assay systems, a beta chain human chorionic gonadotropin radioimmunoassay and a radioreceptor gonadotropin assay, a chorionic gonadotropin-like substance was demonstrated in extracts of liver and colon obtained at autopsy from three patients who died of nonneoplastic disease. In contrast to placental chorionic gonadotropin, colon and liver chorionic gonadotropin was not bound to concanavalin A-Sepharose columns, indicating that this substance possessed little or no carbohydrate. Previous workers demonstrated that desialylated human chorionic gonadotropin possesses little or no bioactivity in vivo but retains full radioreceptor and radioimmunoassay activity in vitro. Our data suggest that the genome responsible for the human chorionic gonadotropin production is not completely suppressed in adult nonendocrine tissues, and that the chorionic gonadotropin produced by colon and liver has little or no bioactivity in vivo because of its low carbohydrate content. Since many normal tissues produce chorionic gonadotropin, bioactivity may be modulated by regulation of carbohydrate content.
Assuntos
Gonadotropina Coriônica/análise , Colo/análise , Fígado/análise , Gonadotropina Coriônica/metabolismo , Cromatografia de Afinidade , Humanos , Masculino , Radioimunoensaio , Receptores de Superfície Celular/metabolismo , Ácidos Siálicos/análise , Relação Estrutura-AtividadeRESUMO
Daily determinations of luteinizing hormone activity in plasma throughout a menstrual cycle in ten young women showed a sharp peak of activity lasting less than 48 hours around midcycle and higher mean values during the follicular phase than during the luteal phase in nine instances.
Assuntos
Hormônio Luteinizante/sangue , Menstruação , Adulto , Feminino , Humanos , ImunoensaioRESUMO
Measurements of serum thyrotropin (TSH) concentrations were conducted in maternal and fetal blood during labor and delivery and the early postnatal and neonatal periods. Mean TSH concentration was significantly higher in cord blood (9.5 muU/ml) than maternal blood (3.9 muU/ml), a finding suggesting a fetal-maternal TSH gradient at term. Serum TSH concentration in the newborn increased rapidly to mean levels of 60 muU/ml at 10 min and 86 muU/ml at 30 min of age. Between 30 min and 3-4 hr serum levels decreased rapidly, then fell more gradually to a mean concentration of 13 muU/ml at 48 hr. The half-time of the decrease in serum TSH concentration between 30 and 90 min was 77 min, a value slightly greater than the half-time of disappearance of radioiodinated TSH measured in adults. This indicates that the early high rate of TSH secretion in the newborn ceases by 30 min, and that the rapid rise and fall in serum TSH concentrations may represent release of stored pituitary TSH. A more chronic TSH hypersecretion persisted throughout the first 24-48 hr of extrauterine life. Measurements of serum PBI concentrations were conducted in a separate group of maternal and cord blood samples and in the newborn infants during the first 48 hr of extrauterine life to relate the TSH and serum hormonal iodine concentration changes. Serum protein-bound iodine (PBI) concentrations were similar in maternal and cord blood, increased significantly by 4 hr of age in the newborn, and peaked at about 24 hr, presumably in response to the TSH hypersecretion. The pattern of TSH hypersecretion was similar in infants delivered vaginally and by caesarean section. Maternal serum TSH concentrations were stable throughout the perinatal period. Warming the infants at 99-103 degrees F during the first 3 hr of life did not prevent the early, acute release of thyrotropin. Cooling of warm infants at room temperature (72-78 degrees F) between 3 and 4 hr resulted in a decrease in mean rectal temperature of 3.3 degrees F and produced a significant increment in serum TSH concentration. These data suggest that the mechanism of the early, acute release of thyrotropin in the newborn may involve a potent stimulus other than cooling. However, the increase in serum TSH stimulated by delayed (3-4 hr) cooling indicates that neonatal hyperthyroxinemia is, at least, augmented by extrauterine cooling. Thus, cold exposure is capable of increasing TSH secretion in humans.
Assuntos
Recém-Nascido , Tireotropina/sangue , Proteínas Sanguíneas , Temperatura Corporal , Parto Obstétrico , Feminino , Humanos , Iodo/sangue , Hipófise/metabolismo , Gravidez , Ligação Proteica , Radioimunoensaio , Tireotropina/metabolismo , Tiroxina/sangue , Cordão Umbilical/irrigação sanguíneaRESUMO
Metabolic clearance rates and production rates of human luteinizing hormone (HLH) were determined in pre- and postmenopausal women by the constant infusion technique. Highly purified HLH-(131)I was infused into the fasting subjects at a constant rate. Serial plasma samples were obtained and the radioactive hormone was precipitated by a double antibody technique. Plasma HLH-(131)I levels reached equilibrium by 4 hr after the infusion started. Metabolic clearance rates were: 24.4 +/- 1.8 (mean +/- SE) ml/min in five normal premenopausal women; 23.3 +/- 1.1 ml/min in five normal women taking norethinodrel and mestranol; and 25.6 +/- 4.1 ml/min in four postmenopausal women. Endogenous plasma HLH levels measured in the same subjects by radioimmunoassay immediately before infusion were 32.0 +/- 9.6 mU/ml in the normal women, 16.8 +/- 3.2 mU/ml in the women on oral contraceptives, and 99.2 +/- 23.2 mU/ml in the postmenopausal women. The corresponding HLH production rates were: 734 +/- 170 mU/min in the normal women: 387 +/- 86 mU/min in the women on norethinodrel and mestranol; and 2400 +/- 410 mU/min in the postmenopausal women. The metabolic clearance rate did not change after ovariectomy in one women, but the production rate rose from 583 to 1420 mU/min. Based on previously reported bioassay values for pituitary content and urinary excretion of HLH, the estimated turnover of HLH in the pituitary is about once per day and less than 5% of the total HLH produced appears in the urine in a biologically active form.
RESUMO
It is not practical to quantitate gonadotropin in the blood of normal men and women by utilizing bioassays. We have developed a method for sensitive, precise, and specific radioimmunoassay of luteinizing hormone (LH) in human serum or plasma. Antisera were developed against human chorionic gonadotropin, and one of these was selected for extensive cross-reaction with human LH. Highly purified LH was radioiodinated by the method of Greenwood, Hunter, and Glover. Separation of antibody-bound from free LH-(131)I was accomplished by a double antibody technique. Dose-response curves for the purifed LH, an impure urinary LH preparation, pituitary powder, and LH in plasma were all identical. Immunoassay and bioassay of impure urinary and pituitary gonadotropin preparations in terms of a common standard resulted in an index of discrimination of close to unity. LH levels in plasma from 32 adult men and 30 women outside the midcycle ranged from 0.6 to 3.2 mmug per ml (1 mmug of our laboratory LH standard is equivalent to 8 mU of the Second International Reference Preparation of Human Menopausal Gonadotropin). Levels were remarkably constant in men from day to day and in women except at midcycle, when a sharp peak occurred lasting less than 24 hours. In all women studied who had a midcycle LH peak, mean plasma LH levels during the follicular phase of the menstrual cycle were higher than mean values obtained during the luteal phase. Prepubertal children had detectable plasma LH, and mean values were only slightly less than in adults. Plasma from castrate men or women or postmenopausal women contained 4.5 to 10.5 mmug per ml. Clomiphene treatment of four men resulted in a doubling of plasma LH in 5 days.
Assuntos
Hormônio Luteinizante/sangue , Bioensaio , Gonadotropina Coriônica/sangue , Clomifeno/uso terapêutico , Feminino , Gonadotropinas Hipofisárias/sangue , Humanos , Soros Imunes , Imunoensaio , Isótopos de Iodo , Hormônio Luteinizante/urina , Masculino , Ovulação , Estilbenos/uso terapêuticoRESUMO
The plasma concentration of a pituitary hormone is determined by the rate of secretion, degradation, and the volume of distribution of that hormone. Using a radioimmunoassay for human thyrotropin (TSH) and human TSH-(131)I, we have estimated the rates of degradation and distribution of TSH in man and calculated the rate of secretion. Either 0.5 or 5 mug of TSH-(131)I with specific activities of 1 to 50 muc per mug was administered intravenously to 12 euthyroid subjects. Serial determinations were made of TSH-(131)I, and the half-time of disappearance (t((1/2))) was thus estimated. The average t((1/2)) in euthyroid subjects was 53.9 minutes with a volume of distribution averaging 5.8% of body weight. The mean endogenous plasma TSH concentration was 1.8 mmug per ml (2.7 muU per ml in terms of the human TSH reference standard A). The mean total TSH pool, excluding the pituitary, was 5.8 mug (8.7 mU). From these data the mean secretion rate of TSH in euthyroid man was calculated to be 110.1 mug per day (165.2 mU). Similar data were estimated for 3 mildly hypothyroid patients. The t((1/2)) were 75.1, 97.1, and 83.6 minutes, with a mean of 85.3 minutes (1.6 times normal). The mean TSH pool was 58.1 mug (10 times normal). The secretion rate was 688.7 mug per day (1,033.1 mU). In other hypothyroid patients, plasma TSH levels ranging from 6 to 230 mmug per ml (9 to 345 muU) have been found. If similar half-times and a normal distribution volume are assumed, the secretion rate of TSH in hypothyroid patients can be estimated to range from about 260 to 15,350 mug per day (390 to 23,025 mU) or from about 2 to 307 times normal. Therefore, the elevated plasma TSH levels found in hypothyroidism are a result of both slower degradation and increase in rate of secretion.
Assuntos
Hipotireoidismo/diagnóstico , Hipófise/fisiologia , Tireotropina/fisiologia , Adolescente , Adulto , Idoso , Feminino , Humanos , Imunoensaio , Masculino , Pessoa de Meia-Idade , Taxa Secretória , Tireotropina/sangueRESUMO
Most of the information concerning secretion changes in follicle-stimulating hormone (FSH) in humans has been gained with relatively insensitive bioassays of concentrates of pools of urine. We have developed a sensitive and specific radioimmunoassay for FSH that is 500-1000 times more sensitive than the rat ovarianweight augmentation assay and which is capable of quantifying FSH in small volumes of serum. Anti-FSH was prepared by immunizing rabbits with an impure FSH preparation. The majority of antisera showed complete inability to distinguish LH, TSH, and FSH, illustrating the immunological similarities of these hormones. One antiserum was specific when used in a radioimmunoassay. Potency estimates by bioassay were in good agreement, with a single exception, with those obtained with the radioimmunoassay for 10 FSH-containing preparations. Highly purified LH gave a higher potency by immunoassay than by bioassay. Sera from eugonadal men contained 5-25 mIU/ml; sera from castrate men contained over 30 mIU/ml. Sera from eugonadal women contained 7-25 mIU/ml during the follicular phase and 5-15 mIU/ml during the luteal phase of the menstrual cycle. Sera from castrate or postmenopausal women contained 40-250 mIU/ml. FSH was measured throughout the menstrual cycle in 19 women. The general pattern that emerged is summarized as follows: there is a small early follicular phase rise in FSH, and then FSH is relatively constant until mid-cycle; in the majority of women a mid-cycle rise of FSH occurs coincidentally to the mid-cycle LH ovulatory peak; during the luteal phase FSH levels are relatively constant and lower than during the follicular phase. Nonsequential oral contraceptives containing estrogen and progestogen abolish these changes and FSH concentrations remain low throughout treatment. Treatment of castrate men and castrate or postmenopausal women with high doses of oral estrogens results in a fall of FSH to levels found in eugonadal men or women, but not to undetectable levels. Children less than 5 yr of age had undetectable FSH (< 5 mIU/ml).
Assuntos
Hormônio Foliculoestimulante/sangue , Adulto , Idoso , Animais , Bioensaio , Castração , Criança , Gonadotropina Coriônica/sangue , Dietilestilbestrol/uso terapêutico , Cães , Estrogênios/uso terapêutico , Feminino , Hormônio Foliculoestimulante/fisiologia , Gonadotropinas/sangue , Humanos , Soros Imunes , Isótopos de Iodo , Hormônio Luteinizante/sangue , Masculino , Menopausa , Menstruação , Métodos , Pessoa de Meia-Idade , Coelhos , Radioimunoensaio , Tireotropina/sangueRESUMO
The development of a species specific radioimmunoassay for rabbit luteinizing hormone (LH) has permitted the direct demonstration of LH feedback control of LH secretion (short-loop feedback control). In previous studies we showed that small bolus injections of human LH (hLH) intravenously administered to castrate female rabbits suppressed rabbit LH for 20-30 min. Human LH had no effect on rabbit follicle-stimulating hormone secretion. This control system was responsive to amounts of hLH estimated to be present in blood of eugonadal men and women. These studies were designed to determine whether this feedback control was exerted at a pituitary or hypothalamic level. Two groups of studies were carried out: (a) in vivo studies: Rabbit LH was quantified in the blood of castrated female New Zealand White rabbits receiving either constant hLH perfusion (2.75 IU/min) or saline perfusion, plus a bolus injection of 0.5, 6, or 20 mug of gonadotropin-releasing hormone (GnRH). Human LH decreased the response to 6 and 20 mug of GnRH by 31 and 36%, respectively, and abolished the response to 0.5 mug, GnRH. (b) in vitro studies: Rabbit pituitary slices were incubated in the presence of medium alone, medium plus hLH (25 mIU/ml), medium plus GnRH (20 mug/ml), and medium plus both GnRH and hLH. hLH decreased basal rabbit LH release into the medium and abolished GnRH-stimulated rabbit LH release. hLH had no effect on rabbit follicle-stimulating hormone release. From these results we conclude that a direct and specific feedback control of LH on LH exists at a pituitary level.
Assuntos
Hormônio Luteinizante/fisiologia , Hipófise/fisiologia , Animais , Castração , Retroalimentação , Feminino , Hormônio Luteinizante/sangue , Hormônio Luteinizante/metabolismo , Hipófise/metabolismo , CoelhosRESUMO
A chorionic gonadotropinlike material is present in blood and urine of normal, nonpregnant humans, is secreted in pulsatile fashion in parallel with luteinizing hormone, is stimulated by gonadotropin-releasing hormone, and is suppressed by GnRH agonists and estrogen. The source is probably the pituitary gland.
RESUMO
In separate studies we have shown that Pseudomonas maltophilia (American Type Culture Collection 13637) possesses an immunoglobulin Fc-binding protein. We have found that this protein prevents the application of immunoassays using monoclonal antibodies to study possible production of a CG-like material by this bacteria. Employing an immunoglobulin saturation technique to block this protein as well as a zwitterion detergent membrane solubilization technique, we now report the isolation of a membrane protein from Pseudomonas maltophilia which shows immunological relationships to the beta-subunit and carboxyl tail of human pregnancy CG. This pseudomonas immunoreactive material produced dose-response curves in the following CG immunoassays: 1) a polyclonal rabbit anti-CG equilibrium assay, 2) carboxyl tail CG equilibrium assay, and 3) two CG equilibrium assays using monoclonal antibodies. The pseudomonas CG-like protein did not react in equilibrium assays for human TSH, human LH, or free alpha-subunit of CG. The pseudomonas CG-like protein was purified by affinity chromatography. The purified protein showed only 0.25% cross-reaction with pregnancy CG in the monoclonal antibody equilibrium assays. Furthermore, the purified protein showed no binding to rat testicular CG/LH receptors, but showed avid binding to the pseudomonas CG-binding protein previously described by Richert and Ryan. The pseudomonas protein showed no binding to Concanavalin-A, which avidly binds pregnancy CG. When assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, this protein had a mol wt of 48,500 daltons, which is larger than the mol wt of the unglycosylated beta-subunit of pregnancy CG. We conclude that pseudomonas contain a protein that has partial homology to the beta-subunit of pregnancy CG. This material does not bind to mammalian CG receptors, but does bind to a pseudomonas CG-binding site. The latter suggests it has a function, as yet unknown, in pseudomonas.
Assuntos
Proteínas de Bactérias/isolamento & purificação , Gonadotropina Coriônica/imunologia , Proteínas de Membrana/isolamento & purificação , Pseudomonas/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/imunologia , Western Blotting , Cromatografia , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Endopeptidases/metabolismo , Humanos , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Peso Molecular , SolubilidadeRESUMO
ACTH activity in glacial acetic acid extracts of normal rat tissues was studied by both ACTH RIA and a sensitive in vitro bioassay. ACTH immunoactivity was found in all tissues: brain, 278 +/- 54 (mean +/- SE; picograms per mg protein); stomach, 59 +/- 4; kidney, 47 +/- 3; colon, 40 +/- 4; small intestine, 37 +/- 4; liver, 18 +/- 2; and heart, 16 +/- 3. Tissue ACTH showed parallelism with ACTH standard in the RIA. No correlation existed between tissue ACTH and plasma ACTH in normal rats. Dexamethasone treatment (0.4 mg/day for 5 days) suppressed plasma ACTH, but did not affect tissue ACTH levels. When tissue extracts were passed through Sephadex G-75-SF columns, ACTH immunoactivity was exclusively eluted in the portion of bigger molecular weight than ACTH standard, except in the brain. Based on this column chromatography, the molecular weight of the main peak of activity was estimated as 26,000. Tissue ACTH-like material contained no detectable biological activity (less than 2 pg/100 ng tissue). However, biological activity was easily detectable after exposure of the tissue extracts to trypsin. When studied by immunoassay and bioassay, this 26,000 mol wt ACTH was digested and cleaved to 4,500 mol wt and biologically active ACTH with trypsin treatment. Tissue ACTH immunoactivity does not seem to be the result of artifacts: 1) extracts were adjusted to pH 8.0 and a common osmolality (150 mosmol/liter) before assay; 2) protein contents in RIA tubes were only 0.1-1.6 mg; 3) tissue extracts incubated with [125I]iodo-ACTH for 48 h produced less than 5% damage of labeled hormone, as assessed by moderate excess of antibody binding; 4) enzyme inhibitors did not modify tissue ACTH levels; and 5) ACTH immunoactivity of tissue extracts was absorbed by anti-ACTH immunocolumns. We conclude that high molecular weight ACTH-like materials are widespread in normal rat extrapituitary tissues and are probably a precursor form of 4500 mol wt ACTH.
Assuntos
Hormônio Adrenocorticotrópico/metabolismo , Hormônio Adrenocorticotrópico/sangue , Animais , Bioensaio , Dexametasona/farmacologia , Peso Molecular , Radioimunoensaio , Ratos , Ratos Endogâmicos , Distribuição Tecidual , TripsinaRESUMO
Studies were designed to determine whether an autoregulation system exists for TSH in the rabbit. For this purpose, a species-specific RIA for rabbit TSH that does not cross-react with human (h) TSH was developed. Hypothyroid animals were studied at varying time periods up to 3 months after either surgical thyroidectomy or propylthiouracil (PTU) treatment. Highly purified hTSH was injected iv at doses of 0 (saline control), 0.1, 0.3, 1,3, and 10 micrograms into unanesthetized rabbits bearing chronically implanted Silastic catheters. Blood samples were obtained at -30, 0, 10, 20, 30, 60, 120, 180, 240, and 300 min and 24 h. Doses between 0.3 and 10 micrograms hTSH produced a prompt fall (10 min) in rabbit TSH in hypothyroid rabbits studied 8-21 days after thyroidectomy. The minimum dose of hTSH that significantly suppressed rabbit TSH was 0.3 micrograms. This dose produced a peak value of hTSH in rabbit serum of 1.3 +/- 0.1 (+/- SEM) ng/ml 10 min after injection, which translates into a bioassay potency of 2.0 microU/ml (close to the physiological level in humans). A dose-response relationship existed between the hTSH dose injected and the duration and magnitude of suppression of rabbit TSH. This response to TSH was specific; 10 micrograms hTSH produced no change in endogenous rabbit serum LH and, conversely, 10 IU hLH produced no change in rabbit serum TSH. In contrast to these striking effects in acute hypothyroid animals, hTSH produced no detectable suppression of rabbit TSH in animals that were hypothyroid for 2-3 months. The sensitivity of the autoregulatory system to the suppressive effects of exogenous hTSH decreased with increasing duration of hypothyroidism; a time-response relationship existed. We conclude that: 1) a sensitive and specific autoregulatory control system for TSH exists in the rabbit; and 2) as the duration of hypothyroidism increases, the sensitivity of the autoregulatory system to the suppressive effects of endogenous TSH changes.
Assuntos
Glândula Tireoide/fisiologia , Tireotropina/metabolismo , Animais , Retroalimentação , Humanos , Hipotireoidismo/fisiopatologia , Propiltiouracila/toxicidade , Coelhos , Tireoidectomia , Tireotropina/sangue , Tireotropina/farmacologia , Tiroxina/sangue , Fatores de TempoRESUMO
In the human and all other mammalian systems studied, LH and hCG bind to a common high affinity receptor with equal affinity. We have recently reported that a unique high affinity binding site in Xanthomonas maltophilia preferentially binds hCG and a native CG-like ligand over LH or other glycoprotein hormones. In the current studies, we have analyzed the effect of hCG or the native ligand on culturing Xanthomonas maltophilia. Both the human and native ligand caused a dose-dependent alteration in the pattern of the growth cycle and a change in the morphology of the bacteria during the stationary phase of the growth cycle. The protein concentration reached during the stationary phase was significantly (P < 0.005) higher in cultures supplemented with hCG or the native ligand. When an aliquot of the culture was diluted and plated on Earl's Martin Balanced agar plates, the number of subsequent colonies was increased (P < 0.02) in the fractions supplemented with the ligands. The increased growth was significant (P < 0.05) to the lowest concentration of 50 ng/ml ligand. When grown under partially anaerobic conditions, the effects of hCG were observed earlier in the growth cycle. The active hormones, hCG and native ligand, also changed bacterial morphology. These data indicate that hCG may have an autocrine and/or paracrine function in bacteria.
Assuntos
Gonadotropina Coriônica/farmacologia , Xanthomonas/metabolismo , Aerobiose , Anaerobiose , Sítios de Ligação , Gonadotropina Coriônica/metabolismo , Gonadotropina Coriônica Humana Subunidade beta , Hormônio Foliculoestimulante/farmacologia , Subunidade alfa de Hormônios Glicoproteicos/farmacologia , Humanos , Cinética , Ligantes , Hormônio Luteinizante/farmacologia , Fragmentos de Peptídeos/farmacologia , Fatores de Tempo , Xanthomonas/efeitos dos fármacos , Xanthomonas/crescimento & desenvolvimentoRESUMO
An approximately 60,000 mol wt glycopeptide has been isolated from acetone-dried human pituitary glands which stimulates production of the adrenal androgen dehydroepiandrosterone, but not cortisol, in acute suspensions of collagenase-dispersed dog adrenal cells. Adrenal androgen secretion has generally been considered, like cortisol, to be under the control of ACTH. This new pituitary glycopeptide, with a molecular weight greater than that of proopiocortin, ACTH, PRL, or LH, may help explain instances during adrenarche, puberty, aging, and stress in which cortisol and adrenal androgen metabolism diverge.
Assuntos
Glândulas Suprarrenais/metabolismo , Androsterona/metabolismo , Glicopeptídeos/farmacologia , Hipófise/análise , Glândulas Suprarrenais/efeitos dos fármacos , Animais , Bioensaio , Cães , Humanos , Hidrocortisona/metabolismo , Masculino , Peso Molecular , Hormônios Adeno-Hipofisários/farmacologiaRESUMO
Studies were designed to assess whether a short loop feedback control for FSH existed in the rabbit. Castrated adult female animals bearing chronically implanted Silastic catheters to permit frequent blood sampling were studied without anesthesia. Ovine FSH was administered as an iv bolus in doses ranging between 0.1-500 micrograms. Endogenous rabbit FSH was quantified using a RIA that did not cross-react with ovine FSH. Blood samples were obtained before and 5, 10, 15, 30, 60, 120, 180, 240, and 300 min after the injection. Each animal was tested at two or more dose levels on different days. Ovine FSH produced suppression of rabbit FSH secretion within 5 min after injection. The minimum effective dose was 1 microgram; maximal suppression occurred with 50-100 micrograms ovine FSH. This short loop feedback control system was specific for FSH; ovine FSH, even at high doses, failed to suppress endogenous rabbit LH. This is the first direct demonstration of a negative short loop feedback control for FSH and the first entirely specific control for the FSH system to be described.
Assuntos
Hormônio Foliculoestimulante/fisiologia , Animais , Especificidade de Anticorpos , Castração , Retroalimentação , Hormônio Luteinizante/sangue , Masculino , Coelhos , Radioimunoensaio , Ovinos , Especificidade da EspécieRESUMO
Studies from our laboratory have demonstrated that human CG (hCG), human LH, (hLH), and an hCG-like protein extracted from Xanthomonas maltophilia were able to induce Candida albicans transition from the blastospore to the germ tube stage. In the present study, we describe the characterization of an hCG-like material extracted from Candida albicans blastospores (CaCGLP), which is potent in inducing transition and presumably represents the endogenous transition-inducing substance. This material was extracted from Candida albicans blastospores with glacial acetic acid and purified by affinity chromatography using a polyclonal rabbit anti-hCG antibody. The product obtained is a 68-kilodalton single band protein, as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. Under reduced conditions a protein smear is seen. Amino acid analysis showed a predominance of glycine (22%), followed by serine (12%), and glutamate (12%). This protein reacted in the following hCG immunoassays: 1) a polyclonal rabbit anti-hCG equilibrium assay, 2) a carboxyl-tail hCG equilibrium assay, 3) two hCG equilibrium assays using monoclonal antibodies (CG no. 4 and CG no. 9), 4) a free alpha-subunit equilibrium assay using a monoclonal antibody, and 5) an ultrasensitive immunoradiometric assay for hCG which does not cross-react with hLH, nor the free beta-subunit of hCG. The CaCGLP showed no reaction in a specific hLH immunoradiometric assay. When CaCGLP was tested in the transition assay, in the presence of 4% rat serum, it was found that this protein was 100 times more potent than hCG in producing Candida albicans transition. We conclude that Candida albicans produces a protein that has certain tertiary structure similarities to hCG and that this material is able to induce germ tube formation. We postulate that CaCGLP has an autocrine/paracrine effect in Candida albicans as a transition factor to control its own pathogenicity.
Assuntos
Candida albicans/química , Gonadotropina Coriônica/isolamento & purificação , Endopeptidases/isolamento & purificação , Proteínas/isolamento & purificação , Aminoácidos/análise , Anticorpos Monoclonais , Western Blotting , Candida albicans/efeitos dos fármacos , Candida albicans/enzimologia , Gonadotropina Coriônica/química , Gonadotropina Coriônica/farmacologia , Eletroforese em Gel de Poliacrilamida , Endopeptidases/química , Endopeptidases/metabolismo , Humanos , Imunoensaio , Peso Molecular , Estrutura Terciária de Proteína , Proteínas/química , Proteínas/farmacologia , Esporos Fúngicos/química , Esporos Fúngicos/enzimologiaRESUMO
Dehydroepiandrosterone (DHA), in large part a measure of adrenal androgen secretion, previously has been measured in urine after hydrolysis or solvolysis of the glucuronide and sulfate conjugates. These procedures are time consuming; they often require several days and variable recoveries are a source of error. A method is presented here for determination of unconjugated DHA in 24-h urine specimens which requires less time, labor, and sample volume than necessary for the assay of DHA derived from conjugates. In 76 men and women, age 20-96 yr, total 24-h urinary unconjugated DHA showed no sex differences. However, the mean unconjugated DHA excretion decreased, which may indicate decreased zona reticularis function with respect to relatively constant zona fasiculata function in advancing age.
Assuntos
Glândulas Suprarrenais/metabolismo , Androgênios/biossíntese , Desidroepiandrosterona/urina , Dexametasona , Glândulas Suprarrenais/efeitos dos fármacos , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Radioimunoensaio/métodosRESUMO
Several groups of investigators have previously reported that small amounts of hCG are present in blood and urine of nonpregnant eugonadal women and men. We have developed highly sensitive and specific, two-monoclonal antibody, sandwich-type assays which can quantify both hCG and LH in sera from postmenopausal women, women at all phases of the menstrual cycle, and men. Using these assays we have also reported that hCG is secreted in a pulsatile fashion in postmenopausal women, is stimulated by GnRH in both men and postmenopausal women, and is suppressed by a GnRH agonist in castrate men. Employing these same sandwich assays, we report herein that hCG is secreted in a pulsatile manner during the follicular and luteal phases of the normal menstrual cycle. During the follicular phase 0.68 +/- 09 (+/- SEM) pulses of hCG occurred each hour, while 0.62 +/- 0.08 pulses of LH occurred. Pulse durations during the follicular phase for hCG and LH, respectively, were 38.3 +/- 4.4 and 62.5 +/- 11.1 min (P less than or equal to 0.05). During the luteal phase there were 0.42 +/- 0.16 pulses/h of hCG and 0.38 +/- 0.08 pulses of LH (P greater than 0.05). Pulse durations were 22.3 +/- 7.5 and 126.4 +/- 19.0 min for hCG and LH, respectively (P less than 0.01). The t1/2 of hCG disappearance was 37.2 +/- 3.8 min during the follicular phase and 22.9 +/- 4.6 min during the luteal phase. The t1/2 values of LH were 82.9 +/- 5.7 and 67.5 +/- 5.12 min during follicular and luteal phases, respectively. The t1/2 of LH was greater than the t1/2 of hCG (P less than 0.01). We conclude that small amounts of hCG are secreted in a pulsatile manner during follicular and luteal phases of the human menstrual cycle.
Assuntos
Gonadotropina Coriônica/metabolismo , Ciclo Menstrual , Adulto , Anticorpos Monoclonais , Gonadotropina Coriônica/sangue , Cromatografia de Afinidade , Feminino , Humanos , Hormônio Luteinizante/sangue , Periodicidade , Progesterona/sangue , Valores de ReferênciaRESUMO
A 33-yr-old male developed typical symptoms and findings of diabetes insipidus. A computed tomographic scan of the hypothalamus/pituitary was normal, and he was diagnosed as having idiopathic diabetes insipidus. At age 38 yr, he developed two 1- to 2-mm reddish papules on his skin. Biopsy revealed infiltrative histiocytes laden with lipid. Bone scans and bone x-rays showed widespread osteolytic and osteoblastic disease. The disease was diagnosed as a rare disseminated histiocytic disorder, xanthoma disseminatum. A classification of histiocytic disease is presented.
Assuntos
Diabetes Insípido/etiologia , Histiocitose de Células não Langerhans/complicações , Adulto , Biópsia , Osso e Ossos/diagnóstico por imagem , Histiocitose de Células não Langerhans/diagnóstico por imagem , Histiocitose de Células não Langerhans/patologia , Humanos , Masculino , Radiografia , Pele/patologiaRESUMO
Recent data from our laboratory show that hCG is secreted in a pulsatile manner in parallel with human LH (hLH) in nonpregnant normal humans. Furthermore, GnRH stimulates hCG and hLH release. Using a monoclonal antibody specific for the beta-chain of hCG and not reacting with hLH, we have identified a heretofore unknown cell type in human pituitaries which stains only for hCG. The light microscopic, immunocytochemical, and ultrastructural features are described. These data coupled to those from multiple earlier studies indicate that hCG is secreted by these cells in normal nonpregnant humans.