RESUMEN
The inhibin content and aromatase inhibitor activity (AIA) of 72 follicular fluids (FF) obtained from 42 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) were studied as a function of IVF ET outcome. Inhibin levels were determined by bioassay (BA) and RIA; AIA was measured by BA. The inhibin content of follicles characterized as immature by their estradiol (E2) levels and E2/progesterone (P) ratios was significantly lower (P less than 0.05) than that of mature follicles (i.e. leading to pregnancy). The mean AIA for mature follicles were significantly lower than AIA in groups where pregnancy was not obtained. AIA for follicles from which a pregnancy was obtained for each ET was also significantly lower than that in FF characterized as immature of hypermature. The highest E2/AIA and inhibin BA/AIA ratios were associated with the highest incidence of successful IVF ET outcome. No correlation was found between AIA and inhibin, on the one hand, and E2, delta 4-androstenedione, E2/P, and PRL, on the other. However, a positive correlation was found between inhibin (RIA and BA) and P, reflecting the production of inhibin by granulosa cells during luteinization. These studies allowed us to conclude that FF inhibin levels do not differ according to IVF ET outcome, but are an index of follicular maturation. AIA not only constitutes an index of follicular maturation and granulosa cell luteinization, but is of predictive value for IVF ET outcome as E2/AIA and inhibin BA/AIA ratios.
Asunto(s)
Inhibidores de la Aromatasa , Líquidos Corporales/análisis , Transferencia de Embrión , Fertilización In Vitro , Inhibinas/análisis , Folículo Ovárico/fisiología , Adulto , Líquidos Corporales/enzimología , Gonadotropina Coriónica/farmacología , Estradiol/análisis , Femenino , Hormona Folículo Estimulante/farmacología , Hormona Liberadora de Gonadotropina/antagonistas & inhibidores , Humanos , Fase Luteínica/fisiología , Oocitos/análisis , Oocitos/efectos de los fármacos , Progesterona/análisis , RadioinmunoensayoRESUMEN
Concentrations of alpha 1 acid glycoprotein, albumin, transferrin, haptoglobin, immunoglobulins G, A, M and apolipoprotein B were measured in serum and suction blister fluid from a group of individuals presenting a biologically proven inflammatory syndrome, and from a control group. Protein values in suction blister fluid did not change from the 2nd to the 3rd h after the beginning of blister formation. The ratio of the concentration of proteins in blister fluid and serum did not differ significantly between the groups. However, a 25% decrease in blister fluid albumin and a 100% increase in blister fluid alpha 1 acid glycoprotein, recorded in the inflammatory group, were worth noting, since they possibly influence the tissular distribution of some protein-binding drugs. Finally, an inverse relationship was established between the blister fluid/serum concentration ratio and the respective molar mass of each protein.
Asunto(s)
Inflamación/metabolismo , Proteínas/análisis , Piel/análisis , Anciano , Vesícula/metabolismo , Líquidos Corporales/análisis , Líquidos Corporales/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Farmacocinética , Unión Proteica , Proteínas/metabolismo , Piel/metabolismoRESUMEN
The HER2/neu proto-oncogene encodes a receptor that belong to the tyrosine-specific protein kinase family. Amplification of the HER2 gene in patients with breast and ovarian cancer has been shown to predict poorer survival rates. In order to understand the role of HER2 in malignant and normal cells, it is necessary to devise assays that can quantitate expression levels of the HER2 gene product (p185HER2) in production samples, biopsy specimens and biological fluids. We have developed a simple, quantitative ELISA that uses two monoclonal antibodies directed against the extracellular domain of the HER2 gene product, p185HER2 (HER2 ECD). The assay has a detection range of 0.25-120 ng/ml, is precise and sensitive. The ability of this assay to detect biologically active rHER2 ECD is demonstrated by its correlation to a growth inhibitory bioassay (r = 0.92). The sandwich ELISA can also accurately quantitate rHER2 ECD in mouse and monkey serum. This assay should be useful for quantitating low levels of circulating rHER2 ECD in animals in which rHER2 ECD is being used as antigen for immunotherapy and in patients which 'shed' receptor.
Asunto(s)
Líquidos Corporales/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Proteínas Oncogénicas Virales/análisis , Anticuerpos Monoclonales , Neoplasias de la Mama/inmunología , División Celular/efectos de los fármacos , Células Cultivadas , Humanos , Técnicas In Vitro , Masculino , Proto-Oncogenes Mas , Receptor ErbB-2RESUMEN
Measurements of specific antibody titres in uterine fluid of mice immunized by different routes indicated that two immunizations in the pelvic presacral space using aluminium hydroxide as adjuvant was a simple and effective way to elicit a significant IgA and IgG response. Higher IgA and IgG titres were produced in uterine fluid by subcutaneous immunization with antigen in Freund's complete adjuvant followed by intravaginal boosting without adjuvant, but this immunization involved both a toxic adjuvant and repeated applications of large doses of antigen in the vagina. Intragastric immunization produced an IgA response in the uterus but no IgG. Local intravaginal priming and boosting with large doses of antigen without adjuvant produced an IgA response in uterine fluid, but was less effective for IgG and was inefficient in terms of time and the amount of antigen used. Hysterectomy reduced the concentration of specific IgA in vaginal fluid of immunized mice to no more than 5% of normal, indicating that most of the IgA in vaginal fluid originates in the uterus. In contrast, IgG titres were not significantly different in hysterectomized and intact mice. IgA titres in vaginal fluid were at least partly restored to normal levels in sham-hysterectomized mice.
Asunto(s)
Anticuerpos/análisis , Ferritinas/inmunología , Inmunoglobulina A/análisis , Inmunoglobulina G/análisis , Útero/inmunología , Animales , Líquidos Corporales/análisis , Femenino , Histerectomía , Inmunización/métodos , Ratones , Ratones Endogámicos ICR , Vagina/inmunologíaRESUMEN
The polar metabolites of polycyclic aromatic compounds (PACs) are of significant oncological interest. In contrast to parent PACs, isomer selective detection of polar PACs by laser excited Shpol'skii spectrometry (LESS) is severely limited by excessively broadened spectra and rapid photodegradation in n-alkane solvents. To minimize these limitations a 10-min derivatization procedure was developed to produce corresponding methoxy compounds, that exhibit the Shpol'skii effect. By use of the extraction and permethylation procedure, the selective detection of a mixture of hydroxy-benz[a]anthracene isomers spiked at the low picogram level into urine and blood matrices was achieved. A detection limit for 1-hydroxybenz[a]anthracene was 0.6 part per trillion, or 12 fg (0.05 fmol) on a 20-microL sample.
Asunto(s)
Benzo(a)Antracenos/análisis , Rayos Láser , Espectrometría de Fluorescencia , Benzo(a)Antracenos/metabolismo , Líquidos Corporales/análisis , Fenómenos Químicos , QuímicaRESUMEN
A colorimetric method for the screening of cyanide in biological specimens is presented. Cyanide in a 200-microL sample is diffused in a Conway 3 compartment dish into the center well where it catalyzes the reaction between p-nitrobenzaldehyde and o-dinitrobenzene. The resulting blue-violet color intensity is proportional to cyanide concentration. The reaction is specific for cyanide, and concentrations as low as 0.05 mg/L can readily be detected.
Asunto(s)
Cianuros/análisis , Líquidos Corporales/análisis , Colorimetría/métodos , Humanos , Sensibilidad y EspecificidadRESUMEN
This prospective, randomized, single-blind study of 121 healthy children aged 2 to 12 yr investigated the effect of clear fluids on gastric contents. Gastric fluid volume and pH were measured immediately following the induction of general anaesthesia and were not significantly affected by the ingestion of unlimited clear fluids up to three hours preoperatively. After a prolonged fast (mean fast 14 hr), gastric fluid volume was 0.39 +/- 0.37 ml.kg-1 and gastric pH was 1.7 +/- 0.4; after unlimited clear fluids (203 +/- 109 ml) up to three hours before surgery gastric fluid volume was 0.34 +/- 0.28 ml.kg-1 and gastric pH was 1.8 +/- 0.7 (mean +/- SD). Gastric fluid volume (ml.kg-1) increased in both the control and study groups as age increased, P less than 0.005. It is concluded that drinking clear fluid up to three hours before scheduled surgery does not have a measurable effect on gastric volume and pH of healthy children of ages 2 to 12 yr.
Asunto(s)
Ingestión de Líquidos , Contenido Digestivo/análisis , Cuidados Preoperatorios , Líquidos Corporales/análisis , Niño , Preescolar , Ayuno , Humanos , Neumonía por Aspiración/epidemiología , Ensayos Clínicos Controlados Aleatorios como Asunto , Factores de RiesgoRESUMEN
Positive-ion electron impact (PIEI), positive-ion chemical ionization (PICI) and negative-ion chemical ionization (NICI) mass spectra of 9 carbamate pesticides are presented. In the PIEI mode, the spectra showed small molecular peaks, intense or base peaks due to M - CH3NHCO + H and peaks at m/z 58 due to CH3NHCO. In the PICI mode, peaks due to M + H, M + C2H5, M - CH3NHCO + 2H, CH3NHCO(m/z 58) and M-28 appeared. The cations at m/z 58 found in both PIEI and PICI modes seem very useful for screening of a carbamate. In the NICI mode, the spectra showed peaks due to M - CH3NHCO and characteristic anions appearing at mass numbers higher than molecular ones, which were probably due to dimerization of [M - CH3NHCO]-followed by hydrogen attachment. Carbamates, which had been added to urine, plasma, whole blood, the liver, kidney and brain, could be rapidly isolated by use of Sep-Pak C18 cartridges with chloroform as an elution solvent. They could be detected by wide-bore capillary gas chromatography with a SPB-5 column, with satisfactory separation from impurities in their underivatized forms.
Asunto(s)
Carbamatos , Medicina Legal/métodos , Insecticidas/análisis , Espectrometría de Masas/métodos , Líquidos Corporales/análisis , Química Encefálica , Cromatografía de Gases y Espectrometría de Masas , Humanos , Riñón/análisis , Hígado/análisis , Estructura Molecular , Peso MolecularRESUMEN
A sensitive enzyme-linked immunosorbent assay (ELISA) for detection of ABO blood substances in human body fluids was devised. The ELISA plates coated with purified human anti-A or anti-B serum effectively captured the blood substances, and these were then analysed by the combination of rabbit anti-A and goat anti-B. This capture ELISA could differentiate the type AB specimen from a mixture of the type A and the type B specimens, and the method was applied to rape cases to make the ABO typing of the criminal.
Asunto(s)
Sistema del Grupo Sanguíneo ABO , Tipificación y Pruebas Cruzadas Sanguíneas/métodos , Líquidos Corporales/análisis , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Medicina Legal/métodos , Humanos , Masculino , Semen/análisis , Vagina/metabolismoRESUMEN
Immune responses in the uterus are depressed when concentrations of progesterone in the blood are elevated. We tested whether this action of progesterone involves induction of secretion of immunosuppressive molecules into the uterine lumen. Uterine fluid from ovariectomized ewes treated with progesterone for 60 days inhibited [3H]thymidine (TdR) incorporation by phytohemagglutinin-stimulated lymphocytes more than uterine flushings from vehicle-treated ewes. Uterine fluid from progesterone-treated ewes also inhibited TdR uptake by other mitogen-stimulated lymphocytes and mixed lymphocyte cultures, but had no effect on the growth of three nonlymphocyte cell lines. A group of ewes immunized with ovalbumin mixed with uterine fluid from progesterone-treated ewes produced an antibody titer that was lower than the titer for ewes immunized with ovalbumin without uterine fluid. Analysis of the physical properties of the active molecules in uterine fluid from progesterone-treated ewes indicated that activity could be separated into two fractions: a large mol wt (Mr; greater than 5000) fraction that was sensitive to pronase and a low Mr (less than 1000) fraction that was more resistant to pronase. There was more inhibitory activity in the high mol wt fraction. Further analysis of this fraction indicated that most activity was of basic isoelectric point (pI greater than 8.2). The most active fraction of basic material eluted at the void volume of a Sepharose CL-6B column (Mr, 4 x 10(6)). In conclusion, progesterone caused accumulation of factors in the uterine lumen that inhibited lymphocyte function in vitro and antibody formation in vivo. These molecules may play an important role in regulating immune responses in the uterus during pregnancy.
Asunto(s)
Tolerancia Inmunológica/efectos de los fármacos , Ovariectomía , Progesterona/farmacología , Útero/inmunología , Animales , Anticuerpos/análisis , Líquidos Corporales/análisis , Líquidos Corporales/inmunología , Femenino , Inmunización , Punto Isoeléctrico , Activación de Linfocitos/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Peso Molecular , Ovalbúmina/inmunología , Fitohemaglutininas/farmacología , Pronasa/metabolismo , Proteínas/aislamiento & purificación , Proteínas/metabolismo , Proteínas/farmacología , Ovinos , Útero/efectos de los fármacos , Útero/metabolismoRESUMEN
Plasma D2O-accumulation profiles (qualitative indices of fluid-absorption rates) were determined in eight subjects after ingestion of 275 mL of five D2O-labeled beverages: a water placebo (W), 6% maltodextrin (6% M), and three solutions containing a 6%, 8%, and 10% glucose-fructose mix (6% GF, 8% GF, and 10% GF). Except for W all beverages contained 20 mmol sodium/L and 3 mmol potassium/L. No differences in plasma D2O accumulation were found. Plasma glucose increased at 20 and 30 min after ingestion of the carbohydrate drinks and returned to baseline (6% GF and 6% M) or below (8% GF and 10% GF) by 60 min. Insulin responded similarly and, except for a slightly lower value at 30 min for 6% GF, no differences were detected. It appears that fluids in drinks containing less than or equal to 8-10% carbohydrate (simple sugars or maltodextrins) are made available for dilution in body fluids at similar rates and should be similar in replenishing body fluids lost in sweat during exercise.
Asunto(s)
Bebidas/análisis , Líquidos Corporales/análisis , Carbohidratos/análisis , Deportes , Adulto , Disponibilidad Biológica , Glucemia/análisis , Carbohidratos/farmacocinética , Deuterio/análisis , Óxido de Deuterio , Humanos , Insulina/sangre , Masculino , Polisacáridos/análisis , Agua/análisisRESUMEN
In order to investigate the role of renin angiotensin in the epididymis, angiotensin-converting enzyme (ACE) activity and angiotensin I (AI) and angiotensin II (AII) concentrations were measured in the male reproductive tract and blood serum of the rat. High ACE activity was detected in the rat epididymis, with a major part of the activity being associated with epididymal spermatozoa. When spermatozoa were prevented from entering the epididymis by efferent duct ligation, the ACE activity in the epididymis was greatly reduced. The epithelial cells lining the epididymal duct were also shown to possess ACE activity which was dependent upon circulating androgens. Treatment of male rats with captopril at a single oral dose (20 mg/kg) significantly inhibited the ACE activity in the blood serum but had no effect on the activity of the epididymal fluid. The intraluminal ACE was protected from the circulating captopril by the blood-epididymis barrier. Long-term treatment with captopril (20 mg/kg per day, 8 weeks), however, caused an increase in blood serum ACE activity but was without effect on intraluminal ACE. The fertility and fecundity of male rats after treatment were apparently normal. The concentrations of AI and AII were high in the epididymal plasma and epididymal cell when compared with the respective concentrations in blood serum. The intraluminal AII concentration found (13 nmol/l) was close to the threshold concentrations that stimulate anion (and fluid) secretion in cultured epididymal epithelium in vitro. The high intraluminal AII concentration could not have been derived from the testicular fluid or spermatozoa since the rete testis fluid and sperm contained little AII.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Epidídimo/fisiología , Peptidil-Dipeptidasa A/fisiología , Sistema Renina-Angiotensina/fisiología , Angiotensina I/análisis , Angiotensina I/sangre , Angiotensina II/análisis , Angiotensina II/sangre , Animales , Líquidos Corporales/análisis , Captopril/farmacología , Epidídimo/metabolismo , Fertilidad/efectos de los fármacos , Masculino , Orquiectomía , Peptidil-Dipeptidasa A/análisis , Peptidil-Dipeptidasa A/sangre , Ratas , Ratas EndogámicasRESUMEN
Specific and sensitive reverse-phase HPLC assays of buprenorphine and its metabolite in biological fluids were developed with sensitivities of 2-6 ng ml-1 using fluorimetric detection. Pharmacokinetics were monitored on acute bolus administration of buprenorphine in 6 dogs within the 0.7-2.6 mg kg-1 dose range. Toxicity was circumvented when terminal plasma concentrations were increased by infusing 3.7-4.8 mg kg-1 doses of buprenorphine over 3 h in six studies in 6 dogs. The terminal rate constants of the IV infusion studies from the triexponential fits of plasma concentration-time data averaged 41.6 +/- 7.5 h with an averaged total body clearance of 191 +/- 19 ml min-1. This terminal rate constant was in contrast to the less than 100 min half-life of the second exponential fitting of the less lipophilic morphine, naloxone, and naltrexone. The apparent volumes of distribution of buprenorphine, referenced to the total plasma concentration, were 33 +/- 61 (Vc, central compartment volume) and 663 +/- 891 (Vd, total body volume), indicative of a highly bound, sequestered or lipophilic drug. Unchanged buprenorphine was insignificantly renally (less than 0.2 per cent of the dose) and biliary (less than 0.6 per cent) excreted. The major route of buprenorphine disposition was by hepatic conjugation to glucuronide which was eliminated into the bile (about 92 per cent) with only small amounts appearing in urine (less than 1 per cent as metabolite). Minor metabolites excreted in the bile accounted for about 3 per cent of the administered dose. Direct IV administration of the metabolite, buprenorphine glucuronide, gave a terminal half-life of 6 h and more than 90 per cent of the systemically circulating metabolite was excreted in bile; only 10 per cent in urine. The oral bioavailability, estimated from the areas under the buprenorphine plasma concentration-time curve following IV and oral administration of buprenorphine in the dogs, was 3-6 per cent. There were no apparent correlations of the buprenorphine time course with cardiovascular parameters such as heart rate, ECG, and blood pressure. Miotic effect was significant. Respiratory depression was observed during the first 4 h after IV bolus injection, but not during the infusion studies.
Asunto(s)
Buprenorfina/farmacocinética , Morfina/farmacocinética , Animales , Bilis/metabolismo , Líquidos Corporales/análisis , Buprenorfina/sangre , Buprenorfina/orina , Cromatografía Líquida de Alta Presión , Perros , Glucuronatos/metabolismo , Hidrólisis , Masculino , Modelos Biológicos , Morfina/sangre , Morfina/orina , Espectrometría de FluorescenciaRESUMEN
Nuclear magnetic resonance spectroscopy is the epitome of the high-technology, expensive diagnostic method. Extrapolation from a limited number of patient examinations and from experiments in animal models predicts a bright future for the method. However, several barriers block widespread clinical application in the near future; technical difficulties still exist but they seem to be resolvable in due course. A more serious problem is the absence of an adequate database from which to interpret the vast array of information produced by nuclear magnetic resonance. The necessary understanding of the pathologic biochemistry of disease will be frustratingly slow to appear as will the routine clinical use of magnetic resonance spectroscopy. The critical need for improved diagnostic methods will stimulate experimentation to resolve these problems.
Asunto(s)
Técnicas de Laboratorio Clínico , Espectroscopía de Resonancia Magnética , Patología Clínica , Líquidos Corporales/análisis , Encéfalo/patología , Humanos , Riñón/patología , Hígado/patología , Espectroscopía de Resonancia Magnética/instrumentación , Músculos/patología , Miocardio/patología , Neoplasias/patologíaRESUMEN
The use of stable isotopically labeled steroids combined with gas chromatography/mass spectrometry (GC/MS) has found a broad application in pharmacologic studies. Initially, stable isotopically labeled steroids served as the ideal analytic internal standard for GC/MS analysis; however, their in vivo use has expanded and has proven to be a powerful pharmacokinetic tool. We have successfully used stable isotope methodology to study the pharmacokinetic/bioavailability of androgens. The primary advantage of the technique is that endogenous and exogenous steroids with the same basic structure can be differentiated by using stable isotopically labeled analogs. The method was used to examine the pharmacokinetics of testosterone and testosterone propionate, and to clarify the influence of endogenous testosterone. Another advantage of the isotope methods is that steroidal drugs can be administered concomitantly in two formulations (e.g., solution and solid dosage). A single set of blood samples serves to describe the time course of the formulations being compared. This stable isotope coadministration technique was used to estimate the relative bioavailability of 17 alpha-methyltestosterone.