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Biomedical subjects

J A Castilla

Publications and source records attributed to J A Castilla.

At least 19 recordsLinked to original sources
PubMed1995

CA 125, CA 15.3, CA 27.29, CEA, beta-hCG and alpha-fetoprotein levels in cyst fluid of breast macrocysts.

OBJECTIVE: To determine the tumoral markers CA 125, CA 15.3, CA 27.29, CEA, beta-hCG and alpha-fetoprotein in serum and breast cystic fluid in women with fibrocystic breast disease. METHODS: The study comprised 50 women diagnosed with fibrocystic breast disease but without breast macrocysts, and 60 women with macrocysts. RESULTS: Significantly higher levels of CA 125, CA 27.29, beta-hCG and CEA were observed in the cyst fluid than in the serum of the same patients. According to their K+/Na+ quotient we found that when K+/Na+ was < 3, cyst fluid CA 125, CA 27.29, beta-hCG and CEA levels were significantly higher than serum levels. However, when K+/Na+ was > 3, only CA 27.29 and beta-hCG levels were significantly higher. Comparing cyst fluid marker levels with respect to the K+/Na+ ratio, the only difference observed was in CA 125 which was significantly greater when K+/Na+ was < 3. A negative correlation was found between CA 125 and the K+/Na+ quotient. CONCLUSION: These results suggest that CA 125 could be used as a marker to identify cyst type.

17-alpha-Hydroxyprogesterone↗
PubMed1995

CD4+ cells in human ejaculates.

Using flow cytometry, we studied the expression of the CD4 antigen within the different cells present in human ejaculate, both in spermatozoa and round cells. In all, 20 samples of semen were obtained from fertile males; in 11 of these, we detected the presence of leukocytes, using the peroxidase test. Swim-up was performed for the analysis of the spermatozoa. From our results it may be concluded that there is no expression of the CD4 antigen on the surface of human spermatozoa or on CD45- ejaculate cells (epithelial and germinal cells). However, we did detect the presence of the CD4 antigen on the surface of the leukocyte cells (CD45+). A better characterization of these CD45+ cells made it apparent that the CD4+ cells of ejaculate are composed of T lymphocytes (helper/inducer T lymphocytes) and monocytes. Thus we may conclude that human spermatozoa do not express the CD4 antigen, the cell surface receptor for human immunodeficiency virus. However, we did detect CD4+ T lymphocytes and CD4+ monocytes in semen.

CD4 Antigens↗
PubMed1995

Influence of incubation on the chromatin condensation and nuclear stability of human spermatozoa by flow cytometry.

Flow cytometry analysis was used for the accurate and objective evaluation of sperm chromatin condensation and chromatin stability of sperm nuclei. It was also possible to determine the influence of incubation on sperm chromatin. Different types of spermatozoa were studied: unprocessed spermatozoa at 1 and 45 min after ejaculation, after swim-up (migrated), spermatozoa incubated for 6 h in non-capacitating conditions (aged), or in B2 medium (capacitated) or B2 medium followed 1 h later with A23187 (reacted). All types of spermatozoa were analysed before and after treatment with various decondensation agents: sodium dodecyl sulphate (SDS), SDS plus EDTA and SDS plus disulphide-reducing agent [dithiotreitol (DTT)]. Sperm nuclei were enzymatically isolated and stained with propidium iodide. Three flow cytometric parameters were then measured: forward light scatter (cellular size), side light scatter (cellular complexity) and fluorescence (uptake of propidium iodide). Fluorescence was the most suitable parameter to study the degree of condensation and resistance to decondensation of DNA in the spermatozoa. Unprocessed spermatozoa 1 min after ejaculation underwent decondensation by all assessed treatments (anionic detergent, chelating or disulphide-reducing agents). Unprocessed spermatozoa 45 min after ejaculation and migrated spermatozoa did not undergo decondensation with SDS treatment, but decondensation occurred after treatment with SDS+EDTA or SDS+DTT. Spermatozoa incubated for 6 h under both non-capacitating (aged spermatozoa) and capacitating conditions (capacitated spermatozoa) and reacted spermatozoa were decondensed only after treatment with SDS+DTT.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcimycin↗
PubMed1995

Undetectable expression of genomic progesterone receptor in human spermatozoa.

The expression of genomic progesterone receptor in human ejaculated spermatozoa was investigated. Spermatozoa from 10 fertile donors who exhibited normal semen parameters were analysed. Indirect immunofluorescence and an enzyme immunoassay using monoclonal antibodies against genomic progesterone receptor were used. Different types of spermatozoa were studied: fresh, post-swim-up (migrated), capacitated and post-artificial induction of the acrosome reaction by calcium ionophore A23187. Progestin receptor-rich T47D human breast cancer cells were used as a positive control, and progestin receptor-poor MDA-MB-231 human breast carcinoma cells were used as a negative control. Genomic progesterone receptor was not detected in fresh, migrated, capacitated and post-acrosome reaction induction human spermatozoa and MDA-MB-231 cells by either indirect immunofluorescence or enzyme immunoassay. However, in T47D cells a mean concentration of 1043.2 +/- 125.2 fmol genomic progesterone receptor/mg protein was observed by enzyme immunoassay, and indirect immunofluorescence results were positive using both flow cytometry and fluorescence microscopy. These findings suggest that the effect of progesterone on human spermatozoa is not mediated by genomic progesterone receptor.

Antibodies, Monoclonal↗
PubMed1995

Thyroid hormones in fibrocystic breast disease.

This study was undertaken to evaluate the role of thyroid hormones in fibrocystic breast disease. The concentrations of thyroid-stimulating hormone (TSH), thyroxine (T4), free T4 and free triiodothyronine (T3) were determined in serum of 50 women with fibrocystic breast disease without macrocysts (cysts of over 3 mm diameter) and in the serum and breast cyst fluid (BCF) of 60 women with fibrocystic breast disease and macrocysts. Possible relationships between thyroid hormones and estradiol, dehydroepiandrosterone sulfate, testosterone, progesterone and 17-hydroxyprogesterone in the BCF also were analyzed. Serum thyroid hormone levels did not differ between the two groups. Free T3 levels were higher in BCF than in serum (p < 0.001), whereas T4, free T4 and TSH concentrations were lower in BCF as compared to serum (p < 0.001). Cysts were divided according to their K+/Na+ ratio because a ratio above 3 represents a predictor of malignant transformation. Free T3 concentrations were higher in BCF than in serum, in both low K+/Na+ cysts and in cysts with a K+/Na+ ratio above 3; those cysts with a high K+/Na+ ratio had the highest free T3 concentration. Free T3 in cysts correlated positively to the K+/Na+ ratio (r = 0.831; p < 0.001). Multiple linear regression analysis demonstrated that the concentration of free T3 in BCF was predicted statistically by the positive regression coefficient for the estradiol concentration. No candidate variable was included in the model to predict concentrations of TSH, free T4 or T4 in BCF. These data suggest an important role of free T3 in the physiology of fibrocystic breast disease.

17-alpha-Hydroxyprogesterone↗
PubMed1994

Serum CA-125 in the diagnosis of acute pelvic inflammatory disease.

OBJECTIVES: To determine the efficiency of different tumor markers (CA-125, carcinoembryonic antigen, CA-15.3, CA-19.9) and insulin-like growth factor I (IGF-I) measurements as a screening procedure for acute pelvic inflammatory disease (PID). METHODS: Peripheral blood samples were obtained at the time of laparoscopy from three groups of women: (1) 50 women who underwent laparoscopic tubal ligation and had no evidence of PID (control group); (2) 20 women admitted because of suspected PID, but at laparoscopy or laparotomy had no signs of PID; (3) 20 patients who underwent acute PID diagnosed by laparoscopy. Serum levels of: CA-125, carcinoembryonic antigen, CA-15.3 and CA-19.9, and plasma IGF-I, were measured by radioimmunoassay. RESULTS: No differences were observed in the levels of CA-15.3, CA-19.9, carcinoembryonic antigen and IGF-I between the three groups studied. Serum levels of CA-125 were significantly higher in patients who had PID. Analysis of receiver operating characteristic curves showed that only CA-125 was useful in diagnosis of acute PID. The cut-off level was 43.7 U/ml for CA-125. CONCLUSIONS: Measurement of serum CA-125 concentrations is recommended as a useful test for acute PID in patients undergoing laparoscopy for pelvic pain.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1994

Reactive sacroiliitis as late sequela after severe pelvic inflammatory disease verified by laparoscopy or laparotomy.

OBJECTIVE: To investigate the frequency of lumbosacral pains and sacroiliitis as late sequela of severe pelvic inflammatory disease (PID) confirmed by laparoscopy or laparotomy. DESIGN: The sacroiliac joints were examined radiographically and scintigraphically to search for signs of reactive sacroiliitis. SETTING: Department of Obstetrics and Gynecology, Hospital General Virgen de las Nieves, Granada, Spain. PATIENTS: Thirty-five out of 70 women admitted with severe clinical symptoms of PID during a five-year period. PID was confirmed by laparoscopy or laparotomy. MEASUREMENTS: The sacroiliac joints were examined radiographically, and bone scintigraphic studies of the pelvic girdle were performed with 99technetium labelled pyrophosphate. RESULTS: Twenty out of 35 patients reported lumbosacral pain. The bone scintigraphic findings were positive for sacroiliitis in 18 out of 33 patients (54.6%), and the radiographic findings were positive in 11 women (33.3%). CONCLUSIONS: Patients with previous severe clinical PID complaining of lumbosacral pains may be offered a bone scintigraphy, a sensitive procedure using low-dose radiation, to search for early signs of reactive sacroiliitis.

Arthritis↗
PubMed1994

Intracystic lipidic profile in fibrocystic breast disease.

This study was designed to investigate the lipidic profile of fibrocystic breast disease. The study entailed measuring total cholesterol, high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), very low-density lipoprotein-cholesterol (VLDL-C) and triglyceride levels in the serum of 50 women with fibrocystic breast disease without macrocysts, and in the serum and breast cyst fluid of 60 women with fibrocystic breast disease and macrocysts. The relationships between the lipids analyzed in the cyst fluid and intracystic estradiol, progesterone, dehydroepiandrosterone sulfate, testosterone and 17 alpha-hydroxyprogesterone were studied. No differences were found for serum lipid levels between the two groups of patients. The levels of total cholesterol and HDL-C were found to be significantly higher, and the levels of triglycerides and VLDL-C significantly lower in breast cyst fluid compared to serum. In cysts with high potassium concentrations, we found a higher concentration of cholesterol and HDL-C and a lower concentration of LDL-C than in cysts with a potassium/sodium ratio < 3. No significant differences were found in cystic levels of triglycerides and VLDL-C. Multiple regression analysis showed that the only intracystic variable, which was, in part, positively responsible for the hormone levels measured, was HDL-C. These data suggest that local steroidogenesis in fibrocystic breast disease uses HDL-C as the substrate.

17-alpha-Hydroxyprogesterone↗
PubMed1993

Follicular fluid alpha-fetoprotein, carcinoembryonic antigen, and CA-125 levels in relation to in vitro fertilization and gonadotropin and steroid hormone concentrations.

OBJECTIVE: To investigate the possible role of alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and CA-125 in the ovarian follicle. DESIGN: alpha-Fetoprotein, CEA, and CA-125 were measured in human follicular fluid (FF) and correlated to IVF outcome and gonadotropin and gonadal steroid hormones. SETTING: The In Vitro Fertilization Program at Virgen de las Nieves Hospital, Granada, Spain. PATIENTS: Thirty-six FF from 12 infertile women with irreparable tubal damage who underwent treatment for ovarian stimulation with clomiphene citrate, hMG, and hCG. INTERVENTIONS: Laparoscopic follicular aspiration followed 48 hours later by ET. MAIN OUTCOME MEASURE: Serum and FF AFP, CEA, and CA-125 levels. RESULTS: alpha-Fetoprotein, CEA, and CA-125 were detected in all samples of serum and FF. There were no significant differences between the serum and FF levels. Follicular fluid AFP, CEA, and CA-125 of fertilized oocytes were similar to those in the unfertilized oocytes group. There were no significant correlations between the FF AFP, CEA, and CA 125 levels and gonadotropin and gonadal steroid hormone levels. CONCLUSIONS: alpha-Fetoprotein, CEA, and CA-125 are present in FF after ovarian stimulation, but the mean intrafollicular levels do not differ significantly regardless of the outcome of oocyte IVF.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1993

Distribution of free amino acids in human preovulatory follicles.

The purpose of the present study was to investigate the possible role of intrafollicular amino acids in the human ovarian follicle. Follicular fluid (FF) and oocytes were obtained from 12 women for in vitro fertilization. Follicular development was induced with clomiphene citrate and human menopausal gonadotropin. Thirty-six FF samples, free of visible blood contamination and containing mature oocytes were used to measure free amino acids, steroids and gonadotropins. The FF were divided into three groups: (1) follicles yielding oocytes that were unfertilized (n = 12); (B) follicles containing oocytes that were fertilized and cleaved to less than four cells (n = 12); (C) follicles yielding oocytes that were fertilized and cleaved to four or more cells (n = 12). The concentrations of histidine, phenylalanine and asparagine in FF were significantly greater than those in plasma at the time of follicle aspiration. However, amino acid concentrations in FF did not differ significantly between the three groups studied. No significant relationships were found between intrafollicular levels of amino acids and those of LH, FSH, estradiol, progesterone and testosterone. These results suggest that intrafollicular amino acids are not involved in the regulation of human preovulatory follicle.

Amino Acids↗
PubMed1993

Lack of expression of HLA antigens on immature germ cells from ejaculates with antisperm antibodies.

PROBLEM: The lack of expression of HLA antigen on immature germ cells from ejaculates with antisperm antibodies has been reported. METHOD: The expression of human leukocyte antigens on immature germ cells from ejaculates with antisperm antibodies (ASA) was investigated by indirect immunofluorescence using a panel of monoclonal antibodies (MAb) and automated flow cytometry. Patients were divided into two groups: fertile (prevasectomic; N = 10), and ejaculates with ASA (10 samples with IgG and IgA ASA, and five semen samples with only IgG ASA). ASA were detected on sperm using the direct immunobead test. After centrifuging semen samples on a Ficoll-Hypaque gradient, round cells obtained at the gradient interface were gated by a flow cytometer. The "immature germ cell window" was defined in terms of cellular volume and granularity. RESULTS: The percentage of gated round cells from semen samples that reacted with anti-CD45 was always less than 5%, and with anti-CD44 less than 3%. This lack of reactivity of gated round cells with MAb specific for leukocytes and epithelial cells suggests that they were immature germ cells. Immature germ cells were unreactive with W6/32 and anti-beta-2-microglobulin MAb, which suggests that these cells do not express HLA class I molecules. Similarly, no reactivity of the immature germ cells with the MAb that recognize HLA class II molecules was found. No significant differences were observed in the expression of HLA molecules on immature germ cells between the different semen samples studied: fertile, and ejaculates with ASA. CONCLUSION: The presence of ASA in ejaculate is not associated with abnormal HLA antigen expression on immature germ cells.

Animals↗
PubMed1993

Diagnosis of pelvic inflammatory disease with 99mtechnetium-hexamethylpropylenamine-oxime-labeled autologous leukocytes and pelvic radionuclide scintigraphy.

In a prospective study, we assessed the performance of pelvic radionuclide scintigraphy after the injection of 99mtechnetium-hexamethylpropylenamine-oxime-labeled autologous leukocytes as a noninvasive tool for the differential diagnosis of pelvic inflammatory disease (PID). The results of radionuclide scintigraphy in 20 women with PID confirmed laparoscopically were compared with the findings in 20 others hospitalized for suspected PID but with PID ruled out later by laparoscopy. The proportion of radionuclide scintigraphic findings demonstrating increased uptake in the genital region, compatible with an inflammatory process, was significantly larger (P < .001) in patients with PID (95%) than in those without PID (15%). The sensitivity of the technique was 95% and specificity was 85%; in all, 90% of the patients were correctly classified. We conclude that radionuclide scintigraphy preceded by the injection of 99mtechnetium-hexamethylpropylenamine-oxime-labeled autologous leukocytes can detect PID in a high proportion of patients.

Female↗
PubMed1992

Adrenal hormones in human follicular fluid.

Considerable evidence indicates that adrenal hormones may affect gonadal function. To assess the role of some adrenal hormones in human follicular fluid and their relationship with the ability of the oocyte to be fertilized and then to cleave in vitro, cortisol and dehydroepiandrosterone sulfate were measured in follicular fluid obtained at the time of oocyte recovery for in vitro fertilization from cycles stimulated by clomiphene citrate, human menopausal gonadotropin and human chorionic gonadotropin. Thirty-six follicular fluid containing mature oocyte-corona-cumulus complexes and free of visible blood contamination were included in this study. There was no significant difference in follicular fluid dehydroepiandrosterone sulfate concentration between follicles with oocytes which did or did not fertilize (5.1 +/- 1.1 vs 5.8 +/- 2.0 mumol/l). However, follicular fluid from follicles whose oocytes were not fertilized had levels of cortisol significantly higher than those in follicular fluid from follicles containing successfully fertilized oocytes (406.0 +/- 75.9 vs 339.2 +/- 37.0 nmol/l; p < 0.005). No significant correlations were found between rates of embryo cleavage and cortisol and dehydroepiandrosterone levels in follicular fluid. We conclude that cortisol levels in follicular fluid may provide an index of fertilization outcome, at least in stimulated cycles by clomiphene citrate, human menopausal gonadotropin and human chorionic gonadotropin.

Adrenal Cortex Hormones↗
PubMed1992

Insulin and insulin-like growth factor I in follicular fluid after induction of ovulation in women undergoing in vitro fertilization.

This study was undertaken to evaluate the relationship between concentrations of insulin and insulin-like growth factor I (IGF-I) in follicular fluid and fertilization and cleavage of human oocytes fertilized in vitro. The concentration of oestradiol, progesterone, luteinizing hormone, follicle-stimulating hormone, testosterone, insulin and IGF-I was determined in 36 follicular fluids, free of visible blood contamination and containing mature oocyte-corona-cumulus complexes, obtained from 12 women undergoing in vitro fertilization. Follicular development was induced by clomiphene citrate and human menopausal gonadotrophin, and follicular aspiration was performed 35 h after an ovulatory dose of human chorionic gonadotrophin. Concentrations of IGF-I were significantly higher in follicular fluids associated with mature oocytes that fertilized and cleaved, than in follicular fluid associated with mature oocytes that did not fertilize (P < 0.001). There was no difference in the concentration of insulin between follicular fluids from which fertilized oocytes were obtained and those with oocytes that remained unfertilized. No significant correlations were found between rates of embryo cleavage, concentrations of insulin and IGF-I. Multiple linear regression analysis demonstrated that the concentrations of IGF-I in follicular fluid were predicted statistically by a negative regression coefficient for the concentration of testosterone, and by a positive regression coefficient for the concentration of progesterone in follicular fluid. No candidate variable was included in the model to predict concentrations of insulin. These data suggest an important role for IGF-I in the mature follicle.

Adult↗
PubMed1992

[Diurnal variations in circulating leukocytes and subsets: relation to plasma cortisol and ACTH].

Surface cellular antigens of leukocytes, lymphocytes and corresponding subpopulations have been analysed by using monoclonal antibodies marked with fluorescein (PITC), parallel to those marked with phycoerythrin (PE). Cortisol and ACTH plasmatics have also been determined through RIA, on two samples at 8am. and 8pm. During this twelve hour evolution, a highly significant dependence of the leukocytes, T, T4 and T8 lymphocytes on the circulating ACTH has also been found. In general during the experiment time leukocytes, lymphocytes and subpopulations, have experimented an increase which is significantly related to the pituitary hormone secretion. The existence of this significant correlation establishes the presence of a possible mechanism that connects the cellular immunity to determined hypothalamus hormones.

Adrenocorticotropic Hormone↗
PubMed1990

Short-lived suppressor cell activity during normal human pregnancy.

Short-lived suppressor cell (SLSC) activity was determined in normal pregnant women. This activity was significantly increased in all three trimesters of pregnancy and during the first week postpartum. When pregnant women were divided into primiparous and multiparous groups, no significant differences were found between the two groups in any of the periods studied. These results suggest that increased SLSC activity may play a role in the materno-fetal tolerance and that parity has no influence on this activity.

Adult↗
PubMed1990

Immunosuppressive properties of human follicular fluid.

Human preovulatory follicular fluids (FF) obtained in the course of stimulated cycles were analyzed for their possible immunologic functions. Different concentrations of FF (20%, 2%, 1%) inhibited the mitogenic response of normal human lymphocytes to concanavalin A (Con A). Lymphocytes were assessed for immunosuppressor activity after preincubation with FF. Lymphocyte mitogenic response to Con A was only suppressed by cells preincubated with FF at concentrations of 2% and 1% for at least 48 hours. No evidence of suppressor cell induction was seen following incubation of lymphocytes with 20% FF, nor was any significant relationship between FF immunosuppressor activity and the outcome of in vitro fertilization observed. We conclude that some factor(s) in FF may be capable of directly inhibiting lymphocyte response and inducing immunosuppressor cell activity in vitro.

Body Fluids↗