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Biomedical subjects

J A Castilla

Publications and source records attributed to J A Castilla.

34 records · Page 2Linked to original sources

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↗

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↗

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↗

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↗

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↗

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↗

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↗

[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↗

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↗

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↗

Decreased levels of circulating CD4+ T lymphocytes during normal human pregnancy.

Peripheral blood mononuclear cell (PBMC) populations during human pregnancy have been investigated by many authors, although the different results obtained, principally in relation to T cells, are very discrepant. In this study we aimed to exclude all the possible causes of these discrepancies: small sample size; diurnal rhythm of CD4+ T cells; smoking habits; haemodilution which occurs during pregnancy and inappropriate statistical analysis; in order to determine whether gestation has a definite effect on PBMC populations. We found that the percentage of CD4+ T lymphocytes decreases in the first and second trimesters, returns to the non-pregnant level in the third trimester and remains there in the postpartum period. The percentages of CD3+ T lymphocytes run parallel to those of CD4+ while CD8+ T lymphocytes do not vary. The proportion of CD16+ cells, which include mature NK cells, diminishes in the second trimester and this reduction is maintained in the third trimester and the puerperium. No variation was found in the other PBMC studied (CD20+ lymphocytes, CD14+ monocytes and D/DR+ cells). When parity was considered no difference was seen between primiparous and multiparous women in any of the cell populations tested.

Adult↗

Mononuclear cell subpopulations in human follicular fluid from stimulated cycles.

The study of lymphocyte subsets from human follicular fluid (FF) provides an opportunity to evaluate immunological features of the ovary. We investigated the mononuclear cell subsets in FF and peripheral blood obtained at the time of laparoscopy from ten in vitro fertilization (IVF) patients. Midcycle nonpregnant peripheral blood was used as the control. A marked increase in the proportion of monocytes (CD14+) was observed in FF. Although FF was enriched with CD8+ lymphocytes, a decrease in the proportion of CD4+ lymphocytes was observed. "Memory" T cells in FF, identified by the CD4+ CD45R- phenotype, predominated over "naive" T cells (CD4+ CD45R+) at a ratio of 2:1, which differs from the ratio yielded by control blood samples (1:1). The percentage of activated T cells (CD3+ HLA-DR+ cell) increased significantly in FF. When lymphocyte subsets were studied in the peripheral blood of IVF patients, changes similar to but less significant than those in FF were found. These data support the concept that lymphocytes play an important role in ovarian physiology.

Antibodies, Monoclonal↗