Biomedical subjects

J A Castilla

Publications and source records attributed to J A Castilla.

At least 19 recordsLinked to original sources

PubMed2005

Influence of analytical and biological variation on the clinical interpretation of seminal parameters.

Quality assurance in semen analysis has been questioned recently in this journal. Based on the limited capacity of seminal parameter in the determination of fertility, the authors advocated abandoning methods of quality assurance in semen analysis for clinical situations. In this article, we explore arguments as to why quality assurance in semen analysis for clinical use is not 'a waste of time'. Imprecision and within-subject biological variations are the two major components involved in the dispersion of seminal parameter results obtained by analysis of a semen sample from an individual. As within-subject biological variation is constant across geography, time and population, imprecision is a very important factor in the quality of laboratory test results. We analyse this influence on various seminal parameters and observe that there is an amount of error that can be tolerated without invalidating the medical usefulness of seminal parameter determination. However, there is a maximum allowable analytical error above which the medical usefulness of seminal parameter results is invalidated. The level of performance required to facilitate clinical decision-making is termed quality specification. We comment on different strategies to define the maximum allowable analytical error.

Clinical Laboratory Techniques↗

PubMed2005

Quality specifications for seminal parameters based on the state of the art.

BACKGROUND: The aim of this study was to calculate the analytical goal for seminal parameters based on the state of the art, and then to compare these specifications with those previously obtained by our group based on biological variation. METHODS: All data used for analysis were derived from the Spanish programme of external quality control on semen analysis. Over 90 laboratories participated from 1999 to 2003. Using graphs of the state of the art, we also determined the numbers of laboratories that achieved quality specifications. RESULTS: The total allowable error calculated using state of the art graphs is similar to that calculated using biological variation for concentration and total motility. However, it is much higher for morphology and rapidly progressive motility. Over 80% of the laboratories achieved the minimum quality specification based on biological variation for concentration, total and progressive motility. However, only approximately 30% of the laboratories achieved the minimum quality specification based on biological variation for morphology and rapidly progressive motility. CONCLUSIONS: The study enabled us to identify the state of the art of analytical performance for seminal parameters, and revealed the difficulty inherent in meeting the quality specifications based on biological variation.

Andrology↗

PubMed2005

External quality control program for semen analysis: Spanish experience.

PURPOSE: Results from an external quality control programme for semen analysis carried out in Spain are analysed. METHODS: Quality control materials were distributed and the following seminal parameters were determined: concentration, total motility, progressive motility, rapid progressive motility, morphology and sperm vitality. The between-laboratories coefficients of variation were assessed on different types of quality control material. RESULTS: The majority of participating laboratories utilised manual versus computer-assisted semen analysis methods. Some between-laboratories coefficients of variation ranges were: 20.8-33.8% for concentration (semen pool suspension); 13.9-19.2% for total motility (videotapes); 54.2-70.2% for sperm morphology (strict criteria using stained smears); and 9.8-41.1% for sperm vitality (stained smears). There was an inverse relation between mean percentage of sperm and coefficients of variation between laboratories for sperm motility, morphology and vitality. CONCLUSIONS: These data highlight the urgent need for improvement in the overall quality of andrology testing.

Clinical Laboratory Techniques↗

PubMed2004

Expression of integrin fraction and adhesion molecules on human granulosa cells and its relation with oocyte maturity and follicular steroidogenesis.

PURPOSE: To study the correlation between the expression of integrin fractions and adhesion molecules on granulosa cells (GC) and follicular development. METHODS: GC and follicular fluid (FF) were obtained at oocyte retrieval for ICSI. Expression of adhesion molecules on GC was studied by flow cytometry. Statistics were evaluated using the Student t test and simple linear regression analysis. RESULTS: alpha5 integrin fraction was significantly (p < 0.01) higher, while alpha6 fraction and CD9 were significantly (p < 0.01 and p < 0.001, respectively) lower in GC from FF with metaphase II oocytes. A direct significant correlation was observed between FF progesterone and the alpha5 expression on GC (r = 0.54). In contrast, an inverse significant correlation was observed between FF progesterone level and the expression of alpha6 and CD9 (r = -0.40 and -0.41, respectively). CONCLUSIONS: The expression pattern for integrin fractions and adhesion molecules may be of predictive value in assessing the state of differentiation of the human follicle.

Estradiol↗

PubMed2003

Biological variation of seminal parameters in healthy subjects.

BACKGROUND: A study was undertaken to assess the components of biological variation of seminal parameters in healthy subjects. METHODS: Twenty donor candidates were included in a 10-week follow-up study. Within- and between-subject biological variation, indices of individuality and heterogeneity, coefficient of reliability, critical differences, analytical goals and the lowest value observed with a <5% probability of having a true value less than the World Health Organization (1999) reference value were estimated for the following seminal parameters: concentration, total motility (WHO grades a + b + c), progressive motility (grades a + b), rapid progressive motility (grade a), sperm morphology and vitality. All analysis was performed by a single technician according to WHO 1999 guidelines for routine semen analysis. Analytical variation was assessed on different types of quality control material (frozen straws, sperm suspension, videotape, and slides) and at different (low, medium, high) quality levels. RESULTS: The analytical variation observed depended on the quality control material used and the level of semen quality. Concentration was the semen parameter with highest within- and between-subject variation, and vitality the lowest. Indices of individuality were all <0.7, and coefficients of reliability were high (0.68-0.84). The critical difference for sequential values significant at P < 0.05 for vitality, progressive motility and morphology (34.4, 49.2 and 58.0% respectively) were lower than for concentration (77.8%). CONCLUSIONS: The study results showed that conventional reference values for seminal parameters have little diagnostic value because of their marked individuality, though seminal parameters can be useful for assessing differences in an individual's serial results, in particular of progressive motility, morphology and vitality.

Adult↗

PubMed2001

Alterations in sperm protein phosphorylation in male infertility.

Protein phosphorylation is involved in sperm capacitation, so the effect of protein phosphatase inhibitors on the capacitation of spermatozoa of males with unexplained infertility was investigated. d-mannose ligand specific receptor expression in fresh, living spermatozoa, capacitated or treated with calyculin A (an inhibitor of protein phosphatases 1 and 2A), was studied in three groups of men: pre-vasectomy (fertile) males, males in couples with male infertility, and males in couples with infertility of unknown aetiology. Flow cytometry showed significant differences between infertile couples with a male factor and fertile couples (P < 0.05), both after capacitation and after treatment with calyculin A. In the group of couples with infertility of unknown aetiology (n = 15), d-mannose receptor expression was diminished in six cases after classical capacitation. However, when the spermatozoa of these six men were treated with calyculin A, five showed an increased specific d-mannose receptor expression. From these results it is suggested that in vitro treatment of spermatozoa with inhibitors of protein phosphatases may be of great value in some cases of unexplained infertility.

Flow Cytometry↗

PubMed2001

Chromatin status in human ejaculated spermatozoa from infertile patients and relationship to seminal parameters.

The aim of this study was to evaluate the chromatin status in different groups of patients. Five groups of men were selected: pre-vasectomy; male factor infertility; varicocele; immunological male infertility; and idiopathic infertility. Chromatin status was evaluated using flow cytometry after staining the DNA with the fluorochrome propidium iodide. Differences were observed in the state of sperm chromatin between the male factor and varicocele groups with respect to the others. These two groups presented poorer quality chromatin, as evidenced fundamentally by a lower degree of condensation. These deficiencies in chromatin status were usually accompanied by alterations in the other standard parameters of semen analysis. Individuals who are infertile due to male factor and those presenting varicocele have spermatozoa with less condensed chromatin which might, in part, explain their sterility.

Chromatin↗

PubMed2001

[Alterations of nuclear maturity in spermatozoids of males with antecedents of cryptorchidism].

The association between infertility and cryptorchidism is an accepted fact, usually attributed to the oligozoosperm, asthenozoosperm or teratozoosperm presented in ejaculation products of males with this antecedent. The nuclear maturity in a sample of men with antecedents of cryptorchidism have been studied and these results have been compared to those of a control group. The results of this work show the deficient transformation of nuclear proteins to protamines in males with antecedents of cryptorchidism compared to the control group, due to the remaining of immature histones. Alterations of nuclear maturity able to contribute to the subfertility of these men were found in spermatozoids of adult males with antecedents of cryptorchidism.

Cryptorchidism↗

PubMed2000

Decreased sperm function of patients with myotonic muscular dystrophy.

Myotonic muscular dystrophy (MMD) is a genetic disease caused by a defective enzyme, myotoninkinase. Male patients with MMD are reported to have reduced fertility. The purpose of this work was to study sperm capacitation and acrosome reaction in the ejaculates of sterile males with MMD and of healthy males (control group). The expression of the specific D-mannose receptors was explored by microscopic examination and by flow cytometry analysis. In addition, the binding patterns of Pisum sativum (PSA) lectin to acrosome content and outer acrosomal membrane in the spermatozoa of each group were analysed. Both the capacitation and the acrosome reaction in the spermatozoa of the MMD group were deficient and these findings strongly suggest that these anomalies may account for the sterility of these patients.

Acrosome Reaction↗

PubMed1999

Tumour necrosis factor-alpha and interleukin-1 and -6 in fibrocystic breast disease.

The risk of developing breast cancer is higher in women presenting gross cystic disease (cysts > 3 mm in diameter) of the breast with intracystic K+/Na+ > 3 as compared with K+/Na+ < 3. The present study reports the levels of tumour necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), and interleukin-6 (IL-6) in the breast cyst fluid of women with gross cystic disease and analyses the relationship between the intracystic concentration of these cytokines, sex steroid hormones, and the K+/Na+ ratio. The concentration of these cytokines, estradiol, testosterone, dehydroepiandrosterone sulfate (DHEA-S), and 17-OH-progesterone were determined in the breast cyst fluid of 54 women with gross cystic disease. No significant differences were found in the cystic levels of IL-1 between cysts with intracystic K+/Na+ < 3 and > 3. However, in cysts with intracystic K+/Na+ > 3 we found a lower concentration of IL-6 and TNF-alpha than in those with intracystic K+/Na+ < 3. Stepwise multiple linear regression analysis demonstrated that the concentration of IL-6 in breast cyst fluid was predicted statistically by a negative regression coefficient for the concentration of estradiol and DHEA-S, and by a positive regression coefficient for the concentration of TNF-alpha. The concentration of TNF-alpha in breast cyst fluid was predicted statistically by a positive regression coefficient for the concentration of IL-6, and by a negative regression coefficient for the concentration of estradiol. No candidate variable was included in the model to predict concentrations of IL-1 in breast cyst fluid. Our results indicate that IL-6 and TNF-alpha could have a local 'protector' role in gross cystic disease, and that they could be used as a marker to identify cyst type.

17-alpha-Hydroxyprogesterone↗

PubMed1998

Increase of large granular lymphocytes in human ejaculate containing antisperm antibodies.

Using flow cytometry, we studied the expression of the CD16 antigen by lymphocytes present in human semen samples from three groups of patients: 60 fertile men attending for vasectomy, 60 sterile patients without antisperm antibodies (ASA) and 18 immunological sterile patients with ASA in their ejaculate. No significant difference was found in the concentration of leukocytes or subpopulations of these cells (monocytes, lymphocytes and granulocytes) between fertile, sterile without ASA and immunological sterile groups. However, we detected a predominance of macrophages/monocytes within the population of seminal leukocytes. No statistically significant difference was found in the absolute number of T and B lymphocytes between the three groups studied. However, a significant increase in the number of CD16+ lymphocytes was observed in the ejaculate of sterile patients with ASA as compared to the other groups. This finding might establish an important parameter in the follow-up and prognosis of patients with immunological sterility.

Adult↗

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↗