The contribution of different sperm parameters to better explain ram semen cryoresistance.

In order to determine an effective procedure for explaining ram sperm cryoresistance and develop a new model for breeders classification, a retrospective study was conducted using sperm analysis data obtained over two consecutive years from a total of 82 sessions of ram semen cryopreservation. In each session, fresh ejaculates from eight males were collected via artificial vagina, pooled and frozen in liquid nitrogen vapors. After thawing, a total of 19,084 sperm tracks and 11,319 morphometric measurements were analysed. Clustering analyses were applied to establish motile and morphometric sperm subpopulations. Additionally, plasma and acrosome membrane integrity, as well mitochondrial activity using flow cytometry immediately after sperm thawing and following hypoosmotic shock test (HOST) was assessed. To develop a Ram Sperm Cryoresistance Index, Principal Component Analyses (PCA) using 22 variables were conducted. In the first PCA, the parameters that best explain cryoresistance include total motility (TM), motile subpopulation 2 (motSP2, which groups slow, very linear spermatozoa with low lateral head displacement), morphometric subpopulation 1 (morphSP1, grouping spermatozoa with the smallest head size and lowest shape values), sperm plasma membrane integrity immediately after thawing and following hypoosmotic shock test. These parameters collectively account for 77.34 % of the accumulated variance. To emphasize their importance, a second PCA was performed, revealing significant higher weighting coefficients for the quantity (TM) and quality (motSP2) of sperm movement after thawing, compared to the head size and shape of the thawed sperm (morphSP1). Furthermore, HOST Viability played a more decisive role than what was observed under isotonic conditions.

Variability in in vitro fertilization outcomes of prepubertal goat oocytes explained by basic semen analyses.

This work analyses the changes that caprine spermatozoa undergo during in vitro fertilization (IVF) of in vitro matured prepubertal goat oocytes and their relationship with IVF outcome, in order to obtain an effective model that allows prediction of in vitro fertility on the basis of semen assessment. The evolution of several sperm parameters (motility, viability and acrosomal integrity) during IVF and their relationship with three IVF outcome criteria (total penetration, normal penetration and cleavage rates) were studied in a total of 56 IVF replicates. Moderate correlation coefficients between some sperm parameters and IVF outcome were observed. In addition, stepwise multiple regression analyses were conducted that considered three grouping of sperm parameters as potential explanatory variables of the three IVF outcome criteria. The proportion of IVF outcome variation that can be explained by the fitted models ranged from 0.62 to 0.86, depending upon the trait analysed and the variables considered. Seven out of 32 sperm parameters were selected as partial covariates in at least one of the nine multiple regression models. Among these, progressive sperm motility assessed immediately after swim-up, the percentage of dead sperm with intact acrosome and the incidence of acrosome reaction both determined just before the gamete co-culture, and finally the proportion of viable spermatozoa at 17 h post-insemination were the most frequently selected sperm parameters. Nevertheless, the predictive ability of these models must be confirmed in a larger sample size experiment.

Prepubertal goat oocytes from large follicles result in similar blastocyst production and embryo ploidy than those from adult goats.

Developmental competence of oocytes from prepubertal females is lower than those from adult females. Oocyte development competence is positively related to follicular diameter. Most of the follicles of prepubertal goat ovaries are smaller than 3 mm. The aim of this study was to compare oocytes of two follicle sizes (< 3 mm and ≥ 3 mm) from prepubertal goats with oocytes from adult goats in relation to their in vitro production and quality of blastocysts. Oocytes from prepubertal goats were obtained from slaughterhouse ovaries and selected according to the follicle diameter whereas oocytes from adult goats were recovered in vivo by LOPU technique without prior selection of follicle size. COCs were IVM for 27 h, IVF at the conventional conditions with fresh semen and presumptive zygotes were cultured in SOF medium for 8 days. Blastocysts obtained were vitrified and after warming their blastocoele re-expansion and the ploidy by FISH technique were assessed. We found significant differences between blastocysts yield of oocytes recovered from follicles smaller than 3 mm of prepubertal goats compared to those from adult goats (5.45% vs 20. 83%, respectively) however, these differences disappear if oocytes were recovered form large follicles (18.07%). A total of 28 blastocysts were analysed and 96.43% showed mixoploidy. Age did not affect the number of embryos with abnormal ploidy or blastocyst re-expansion after warming. Furthermore, the percentage of diploid blastomeres per embryo was similar in the 3 groups studied, adult, prepubertal from follicles ≥ 3 mm and < 3 mm (68.6%, 80.8% and 73.6%, respectively). In conclusion, IVP of blastocysts coming from follicles larger than 3 mm of goats 45 days old were not different to the blastocysts produced from adult goats, both in terms of quantity and quality.

Oocyte secreted factors improve embryo developmental competence of COCs from small follicles in prepubertal goats.

Oocytes secrete soluble paracrine factors called Oocyte Secreted Factors (OSFs) which regulate the cumulus cell phenotype. Follicle populations in ovaries from prepubertal females have smaller diameters than their adult counterparts. Oocytes from small follicles are less competent than those from large follicles. The aim of this study was to investigate, in prepubertal goats, the effect of OSFs secreted by denuded oocytes (DOs) from small (<3 mm) or large (>or=3 mm) follicles during IVM on embryo development and the blastocyst quality of cumulus-oocyte complexes (COCs) from small follicles and to determine if GDF9 participates in this process. Treatment groups were: (A) COCs non selected by their follicle size (control group); (B) cumulus oocytes complexes from small follicles (SFCOCs), (C) cumulus oocytes complexes from small follicles co-cultured with denuded oocytes from small follicles (SFCOCs + SFDOs), and (D) cumulus oocytes complexes from small follicles co-cultured with denuded oocytes from large follicles (SFCOCs + LFDOs). The effect of the addition of kinase inhibitor SB-431542, which antagonizes GDF9, was tested in A, C, and D treatment groups. Co-cultured SFCOCs with SFDOs or LFDOs significantly augmented the blastocyst rate in comparison to SFCOCs alone (15.77%, 17.39% vs. 10.31%, respectively). Blastocysts from SFCOCs + LFDOs group showed higher rates of tetraploid nuclei than blastocysts from SFCOCs and the control group (14.43% vs. 5.45% and 5.24%, respectively; P < 0.05). However, we did not observe differences in the hatching rate, mean cell number or embryo cryotolerance (P > 0.05) between the four treatment groups. The addition of SB-431542 during IVM did not have any effect on blastocyst rate (P > 0.05). In conclusion, in prepubertal goats, COCs with a low embryo developmental competence as a consequence of follicle size can be improved by coculturing them with denuded oocytes from both small and large follicles. GDF9 does not seem play a role in this improvement.

The influence of sperm concentration, length of the gamete co-culture and the evolution of different sperm parameters on the in vitro fertilization of prepubertal goat oocytes.

The aims of the present study were: (1) to evaluate the influence of sperm concentration (ranging from 0.5 × 10(6) to 4 × 10(6) spermatozoa/ml) and length of the gamete co-incubation time (2, 4, 6, 8, 10, 12, 16, 20, 24 or 28 h) on in vitro fertilization (IVF), assessing the sperm penetration rate; (2) to investigate the kinetics of different semen parameters as motility, viability and acrosome status during the co-culture period; and (3) to analyse the effect of the presence of cumulus-oocytes complexes (COCs) on these parameters. To achieve these objectives, several experiments were carried out using in vitro matured oocytes from prepubertal goats. The main findings of this work are that: (1) in our conditions, the optimum sperm concentration is 4 × 10(6) sperm/ml, as this sperm:oocyte ratio (approximately 28,000) allowed us to obtain the highest penetration rate, without increasing polyspermy incidence; (2) the highest percentage of viable acrosome-reacted spermatozoa is observed between 8-12 h of gamete co-culture, while the penetration rate is maximum at 12 h of co-incubation; and (3) the presence of COCs seems to favour the acrosome reaction of free spermatozoa on IVF medium, but not significantly. In conclusion, we suggest that a gamete co-incubation for 12-14 h, with a concentration of 4 × 10(6) sperm/ml, would be sufficient to obtain the highest rate of penetration, reducing the exposure of oocytes to high levels of reactive oxygen species produced by spermatozoa, especially when a high sperm concentration is used to increase the caprine IVF outcome.

[Chemosensitivity test on brain tumors]. / Ensayos de quimiosensibilidad en cultivos primarios de tumores cerebrales.

UNLABELLED: During last 50 years chemotherapy has played a very important part in the cancer treatment. However, success or failures of news drugs in one particular cancer its difficult to predict. In vitro chemosensitivity is an attractive method for knowing about responses of a tumor to ChT treatment and assess the best dose in the patient with cancer. OBJECTIVE: To know brain tumors sensitivity against antineoplastic drugs. METHODS: Five different drugs (carmustin, camptotecin, taxol, hydroxyurea and tamoxifen) were tested on short-term cultures from 7 patients with Glioblastoma multiforme, 15 patients with meningiomas and one patient with meduloblastoma. For testing chemosensitivity we used MTT assay, and we measured optic density by spectophotometry to 450 nm. RESULTS. A total of 49 measurement were done, getting 44 valid dose-result curves. For each drug we used from 10-2 M to 10-12 M gap, and IC50 result was representative of tumor sensitivity to the drug. CONCLUSION: our data support MTT assays like valid method for measuring in vitro chemosensitivity in brain tumors to news drugs.

Tungstate administration improves the sexual and reproductive function in female rats with streptozotocin-induced diabetes.

BACKGROUND: Diabetes induces great alterations in female reproductive function. We analyzed the effects of tungstate, an anti-diabetic agent, on the reproductive function of healthy and diabetic female rats. METHODS: Healthy and streptozotocin-induced diabetic rats were treated with sodium tungstate (2 mg/ml in their drinking water) for 12 weeks. Markers of reproductive function and diabetes were measured in serum, and in uterus and ovaries by Western blot or RT-PCR. Reproductive function was also assessed by mating. RESULTS: Diabetic rats showed great impairment of libido, which was accompanied by a total loss of fertility (P < 0.05) and a decrease in the serum levels of FSH (P < 0.05) and LH (P < 0.05) compared with healthy rats. Tungstate treatment of diabetic rats partially recovered libido while fertility rate increased to 66.6%. This improvement was accompanied by a recovery of serum FSH (to a level higher than healthy rats) and LH. Moreover, tungstate treatment normalized ovarian expression of GLUT 3 hexose transporter, and estrogen, progesterone and FSH receptors, whereas only GLUT 3 and FSH receptors were normalized in the uterus. CONCLUSIONS: Our results indicate that the alterations in female reproduction in diabetes were partially reversed after tungstate treatment by a mechanism(s) involving the normalization of serum FSH and LH levels, and ovarian and uterine expression of FSH receptors and GLUT3.

[Analysis and predictor of delays in the suspicion and treatment among hospitalized patients with pulmonary tuberculosis]. / Análisis y factores predictores del retraso en la sospecha diagnóstica y tratamiento de los pacientes hospitalizados con tuberculosis pulmonar.

BACKGROUND AND OBJECTIVES: Sometimes tuberculosis diagnosis is missed at hospital admission. Delayed diagnosis of active pulmonary tuberculosis among hospitalized patients could to contribute to nosocomial transmission. The objective of this study was to define the occurrence and associated patient risk factors among hospitalized patients with delayed diagnosis of respiratory tuberculosis. METHODS AND PATIENTS: A retrospective chart review was undertaken between 1995 and 2002 on all patients with pulmonary tuberculosis. Time intervals between admission, diagnosis and treatment of tuberculosis were determined. Epidemiological and clinical features were evaluated for their effect on these time intervals. RESULTS: Among 149 patients newly diagnosed to have active pulmonary TB, the diagnosis was initially missed in 102 (68% [95% CI, 61 to 75%]) of all hospitalized patients, of whom 66 (65% [95% CI, 56 to 74%]) were smear positive. Treatment was initiated after a week or more in 62 (42% [95% CI, 34% to 50%]) of all patients, of whom 34 (55% [95% CI, 43 to 67%])) were smear positive. Age >or= 60 years (OR 3.17 [95% CI; 1.27 to 7.87]; p = 0.013), presence of chronic lung disease (OR 2.99 [95% CI; 1.21 to 7.38]; p = 0.017), negative sputum AFB smear (OR 4.51 [95% CI; 1.34 to 15.16]; p = 0.015) and absence of hemoptysis or melanoptysis (OR 2.96 [95% CI; 1.18 a 7.41]; p = 0.020), were independently associated with delays. CONCLUSIONS: The diagnosis and treatment of hospitalized patients with pulmonary tuberculosis is often delayed because absence of clinic suspicion owing to old age, chronic lung disease or atypical presentations and slow confirmation by culture. Improved clinical acumen, development of rapid diagnostic tests, and the institution of early empiric therapy are desirable objectives to improve the tuberculosis control.

Modeling photosynthesis in olive leaves under drought conditions.

We quantified parameters for a model of leaf-level photosynthesis for olive, and tested the model against an independent dataset. Specific temperature-dependence parameters of the model for olive leaves were measured, as well as the relationship of the model parameters with area-based leaf nitrogen (N) content. The effect of soil water deficit on leaf photosynthesis was examined by applying two irrigation treatments to 29-year-old trees growing in a plantation: drip irrigation sufficient to meet the crop water requirements (I) and dry-farming (D). In both treatments, leaves had a higher photosynthetic capacity in April than in August. In August, photosynthetic capacity was lower in D trees than in I trees. Leaf photosynthetic capacity was linearly and positively related to leaf N content on an area basis (N(a)) and to leaf mass per unit area (LMA), and the regression slope varied with irrigation treatment. The seasonal reduction in N(a) was used in the model to predict photosynthesis under drought conditions. Olive leaves showed a clear limitation of photosynthesis by triose phosphate utilization (TPU) even at 40 degrees C, and the data suggest that olive invests fewer resources in TPU than other species. The seasonal decrease in photosynthetic capacity moderated the stomatal limitation to carbon dioxide (CO(2)) fixation as soil water deficit increased. Further, it enabled leaves to operate close to the transition point between photosynthetic limitation due to RuBP carboxylation capacity and that due to RuBP regeneration capacity, and resulted in a near constant value of internal CO(2) concentration from April to August. Under well watered conditions, N-use efficiency of the olive leaves was enhanced at the expense of reduced water-use efficiency.

Calibration of sap flow estimated by the compensation heat pulse method in olive, plum and orange trees: relationships with xylem anatomy.

The compensation heat pulse method is widely used to estimate sap flow in conducting organs of woody plants. Being an invasive technique, calibration is crucial to derive correction factors for accurately estimating the sap flow value from the measured heat pulse velocity. We compared the results of excision and perfusion calibration experiments made with mature olive (Olea europaea L. 'Manzanilla de Sevilla'), plum (Prunus domestica L. 'Songal') and orange (Citrus sinensis (L.) Osbeck. 'Cadenero') trees. The calibration experiments were designed according to current knowledge on the application of the technique and the analysis of measured heat pulse velocities. Data on xylem characteristics were obtained from the experimental trees and related to the results of the calibration experiments. The most accurate sap flow values were obtained by assuming a wound width of 2.0 mm for olive and 2.4 mm for plum and orange. Although the three possible methods of integrating the sap velocity profiles produced similar results for all three species, the best results were obtained by calculating sap flow as the weighted sum of the product of sap velocity and the associated sapwood area across the four sensors of the heat-pulse-velocity probes. Anatomical observations showed that the xylem of the studied species can be considered thermally homogeneous. Vessel lumen diameter in orange trees was about twice that in the olive and plum, but vessel density was less than half. Total vessel lumen area per transverse section of xylem tissue was greater in plum than in the other species. These and other anatomical and hydraulic differences may account for the different calibration results obtained for each species.

Effects of exposing boars to different artificial light regimens on semen plasma markers and "in vivo" fertilizing capacity.

This study was designed to assess the effects of exposing boars to an artificial photoperiod on semen quality in terms of sperm concentration, sperm vitality, sperm motility and acrosome integrity. We also determined biochemical semen plasma variables, such as total protein concentration, phosphorylated tyrosine residues and fructose, glucose and sorbitol contents, along with their effects on the fertility, prolificacy and libido of the boars. Three groups of 10 males were kept for 3 months under experimental conditions of 24, 12 and 0 h of artificial light, and a constant temperature of 21 +/- 1 degrees C and 60-75% humidity. The animals were fed a nutritious diet and subjected to semen collection twice per week. Semen samples were analyzed throughout the entire experimental period. Our results indicate that, while the extreme photoperiods (0 and 24 h of light) affected semen quality in terms of sperm concentration, acrosome integrity and semen volume, its fertilizing capacity was only significantly reduced under conditions of absolute darkness. Sperm motility was found to be a poor indicator of fertilizing ability, while other sperm factors, such as acrosome integrity or other functional variables seemed to behave better. The photoperiod was found to affect the production of accessory sex gland secretions more than their composition. In addition, light effects on fertility, prolificacy and libido seemed to be achieved through independent mechanisms.

Multiple pulmonary neoplasia positive for thyroglobulin.

We present, for the first time, a case of carcinoid metastatic pulmonary tumor positive for thyroglobulin. A 73-yr-old woman diagnosed with metastases of probable thyroid carcinoma, manifested by a mediastinic mass and multiple pulmonary nodules positive for radioiodine (I131). Histological analysis showed typical findings of carcinoid tumor with positive immunohistochemistry for neuron-specific enolase, synaptophysin and thyroglobulin, and negative for chromogranin and calcitonin. We underline the exceptional finding of positive reaction for thyroglobulin, and discuss difficulties in differential diagnosis between metastatic differentiated thyroid and carcinoid neoplasms.

[Medulloblastoma presenting as an extra-axial tumor in the cerebellopontine angle]. / Meduloblastoma de presentación extra-axial en el ángulo pontocerebeloso.

Differences in the imaging characteristic of adult medulloblastomas have been reported, including involvement of lateral cerebellar hemispheres with an extra-axial appearance. We present a case report of this rare circumstance: a 40 year old man presented with a 3 weeks history of headache, morning vomiting and left hearing difficulties. Magnetic resonance imaging revealed a left cerebellopontine angle (CPA) tumor, like a well circumscribed homogeneously enhancing mass. Through a left suboccipital craniectomy the tumor was totally removed. It presented as a cerebellopontine angle tumor, like a meningioma, and not as an intra-axial tumor. Histological analysis revealed that the tumor was composed of densely packed with highly proliferative cells that produce a dense intercellular reticulin fiber network. Inmunohistochemical analysis showed positive expression to synaptophysin, specific neuronal enolase and cromogranin. Histological diagnosis was crucial to define it as a desmoplastic medulloblastoma the present case and to perform postoperative adjuvant therapy. Neurosurgeons should be aware of the possibility that a CPA tumor is of intraaxial origin, because this increase the variability on pathological diagnosis.

In vitro capacitation and acrosome reaction of dog spermatozoa can be feasibly attained in a defined medium without glucose.

Incubation of dog spermatozoa in a medium without glucose and in the presence of lactate and pyruvate (l-CCM) for 4 h at 38.5 degrees C in a 5% CO(2) atmosphere induced in vitro capacitation of these cells. This was verified after the combined specific capacitation-like changes in percentages of viability and altered acrosomes, motility characteristics, sperm location of reactivity against Pisum sativum, Arachis hypogaea and Helix pomatia lectins and the tyrosine phosphorylation pattern. Furthermore, a feasible acrosome reaction (AR) was induced when spermatozoa incubated in l-CCM for 4 h were further co-incubated for 1 h with canine oocytes. This was demonstrated by AR-like changes in percentages of viability, altered acrosomes, motility characteristics and sperm location of reactivity against P. sativum, A. hypogaea and H. pomatia lectins. All these results clearly indicate that in vitro capacitation, and subsequent AR, can be feasibly achieved without the presence of sugars. This ability can be related to the specific characteristics of energy-metabolism regulation reported in dog spermatozoa.

Differential effects of glucose and fructose on hexose metabolism in dog spermatozoa.

Incubation of dog spermatozoa with 10 mmol l(-1) glucose or fructose rapidly increased the intracellular content of glucose 6-phosphate and fructose 6-phosphate, although the effect of fructose was greater. These effects were correlated with increases in ATP, ribose 5-phosphate and glycogen contents, and in the rates of formation of L-lactate and CO2. In all cases, except for ATP and glycogen, the effect of fructose was greater than that of glucose. The total hexokinase activity of the crude extracts of dog spermatozoa was more sensitive to fructose than to glucose at lower concentrations (0.1-3.0 mmol l(-1)). Both monosaccharides induced a fast and intense increase in the overall tyrosine phosphorylation of dog spermatozoa, although their specific induced-phosphorylation patterns differed slightly. Glut 3 and Glut 5 hexose transporters were the main hexose transporters in dog spermatozoa; however, other possible SGLT family-related hexose transporters were also localized. These data indicate that, at concentrations from 1 mmol l(-1) to 10 mmol l(-1), fructose has a stronger effect than glucose on hexose metabolism of dog spermatozoa. These differences appear to be related to variations in the sensitivity of hexokinase activity. Moreover, the differential hexose metabolism induced by the two sugars had distinct effects on the function of dog spermatozoa, as revealed by the diverse patterns of tyrosine phosphorylation.

Effect of semen preparation on IVF of prepubertal goat oocytes.

The aim of these experiments was to study the effects of different methods of washing and selection of spermatozoa on the IVF of IVM oocytes from prepubertal goats. Fresh ejaculates from 3 males of proven fertility were processed according to the following treatments: 1) centrifugation in TALP, 2) centrifugation in sucrose-based Ficoll medium, 3) centrifugation in Percoll gradients at 40 and 80%, 4) by swim-up and 5) by dilution of spermatozoa (1:40) in (1:1) TALP. In all 5 treatments spermatozoa were incubated for 45 min with 100 microg/mL of heparin and then added to Fert-TALP. Oocytes were matured for 27 h in TCM-199 supplemented with 20% estrous goat serum (EGS), FSH, LH and estradiol-17beta. Spermatozoa (4x10(6) cells/mL) were coincubated with oocytes in 100 microL of Fert-TALP with hypotaurine for 24 h, after which the oocytes were transferred to a granulosa cells monolayer in TCM-199 plus 10% of EGS for 24 h (48 h post insemination). At 17 h post insemination a sample of sperm-exposed oocytes was taken and stained in lacmoid to observe sperm penetration and the formation of pronuclei. At 48 h post insemination the cleavage rate of oocytes was evaluated. Motility, viability and acrosome status of the spermatozoa were evaluated immediately after the mixing of the ejaculates, after washing and selection treatments, and after incubation with heparin and at 17 h post insemination. The different ejaculate treatments did not affect the penetration and cleavage rates of oocytes. At 48 h post insemination the cleavage rate was 46.9, 36.6 and 29.0% for dilution, Ficoll and swim-up preparations, respectively. Only the swim-up protocol improved sperm motility and viability compared with that of the initial semen sample and with the other sample treatments. At 17 h post insemination the semen parameters were the same for all sperm sample treatments.

Effect of sperm capacitation and fertilization media on IVF and early embryo development of prepubertal goat oocytes.

Experiments were carried out to develop an improved IVF system for prepubertal goat oocytes matured in vitro. Cumulus oocyte complexes (COC) were obtained by slicing ovaries from slaughtered prepubertal goats. Oocytes were matured in TCM199 supplemented with 20% estrous goat serum (EGS) + 10 micrograms/mL FSH + 10 micrograms/mL LH + 1 microgram/mL estradiol 17 beta for 27 h at 38.5 degrees C in 5% CO2 in air. In Experiments 1 and 2, freshly ejaculated spermatozoa were capacitated in 1 of 3 media: TALP/H, modified Defined Medium (mDM) and mH-M199 with 50 micrograms/mL heparin for 45 min. Matured oocytes were fertilized in TALP, mDM or mH-M199 in Experiment 1 and in TALP in Experiment 2. In Experiment 3, three media were used for sperm capacitation and fertilization: Treatment A (control group): spermatozoa were capacitated in mDM with 50 micrograms/mL heparin for 45 min and fertilized in TALP medium with 1 microgram/mL hypotaurine; Treatment B: spermatozoa were capacitated in mDM with 50 micrograms/mL heparin + 388 micrograms/mL caffeine for 30 min and fertilized in TALP medium without hypotaurine; Treatment C: spermatozoa were capacitated in mDM with 50 micrograms/mL heparin for 45 min and fertilized in TALP medium with PHE (20 microM penicillamine, 10 microM hypotaurine and 2 microM epinephrine). At 24 h post insemination, the ova were transferred to a granulosa cell monolayer, and early embryo development was evaluated until Day 8. In experiment 2, the results show, that mDM plus heparin for sperm capacitation and TALP medium with hypotaurine for oocyte fertilization provided the highest proportion of penetrated oocytes, both total number (79.6%) and normal fertilization (55.1%), whereas the use of caffeine (44.6 and 31.2%, total and normal fertilization rate, respectively) and PHE (31.8 and 20.6%, total and normal fertilization rate, respectively) as motility enhancers did not improve the results obtained in the control group (48.7% and 37.2%, total and normal fertilization rate, respectively). These were no differences for the results of morulae and blastocysts.

Isolation of Hanseniaspora uvarum (Kloeckera apiculata) in humans.

Isolation of Hanseniaspora uvarum, a yeast of the ascomycetes group, whose anamorph corresponds to Kloeckera apiculata, obtained from stool and two ungual specimens from three patients, is reported. This yeast has been found in soil, water, various fruits, bivalve molluscs, crabs, prawns and fruit flies; in Spain, it has been described in the fermentation processes of some wines. In our region, it has also been found in the intestine of mackerel (Scomber scombrus). Its finding in humans constitutes a clinical rarity.

[Small intestine adenocarcinoma with Crohn's disease]. / Adenocarcinoma de intestino delgado y enfermedad de Crohn.

We report a 57-year-old male with a 11-years history of Crohn's disease who developed small bowel adenocarcinoma. The patient was admitted with intestinal partial obstruction and he underwent a laparotomy because an abscess in right lower quadrant was suspected. He was successfully treated with resection and adjuvant chemotherapy, without evidence of recurrence after a 2-year follow-up. A review of the literature reveals that about 100 cases have been reported. We describe the distinguishing features of small bowel adenocarcinoma arising in Crohn's disease and we emphasize the difficulty in making an early diagnosis.