Molecular phylogeny of Cosmetocleithrum Kritsky, Thatcher & Boeger, 1986 (Monogenoidea: Dactylogyridae), gill parasites of Neotropical catfishes (Siluriformes).

Cosmetocleithrum Kritsky, Thatcher & Boeger, 1986 (Dactylogyridae) represents one of the most species-rich groups (22 species currently recognized as valid) of all dactylogyrid parasites infecting Neotropical catfishes. Species of Cosmetocleithrum exhibit a remarkable affinity towards catfishes of the Doradidae and the Auchenipteridae. However, phylogenetic relationships between members of this genus have not been yet analysed. This study analysed newly obtained partial sequences of the 28S ribosomal RNA gene of seven species of Cosmetocleithrum, including its type species C. gussevi Kritsky, Thatcher & Boeger, 1986, along with several other dactylogyrids infecting siluriform, gymnotiform, perciform and characiform fishes. Cosmetocleithrum appeared as an evolutionary recent group, composed of two well-defined lineages: lineage 1 includes parasites of doradids - namely, C. bulbocirrus, C. confusum, C. parvum and C. bifurcum - whereas lineage 2 is composed of species from doradids - that is, C. rarum, C. gussevi, C. gigas, C. trachydorasi and C. falsunilatum - together with parasites of auchenipterids - namely, C. laciniatum and C. baculum. The search for synapomorphies to characterize taxonomic groups within Cosmetocleithrum appears challenging, since the morphology of their haptoral elements is quite conservative, and that of the copulatory complex is highly variable between species. The results of the present study support the recent synonymization of Paracosmetocleithrum Acosta, Scholz, Blasco-Costa, Alves & Silva, 2018 with Cosmetocleithrum. Whereas the 28S ribosomal DNA data resolved Cosmetocleithrum as monophyletic, the statistical support for the lineage was low, rendering its phylogenetic position between other Neotropical dactylogyrids yet undefined.

Trinigyrus spp. (Monogenea: Dactylogyridae) from Brazilian catfishes: new species, molecular data and new morphological contributions to the genus.

This study describes two new species, Trinigyrus anthus n. sp. and Trinigyrus carvalhoi n. sp., from gills of Hypostomus spp. from the Upper Paraná River basin, Brazil. Trinigyrus peregrinus is redescribed based on examination of its holotype, paratypes and new material of specimens parasitizing Pterygoplichthys ambrosettii, also from the Upper Paraná River basin, Brazil. New morphological features were included in the diagnosis of the genus, such as the presence of a sclerotized border on the anchor base, and a weakly sclerotized fringe on the base of the male copulatory organ (MCO). Trinigyrus anthus n. sp. differs from other congeners by the shape of the MCO, presenting an enlarged base with sclerotized fringes resembling flower petals. Trinigyrus carvalhoi n. sp. and T. peregrinus are similar but can be differentiated from each other mainly by the sclerotization of the vagina (absent in the new species), and the morphology of the MCO (C-shaped versus one counterclockwise circle, respectively). For the first time, gene sequences of Trinigyrus spp. from Brazil were obtained (partial ribosomal 28S and mitochondrial cytochrome c oxidase I (mtCOI)). The genetic divergences among the new species and T. peregrinus varied from 2 to 3% (6‒18 pb) based on sequences of 28S ribosomal DNA (rDNA), and 6-7% (83‒92 pb) using mtCOI. Phylogenetic analyses based on partial 28S rDNA revealed that Trinigyrus, Heteropriapulus and Unilatus formed a monophyletic and well-supported clade of monogeneans from Neotropical freshwater loricariids, suggesting a closer relationship among these dactylogyrids and their hosts.

First record of Hysterothylacium sp. Moravec, Kohn et Fernandes, 1993 larvae (Nematoda: Anisakidae) infecting the ornamental fish Hyphessobrycon eques Steindachner, 1882 (Characiformes, Characidae).

This study reports for the first time infection with Hysterothylacium sp. larvae in the ornamental fish Hyphessobrycon eques from the Paranapanema River, Jurumirim Reservoir, São Paulo State, Brazil. A sample of 33 specimens of H. eques was collected in October, 2011. Four specimens of H. eques were parasitized by Hysterothylacium sp. larvae in the intestine and coelomic cavity, with prevalence of 12.1%, mean intensity of infection of 1, and mean abundance of 0.121 ± 0.05. A total of 40 unidentified free-living nematodes were found in the stomach content of 17 fish. This fish species is introduced in the Paranapanema River. Invasive species may affect the native fauna given the introduction of pathogens and parasites. This study also complements data on the diet of H. eques due to the records of free-living nematode as part of the stomach content. Infections with Hysterothylacium sp. larvae may affect the biology of this fish and bring about profit losses to aquarists.

[Resident of emergency attitude towards patients-physician relationship at the Hospital de Clínicas- Asunción, Paraguay]. / Actitud de residentes de urgencias hacia la relación médico-paciente en el Hospital de Clínicas- Asunción, Paraguay.

INTRODUCTION: The doctor-patient relationship is reaching great importance in recent times, is highlighted their importance in areas as varied as satisfaction, compliance, perception of professional competence, the frequency of legal issues relating to malpractice and even the prognosis of the disease or the general health of the patient. OBJECTIVE: To evaluate the doctor-patient relationship from the point of view of residents of emergency unit. METHODS: An observational, descriptive study. The sample consisted of 36 doctors from different areas of the Emergency Rooms of the Hospital de Clínicas- Asunción, Paraguay. The patient-physician relationship was evaluated using an instrument developed by RA Chavarria-Islas et al. with four indicators: Respect, Information, Consent and Dedication. RESULTS: 69.4% of residents have a regular patient-physician relationship; despite the 2.78% has a good relationship, 25% bad relationship and 2.78% a very bad relationship. CONCLUSION: Gaps in doctor-patient relationship were found in this study.. It is interesting to invest greater efforts to enhance the doctor-patient relationship as one of the edges to improve health care, which is vital in emergency care.

Se estudiaron retrospectivamente pacientes con diagnóstico de lupus eritematoso sistémico (LES) de acuerdo a criterios ACR 1982, con nefritis lúpica (NL) durante el período comprendido desde 2005 al 2012 y que fueran sometidos a una biopsia renal repetida. El número total de pacientes con NL atendidos fue de 120, de los cuales 18 (15%) pacientes fueron sometidos a biopsia renal repetida, 18 con 2 biopsias renales y 6 con 3 biopsias. 3 (16,7%) de los pacientes fueron fumadores; 1 (5,6%) poseía antecedentes de DBT previa, 2 (11,1%) poseían antecedentes de HTA; y 3 (16,7%) pacientes tenían obesidad previa. El tiempo de diagnóstico de LES al momento del estudio fue de 96 meses ± 15; el tiempo transcurrido entre la 1° y la 2° biopsia fue de 45 ± 11 meses y el tiempo entre la 2° y 3° biopsia fue de 56 ± 12 meses. Las indicaciones de la biopsia repetida fueron proteinuria en 10 biopsias (41,6%); proteinuria con alteración de la función renal en 2 biopsias (8,3%); proteinuria con sedimento patológico en 8 biopsias (33,3%); y proteinuria con sedimento patológico y alteración de la función renal en 4 biopsias (16,6%). Los cambios histológicos más frecuentes encontrados entre las primeras y las biopsias repetidas fueron: de clase IV a clase III: 2 (8,2%); clase IV a clase IV: 8 (33,3%), clase IV a clase III+V: 2 (8,2%); clase IV a clase IV+V: 3 (12,5%); clase IV a clase V: 2 (8,2%). Los cambios en las biopsias de NL proliferativas con índices de actividad y cronicidad (A/C) fueron: de A a A/C: 7 (29,1%), A/C a A/C: 7 (29,1%). La intensidad de la terapia inmunosupresora aumentó en 79,1%, se mantuvo el tratamiento inmunosupresor en 16.6%. Con respecto al cambio de medicación 7 (20%) pacientes recibieron Ciclofosfamida 1 gr cada 30 días, 9 (26%) Ciclofosfamida 500 mg cada 15 días, 8 (23%) tratamiento de reinducción con Micofenolato mofetil; Rituximab 8 (23%); y 3 (8%) Ciclosporina A. El tratamiento de mantenimiento se realizó con micofenolato mofetil en 23 casos (55%); con azatioprina en 11 (26%) casos; ciclosporina en 3 (7%) oportunidades y rituximab en 5 (12%). En todos los casos se utilizó hidroxicloroquina.

Helminth fauna of Astyanax fasciatus Cuvier, 1819, in two distinct sites of the Taquari River, São Paulo State, Brazil.

This study assessed the helminth fauna of Astyanax fasciatus in two distinct sites of the Taquari River, São Paulo State, with 30 individuals sampled in a lotic site and 30 in a lentic site, recording the monogeneans: Cacatuocotyle paranaensis, Characithecium costaricensis, Diaphorocleidus kabatai, Jainus sp., Notozothecium sp. and Gyrodactylus sp., the digenean Antorchis lintoni and no-identified metacercariae; the nematode Procamallanus (Spirocamallanus) inopinatus and no-identified larvae. The mean abundances of total monogeneans (U = 1053; p = 0.042) and C. costaricensis (U = 1107; p = 0.005) were higher in the lotic site. This difference may be due to the higher density of the host population in the lotic site, and the water transparence in lentic environments that prevents A. fasciatus to form shoals, precluding the exchange of parasites with direct cycle within a host population. This study is the first report of the helminth fauna of A. fasciatus in the Taquari River, with ten taxa recorded, and reports A. fasciatus as a new host for Notozothecium sp. and C. paranaensis.

Complete mitochondrial genomes of ancient canids suggest a European origin of domestic dogs.

The geographic and temporal origins of the domestic dog remain controversial, as genetic data suggest a domestication process in East Asia beginning 15,000 years ago, whereas the oldest doglike fossils are found in Europe and Siberia and date to >30,000 years ago. We analyzed the mitochondrial genomes of 18 prehistoric canids from Eurasia and the New World, along with a comprehensive panel of modern dogs and wolves. The mitochondrial genomes of all modern dogs are phylogenetically most closely related to either ancient or modern canids of Europe. Molecular dating suggests an onset of domestication there 18,800 to 32,100 years ago. These findings imply that domestic dogs are the culmination of a process that initiated with European hunter-gatherers and the canids with whom they interacted.

Incidence of sperm-tail tyrosine phosphorylation and hyperactivated motility in normozoospermic and asthenozoospermic human sperm samples

Our objective was to study the incidence of sperm-tail phosphotyrosine immunoreactivity in normozoospermic and asthenozoospermic human sperm samples, its association with sperm motion parameters, particularly hyperactivated motility, and its potential involvement in the pathogenesis of asthenozoospermia. The work was conducted as a prospective experimental study in the Sperm Biology and Andrology laboratories of the Jones Institute, a medical school-based fertility center. The study subjects were healthy fertile male donors (normozoospermic samples) and infertile patients (asthenozoospermic samples) attending the center. Recently ejaculated semen samples were washed twice to eliminate seminal plasma and a swim-up was performed to select the motile population which, in turn, was incubated up to 18 h at 37 degrees C in 3.5% human serum albumin-supplemented Ham's F10 to allow for capacitation. For evaluation, sperm aliquots were taken pre-swim-up (T0), immediately post swim-up (T1), at 6 h (T6), and 18 h (T18) of incubation. The main outcome measures were computer-analyzed sperm motion parameters and hyperactivated motility, and immunodetection of phosphotyrosine (PY)-containing proteins. During the capacitating incubation, normozoospermic samples displayed maximum motility, velocity, and hyperactivation at T6, significantly decreasing their values at T18. PY-proteins were located both at the tail and head of spermatozoa. Their expression increased progressively during the incubation, being present in about 70% of the sperm tails at T18. Asthenozoospermic samples showed an inability to respond to capacitation with an increase in motion parameters and PY-phosphorylation. At T6, both hyperactivation and PY-phosphorylation were significantly lower than in normal samples. Our results suggest that PY-phosphorylation of tail proteins is highly conspicuous in human spermatozoa, and increases its incidence in a time-dependent manner, as more sperm become capacitated. Asthenozoospermic samples displaying low percentages of motile sperm and altered motion characteristics showed a decreased incidence of PY-phosphorelated sperm. Tail protein PY-phosphorylation may be related to sperm movement, especially to hyperactivated motility and its deficiency may be associated to asthenozoospermia.

Incidence of sperm-tail tyrosine phosphorylation and hyperactivated motility in normozoospermic and asthenozoospermic human sperm samples

Our objective was to study the incidence of sperm-tail phosphotyrosine immunoreactivity in normozoospermic and asthenozoospermic human sperm samples, its association with sperm motion parameters, particularly hyperactivated motility, and its potential involvement in the pathogenesis of asthenozoospermia. The work was conducted as a prospective experimental study in the Sperm Biology and Andrology laboratories of the Jones Institute, a medical school-based fertility center. The study subjects were healthy fertile male donors (normozoospermic samples) and infertile patients (asthenozoospermic samples) attending the center. Recently ejaculated semen samples were washed twice to eliminate seminal plasma and a swim-up was performed to select the motile population which, in turn, was incubated up to 18 h at 37 degrees C in 3.5% human serum albumin-supplemented Hams F10 to allow for capacitation. For evaluation, sperm aliquots were taken pre-swim-up (T0), immediately post swim-up (T1), at 6 h (T6), and 18 h (T18) of incubation. The main outcome measures were computer-analyzed sperm motion parameters and hyperactivated motility, and immunodetection of phosphotyrosine (PY)-containing proteins. During the capacitating incubation, normozoospermic samples displayed maximum motility, velocity, and hyperactivation at T6, significantly decreasing their values at T18. PY-proteins were located both at the tail and head of spermatozoa. Their expression increased progressively during the incubation, being present in about 70% of the sperm tails at T18. Asthenozoospermic samples showed an inability to respond to capacitation with an increase in motion parameters and PY-phosphorylation. At T6, both hyperactivation and PY-phosphorylation were significantly lower than in normal samples. Our results suggest that PY-phosphorylation of tail proteins is highly conspicuous in human spermatozoa, and increases its incidence in a time-dependent manner, as more sperm become capacitated. Asthenozoospermic samples displaying low percentages of motile sperm and altered motion characteristics showed a decreased incidence of PY-phosphorelated sperm. Tail protein PY-phosphorylation may be related to sperm movement, especially to hyperactivated motility and its deficiency may be associated to asthenozoospermia. (AU)

Male antisperm antibodies: association with a modified sperm stress test and lipid peroxidation.

We previously reported a modified sperm stress test (MOST), low scores (< 0.39) in which were associated with sperm-related abnormal in vitro fertilization. Preliminary observations suggested that the presence of male sperm antibodies (ASA) could give low MOST scores. It was therefore decided to undertake a study to verify this possible association and also to ascertain if such a relationship was causal in nature. Six hundred and fifty semen samples from patients consulting for infertility were assessed for basic seminal characteristics, motion parameters (CASA), ASA and MOST. Thirty-nine samples (6%) were ASA-positive. Samples with and without ASA showed similar characteristics, except for percentage of normal forms and MOST scores (0.35 +/- 0.03 vs. 0.67 +/- 0.01, P < 0.001, for ASA-positive and -negative, respectively). There was a strong statistical association between presence of ASA and low MOST scores (P < 0.0001). One-hundred per cent of ASA-positive samples displayed low MOST scores. To verify the nature of this relationship, we incubated ASA-free spermatozoa with ASA-positive and -negative (control) sera. Despite an increase in the percentage of ASA-bearing spermatozoa in those aliquots incubated with ASA-positive serum, their original (pre-incubation) MOST scores remained unchanged. Furthermore, the rate of lipid peroxidation, indirectly reflected in MOST scores, was not different in the aliquots incubated with ASA. In conclusion, there seems to be a strong association between presence of ASA and low MOST values in semen samples of infertile patients; however, the relationship does not appear to be causal.

Serum inhibin B may be a reliable marker of the presence of testicular spermatozoa in patients with nonobstructive azoospermia.

OBJECTIVE: To establish the predictive value of serum inhibin B levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia, compared with the traditional serum FSH marker. DESIGN: Prospective study. SETTING: Private high-complexity reproductive center with university affiliation. PATIENT(S): Seventy-eight patients with nonobstructive azoospermia, 15 patients with obstructive azoospermia, and 10 fertile volunteers. INTERVENTION(S): Blood samples, testicular sperm extraction, percutaneous epididymal sperm aspiration, and semen collection. MAIN OUTCOME MEASURE(S): Serum levels of inhibin B and FSH and presence of spermatozoa on TESE, PESA, or regular semen analysis. RESULT(S): Patients with nonobstructive azoospermia has significantly higher levels of serum FSH and significantly lower levels of inhibin B. Mean inhibin B serum levels were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa were found (89.31 +/- 73.24 pg/mL vs. 19.23 +/- 22.34 pg/mL), but mean FSH serum levels did not have similar predictive power (21.37 +/- 12.92 IU/mL vs. 19.27 +/- 10.28 IU/mL). The cut-off level of inhibin B separating both groups, as determined by the receiver-operating characteristic curves, was >53 pg/mL. CONCLUSION(S): Serum inhibin B level seems to be more accurate than serum FSH level in prediction of the presence of testicular spermatozoa in patients with nonobstructive azoospermia.

Effects of long-term in vitro incubation of human spermatozoa: functional parameters and catalase effect.

Prolonged incubation of human spermatozoa can have deleterious effects on sperm function. The aim of this paper was to describe the effects of a prolonged in vitro incubation, under similar conditions to those employed in human assisted reproduction, on various sperm functional parameters, and to investigate the effect of an antioxidant (catalase) on this system. Freshly collected ejaculates from 20 healthy donors were studied. Samples were divided into two aliquots: the first was incubated with Ham's F10 containing 3.5% HAS, and the second was incubated in the same medium plus catalase (100 units ml-1). All experiments were carried out with spermatozoa isolated using the swim-up technique. Spermatozoa recovered from the supernatant after 1 h (T1) of incubation in 5% CO2 in air at 37 degrees C, and after 5 h (T6), 23 h (T24) and 47 h (T48), were evaluated for concentration, motion parameters including hyperactivation (computer-assisted analysis), viability, ATP concentration, reactive oxygen species (ROS) generation, DNA integrity (acridine orange), and acrosome reaction (AR). The major alteration observed in sperm function during the prolonged in vitro incubation was a reduction in the number of motile spermatozoa, together with an impairment in the quality of sperm movement. ROS levels increased with the incubation time. No substantial modifications of sperm viability, chromatin condensation and AR inducibility were observed. The addition of catalase to the medium, while keeping ROS values within baseline levels, did not prevent the loss of motility or the corresponding increase in ATP.

Incidence of tail structure distortions associated with dysplasia of the fibrous sheath in human spermatozoa.

Dysplasia of the fibrous sheath (DFS) is an anomaly found in spermatozoa of severe asthenozoospermic patients. Marked hypertrophy and hyperplasia of the fibrous sheath is the common characteristic. Immunocytochemistry allowed us to visualize the distortions and incidence of tail structure abnormalities associated with this phenotype in six patients; four with a complete form and two with an incomplete form of this pathology previously diagnosed and studied by electron microscopy. Microtubules and fibrous sheaths were studied using monoclonal antibodies against alpha-acetylated tubulin and anti-FSC1 (the major protein component of the fibrous sheath). Mitochondrial sheaths were visualized using the mitochondrion-specific vital dye MitoTracker green FM(TM). Phase contrast and fluorescent microscopy of semen samples showed large numbers of spermatozoa with short, rigid, thick and irregular tails. As expected, anomalous and completely distorted fibrous sheaths, severe alterations of the axonemal microtubules and different patterns of mitochondrial sheath configurations were found. While ultrastructural studies of thin sections allow an in-depth knowledge of the internal organization of the sperm tail, fluorescence labelling of selected sperm components affords a unique view of the whole flagellum including topographical relationships of various organelles. The combination of these different approaches is essential for a comprehensive understanding of this particular pathology.

Puntaje embrionario promedio: un nuevo criterio para la evaluación morfológica de embriones en un programa de fertilización in vitro / Average embryo score: a new criterion for the evaluation of embryos in a in vitro fertilization program

En 1992, Steer y cols. (Hum Reprod 1992; 7:117-9) propusieron el "Puntaje Embrionario Acumulado" (PEA), criterio de evaluación embrionaria que asigna un puntaje a cada embrión (PE), igual a su calidad por su número de células, y a cada transferencia (PEA), igual a la suma de los PE de los embriones transferidos. Concluyeron que un valor de PEA elevado aumenta la chance de embarazo independientemente del número de embriones transferidos, pese a que sus valores de PEA aumentaban con éste. En base a esta observación, en este trabajo se propone un nuevo criterio, el puntaje embrionario promedio (PEP), para añadir al PEA la variable número de embriones (N) como denominador. Objetivo: Evaluar el valor pronóstico, en un programa de FIV, del criterio PEP propuesto, y compararlo con el del PEA y con el de la combinación descriptiva de la calidad embrionaria y el número de células. Materiales y Metodos: Se estudiaron retrospectivamente ciclos de FIV-TE en los que se transfirieron de 2 a 6 embriones entre las 41 y 50 horas post-inseminación. Se analizaron los criterios: CGDE (calidad embrionaria y número de células), PEA y PEP (para PEP=PEA/N). En cada criterio se utilizaron curvas ROC en base a implantación y embarazo clínico para determinar valores umbrales de predicción que delimitaron grupos de prónosticos bueno (+) y malo (-). Se usó el coeficiente r de Pearson para evaluar la correlación de los criterios con el número de embriones transferidos. Resultados: En los 122 ciclos estudiados se obtuvieron 35 embarazos (28.59 por ciento), implantando un total de 45 embriones de los 473 transferidos (9.51 por ciento). Los valores de correlación con el número de embriones transferidos fueron de 0.13 para PEP, 0.61 para CGDE y 0.79 para PEA (p<0.001). Los umbrales de predicción determinados para los criterios fueron de: PEP=11, PEA=34 y CGDE=2 embriones de clase III o IV y 4 o más células. Los tres criterios encontraron diferencias significativas (p<0.001) entre los grupos de prónostico (+) y (-) para las tasas de embarazo clínico e implantación: PEP (+): 38.3 por ciento y 12.3 por ciento, PEP(-): 9.8 por ciento y 2.9 por ciento; PEA(+): 38.4 por ciento y 11.2 por ciento, PEA(-): 5.6 por ciento y 2.3 por ciento; CGDE (+): 38.8 por ciento y 11.7 por ciento y CGDE (-): 9.5 por ciento y 3.3 por ciento...

Puntaje embrionario promedio: un nuevo criterio para la evaluación morfológica de embriones en un programa de fertilización in vitro / Average embryo score: a new criterion for the evaluation of embryos in a in vitro fertilization program

En 1992, Steer y cols. (Hum Reprod 1992; 7:117-9) propusieron el "Puntaje Embrionario Acumulado" (PEA), criterio de evaluación embrionaria que asigna un puntaje a cada embrión (PE), igual a su calidad por su número de células, y a cada transferencia (PEA), igual a la suma de los PE de los embriones transferidos. Concluyeron que un valor de PEA elevado aumenta la chance de embarazo independientemente del número de embriones transferidos, pese a que sus valores de PEA aumentaban con éste. En base a esta observación, en este trabajo se propone un nuevo criterio, el puntaje embrionario promedio (PEP), para añadir al PEA la variable número de embriones (N) como denominador. Objetivo: Evaluar el valor pronóstico, en un programa de FIV, del criterio PEP propuesto, y compararlo con el del PEA y con el de la combinación descriptiva de la calidad embrionaria y el número de células. Materiales y Metodos: Se estudiaron retrospectivamente ciclos de FIV-TE en los que se transfirieron de 2 a 6 embriones entre las 41 y 50 horas post-inseminación. Se analizaron los criterios: CGDE (calidad embrionaria y número de células), PEA y PEP (para PEP=PEA/N). En cada criterio se utilizaron curvas ROC en base a implantación y embarazo clínico para determinar valores umbrales de predicción que delimitaron grupos de prónosticos bueno (+) y malo (-). Se usó el coeficiente r de Pearson para evaluar la correlación de los criterios con el número de embriones transferidos. Resultados: En los 122 ciclos estudiados se obtuvieron 35 embarazos (28.59 por ciento), implantando un total de 45 embriones de los 473 transferidos (9.51 por ciento). Los valores de correlación con el número de embriones transferidos fueron de 0.13 para PEP, 0.61 para CGDE y 0.79 para PEA (p<0.001). Los umbrales de predicción determinados para los criterios fueron de: PEP=11, PEA=34 y CGDE=2 embriones de clase III o IV y 4 o más células. Los tres criterios encontraron diferencias significativas (p<0.001) entre los grupos de prónostico (+) y (-) para las tasas de embarazo clínico e implantación: PEP (+): 38.3 por ciento y 12.3 por ciento, PEP(-): 9.8 por ciento y 2.9 por ciento; PEA(+): 38.4 por ciento y 11.2 por ciento, PEA(-): 5.6 por ciento y 2.3 por ciento; CGDE (+): 38.8 por ciento y 11.7 por ciento y CGDE (-): 9.5 por ciento y 3.3 por ciento... (AU)

Present and future of maternal mortality in Latin America.

Twenty-three thousand maternal deaths per year in Latin America and the Caribbean reflects clearly the critical situation in this part of the world. Although we don't know exactly how many women die every year from pregnancy and childbirth, it is clear that the majority of these women are from low socioeconomic backgrounds, live in remote places and have a low level of education. Hemorrhages, infections and hypertension induced by pregnancy are the most common causes of maternal mortality. These pathologies can in many cases be prevented if there is a will for a positive change that involves different sectors related with health. The role of the scientific societies, and the role of FIGO through the 'Save the Mothers' Project are of extreme importance, assuming a chief role and compromise that can help in the right way to be able to revert this situation.

Cytoskeletal organization defects and abortive activation in human oocytes after IVF and ICSI failure.

In this study, we analysed the distribution of beta tubulins to detect spindle and cytoplasmic microtubules, alpha acetylated tubulins for sperm microtubules and chromatin configuration in oocytes showing fertilization failure after conventional IVF or intracytoplasmic sperm injection (ICSI). A total of 450 human oocytes that failed to fertilize were studied 20-40 h after IVF or ICSI. In all, 287 oocytes were stained for immunofluorescence and chromosomal spreads were performed by Tarkowski's air-drying method in 163 IVF or ICSI oocytes that did not develop pronuclei after the extrusion of a second polar body. Immunofluorescence analysis showed that the main reason of fertilization failure after IVF was no sperm penetration (55.5%). The remaining oocytes showed different abnormal patterns, e.g. oocyte activation failure (15.1%) and defects in pronuclei apposition (19.2%). On the other hand, fertilization failure after ICSI was mainly associated to incomplete oocyte activation (39.9%), and to a lesser extent with defects in pronuclei apposition (22.6%) and failure of sperm penetration (13.3%). A further 13.3% of the ICSI oocytes arrested their development at the metaphase of the first mitotic division. The chromosomal spreads allowed the analysis of abortive activations, in which no pronuclei formed but a second polar body was extruded. Immunofluorescence and cytogenetic analysis provided a useful tool to improve infertility diagnosis and prognosis in each particular case.

Endometrial dating and determination of the window of implantation in healthy fertile women.

OBJECTIVE: To reassess endometrial morphological criteria of normality identifying the best morphological and molecular "implantation window" indicators in normal women. DESIGN: Prospective clinical study. SETTING: Assisted reproductive unit. PATIENT(S): Fourteen healthy volunteers. INTERVENTION(S): Blood sampling for LH, E(2), and progesterone (P4) determinations. Daily vaginal ultrasounds. Two endometrial biopsies per volunteer, 7 days apart, during luteal phase. MAIN OUTCOME MEASURE(S): Endometrial dating, pinopodes formation, immunohistochemical determination of integrins (alphavbeta3, alpha4beta1), leukemia inhibitory factor (LIF), interleukin-1 receptor type I (IL-1R tI), mouse ascites Golgi (MAG), the transmembrane mucin (MUC-1), and P4 receptor expression. RESULT(S): In 26 of 28 biopsies observers agreed; in two biopsies there was a discrepancy (difference of 72 hours). With use of LH peak, 24 of 26 samples were in phase, and 2 were 3 days behind. Pinopodes appeared on days 20-21 and persisted through day 28 in small groups or larger areas. beta3 Integrin was highly expressed in luminal and glandular epithelium from day 22 through 28; 48 hours thereafter pinopodes appeared. alpha4 Subunit exhibited luminal epithelium reaction positivity on days 22-23 and glands on days 18-23. LIF and IL-1R tI showed weak, erratic expression. MAG antibodies showed luminal epithelium expression up to day 22 and glands up to day 25. MUC-1 showed positivity during the whole luteal phase. P4 receptors were positive through day 20 and at the end of the luteal phase. CONCLUSION(S): The three most cited markers that frame the window of implantation do not correlate in our material. Pinopodes are present from day 20 on; beta3 and alpha4 integrin subunits indicate a window opening on days 22-23.

Pregnancies established through intracytoplasmic sperm injection (ICSI) using spermatozoa with dysplasia of fibrous sheath.

AIM: Dysplasia of the fibrous sheath (DFS) is an anomaly found in asthenozoospermic patients with extremely low or absent motility. In order to determine the efficacy of ICSI in these patients, a retrospective analysis of ICSI results in DFS patients has been done. METHODS: Ten ICSI attempts were performed in 6 patients with diagnosis of Dysplasia of the Fibrous Sheath studied by transmission and scanning electron microscopy. RESULTS: In the cases studied, sperm concentration was (29.62 +/- 18.05) x 10(6)/mL, total motility was 1.14 +/- 1.31%. Progressive motility was 0% except for one case with 0.1% . One hundred and three preovulatory oocytes were obtained and 94 metaphase II oocytes were injected. Sixty-nine of them showed two pronuclei (fertilization rate: 73.4%). Forty-nine embryos were obtained and 34 were transferred (mean: 3.4 embryos per transfer). Five pregnancies were diagnosed by beta-hCG plasma level determinations that resulted to be one preclinical abortion, one clinical abortion and three deliveries. Another pregnancy (ongoing) was achieved from a cryopreserved embryo transfer. CONCLUSION: These results showed that ICSI provides a suitable solution for patients suffering from irreversible sperm defects such as DFS. Nevertheless, it is mandatory to inform couples of possible transmission risks to offspring, which are unknown at present. Only when the etiology of this problem is disclosed, it will be possible to assess the real genetic risk.

Birth of twin males with normal karyotype after intracytoplasmic sperm injection with use of testicular spermatozoa from a nonmosaic patient with Klinefelter's syndrome.

OBJECTIVE: To report the birth of healthy twin males after the use of testicular spermatozoa from a nonmosaic patient with Klinefelter's syndrome. DESIGN: Case report. SETTING: Private reproduction center with university affiliation. PATIENT(S): A couple undergoing intracytoplasmic sperm injection (ICSI) combined with testicular sperm extraction because of the husband's secretory azoospermia and a nonmosaic 47,XXY peripheral blood karyotype. The wife, a healthy female, presented with a history of oligomenorrhea. INTERVENTION(S): ICSI was performed using testicular spermatozoa; 3 mM pentoxifylline solution was used to induce sperm motility because the spermatozoa recovered were all immotile. MAIN OUTCOME MEASURE(S): Normal fertilization, embryo cleavage, pregnancy outcome, and peripheral blood karyotype of the newborns. RESULT(S): Thirteen metaphase II oocytes were injected. Seven of them fertilized normally and six did not fertilize. Three good-quality embryos (4-cell stage class II) were transferred, and four were cryopreserved at the two-cell and four-cell stages using a slow freezing protocol. Twelve days after ET, a beta-hCG determination was positive. Ultrasonographic examination revealed three intrauterine fetal sacs, but one of them showed a fetal pole without cardiac activity and vanished in subsequent ultrasonographic examinations. The patient delivered twins with normal male peripheral blood karyotypes. CONCLUSION(S): Normal outcome after the use of testicular sperm extraction and ICSI in a nonmosaic patient with Klinefelter's syndrome reaffirms the notion of low transmission risk of this gonosomal aneuploidy.