RESUMEN

Antimicrobial resistance is one of the major health problems we face in the 21st century. Nowadays we cannot understand global health without the interdependence between the human, animal and environmental dimensions. It is therefore logical to adopt a "One Health" approach to address this problem. In this review we show why a collaboration of all sectors and all professions is necessary in order to achieve optimal health for people, animals, plants and our environment.

Asunto(s)

Antibacterianos , Salud Única , Animales , Humanos , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana , Salud Global

RESUMEN

Improvements in cost and speed of next generation sequencing (NGS) have provided a new pathway for delivering disease diagnosis, molecular typing, and detection of antimicrobial resistance (AMR). Numerous published methods and protocols exist, but a lack of harmonisation has hampered meaningful comparisons between results produced by different methods/protocols vital for global genomic diagnostics and surveillance. As an exemplar, this study evaluated the sensitivity and specificity of five well-established in-silico AMR detection software where the genotype results produced from running a panel of 436 Escherichia coli were compared to their AMR phenotypes, with the latter used as gold-standard. The pipelines exploited previously known genotype-phenotype associations. No significant differences in software performance were observed. As a consequence, efforts to harmonise AMR predictions from sequence data should focus on: (1) establishing universal minimum to assess performance thresholds (e.g. a control isolate panel, minimum sensitivity/specificity thresholds); (2) standardising AMR gene identifiers in reference databases and gene nomenclature; (3) producing consistent genotype/phenotype correlations. The study also revealed limitations of in-silico technology on detecting resistance to certain antimicrobials due to lack of specific fine-tuning options in bioinformatics tool or a lack of representation of resistance mechanisms in reference databases. Lastly, we noted user friendliness of tools was also an important consideration. Therefore, our recommendations are timely for widespread standardisation of bioinformatics for genomic diagnostics and surveillance globally.

Asunto(s)

Antibacterianos , Infecciones por Escherichia coli , Antibacterianos/farmacología , Biología Computacional/métodos , Farmacorresistencia Bacteriana/genética , Escherichia coli , Infecciones por Escherichia coli/diagnóstico , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Pruebas de Sensibilidad Microbiana

RESUMEN

Bacterial antimicrobial resistance (AMR) is constantly evolving and horizontal gene transfer through plasmids plays a major role. The identification of plasmid characteristics and their association with different bacterial hosts provides crucial knowledge that is essential to understand the contribution of plasmids to the transmission of AMR determinants. Molecular identification of plasmid and strain genotypes elicits a distinction between spread of AMR genes by plasmids and dissemination of these genes by spread of bacterial clones. For this reason several methods are used to type the plasmids, e.g. PCR-based replicon typing (PBRT) or relaxase typing. Currently, there are 28 known plasmid types in Enterobacteriaceae distinguished by PBRT. Frequently reported plasmids [IncF, IncI, IncA/C, IncL (previously designated IncL/M), IncN and IncH] are the ones that bear the greatest variety of resistance genes. The purpose of this review is to provide an overview of all known AMR-related plasmid families in Enterobacteriaceae, the resistance genes they carry and their geographical distribution.

Asunto(s)

Farmacorresistencia Bacteriana , Enterobacteriaceae/efectos de los fármacos , Enterobacteriaceae/genética , Transferencia de Gen Horizontal , Genes Bacterianos , Plásmidos/análisis , Plásmidos/clasificación , Enterobacteriaceae/clasificación , Genotipo , Humanos

RESUMEN

Objectives: The mobile colistin resistance gene mcr-1 has been identified worldwide in human and animal sources, while its occurrence in the environment is still largely unknown. The aim of this study was to investigate the presence of mcr-1 -harbouring Enterobacteriaceae in water samples obtained from rivers and waste water treatment plants in the area of Barcelona, Spain. Methods: The presence of mcr-1 was detected by PCR. Bacterial identification was performed via MALDI-TOF MS. Resistance to colistin was determined by a broth dilution method. The epidemiological relationship between the positive isolates was assessed with PFGE and ST was determined by MLST. Plasmid characterization was performed by transformation experiments, antimicrobial susceptibility testing and incompatibility group PCR. Results: Thirty MDR isolates bearing mcr-1 , 29 Escherichia coli (ST632 and ST479) and 1 Klebsiella pneumoniae (ST526), were identified in sewage from two different waste water treatment plants, whereas the gene was not found in river water. All isolates, including the K. pneumoniae , harboured bla CTX-M-55 and bla TEM-1 . mcr-1 was in all cases associated with an IncI2 plasmid, which only conferred resistance to colistin. mcr-1 was harboured by two predominant E. coli clones that were found in both waste water treatment plants. Conclusions: This study showed a high occurrence of mcr-1 in the sewage of Barcelona, mainly due to the dissemination of two E. coli pulsotypes that are circulating in the population. The presence of mcr-1 in the environment is a cause for concern, and suggests high prevalence of mcr-1 in the community.

Asunto(s)

Proteínas de Escherichia coli/genética , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/aislamiento & purificación , Aguas del Alcantarillado/microbiología , Microbiología del Agua , Animales , Antibacterianos/farmacología , Colistina/farmacología , Farmacorresistencia Bacteriana , Electroforesis en Gel de Campo Pulsado , Escherichia coli/química , Escherichia coli/clasificación , Humanos , Klebsiella pneumoniae/química , Klebsiella pneumoniae/clasificación , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Plásmidos/análisis , Plásmidos/clasificación , Reacción en Cadena de la Polimerasa , España , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Transformación Bacteriana

RESUMEN

The aim of this study was to examine whether there is an association among genetic variability in leptin (LEP) and leptin receptor (LEPR) genes and male infertility. We performed a case-control study and were searching for an association between polymorphisms of LEP and LEPR genes and male infertility. The study group consisted of 317 patients with idiopathic infertility and a control group of 241 fertile men from Slovenia. Four single nucleotide polymorphisms (SNPs) in LEP gene and four single nucleotide polymorphisms (SNPs) in LEPR gene were chosen and genotyped. Statistically significant SNP was further validated in additional 255 infertile patients and 168 controls from Serbia and Macedonia. In the Slovenian population, we found a statistically significant difference in genotype distribution for rs10244329 polymorphism in LEP gene (recessive genotype model, p value = 0.048). The trend toward statistically significant difference in genotype distribution for rs10244329 polymorphism was confirmed in the Serbian and Macedonian populations (p value = 0.07). Our data suggest that genetic variability in the LEP gene might be associated with male infertility warranting further confirmation and mechanistic investigations.

Asunto(s)

Predisposición Genética a la Enfermedad , Infertilidad Masculina/genética , Leptina/genética , Receptores de Leptina/genética , Adulto , Alelos , Estudios de Casos y Controles , Frecuencia de los Genes , Estudios de Asociación Genética , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , República de Macedonia del Norte , Factores de Riesgo , Serbia , Recuento de Espermatozoides

RESUMEN

BACKGROUND The global obesity epidemic has paralleled a decrease in semen quality. Yet, the association between obesity and sperm parameters remains controversial. The purpose of this report was to update the evidence on the association between BMI and sperm count through a systematic review with meta-analysis. METHODS A systematic review of available literature (with no language restriction) was performed to investigate the impact of BMI on sperm count. Relevant studies published until June 2012 were identified from a Pubmed and EMBASE search. We also included unpublished data (n = 717 men) obtained from the Infertility Center of Bondy, France. Abstracts of relevant articles were examined and studies that could be included in this review were retrieved. Authors of relevant studies for the meta-analysis were contacted by email and asked to provide standardized data. RESULTS A total of 21 studies were included in the meta-analysis, resulting in a sample of 13 077 men from the general population and attending fertility clinics. Data were stratified according to the total sperm count as normozoospermia, oligozoospermia and azoospermia. Standardized weighted mean differences in sperm concentration did not differ significantly across BMI categories. There was a J-shaped relationship between BMI categories and risk of oligozoospermia or azoospermia. Compared with men of normal weight, the odds ratio (95% confidence interval) for oligozoospermia or azoospermia was 1.15 (0.93-1.43) for underweight, 1.11 (1.01-1.21) for overweight, 1.28 (1.06-1.55) for obese and 2.04 (1.59-2.62) for morbidly obese men. CONCLUSIONS Overweight and obesity were associated with an increased prevalence of azoospermia or oligozoospermia. The main limitation of this report is that studied populations varied, with men recruited from both the general population and infertile couples. Whether weight normalization could improve sperm parameters should be evaluated further.

Asunto(s)

Azoospermia/epidemiología , Índice de Masa Corporal , Obesidad/epidemiología , Oligospermia/epidemiología , Recuento de Espermatozoides , Espermatozoides/citología , Peso Corporal , Fertilidad , Francia , Humanos , Masculino , Oportunidad Relativa , Sobrepeso/epidemiología , Análisis de Semen

RESUMEN

A 45-year-old male with azoospermia was presented to us with primary infertility. We found that he had been taking testosterone due to erectile dysfunction. Upon its discontinuation, the normozoospermia was restored. This led to a pregnancy. In men aged 40-50, the diagnostic work up of testosterone deficiency should be in accordance with the current guidelines. Moreover, no testosterone prescription should be made without having inquired about parenthood desires. In men of the reproductive age with recent-onset azoospermia, consider the possibility of an iatrogenic cause, primarily due to the testosterone replacement therapy. The treatment is simple: to discontinue the therapy.

Asunto(s)

Instituciones de Atención Ambulatoria , Azoospermia/terapia , Testosterona/efectos adversos , Adulto , Azoospermia/inducido químicamente , Femenino , Humanos , Masculino , Persona de Mediana Edad , Embarazo

RESUMEN

To elucidate the effects of inflammation on sperm quality, this study analysed classical sperm characteristics, leukocytes and elastase in neat semen, and sperm apoptotic markers, i.e. changes in plasma membrane phospholipid asymmetry, mitochondrial membrane potential (MMP), DNA integrity and intracellular reactive oxygen species (ROS), in semen prepared by density gradient using flow cytometry from 348 men of infertile couples. Increased leukocytes (> or = 0.1 x 10(6)/ml) were associated with a decreased sperm concentration, motility and normal morphology (P < or = 0.001). Sperm necrosis and DNA denaturation were increased (31.3 versus 26.6%, P=0.020; 15.5 versus 11.5%, P = 0.011, respectively), whereas spermatozoa with normal MMP were decreased (64.1 versus 70.0%, P=0.004). High leukocyte levels ((> or = 1 x 10(6)/ml) were not associated with any of the observed sperm parameters. At low elastase concentration (100-290 microg/l), DNA denaturation was higher (16.1 versus 10.5%, P = 0.024) compared with very low elastase concentration (< 100 microg/l). A high elastase concentration (290-1000 microg/l) was associated with higher ROS index compared with low elastase concentration (1.28 versus 1.01, P=0.016). Slightly increased leukocytes and elastase are associated with slightly poorer sperm characteristics and/or increased sperm necrosis, DNA denaturation and intracellular ROS and decreased MMP.

Asunto(s)

Apoptosis/fisiología , Infertilidad Masculina/fisiopatología , Potencial de la Membrana Mitocondrial/fisiología , Desnaturalización de Ácido Nucleico , Elastasa Pancreática/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Semen/citología , Espermatozoides/patología , Adulto , Biomarcadores/metabolismo , ADN/química , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Necrosis/patología , Recuento de Espermatozoides , Motilidad Espermática , Espermatozoides/metabolismo

RESUMEN

In this study we sought to evaluate whether psychological factors in males affect semen quality and pregnancy. In 1076 men of infertile couples, psychological factors, i.e. exposure to acute stress, coping with stress, the WHO (five) Well-Being Index and the Zung's Anxiety Scale Inventory scores were assessed by a questionnaire at the time of semen analysis. Relationships between psychological factors and semen quality (sperm concentration, rapid and progressive motility and normal morphology) were assessed. In 353 men with infertility duration of < or =1.5 years, sperm concentration > or =5 x 10(6) sperm/mL and a female partner with a laparoscopically confirmed tubal patency, we looked prospectively for relations between psychological factors and the occurrence of a natural pregnancy at a 6-month follow-up (n = 124), and first-trimester loss (n = 18). Anxiety trait, found in 19% of men, was related to previous in vitro fertilization/intracytoplasmic sperm injection attempts (p = 0.014), cigarette intake (p = 0.006), alcohol intake (p = 0.026) and sexual difficulties (p < 0.001). Regression analyses indicated a significant positive relationship between the level of sperm concentration and the WHO (five) Well-Being Index score, each successive score number accounting for a 7.3% increase in sperm concentration (p = 0.039), whereas no correlation was found between psychological factors and sperm rapid progressive motility and normal morphology. Poorer coping with stress was related to the occurrence of a first-trimester miscarriage (p = 0.016) in the female partner. Possible depression in males is related to decreased sperm concentration, and poor coping with stress is associated with increased occurrence of early miscarriage.

Asunto(s)

Aborto Espontáneo/psicología , Infertilidad Masculina/psicología , Análisis de Semen , Aborto Espontáneo/etiología , Adaptación Psicológica , Adulto , Ansiedad/complicaciones , Depresión/complicaciones , Femenino , Humanos , Infertilidad Masculina/etiología , Masculino , Persona de Mediana Edad , Embarazo , Primer Trimestre del Embarazo , Estudios Prospectivos , Semen/citología , Recuento de Espermatozoides , Estrés Psicológico/complicaciones , Estrés Psicológico/psicología

RESUMEN

In male mice heterozygous for a null apolipoprotein B (apoB), allele infertility was noticed. These data led us to investigate a possible role of APOB gene polymorphism and male infertility in humans. In this case-control study, we searched for an association between the insertion/deletion (I/D) polymorphism of the APOB gene and male infertility in 560 Slovene Caucasian men. The study group consisted of 310 infertile patients: 115 with azoospermia and 195 with oligoasthenoteratozoospermia (OAT) and a control group of 250 fertile men. We found a statistically significant difference in the genotype distribution between the two groups (chi2 = 6.315, P = 0.043). A separate analysis of azoospermic and OAT patients demonstrated that significant differences in genotype distribution were limited to the OAT group (chi2 = 7.011, P = 0.030). The presence of the D allele (DD or ID genotypes) conferred a 1.6 risk [chi2 = 6.089, P = 0.014, 95% confidence interval (95% CI) = 1.102-2.347] for male infertility in the OAT group of patients. We did not find a correlation between the I/D polymorphism genotypes and the clinical characteristics of infertile men: sperm concentration (P = 0.102), rapid progressive motility (P = 0.449), normal morphology (P = 0.085) and Johnsen score (P = 0.531). These data suggest that genetic variation in the signal peptide of the APOB gene (I/D polymorphism) might be a risk factor for the development of male infertility.

Asunto(s)

Apolipoproteínas B/genética , Astenozoospermia/genética , Mutagénesis Insercional , Oligospermia/genética , Señales de Clasificación de Proteína/genética , Eliminación de Secuencia , Adulto , Astenozoospermia/epidemiología , Astenozoospermia/patología , Azoospermia/epidemiología , Azoospermia/genética , Azoospermia/patología , Estudios de Casos y Controles , Hormona Folículo Estimulante/sangre , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Oligospermia/epidemiología , Tamaño de los Órganos , Polimorfismo Genético , Células de Sertoli/patología , Eslovenia/epidemiología , Motilidad Espermática , Espermatogénesis , Testículo/patología

RESUMEN

The aim of this retrospective study was to evaluate the efficiency of testicular biopsy and intracytoplasmic sperm injection (ICSI) in patients with aspermia or non-obstructive azoospermia (NOA) after cancer treatment. From 1996 to 2003, 30 men with a history of cancer, affected by aspermia or NOA and without sperm cryopreserved before cytotoxic treatment underwent testicular sperm extraction (TESE). In these men, clinical, hormonal and histological characteristics were compared; 13 underwent 39 TESE-ICSI cycles using frozen-thawed testicular spermatozoa (TESE-ICSI group). In the same period, 31 ICSI cycles were performed in 20 men with aspermia or NOA using ejaculated sperm frozen before cancer treatment (ejaculated sperm-ICSI group). Fertilization, blastocyst development, pregnancy and miscarriage rates were compared between the groups. Testicular volume, serum follicle-stimulating hormone level and Johnsen score indicated complete although reduced spermatogenesis in men with aspermia and abnormal spermatogenesis in men with NOA. After TESE, sperm retrieval was positive in 92% of men with aspermia and 58% of men with NOA. In TESE-ICSI patients with NOA a significantly lower proportion of embryos developed to the blastocyst stage than in patients with aspermia and in those after ICSI with frozen-thawed ejaculated sperm (23% vs. 43% and 47%, p = 0.03 and p < 0.01 respectively). In all groups the miscarriage rates were high; in patients with aspermia and NOA, characterized by increased age, the miscarriage rate tended to be higher in spite of similar female age and female indications of infertility. In patients affected by aspermia or NOA after cancer treatment and without sperm cryopreserved before treatment, TESE-ICSI using testicular sperm provide a chance to father a child.

Asunto(s)

Neoplasias/complicaciones , Oligospermia , Inyecciones de Esperma Intracitoplasmáticas , Separación Celular/métodos , Eyaculación , Femenino , Humanos , Infertilidad Femenina , Masculino , Neoplasias/terapia , Oligospermia/etiología , Embarazo , Resultado del Embarazo , Estudios Retrospectivos , Preservación de Semen , Espermatozoides , Testículo/citología

RESUMEN

BACKGROUND: Deletions of the AZFb region on the long arm of the human Y chromosome cause male infertility. However, the reciprocal products of these deletion events, AZFb duplications, have not been reported to date. Furthermore, it is not known whether potential AZFb duplications represent a risk factor for spermatogenic failure. METHODS: A total of 150 patients with male idiopathic subfertility (79 non-obstructive azoospermics and 71 oligozoospermics) and 150 fertile men were analysed for deletion/duplication of the sY125 locus and of the JARID1D gene using real-time PCR. RESULTS: Three azoospermic men had deletion of the sY125 locus and of the JARID1D gene. No duplication was detected. CONCLUSIONS: In our limited sample, AZFb duplications do not appear to be associated with male infertility.

Asunto(s)

Cromosomas Humanos Y/genética , Reacción en Cadena de la Polimerasa/métodos , Secuencia de Bases , Mapeo Cromosómico , Cartilla de ADN , Duplicación de Gen , Humanos , Infertilidad Masculina/genética , Masculino , Oligospermia/genética

RESUMEN

The cAMP-responsive element modulator (CREM) gene plays a pivotal role in the mouse spermatogenesis, but its role in the human infertility has not been fully established. We performed a mutation screening in 13 Slovenian men with round spermatid arrest and in six controls. Eleven genetic changes have been identified in the human CREM gene, three novel single-nucleotide polymorphisms [within the promoters P1, P3 and intervening sequence 1 (IVS1)], one insertion (IVS2) and one non-sense mutation (exon gamma). Some infertile patients seem to accumulate potentially harmful genetic changes. We identified a patient with no CREM immunoreactive protein that was homozygous for the nucleotide changes in all promoters, IVS 1, 2, 6, and was heterozygous for the mutation in exon gamma. Interestingly, insertion in IVS2 (IVS2-58_55insT) results in a four-fold decrease in binding of nuclear proteins. Computer predictions suggested the presence of a potential novel CREM promoter, however, random amplification of cDNA ends from the human testis cDNA library was not successful in confirming a novel transcription start site of the CREM gene. Screening of a larger number of patients and controls is required to elucidate whether the observed combinations of genetic changes in the CREM gene can explain some forms of male infertility.

Asunto(s)

Modulador del Elemento de Respuesta al AMP Cíclico/genética , AMP Cíclico/fisiología , Infertilidad Masculina/genética , Elementos de Respuesta/genética , Adulto , Secuencia de Bases , Cromosomas Artificiales Bacterianos , Modulador del Elemento de Respuesta al AMP Cíclico/metabolismo , Marcadores Genéticos , Haplotipos , Humanos , Masculino , Datos de Secuencia Molecular , Mutación , Alineación de Secuencia , Análisis de Secuencia de ADN , Testículo/metabolismo

RESUMEN

Genital tract inflammation was for a long time suspected of inducing sterility in the male. The role of leukocytes in sperm changes is still debated. The identification of inflammation is based on the presence of activated leukocytes and the elastase measurement. Residual inflammation is evaluated by the reactive oxygen species activity. The true damage caused by inflammation on spermatozoa is reflected in negative changes observed during sperm apoptosis. Increased sperm DNA fragmentation is frequently observed while classical sperm characteristics are not modified automatically. The diagnosis of genital tract inflammation, follow-up after treatment and prognosis in terms of fertility require the use of different and complementary parameters.

Asunto(s)

Biomarcadores/análisis , Enfermedades de los Genitales Masculinos/enzimología , Inflamación/enzimología , Espermatozoides/química , Apoptosis , Fragmentación del ADN , Enfermedades de los Genitales Masculinos/patología , Enfermedades de los Genitales Masculinos/terapia , Humanos , Infecciones/patología , Infertilidad Masculina/etiología , Inflamación/patología , Inflamación/terapia , Leucocitos , Masculino , Elastasa Pancreática , Pronóstico , Espermatozoides/ultraestructura

RESUMEN

Benign prostatic hyperplasia (BPH) involves proliferation of smooth muscle cells and increased deposition of extracellular matrix (ECM). We recently found that pentosan polysulfate (PPS) has marked effects on growth and ECM of smooth muscle cells derived from vascular tissues. We examined smooth muscle cells cultured from human prostates and the effects of PPS on their growth and ECM production. Fragments of surgical prostatectomy specimens were diced, digested with collagenase (0.01%), and placed in culture medium supplemented with 20% fetal bovine serum. Outgrowths of elongated cells were characterized by light microscopic examination and immunohistochemical techniques by the presence of F-actin, alpha-smooth muscle actin, and myosin, which is a characteristic of smooth muscle cells. Two independent isolates were propagated, and growth curves and ECM production were assessed in the presence and absence of PPS (10 or 100 microg/ml). PPS decreased cell number beginning at day 1 and throughout the incubation period, up to 4 days. The amount of the ECM degradative enzymes, metallo-proteinases MMP-9 and MMP-2, was examined by zymography. PPS did not alter the amount of MMP-2 in the supernatants but MMP-9 was increased 234.4 +/- 17.23-fold over control cells. Tissue inhibitor of MMP (TIMPS), examined by reverse zymography, increased 200% over control. The amount of alpha I type (IV) and alpha I type (I) collagen released in the supernatant, measured by ELISA, significantly decreased in PPS-treated cultures. In conclusion, we found that the administration of PPS decreased proliferation as well as ECM production in prostate smooth muscle. Since smooth muscle proliferation and ECM are involved in the pathophysiology of BPH, PPS may have therapeutic potential.

Asunto(s)

División Celular/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Matriz Extracelular/metabolismo , Músculo Liso/efectos de los fármacos , Músculo Liso/crecimiento & desarrollo , Poliéster Pentosan Sulfúrico/farmacología , Próstata/fisiología , Hiperplasia Prostática/tratamiento farmacológico , Técnicas de Cultivo de Célula , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunohistoquímica , Masculino , Hiperplasia Prostática/fisiopatología , Neoplasias de la Próstata/fisiopatología

RESUMEN

BACKGROUND: The aim of this study was to evaluate the role of blastocyst culture in patients with azoospermia. METHODS: In 98 cycles embryos were cultured for 2 days and in 128 cycles for 5 days to reach the blastocyst stage; a maximum of two of the most developed embryos were transferred in each group. RESULTS: There was a negative correlation between a high (>/=20 IU/l) male serum FSH and embryo development, manifested as embryos not reaching the morula stage on day 5 (r = 0.387; P < 0.05). After prolonged culture, 23% of embryos reached the blastocyst stage. The pregnancy rates per transfer, and the abortion rates were approximately the same in the day 2 group and the day 5 group (20 versus 20% and 19 versus 18% respectively). After blastocyst transfer, a high clinical pregnancy rate (55%) and a low abortion rate (6%) were achieved, whereas the transfer of arrested embryos provided a low pregnancy rate (2%) and a high abortion rate (100%). If only blastocysts had been transferred on day 5, the clinical pregnancy rate per started cycle would have been approximately the same in both groups (13 versus 16%). CONCLUSIONS: Blastocyst formation is a good indicator of clinical results after ICSI with testicular sperm.

Asunto(s)

Blastocisto/fisiología , Infertilidad Masculina/terapia , Oligospermia/complicaciones , Inyecciones de Esperma Intracitoplasmáticas , Espermatozoides , Testículo , Aborto Espontáneo/epidemiología , Técnicas de Cultivo , Transferencia de Embrión , Desarrollo Embrionario y Fetal , Femenino , Humanos , Incidencia , Infertilidad Masculina/etiología , Masculino , Embarazo , Índice de Embarazo , Pronóstico , Resultado del Tratamiento

RESUMEN

BACKGROUND: The objective of this study was to estimate the frequency of Y chromosome microdeletions in the Slovenian population of infertile men and to analyse the consequences of mutation in respect to clinical severity and prognosis. METHODS: In a controlled clinical study at the university-based medical genetics service and infertility clinic, 226 infertile men undergoing ICSI were tested. The main outcome measures included polymerase chain reaction amplification of 16 genes and gene families and 42 sequence-tagged sites in the non-recombining region of the Y chromosome, semen, testicular volume and testicular histological analysis, serum FSH concentrations, fertilization and respective pregnancy rates. RESULTS: The incidence of deletions was 4.4%: 8.6% in men with azoospermia and 1.5% in men with oligoasthenoteratozoospermia. Isolated gene deletions were not identified. No statistically significant differences in clinical outcome measures were found in patients with mutations versus patients without mutations. High fertilization (49%) and pregnancy (43%) rates with sperm of patients with Y chromosome deletions were obtained. CONCLUSIONS: Testing for gene-specific microdeletions does not contribute significantly to the sensitivity of microdeletion test. Fertilization and pregnancy rates obtained using sperm of patients with Y chromosome deletions were comparable with those achieved in conventional IVF.

Asunto(s)

Eliminación de Gen , Infertilidad Masculina/genética , Aberraciones Cromosómicas Sexuales , Cromosoma Y/genética , Femenino , Fertilización In Vitro , Hormona Folículo Estimulante/sangre , Humanos , Infertilidad Masculina/patología , Masculino , Mosaicismo , Reacción en Cadena de la Polimerasa , Embarazo , Resultado del Embarazo , Semen , Eslovenia , Inyecciones de Esperma Intracitoplasmáticas , Testículo/patología

RESUMEN

We assessed the role of the actin-polymerizing protein, ActA, in host cell invasion by Listeria monocytogenes. An in frame DeltaactA mutant was constructed in a hyperinvasive strain of prfA* genotype, in which all genes of the PrfA-dependent virulence regulon, including actA, are highly expressed in vitro. Loss of ActA production in prfA* bacteria reduced entry into Caco-2, HeLa, MDCK and Vero epithelial cells to basal levels. Reintroduction of actA into the DeltaactA prfA* mutant fully restored invasiveness, demonstrating that ActA is involved in epithelial cell invasion. ActA did not contribute to internalization by COS-1 fibroblasts and Hepa 1-6 hepatocytes. Expression of actA in Listeria innocua was sufficient to promote entry of this non-invasive species into epithelial cell lines, but not into COS-1 and Hepa 1-6 cells, indicating that ActA directs an internalization pathway specific for epithelial cells. Scanning electron microscopy of infected Caco-2 human enterocytes suggested that this pathway involves microvilli. prfA* bacteria, but not wild-type bacteria (which express PrfA-dependent genes very weakly in vitro) or prfA* DeltaactA bacteria, efficiently invaded differentiated Caco-2 cells via their apical surface. Microvilli played an active role in the phagocytosis of the prfA* strain, and actA was required for their remodelling into pseudopods mediating bacterial uptake. Thus, ActA appears to be a multifunctional virulence factor involved in two important aspects of Listeria pathogenesis: actin-based motility and host cell tropism and invasion.

Asunto(s)

Proteínas Bacterianas/fisiología , Listeria monocytogenes/fisiología , Proteínas de la Membrana/fisiología , Secuencia de Aminoácidos , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Células COS , Células CACO-2 , Línea Celular , Chlorocebus aethiops , ADN Bacteriano , Perros , Células Epiteliales/citología , Células Epiteliales/metabolismo , Células Epiteliales/microbiología , Expresión Génica , Células HeLa , Humanos , Listeria monocytogenes/aislamiento & purificación , Listeria monocytogenes/metabolismo , Listeria monocytogenes/ultraestructura , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Microscopía Electrónica de Rastreo/métodos , Datos de Secuencia Molecular , Mutagénesis , Factores de Terminación de Péptidos , Transactivadores , Células Vero

RESUMEN

BACKGROUND: Overactive bladder (OAB) affects >17 million individuals in the United States, but the symptoms of OAB are frequently underreported by patients and therefore untreated by physicians. OBJECTIVE: The purpose of this observational study was to investigate the demographic and clinical factors associated with the decision to treat OAB pharmacologically and identify factors associated with physicians' assessment of symptom severity. METHODS: We studied 31 physicians treating 217 patients with OAB and collected data on patient demographic characteristics, OAB symptoms, previous management strategies, physicians' assessments of OAB severity, and treatment prescribed. Stepwise logistic regression was used to identify factors associated with selecting pharmacologic treatment and with physician assessment of severity of OAB symptoms. RESULTS: The mean age of the patients was 61.3 years; approximately 82% were female, and approximately 73% were white. Participants with urinary incontinence were significantly (P < 0.001) more likely to be treated with medication than were those with only symptoms of nocturia or urinary frequency. Other significant factors associated with pharmacologic treatment were being white (odds ratio [OR], 9.5; 95% CI, 2.9-30.8); being black (OR, 5.9; 95% CI, 1.2-29.7); physician's clinical assessment of OAB as moderate (OR, 3.5; 95% CI, 1.5-8.2) or severe (OR, 3.8; 95% CI, 1.1-13.7); previous use of medication (OR, 2.9; 95% CI, 1.1-8.1); and number of incontinence episodes in the last 24 hours (OR, 1.2; 95% CI, 1.0-1.5). Factors associated with physician assessment of OAB severity included distress due to OAB symptoms (OR, 2.1; 95% CI, 1.3-3.2), number of incontinence episodes in the last 24 hours (OR, 1.2; 95% CI, 1.1-1.4), and use of previous treatment(s) (OR, 0.4; 95% CI, 0.2-0.8). CONCLUSIONS: Both demographic and clinical symptoms of OAB were associated with the decision to treat OAB with medication, whereas physicians' assessment of OAB severity was associated only with clinical symptoms. Urinary incontinence was the key symptom associated with the decision to treat patients with medication and with the assessment of OAB symptom severity.

Asunto(s)

Pautas de la Práctica en Medicina , Incontinencia Urinaria/diagnóstico , Incontinencia Urinaria/terapia , Anciano , Humanos , Masculino , Persona de Mediana Edad

RESUMEN

Apoptosis is a widespread phenomena during development. It represents a form of cell death and has a crucial role in tissue homeostasis. Apoptosis is also involved in a number of pathological conditions. Spermatogenesis is a dynamic process of germ cell proliferation and differentiation. During regular spermatogenesis, the number of testicular germ cells degenerate by an apoptotic process. The significance of regulating cell population by apoptosis is more apparent when sperm production is halted. The presence and frequency of apoptosis in germ cells of human testis biopsy specimens were tested. The results confirm the presence of germ cell apoptosis but not the apoptosis of Sertoli cells. The increased apoptotic index was observed in patients with azoospermia in comparison with normal but obstructed spermatogenesis.

Asunto(s)

Apoptosis , Infertilidad Masculina/patología , Túbulos Seminíferos/patología , Espermatozoides/patología , Biopsia , Humanos , Etiquetado Corte-Fin in Situ , Masculino , Espermatogénesis