RESUMEN
BACKGROUND: Venous thromboembolism (VTE) is a common complication in patients with anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAV) and confers significant morbidity and mortality. Both acute and past cytomegalovirus (CMV) infection have been identified as risk factors for VTE in immunocompetent and immunosuppressed individuals. Here, we examine whether past exposure to CMV is a risk factor for VTE amongst patients with AAV. METHODS: We retrospectively analysed outcomes of patients with a new diagnosis of AAV from a UK cohort. All confirmed cases of VTE where CMV IgG serology was available were recorded. Retrospective collection of the same data for patients at a North American centre was used as a validation cohort. RESULTS: VTE was common with 12% of patients from the study cohort (total 259 patients) developing an event during the median follow-up period of 8.5 years of which 60% occurred within the first 12 months following diagnosis. Sixteen percent of CMV seropositive patients developed a VTE compared with 5% of patients who were seronegative (p = 0.007) and CMV seropositivity remained an independent predictor of VTE in multivariable analysis (HR 2.96 [1.094-8.011] p = 0.033). CMV seropositivity at diagnosis was confirmed as a significant risk factor for VTE in the American validation cohort (p = 0.032). CONCLUSIONS: VTE is common in patients with AAV, especially within the first year of diagnosis. Past infection with CMV is an independent risk factor associated with VTE in AAV.
Asunto(s)
Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos , Infecciones por Citomegalovirus , Tromboembolia Venosa , Citomegalovirus , Infecciones por Citomegalovirus/complicaciones , Infecciones por Citomegalovirus/diagnóstico , Infecciones por Citomegalovirus/epidemiología , Humanos , Estudios Retrospectivos , Factores de Riesgo , Tromboembolia Venosa/diagnóstico , Tromboembolia Venosa/epidemiología , Tromboembolia Venosa/etiologíaRESUMEN
INTRODUCTION: The effect of SARS-CoV-2 severity or the trimester of infection in pregnant mothers, placentas, and infants is not fully understood. METHODS: A retrospective, observational cohort study in Chapel Hill, NC of 115 mothers with SARS-CoV-2 and singleton pregnancies from December 1, 2019 to May 31, 2021 via chart review to document the infants' weight, length, head circumference, survival, congenital abnormalities, hearing loss, maternal complications, and placental pathology classified by the Amsterdam criteria. RESULTS: Of the 115 mothers, 85.2% were asymptomatic (n = 37) or had mild (n = 61) symptoms, 13.0% had moderate (n = 9) or severe (n = 6) COVID-19, and 1.74% (n = 2) did not have symptoms recorded. Moderate and severe maternal infections were associated with increased C-section, premature delivery, infant NICU admission, and were more likely to occur in Type 1 (p = 0.0055) and Type 2 (p = 0.0285) diabetic mothers. Only one infant (0.870%) became infected with SARS-CoV-2, which was not via the placenta. Most placentas (n = 63, 54.8%) did not show specific histologic findings; however, a subset showed mild maternal vascular malperfusion (n = 26, 22.6%) and/or mild microscopic ascending intrauterine infection (n = 28, 24.3%). The infants had no identifiable congenital abnormalities, and all infants and mothers survived. DISCUSSION: Most mothers and their infants had a routine clinical course; however, moderate and severe COVID-19 maternal infections were associated with pregnancy complications and premature delivery. Mothers with pre-existing, non-gestational diabetes were at greatest risk of developing moderate or severe COVID-19. The placental injury patterns of maternal vascular malperfusion and/or microscopic ascending intrauterine infection were not associated with maternal COVID-19 severity.
Asunto(s)
COVID-19 , Complicaciones Infecciosas del Embarazo , Nacimiento Prematuro , Femenino , Humanos , Inmunoglobulina G , Lactante , Transmisión Vertical de Enfermedad Infecciosa , Madres , Placenta/patología , Embarazo , Complicaciones Infecciosas del Embarazo/patología , Nacimiento Prematuro/epidemiología , Nacimiento Prematuro/patología , Estudios Retrospectivos , SARS-CoV-2RESUMEN
Medium-chain acyl CoA dehydrogenase deficiency (MCADD) and other inborn errors of metabolism are common causes of Sudden Unexpected Deaths in Infancy (SUDI). If identified early or before metabolic decompensation, MCADD is manageable. In the US and other countries, identification of MCADD has improved through the routine use of newborn screening (NBS), which is able to identify most cases. This case study presented here occurred before NBS was implemented in Ohio for MCADD and outlines the typical clinical presentation, pathological features, and relevant biochemical and molecular markers for identifying MCADD. Genetic counselling should be sought for the family if MCADD is identified.
Asunto(s)
Errores Innatos del Metabolismo Lipídico , Recién Nacido , Lactante , Humanos , Acil-CoA Deshidrogenasa/genética , Errores Innatos del Metabolismo Lipídico/diagnóstico , Errores Innatos del Metabolismo Lipídico/genética , Tamizaje Neonatal , BiomarcadoresRESUMEN
Pathogenesis of anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis is B cell-dependent, although how particular B cell subsets modulate immunopathogenesis remains unknown. Although their phenotype remains controversial, regulatory B cells (Bregs ), play a role in immunological tolerance via interleukin (IL)-10. Putative CD19(+) CD24(hi) CD38(hi) and CD19(+) CD24(hi) CD27(+) Bregs were evaluated in addition to their CD5(+) subsets in 69 patients with ANCA-associated vasculitis (AAV). B cell IL-10 was verified by flow cytometry following culture with CD40 ligand and cytosine-phosphate-guanosine (CpG) DNA. Patients with active disease had decreased levels of CD5(+) CD24(hi) CD38(hi) B cells and IL-10(+) B cells compared to patients in remission and healthy controls (HCs). As IL-10(+) and CD5(+) CD24(hi) CD38(hi) B cells normalized in remission within an individual, ANCA titres decreased. The CD5(+) subset of CD24(hi) CD38(hi) B cells decreases in active disease and rebounds during remission similarly to IL-10-producing B cells. Moreover, CD5(+) B cells are enriched in the ability to produce IL-10 compared to CD5(neg) B cells. Together these results suggest that CD5 may identify functional IL-10-producing Bregs . The malfunction of Bregs during active disease due to reduced IL-10 expression may thus permit ANCA production.
Asunto(s)
Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/inmunología , Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Linfocitos B Reguladores/inmunología , Regulación de la Expresión Génica/inmunología , Interleucina-10/inmunología , ADP-Ribosil Ciclasa 1/sangre , ADP-Ribosil Ciclasa 1/inmunología , Adyuvantes Inmunológicos/farmacología , Adulto , Anciano , Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/sangre , Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/patología , Vasculitis Asociada a Anticuerpos Citoplasmáticos Antineutrófilos/terapia , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Linfocitos B Reguladores/metabolismo , Linfocitos B Reguladores/patología , Antígeno CD24/sangre , Antígeno CD24/inmunología , Antígenos CD40/sangre , Antígenos CD40/inmunología , Antígenos CD5/sangre , Antígenos CD5/inmunología , Femenino , Citometría de Flujo , Humanos , Interleucina-10/sangre , Masculino , Glicoproteínas de Membrana/sangre , Glicoproteínas de Membrana/inmunología , Persona de Mediana Edad , Oligodesoxirribonucleótidos/farmacología , Inducción de RemisiónRESUMEN
Myeloperoxidase (MPO) is one of the major target antigens of antineutrophil cytoplasmic autoantibodies (ANCA) found in patients with small-vessel vasculitis and pauci-immune necrotizing glomerulonephritis. To date, the target epitopes of MPO-ANCA remain poorly defined. Human MPO-ANCA do not typically bind mouse MPO. We utilized the differences between human and mouse MPO to identify the target regions of MPO-ANCA. We generated five chimeric MPO molecules in which we replaced different segments of the human or mouse molecules with their homologous counterpart from the other species. Of serum samples from 28 patients screened for this study, 43 samples from 14 patients with MPO-ANCA-associated vasculitis were tested against recombinant human and mouse MPO and the panel of chimeric molecules. Sera from 64 and 71% of patients bound to the carboxy-terminus of the heavy chain, in the regions of amino acids 517-667 or 668-745, respectively. No patient serum bound the MPO light chain or the amino-terminus of the heavy chain. All sera bound to only one or two regions of MPO. Although the pattern of MPO-ANCA binding changed over time (4-27 months) in 6 of 10 patients with several serum samples, such changes were infrequent. Other target regions of MPO-ANCA may not have been detected due to conformational differences between the native and recombinant forms of MPO. MPO-ANCA do not target a single epitope, but rather a small number of regions of MPO, primarily in the carboxy-terminus of the heavy chain.
Asunto(s)
Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Mapeo Epitopo , Epítopos/análisis , Peroxidasa/genética , Peroxidasa/inmunología , Adulto , Anciano , Secuencia de Aminoácidos , Animales , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Femenino , Glomerulonefritis/sangre , Glomerulonefritis/enzimología , Glomerulonefritis/inmunología , Glomerulonefritis/fisiopatología , Humanos , Masculino , Ratones , Persona de Mediana Edad , Modelos Moleculares , Datos de Secuencia Molecular , Peroxidasa/sangre , Peroxidasa/química , Estructura Cuaternaria de Proteína , Proteínas Recombinantes/sangre , Proteínas Recombinantes/química , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Homología de Secuencia de Aminoácido , Vasculitis/inmunologíaRESUMEN
Women who inherit mutations in the BRCA2 cancer susceptibility gene have an 85% chance of developing breast cancer. The function of the BRCA2 gene remains elusive, but there is evidence to support its role in transcriptional transactivation, tumor suppression, and the maintenance of genomic integrity. Individuals with identical BRCA2 mutations display a different distribution of cancers, suggesting that there are low-penetrance genes that can modify disease outcome. We hypothesized that genetic background could influence embryonic survival of a Brca2 mutation in mice. Brca2-null embryos with a 129/SvEv genetic background (129(B2-/-)) died before embryonic day 8. 5. Transfer of this Brca2 mutation onto the BALB/cJ genetic background (BALB/c(B2-/-)) extended survival to embryonic day 10.5. These results indicate that the BALB/c background harbors genetic modifiers that can prolong Brca2-null embryonic survival. The extended survival of BALB/c(B2-/-) embryos enabled us to ask whether transcriptional regulation of the Brca1 and Brca2 genes is interdependent. The interdependence of Brca1 and Brca2 was evaluated by studying Brca2 gene expression in BALB/c(B1-/-) embryos and Brca1 gene expression in BALB/c(B2-/-) embryos. Nonisotopic in situ hybridization demonstrated that Brca2 transcript levels were comparable in BALB/c(B1-/-) embryos and wild-type littermates. Likewise, reverse transcriptase-polymerase chain reactions confirmed Brca1 mRNA expression in embryonic day 8.5 BALB/c(B2-/-) embryos that was comparable to Brca2-heterozygous littermates. Thus, the Brca1 and Brca2 transcripts are expressed independently of one another in Brca1- and Brca2-null embryos. Mol. Carcinog. 28:174-183, 2000.
Asunto(s)
Muerte Fetal/genética , Regulación del Desarrollo de la Expresión Génica/genética , Ratones Endogámicos BALB C/genética , Proteínas de Neoplasias/fisiología , Factores de Transcripción/fisiología , Animales , Proteína BRCA1/deficiencia , Proteína BRCA1/fisiología , Proteína BRCA2 , Secuencia de Bases , Desarrollo Embrionario y Fetal/genética , Femenino , Genes BRCA1 , Genes Letales , Predisposición Genética a la Enfermedad , Genotipo , Ratones , Ratones Endogámicos BALB C/embriología , Ratones Noqueados , Datos de Secuencia Molecular , Proteínas de Neoplasias/deficiencia , Proteínas de Neoplasias/genética , Factores de Transcripción/deficiencia , Factores de Transcripción/genética , Activación Transcripcional/genéticaRESUMEN
Although the process of glycolysis is highly conserved in eukaryotes, several glycolytic enzymes have unique structural or functional features in spermatogenic cells. We previously identified and characterized the mouse complementary DNA (cDNA) and a gene for 1 of these enzymes, glyceraldehyde 3-phosphate dehydrogenase-s (Gapds). This gene is expressed only in spermatids. The enzyme appears to have an essential role in energy production required for fertilization, and it is reported to be susceptible to inhibition by certain environmental chemicals. We have now cloned and sequenced the cDNA for the human homologue of glyceraldehyde 3-phosphate dehydrogenase (GAPD2) and determined the structure of the gene. The messenger RNA (mRNA) was detected in testis, but not in 15 other human tissues analyzed by Northern blot technique. The deduced GAPD2 protein contains 408 amino acids and is 68% identical with somatic cell GAPD. GAPD2 has a 72-amino acid segment at the amino terminal end that is not present in somatic cell GAPD. This segment is proline-rich but contains smaller stretches of polyproline and is 30 amino acids shorter than the comparable segment of mouse GAPDS. The structure of the human GAPD2 gene was determined by polymerase chain reaction (PCR) to identify exon-intron junctions in a genomic clone and in total genomic DNA. The locations of these junctions in the GAPD2 gene corresponded precisely to those of the 11 exon-intron junctions in the mouse Gapds gene. Immunohistochemical studies found that GAPD2 is located in the principal piece of the flagellum of human spermatozoa, as are GAPDS in mouse and rat spermatozoa. GAPD2 extracted from human spermatozoa and analyzed by Western blot technique migrated with an apparent molecular weight of approximately 56,000, although the calculated molecular weight is 44 501. The conserved nature of the mouse, rat, and human enzymes suggests that they serve similar roles in these and other mammalian species.
Asunto(s)
Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Espermatozoides/enzimología , Secuencia de Aminoácidos , Secuencia de Bases , Western Blotting , Mapeo Cromosómico , Cromosomas Humanos Par 19 , ADN Complementario , Humanos , Masculino , Datos de Secuencia Molecular , Homología de Secuencia de AminoácidoRESUMEN
Mice lacking estrogen receptors alpha and beta were generated to clarify the roles of each receptor in the physiology of estrogen target tissues. Both sexes of alphabeta estrogen receptor knockout (alphabetaERKO) mutants exhibit normal reproductive tract development but are infertile. Ovaries of adult alphabetaERKO females exhibit follicle transdifferentiation to structures resembling seminiferous tubules of the testis, including Sertoli-like cells and expression of Müllerian inhibiting substance, sulfated glycoprotein-2, and Sox9. Therefore, loss of both receptors leads to an ovarian phenotype that is distinct from that of the individual ERKO mutants, which indicates that both receptors are required for the maintenance of germ and somatic cells in the postnatal ovary.
Asunto(s)
Trastornos del Desarrollo Sexual , Chaperonas Moleculares , Ovario/anatomía & histología , Ovario/fisiología , Receptores de Estrógenos/fisiología , Animales , Hormona Antimülleriana , Diferenciación Celular , Clusterina , Estradiol/fisiología , Receptor alfa de Estrógeno , Receptor beta de Estrógeno , Femenino , Marcación de Gen , Glicoproteínas/análisis , Inhibidores de Crecimiento/análisis , Proteínas del Grupo de Alta Movilidad/análisis , Hormona Luteinizante/sangre , Masculino , Ratones , Ratones Noqueados , Ovario/citología , Ovario/crecimiento & desarrollo , Receptores de Estrógenos/genética , Factor de Transcripción SOX9 , Túbulos Seminíferos/anatomía & histología , Túbulos Seminíferos/citología , Células de Sertoli/citología , Transducción de Señal , Hormonas Testiculares/análisis , Testículo/anatomía & histología , Testículo/citología , Testículo/crecimiento & desarrollo , Testículo/fisiología , Factores de Transcripción/análisisRESUMEN
Ovarian-derived estradiol plays a critical endocrine role in the regulation of gonadotropin synthesis and secretion from the hypothalamic-pituitary axis. In turn, several para/autocrine effects of estrogen within the ovary are known, including increased ovarian weight, stimulation of granulosa cell growth, augmentation of FSH action, and attenuation of apoptosis. The estrogen receptor-alpha (ERalpha) is present in all three components of the hypothalamic-pituitary-ovarian axis of the mouse. In contrast, estrogen receptor-beta (ERbeta) is easily detectable in ovarian granulosa cells but is low to absent in the pituitary of the adult mouse. This distinct expression pattern for the two ERs suggests the presence of separate roles for each in the regulation of ovarian function. Herein, we definitively show that a lack of ERalpha in the hypothalamic-pituitary axis of the ERalpha-knockout (alphaERKO) mouse results in chronic elevation of serum LH and is the primary cause of the ovarian phenotype of polycystic follicles and anovulation. Prolonged treatment with a GnRH antagonist reduced serum LH levels and prevented the alphaERKO cystic ovarian phenotype. To investigate a direct role for ERalpha within the ovary, immature alphaERKO females were stimulated to ovulate with exogenous gonadotropins. Ovulatory capacity in the immature alphaERKO female was reduced compared with age-matched wild-type (14.5+/-2.9 vs. 40.6+/-2.6 oocytes/animal, respectively); however, oocytes collected from the alphaERKO were able to undergo successful in vitro fertilization. A similar discrepancy in oocyte yield was observed after superovulation of peripubertal (42 days) wild-type and alphaERKO females. In addition, ovaries from immature superovulated alphaERKO females possessed several ovulatory but unruptured follicles. Investigations of the possible reasons for the reduced number of ovulations in the alphaERKO included ribonuclease protection assays to assess the mRNA levels of several markers of follicular maturation and ovulation, including ERbeta, LH-receptor, cyclin-D2, P450-side chain cleavage enzyme, prostaglandin synthase-2, and progesterone receptor. No marked differences in the expression pattern for these mRNAs during the superovulation regimen were observed in the immature alphaERKO ovary compared with that of the wild-type. Serum progesterone levels just before ovulation were slightly lower in the alphaERKO compared with wild-type. These studies indicate that treatment of alphaERKO females with a GnRH antagonist decreased the serum LH levels to within the wild-type range and concurrently prevented development of the characteristic ovarian phenotype of cystic and hemorrhagic follicles. Furthermore, a lack of functional ERalpha within the ovary had no effect on the regulation of several genes required for follicular maturation and ovulation. However, the reduced numbers of ovulations following the administration of exogenous gonadotropins in the alphaERKO suggests an intraovarian role for ERalpha in follicular development and ovulation.
Asunto(s)
Ovulación , Síndrome del Ovario Poliquístico/genética , Receptores de Estrógenos/deficiencia , Receptores de Estrógenos/genética , Animales , Gonadotropina Coriónica/farmacología , Receptor alfa de Estrógeno , Femenino , Fertilización In Vitro , Hormona Liberadora de Gonadotropina/antagonistas & inhibidores , Gonadotropinas Equinas/farmacología , Hipotálamo/fisiopatología , Hormona Luteinizante/sangre , Ratones , Ratones Noqueados , Oocitos/fisiología , Ovario/patología , Fenotipo , Hipófisis/fisiopatología , Síndrome del Ovario Poliquístico/patología , Síndrome del Ovario Poliquístico/fisiopatología , Progesterona/sangre , ARN Mensajero/metabolismo , Receptores de Estrógenos/fisiología , SuperovulaciónAsunto(s)
Terapia Respiratoria/economía , Instituciones de Cuidados Especializados de Enfermería/economía , Enfermedad Aguda , Anciano , Centers for Medicare and Medicaid Services, U.S. , Humanos , Tiempo de Internación , Medicare/economía , Competencia Profesional , Sistema de Pago Prospectivo , Terapia Respiratoria/estadística & datos numéricos , Estados UnidosRESUMEN
Fertilin, a member of the ADAM family, is found on the plasma membrane of mammalian sperm. Sperm from mice lacking fertilin beta were shown to be deficient in sperm-egg membrane adhesion, sperm-egg fusion, migration from the uterus into the oviduct, and binding to the egg zona pellucida. Egg activation was unaffected. The results are consistent with a direct role of fertilin in sperm-egg plasma membrane interaction. Fertilin could also have a direct role in sperm-zona binding or oviduct migration; alternatively, the effects on these functions could result from the absence of fertilin activity during spermatogenesis.
Asunto(s)
Glicoproteínas de Membrana/fisiología , Metaloendopeptidasas/fisiología , Interacciones Espermatozoide-Óvulo , Espermatozoides/fisiología , Proteínas ADAM , Animales , Calcio/metabolismo , Adhesión Celular , Membrana Celular/fisiología , Trompas Uterinas , Femenino , Fertilinas , Masculino , Fusión de Membrana , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Metaloendopeptidasas/genética , Metaloendopeptidasas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Óvulo/fisiología , Capacitación Espermática , Espermatogénesis , Espermatozoides/química , Zona Pelúcida/fisiologíaRESUMEN
The spermatogenic cell-specific isoform of glyceraldehyde 3-phosphate dehydrogenase (GAPD-S) may regulate glycolysis and energy production required for sperm motility. Although the steady-state level of Gapd-s mRNA is maximal at step 9 of mouse spermatogenesis, GAPD-S protein was not detected by immunohistochemistry until steps 12-13. This result suggests that Gapd-s is translationally regulated. Western blot analysis of isolated germ cells confirmed that GAPD-S is not detected in pachytene spermatocytes or round spermatids. A major immunoreactive protein migrating with a molecular weight (M(r)) of 69,200 was observed in condensing spermatids and cauda sperm. Additional minor proteins that migrated at M(r) 55,200, 32,500, and 27,500 were detected in sperm. The molecular weight of GAPD-S is higher than the predicted molecular weight of 47,445, apparently due to a proline-rich 105-amino acid domain at the N-terminus. Recombinant GAPD-S protein lacking the proline-rich region migrated at M(r) 38,250, comparably to somatic GAPD, which also lacks the proline-rich domain. Indirect immunofluorescence demonstrated that GAPD-S is restricted to the principal piece in the sperm flagellum. Western blot analysis indicated that GAPD-S is tightly associated with the fibrous sheath of the flagellum, consistent with a potential role in regulating sperm motility.
Asunto(s)
Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Isoenzimas/metabolismo , Espermatogénesis/fisiología , Animales , Secuencia de Bases , Cartilla de ADN/genética , Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Gliceraldehído-3-Fosfato Deshidrogenasas/inmunología , Inmunohistoquímica , Isoenzimas/genética , Isoenzimas/inmunología , Masculino , Ratones , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Cola del Espermatozoide/enzimología , Espermatogénesis/genéticaRESUMEN
The mouse homologues of the breast cancer susceptibility genes, Brca1 and Brca2, are expressed in a cell cycle-dependent fashion in vitro and appear to be regulated by similar or overlapping pathways. Therefore, we compared the non isotopic in situ hybridization expression patterns of Brca1 and Brca2 mRNA in vivo in mitotic and meiotic cells during mouse embryogenesis, mammary gland development, and in adult tissues including testes, ovaries, and hormonally altered ovaries. Brca1 and Brca2 are expressed concordantly in proliferating cells of embryos, and the mammary gland undergoing morphogenesis and in most adult tissues. The expression pattern of Brca1 and Brca2 correlates with the localization of proliferating cell nuclear antigen, an indicator of proliferative activity. In the ovary, Brca1 and Brca2 exhibited a comparable hormone-independent pattern of expression in oocytes, granulosa cells and thecal cells of developing follicles. In the testes, Brca1 and Brca2 were expressed in mitotic spermatogonia and early meiotic prophase spermatocytes. Northern analyses of prepubertal mouse testes revealed that the time course of Brca2 expression was delayed in spermatogonia relative to Brca1. Thus, while Brca1 and Brca2 share concordant cell-specific patterns of expression in most proliferating tissues, these observations suggest that they may have distinct roles during meiosis.
Asunto(s)
Genes BRCA1 , Meiosis/genética , Mitosis/genética , Proteínas de Neoplasias/genética , Factores de Transcripción/genética , Animales , Proteína BRCA2 , Desarrollo Embrionario y Fetal/genética , Femenino , Regulación del Desarrollo de la Expresión Génica , Hibridación in Situ , Masculino , Glándulas Mamarias Animales/embriología , Glándulas Mamarias Animales/metabolismo , Ratones , Morfogénesis , Ovario/metabolismo , Embarazo , Testículo/embriología , Testículo/metabolismoRESUMEN
Extensive in vivo gene transfer studies in animal models and human gene therapy clinical trials with E1-deleted adenovirus vectors have demonstrated transience of transgene expression due to direct cytopathic effects of the vectors and host immune response to virally expressed proteins. In order to overcome these difficulties, we have recently developed packaging cell lines which support the growth of adenovirus vectors containing lethal deletions in both E1 and E4 gene regions. Here we demonstrate that use of E1/E4-deleted adenovirus vectors leads to prolonged in vivo transgene expression due to elimination of cytopathic effects and significant reduction of virus-specific immune response.
Asunto(s)
Adenoviridae , Proteínas E1 de Adenovirus/genética , Eliminación de Gen , Técnicas de Transferencia de Gen , Vectores Genéticos , Proteínas E4 de Adenovirus/genética , Animales , Linfocitos T CD8-positivos/inmunología , Expresión Génica , Inmunidad Celular , Hígado/citología , Hígado/inmunología , Ratones , Ratones Endogámicos CBA , TransgenesRESUMEN
The reproductive system of male mice homozygous for a mutation in the estrogen receptor (ER) gene (ER knock-out; ERKO) appears normal at the anatomical level. However, these males are infertile, indicating an essential role for ER-mediated processes in the regulation of male reproduction. Adult ERKO male mice have significantly fewer epididymal sperm than heterozygous or wild-type males. Although spermatogenesis is occurring in some seminiferous tubules of 3- to 5-month-old ERKO males, other tubules either have a dilated lumen and a disorganized seminiferous epithelium with few spermatogenic cells or lack a lumen and contain mainly Sertoli cells. There are no obvious differences in seminiferous tubules at 10 days of age between wild-type and ERKO mice, but the lumen in ERKO males is dilated in all seminiferous tubules by 20 days. However, spermatogenesis progresses and similar numbers of sperm are present in the cauda epididymis of ERKO and wild-type males until 10 weeks of age. Disruption of spermatogenesis and degeneration of the seminiferous tubules become apparent after 10 weeks in the caudal pole of the testis and progresses in a wave to the cranial pole by 6 months. However, the seminal vesicles, coagulating glands, prostate, and epididymis do not appear to be altered morphologically in ERKO mice. Serum testosterone levels are somewhat elevated, but LH and FSH levels are not significantly different from those in wild-type males. Sperm from 8- to 16-week-old mice have reduced motility and are ineffective at fertilizing eggs in vitro. In addition, ERKO males housed overnight with hormone-primed wild-type females produce significantly fewer copulatory plugs than do heterozygous or wild-type males. These results suggest that estrogen action is required for fertility in male mice and that the mutation of the ER in ERKO males leads to reduced mating frequency, low sperm numbers, and defective sperm function.
Asunto(s)
Infertilidad Masculina/genética , Receptores de Estrógenos/genética , Espermatogénesis/genética , Animales , Copulación , Epidídimo/anatomía & histología , Epidídimo/patología , Epidídimo/fisiopatología , Epitelio/patología , Femenino , Hormona Folículo Estimulante/sangre , Heterocigoto , Homocigoto , Infertilidad Masculina/patología , Infertilidad Masculina/fisiopatología , Tamaño de la Camada , Hormona Luteinizante/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Tamaño de los Órganos , Reproducción , Túbulos Seminíferos/patología , Túbulos Seminíferos/fisiopatología , Testículo/anatomía & histología , Testosterona/sangre , Conducto Deferente/anatomía & histologíaRESUMEN
The standard implementation of enzyme-linked immunosorbent assay (ELISA) for single analytes can lead to false conclusions if cross reacting compounds are present in the sample. This paper discusses the extension of the usual four-parameter logistic model for ELISA to the case of multiple cross-reacting analytes. The use of the extended model in multianalyte analysis (MELISA) is illustrated and compared with a more simplistic approach. Data on the analysis of a binary mixture of s-triazines suggests the superiority of the proposed model. This model is also suitable for other forms of immunoassay that use the four-parameter logistic curve.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Reacciones Cruzadas , Relación Dosis-Respuesta Inmunológica , Triazinas/inmunologíaRESUMEN
This study investigated how 99 children who were examined for suspected sexual abuse (SSA) perceived their own medical evaluation experiences. Each child was interviewed about the degree of pain and fear associated with the experience, the kindness of the doctor, general fear of doctor visits, and degree of fear associated with a hypothetical second examination. The majority of children did not perceive their SSA examination to be strongly negative. However children did report greater fear associated with the SSA evaluation compared to an ordinary doctor visit. Using multiple regression, general fear of doctor visits and fear and pain associated with the SSA examination contributed to the prediction of intensity of fear about a hypothetical second SSA evaluation. Perceived kindness of the doctor, patient sex and age, and physician sex and age did not contribute to the regression equation. The relatively low reported rate of intense distress associated with medical evaluation of SSA suggests that fear and pain can be minimized and effectively managed for many children. The results of the regression analysis suggest that previous negative medical experiences may play an important role in determining how a child interprets the experience of an SSA medical evaluation.
Asunto(s)
Actitud Frente a la Salud , Abuso Sexual Infantil/diagnóstico , Abuso Sexual Infantil/psicología , Examen Físico/psicología , Psicología Infantil , Adolescente , Niño , Preescolar , Estudios Transversales , Miedo , Femenino , Humanos , Masculino , Dolor/etiología , Examen Físico/efectos adversos , Relaciones Médico-Paciente , Análisis de RegresiónRESUMEN
A 95-kDa mouse sperm protein has been previously identified as a putative receptor involved in the sperm-egg interactions that lead to fertilization. The ligand for this receptor is the zona pellucida glycoprotein ZP3. This constituent of the oocyte-specific extracellular matrix mediates not only sperm binding to the zona but also triggers acrosomal exocytosis. The latter, also termed the acrosome reaction, is a key regulatory event upon which fertilization is absolutely dependent. Previously, we showed that the 95-kDa protein that binds ZP3 is a substrate for tyrosine kinase, and its phosphotyrosine content increases after sperm-zona pellucida binding. Here, we show the presence of protein tyrosine kinase activity in sperm plasma membranes and in electroeluted 95-kDa protein. The tyrosine kinase activity of the isolated protein is stimulated by solubilized zona pellucida and inhibited by tyrphostin RG-50864, a membrane-permeable tyrosine kinase inhibitor. Furthermore, tyrphostin inhibits zona-triggered acrosomal exocytosis in a dose-dependent manner. These findings indicate that the 95-kDa protein participates in a critical regulatory event of gamete interaction; moreover, our experiments suggest that sperm protein tyrosine kinase may be an excellent target for the control of fertility.
Asunto(s)
Catecoles/farmacología , Proteínas del Huevo , Glicoproteínas de Membrana/metabolismo , Nitrilos/farmacología , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Receptores de Superficie Celular/fisiología , Interacciones Espermatozoide-Óvulo , Tirfostinos , Acrosoma/fisiología , Animales , Membrana Celular/metabolismo , Femenino , Ligandos , Masculino , Ratones , Fosfoproteínas/metabolismo , Fosforilación , Fosfotirosina , Transducción de Señal , Tirosina/análogos & derivados , Tirosina/metabolismo , Glicoproteínas de la Zona PelúcidaRESUMEN
The impact of the acquired immunodeficiency syndrome (AIDS) epidemic on a group of adolescents was investigated by surveying 197 sexually active, predominantly African-American, urban high school students. Reported sexual behavior changes were evaluated in relation to AIDS-related knowledge and attitudes. Over 50% of the students decreased their frequency of sexual activity, increased their condom use, and/or decreased their number of partners. These students had significantly higher scores on a measure of worry about vulnerability to human immunodeficiency virus (HIV) infection than those whose behavior had not changed. AIDS knowledge, AIDS beliefs, and AIDS-related anxiety interacted with gender to affect sexual behavior change. Male students reporting decreased frequency of sexual activity, for example, had more accurate beliefs about AIDS than males reporting no decrease. Among female students, however, those reporting decreased frequency had less accurate beliefs than those reporting no decrease. These results highlight the importance of considering gender and specific sexual behaviors when designing AIDS education interventions.