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BACKGROUND: Little information exists to guide monitoring and treatment of malnourishment during pregnancy after bariatric surgery. Here we present a case with severe deficiencies and recommendations for testing and treatment. CASE: Our patient underwent a duodenal switch procedure resulting in significant weight loss and numerous deficiencies. She then experienced a neonatal demise with multiple congenital abnormalities, including diaphragmatic hernia, possibly related to severe vitamin A deficiency. After high doses of oral and parenteral replacement, pancreatic enzymes, and total parenteral nutrition, she delivered an anatomically normal but growth-restricted neonate in a subsequent pregnancy. CONCLUSION: Bariatric procedures may result in nutritional deficiencies that affect pregnancy outcome. Women with severe deficiencies require pre-pregnancy counselling, monitoring, aggressive treatment, and a multidisciplinary approach to care.
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Cirugía Bariátrica/efectos adversos , Desnutrición/etiología , Complicaciones del Embarazo/etiología , Aborto Espontáneo , Adulto , Avitaminosis/diagnóstico , Avitaminosis/etiología , Femenino , Humanos , Recién Nacido , Obesidad Mórbida/cirugía , Embarazo , Resultado del EmbarazoRESUMEN
Background Drug-induced liver injury is a common cause of transaminitis, occurring in up to 5% of patients who are hospitalized for liver failure. In pregnancy, transaminitis is seen in conditions which may require expedited delivery. Case A 39-year-old G2P0010 at 27 2/7 weeks' gestation with chronic hypertension on labetalol was found to have elevated transaminases. Evaluation for preeclampsia, acute fatty liver, nonalcoholic steatohepatitis, cholelithiasis, infections, and autoimmune conditions were all negative. Labetalol was then discontinued, and liver biopsy was performed. After discontinuation of labetalol, her hepatitis improved, and she was discharged on hospital day 12 and went on to deliver at term. Conclusion Labetalol-induced hepatitis should be considered in the differential for transaminitis during pregnancy to prevent iatrogenic preterm delivery.
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Toll-like receptors (TLRs) are involved in the innate immune response against microbial pathogens in vertebrates and insects. The extracellular region of a TLR recognizes pathogen-associated molecules, while the intracellular region initiates the signaling pathway leading to immune response. Membrane-bound TLRs have been found in most vertebrates, but few soluble forms have been reported. A novel transcript corresponding to a portion of a soluble TLR was identified in liver of infected Atlantic salmon. The complete coding sequence of this TLR was obtained and BLASTN analysis showed the highest sequence identity to a recently described full-length cDNA sequence of a soluble TLR5 from rainbow trout (GenBank Accession No.: ). The deduced protein is 40% identical to the mammalian counterpart of the leucine-rich repeat (LRR)/LRR-like motifs of TLR5. Based on the structure of human TLRs, it contains 21 LRRs with conserved LxxLxLxxNx*xx*xxxxFxxL pattern. Since TLR5 is essential for the recognition of bacterial flagellins, we hypothesize that flagellin and perhaps some other pathogen-derived factors from Aeromonas salmonicida bind to this soluble TLR through an unknown binding domain within the LRR.
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Aeromonas salmonicida/inmunología , Enfermedades de los Peces/microbiología , Forunculosis/veterinaria , Infecciones por Bacterias Gramnegativas/veterinaria , ARN Mensajero/genética , Salmo salar/genética , Receptor Toll-Like 5/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Enfermedades de los Peces/genética , Enfermedades de los Peces/inmunología , Forunculosis/inmunología , Forunculosis/microbiología , Infecciones por Bacterias Gramnegativas/genética , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Datos de Secuencia Molecular , ARN Bacteriano/química , ARN Bacteriano/genética , ARN Mensajero/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Alineación de Secuencia , Receptor Toll-Like 5/inmunologíaRESUMEN
OBJECTIVE: We previously reported that maternal plasma levels of nonesterified polyunsaturated fatty acids were decreased in women with preeclampsia as compared to women with normal pregnancies. Polyunsaturated fatty acids of the n-6 and n-3 families are essential dietary fatty acids. The n-6 polyunsaturated fatty acids are involved in inflammatory reactions, whereas n-3 polyunsaturated fatty acids protect against inflammation. METHODS: In this study, we investigated the composition of nonesterified polyunsaturated fatty acids in placental tissue of normal and preeclamptic pregnancies. Linoleic (18:2) and arachidonic (20:4, AA) acids of the n-6 family and linolenic (18:3), eicosapentaenoic (20:5, EPA) and docosahexaenoic (22:6, DHA) acids of the n-3 family were analyzed. CONCLUSIONS: We found that total concentrations of nonesterified polyunsaturated fatty acids were lower in placental tissues from preeclamptic pregnancies than from normal pregnancies. Both n-6 and n-3 polyunsaturated fatty acids were decreased. The decrease in n-6 polyunsaturated fatty acids was due to a decrease in AA. The concentration of linoleic acid was not altered. For n-3 polyunsaturated fatty acids, DHA was decreased, with no change in linolenic acid. The percentage decrease in EPA and DHA was greater than for AA, so the ratio of n-6 to n-3 polyunsaturated fatty acids was two-fold higher in preeclampsia than normal pregnancy. Because the levels of essential fatty acids, linoleic (18:2) and linolenic (18:3), are preserved but levels of metabolites (AA and DHA) are reduced, we speculate that placental oxidative stress and increased conversion of AA to thromboxane are responsible for the decreased concentrations of polyunsaturated fatty acids in preeclampsia.
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Ácidos Grasos no Esterificados/metabolismo , Ácidos Grasos Insaturados/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Adulto , Cromatografía Líquida de Alta Presión , Femenino , Humanos , Embarazo , Estadísticas no ParamétricasRESUMEN
Vasoconstriction was observed in the fetal middle cerebral and umbilical arteries by Doppler assessment at 27 weeks gestation in a patient requiring continuous morphine infusion for pain control. Fetal heart tracings were also concerning. Fetal status improved after a change to fentanyl infusion, a shorter acting opioid. Caution is recommended when long-term chronic narcotic infusion is used in pregnancy.
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Feto/irrigación sanguínea , Morfina/efectos adversos , Placenta/irrigación sanguínea , Vasoconstricción/efectos de los fármacos , Adulto , Analgesia Obstétrica , Femenino , Edad Gestacional , Frecuencia Cardíaca Fetal , Humanos , Mordeduras y Picaduras de Insectos/complicaciones , Flujometría por Láser-Doppler , Intercambio Materno-Fetal , Arteria Cerebral Media/efectos de los fármacos , Arteria Cerebral Media/embriología , Morfina/administración & dosificación , Embarazo , Complicaciones del Embarazo , Arterias Umbilicales/efectos de los fármacos , Arterias Umbilicales/embriologíaRESUMEN
OBJECTIVE: There is more to be learned about the epidemiology of group B beta-hemolytic streptococci infections in pregnancy. In this study, we investigated the discriminating capabilities of pulsed-field gel electrophoresis of group B streptococci strains from pregnant patients and mother/infant pairs of patients compared with serotyping. METHODS: Forty-two vaginal strains of group B streptococci cultured from pregnant patients in the third trimester and strains from 20 mother/infant pairs with documented newborn group B streptococci infection were studied. Isolates were serotyped by the Lancefield capillary precipitin method and molecularly characterized by counterclamped homogeneous electrical field pulsed-field gel electrophoresis with rarely cutting restriction enzymes. RESULTS: Nine of the 13 serotypes of group B streptococci identified thus far in the scientific literature (Ia, Ia/c, Ib, Ib/c, II, IIc, III, V, and NT/c) were represented among the 62 isolates. Among the 42 maternal isolates, eight serotypes were represented, and among the 20 mother/infant isolates, six serotypes were represented. Serotypes of mother/infant isolates matched in nine of the ten pairs. Restriction endonuclease profiles, or digests, from the 42 maternal isolates resulted in 25 unique profiles that were arranged into five major groups based on the overall relatedness. Each group was comprised of one predominant serotype. The 20 mother/infant paired isolates displayed nine unique restriction endonuclease profiles and nine of the ten paired isolates showed indistinguishable restriction endonuclease profiles between mother and infant. CONCLUSION: Deoxyribonucleic acid profiling using pulsed-field gel electrophoresis is more discriminating of group B streptococci strains than serotyping because of the different yet closely related patterns within each restriction endonuclease profile group that are linked to one specific serotype. Pulsed-field gel electrophoresis can refine our epidemiologic studies of group B streptococci transmission and acquisition.
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Dermatoglifia del ADN/métodos , Electroforesis en Gel de Campo Pulsado/métodos , Complicaciones Infecciosas del Embarazo/microbiología , Pruebas Serológicas/métodos , Infecciones Estreptocócicas/diagnóstico , Streptococcus agalactiae/aislamiento & purificación , ADN Bacteriano/análisis , Femenino , Humanos , Recién Nacido , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Sensibilidad y Especificidad , Vagina/microbiologíaRESUMEN
OBJECTIVE: Third-trimester human placentas from normal and preeclamptic pregnancies were evaluated for possible changes in gene expression patterns by microarray analysis. METHODS: Placentas from four normal pregnancies and four pregnancies complicated by preeclampsia were studied. In a preliminary effort to identify possible differences between the two groups, complementary DNA (cDNA) probes were prepared from pooled total RNA by reverse transcription in the presence of (33)P-dCTP. After hybridization to human GeneFilter cDNA microarrays (GF211; Research Genetics, Huntsville, AL), 319 positive signals were detected above background out of a possible 4131 human cDNAs spotted on the filters. RESULTS: Ten most highly expressed mRNA species, ten most up-regulated, and ten most down-regulated genes in placentas from both groups of patients were identified for future studies. Of the 319 positive hybridizations, one transcript was clearly elevated in preeclamptic pregnancy. This cDNA encodes the muscle subtype of glycogen phosphorylase (GP-M) and was increased more than 2.8-fold (P <.05) in the preeclamptic placentas. In contrast, cDNA for glycogen synthase (muscle and liver isoforms) was not significantly increased, being near the limits of detection. The preeclampsia-induced increase of placental GP-M mRNA expression (approximately 3.5-fold) was confirmed by northern blot analysis. CONCLUSION: We conclude that microarray analysis can detect trends in mRNA and gene expression in placentas from normal and preeclamptic pregnancies that may be further studied in a more targeted fashion. We found that placental GP-M mRNA level is up-regulated in preeclampsia, which is consistent with previous reports of increased glycogen phosphorylase activity, and we suggest that it may be largely regulated at the level of transcription. Further studies may determine whether such up-regulation might be a response to hypoxia.
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Glucógeno Fosforilasa de Forma Muscular/genética , Placenta/fisiología , Preeclampsia/genética , Femenino , Perfilación de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Preeclampsia/enzimología , Embarazo , Tercer Trimestre del Embarazo , Valores de Referencia , Transcripción Genética , Regulación hacia ArribaRESUMEN
CONTEXT: Questionnaire studies linked symptoms of obstructive sleep apnea (OSA) to the risk of gestational diabetes mellitus (GDM). Whether this association is present when OSA is assessed objectively by polysomnography is not known. OBJECTIVE: The objective of the study was to assess the relationship between pregnancy, OSA, and GDM. DESIGN, SETTING, AND PARTICIPANTS: We conducted observational case-control studies using polysomnography in 15 nonpregnant, nondiabetic women (NP-NGT), 15 pregnant women with normal glucose tolerance (P-NGT), and 15 pregnant women with GDM (P-GDM). The groups were frequency matched for age and race/ethnicity. Pregnant women were studied during the late second to early third trimester. MAIN OUTCOME MEASURES: Comparisons of OSA diagnosis and sleep parameters between NP-NGT and P-NGT to assess the impact of pregnancy and between P-NGT and P-GDM to explore the association between GDM and OSA were measured. RESULTS: Compared with NP-NGT, P-NGT women had a higher apnea hypopnea index (AHI) (median 2.0 vs 0.5, P = .03) and more disrupted sleep as reflected by a higher wake time after sleep onset (median 66 vs 21 min, P < .01) and a higher microarousal index (median 16.4 vs 10.6, P = .01). Among the pregnant women, P-GDM had markedly lower total sleep time (median 397 vs 464 min, P = .02) and a higher AHI (median 8.2 vs 2.0, P = .05) than P-NGT women. OSA was more prevalent in P-GDM than in P-NGT women (73% vs 27%, P = .01). After adjustment for prepregnancy body mass index, the diagnosis of GDM was associated with a diagnosis of OSA [odds ratio 6.60 (95% confidence interval 1.15-37.96)]. In pregnancy, after adjusting for prepregnancy body mass index, higher microarousal index significantly associated with higher hemoglobin A1c and fasting glucose levels. Higher oxygen desaturation index was associated with higher fasting glucose levels. CONCLUSION: Pregnancy is associated with sleep disturbances. Sleep is more disturbed in GDM than in P-NGT women. There is a strong association between GDM and OSA.
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Diabetes Gestacional/fisiopatología , Complicaciones del Embarazo/fisiopatología , Apnea Obstructiva del Sueño/complicaciones , Apnea Obstructiva del Sueño/fisiopatología , Adulto , Glucemia , Índice de Masa Corporal , Estudios de Casos y Controles , Femenino , Humanos , Resistencia a la Insulina , Obesidad/complicaciones , Obesidad/fisiopatología , Embarazo , Tercer Trimestre del EmbarazoRESUMEN
OBJECTIVE: The purpose of this study was to compare the risk of adverse neonatal outcomes between women with placenta accreta and placenta increta or percreta. METHODS: This was a single institution retrospective cohort study of women with abnormal placentation (placenta accreta, increta, and percreta) who delivered from 1982-2002. Cases were divided into superficial invasion (placenta accreta) and deep invasion (placenta increta or percreta), and compared. The primary outcomes studied were gestational age at delivery, birth weight, and size for gestational age. RESULTS: 103 viable pregnancies with abnormal placentation were observed (1.6/1000 pregnancies). Cases of deep invasion had higher parity and were more likely to have had a prior cesarean delivery. The mean gestational age at delivery was 33 5/7 weeks with deep placental invasion and 35 2/7 weeks in the superficial invasion group (p = 0.18). Rates of preterm birth were 64.7% and 52.3% (p = 0.43) and low birthweight were 24% and 29% (p = 0.76) in the deep and superficial invasion groups respectively. There were no differences in the remaining outcomes. CONCLUSIONS: Neonatal outcomes of pregnancies complicated by placenta increta and percreta are not different than those with placenta accreta.
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Peso al Nacer , Edad Gestacional , Placenta Accreta/patología , Nacimiento Prematuro/etiología , Adolescente , Adulto , Estudios de Cohortes , Femenino , Humanos , Recién Nacido de Bajo Peso , Recién Nacido , Recien Nacido Prematuro , Recién Nacido Pequeño para la Edad Gestacional , Embarazo , Nacimiento Prematuro/epidemiología , Estudios Retrospectivos , Adulto JovenRESUMEN
OBJECTIVE: To estimate trends in patient characteristics and obstetric complications in an 8-year cohort of patients undergoing cesarean delivery and to use time series analysis to estimate the effect of infection prevention interventions and secular trends in patient characteristics on postcesarean delivery surgical site infections. METHODS: A multivariable autoregressive integrated moving average model was used to perform time series analysis on a 96-month retrospective cohort of patients who underwent cesarean delivery (January 2003-December 2010) in a U.S. tertiary care hospital. RESULTS: We identified 8,668 women who underwent cesarean delivery. Median age was 26 years (range 12-53 years), 3,093 (35.7%) of patients had body mass indexes (BMIs) of 35 or greater, 2,561 (29.5%) were of white race, and 303 (3.5%) had a surgical site infection. Over the study period, there was a significant increase in the proportion of patients who underwent cesarean delivery who had BMIs of 35 or higher, hypertension or mild preeclampsia, and severe preeclampsia or eclampsia. A nonseasonal autoregressive integrated moving average model with a linear trend and no autocorrelation was identified. In the multivariable autoregressive integrated moving average model of postcesarean surgical site infections, implementation of a policy to administer prophylactic antibiotics within 1 hour before incision, instead of at the time of cord clamp, led to a 48% reduction in cesarean delivery surgical site infections (Δ=-5.4 surgical site infections per 100 cesarean deliveries; P<.001). CONCLUSION: A change in policy to administer prophylactic antibiotics before incision resulted in a significant reduction in postcesarean surgical site infections. LEVEL OF EVIDENCE: III.
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Profilaxis Antibiótica , Cesárea/efectos adversos , Control de Infecciones , Infección de la Herida Quirúrgica/prevención & control , Adolescente , Adulto , Niño , Femenino , Humanos , Persona de Mediana Edad , Embarazo , Estudios Retrospectivos , Infección de la Herida Quirúrgica/etiología , Adulto JovenRESUMEN
OBJECTIVE: To explore relationships among sleep disturbances, glucose tolerance, and pregnancy outcomes. RESEARCH DESIGN AND METHODS: Four validated sleep questionnaires were administered to 169 pregnant women at the time of 50-g oral glucose tolerance testing (OGTT) during the second trimester. Pregnancy outcomes were analyzed in 108 women with normal glucose tolerance (NGT). RESULTS: Of the participants, 41% had excessive daytime sleepiness (Epworth Sleepiness Scale [ESS] >8); 64% had poor sleep quality; 25% snored frequently; 29% had increased risk of sleep-disordered breathing (SDB); 52% experienced short sleep (SS); 19% had both increased SDB risk and SS (SDB/SS); and 14% had daytime dysfunction. Reported sleep duration inversely correlated with glucose values from 50-g OGTT (r = -0.21, P < 0.01). Each hour of reduced sleep time was associated with a 4% increase in glucose levels. Increased likelihood of gestational diabetes mellitus (GDM) was found in subjects with increased SDB risk (odds ratio 3.0 [95% CI 1.2-7.4]), SS (2.4 [1.0-5.9]), SDB/SS (3.4 [1.3-8.7]), and frequent snoring (3.4 [1.3-8.8], after adjustment for BMI). Among NGT subjects, preterm delivery was more frequent in those with increased ESS (P = 0.02), poor sleep quality (P = 0.02), and SS (P = 0.03). Neonatal intensive care unit admissions were associated with increased ESS (P = 0.03), SDB/SS (P = 0.03), and daytime dysfunction (P < 0.01) in mothers. CONCLUSIONS: Pregnant women experience significant sleep disturbances that are associated with increased risk of GDM and unfavorable pregnancy outcomes. Pregnant women with increased SDB risk, frequent snoring, and sleep duration of <7 h/night have increased risk of developing GDM.
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Glucemia/metabolismo , Diabetes Gestacional/fisiopatología , Intolerancia a la Glucosa , Resultado del Embarazo , Trastornos del Sueño-Vigilia/complicaciones , Adulto , Estudios de Casos y Controles , Diabetes Gestacional/epidemiología , Femenino , Prueba de Tolerancia a la Glucosa , Humanos , Modelos Logísticos , Embarazo , Segundo Trimestre del Embarazo , Factores de Riesgo , Encuestas y Cuestionarios , Factores de Tiempo , Adulto JovenRESUMEN
The aim of this study was to identify differentially expressed genes by suppression subtractive hybridization (SSH) in HELLP placentas. Two cDNA libraries were constructed; HSI (HELLP subtracted induced or upregulated) and HSS (HELLP subtracted suppressed or downregulated). Two hundred eighty-eight cDNA clones were sequenced; 37 were matched to GenBank entries and included genes in cell communication and organization, cellular processes, genetic information processing, and metabolic processes. A subgroup of 11 genes of interest was further selected for real-time quantitative polymerase chain reaction confirmation. Results showed no differences in expression of chosen upregulated genes between HELLP and non-HELLP placentas; 6 HELLP downregulated genes were significantly suppressed. Two genes related to production of secreted proteins, CTHRC1 and SERPINE2. SERPINE2 (PAI-1) is a soluble protease inhibitor and is a potential biomarker by Western blot analysis, and the protein is significantly decreased in HELLP placentas. SERPINE2 might be tested clinically in patients for early diagnosis of HELLP syndrome.
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Perfilación de la Expresión Génica , Síndrome HELLP/genética , Placenta/química , ARN Mensajero/análisis , Adulto , Precursor de Proteína beta-Amiloide/análisis , Precursor de Proteína beta-Amiloide/genética , Biomarcadores/análisis , Western Blotting , Proteínas de la Matriz Extracelular/análisis , Proteínas de la Matriz Extracelular/genética , Femenino , Perfilación de la Expresión Génica/métodos , Biblioteca de Genes , Síndrome HELLP/metabolismo , Humanos , Reacción en Cadena de la Polimerasa , Embarazo , Nexinas de Proteasas , Receptores de Superficie Celular/análisis , Receptores de Superficie Celular/genética , Reproducibilidad de los Resultados , Serpina E2RESUMEN
OBJECTIVE: Ethanol exposure during pregnancy may result in fetal alcohol syndrome (FAS). The mechanism by which this occurs is unknown. Recent studies in several organ systems, including the placenta, suggest that oxidative stress is involved. In this study we investigated the presence and levels of three oxidative stress markers in placental villous tissue exposed to ethanol. METHODS: Villous tissues from normal placentas were perfused with Dulbeco's modified Eagle's medium (DMEM) with HEPES buffer, sodium bicarbonate, and glucose at pH 7.4. After stabilization, 100 mM ethanol was added to the perfusate. After 2 hours of perfusion, the tissue was removed, fixed and stained for nitrotyrosine, 4-hydroxy-2-nonenal (4HNE) and 8-hydroxyguanosine (8-OHDG). Staining within the trophoblasts was quantified with densitometry. RESULTS: Nitrotyrosine and 4HNE immunostaining was seen in the trophoblasts. 4HNE was also seen in the stroma. In contrast, 8-OHDG was seen only in the stroma and endothelial cells in the fetal circulation. Ethanol exposure significantly increased nitrotyrosine levels in the trophoblasts beyond levels in the control tissue. Nitrotyrosine and 8-OHDG levels were also increased in stroma. CONCLUSION: Within the placental villi, markers of oxidative stress are present in the trophoblasts and stroma after a short period of ethanol exposure. There is an increase in oxidative stress, primarily involving the nitric oxide pathway, in the trophoblasts as well as DNA damage in the stroma. Lipid peroxidation is not acutely changed in our 2-hour exposure window.
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Biomarcadores/análisis , Etanol/farmacología , Estrés Oxidativo , Placenta/química , Placenta/efectos de los fármacos , Aldehídos/análisis , Femenino , Guanosina/análogos & derivados , Guanosina/análisis , Humanos , Embarazo , Trofoblastos/química , Tirosina/análogos & derivados , Tirosina/análisisRESUMEN
The risk of cervical cancer, one of the most prevalent cancers in the world, is determined by two viruses. Human papillomavirus (HPV) is the main risk factor for developing cervical cancer. However, although little known, it is well substantiated that the human Parvovirus adeno-associated virus type 2 (AAV), and its encoded Rep78 protein, interacts with HPV and lowers the risk of cervical cancer. HPV also contributes to AAV inhibition by serving as a helper virus for AAV and stimulating higher AAV replication levels. Here we surveyed four HPV-16 early genes, E1, E2, E6 and E7, for their ability to increase/decrease the basal level of AAV replication in stratifying squamous epithelium (the epithelial raft culture system). It was found that the HPV-16 E1, E2 and E6 genes were able to help/enhance AAV-2 replication in epithelial raft cultures. Under these conditions, with all the HPV genes being expressed from the AAV p5 promoter, E1 appeared to have the strongest enhancing effect on AAV DNA replication (Southern blot), RNA expression (RT-PCR), protein expression (Western blot) and AAV virion production (2 plate-Southern blot). Further study of E1 mutants showed that the carboxy-half of E1, the putative helicase/ATPase domain, was the main contributor of helper activity. These data are important for understanding the HPV-AAV interaction and its effect on modifying cervical cancer risk. These data also suggest the possibility that the identified HPV helper genes may be useful in the generation of recombinant (r)AAV virions for gene therapy, as rAAV is increasing in popularity for such purposes.
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Dependovirus/fisiología , Genes Virales/genética , Papillomaviridae/genética , Proteínas Virales/genética , Proteínas Virales/metabolismo , Replicación Viral , Proteínas de la Cápside/biosíntesis , Células Cultivadas , Dependovirus/genética , Humanos , Queratinocitos/virología , Mutación , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Viral/genética , ARN Viral/metabolismo , Proteínas Virales/químicaRESUMEN
Chronic hepatitis C virus (HCV) infection cases resistant to conventional therapies might be treated by immunotherapy as cytotoxic T lymphocytes (CTL) are the main mechanism through which viral infections are cleared. The HCV core gene, with the highest homology between HCV types, deleted of its autoimmune-stimulating regions (pseudo-GOR and pseudo-p450), may be an appropriate antigen for targeting HCV-infected cells. Two recombinant adeno-associated virus (rAAV) vectors, carrying either the full length (aa 1-190) or truncated (aa 49-180, deleted of the pseudo-GOR and pseudo-p450 sequences) versions of core, were generated. Both AAV/core (l-190) and AAV/core (49-180) were used to transduce/load dendritic cells (DC) at high levels (88-95%). These two genetically altered DC types then stimulated anti-core CTL. The DC and CTL were analyzed by FACS and for killing efficiency (percent target killing). The rAAV-altered DC displayed higher levels of CD80, CD83, CD86, and CD 1a than control DC. The truncated core (aa 49-180) gene stimulated equivalent and strong killing of synthetic core-positive autologous peripheral blood lymphocyte (PBL) targets to that stimulated by the full length core gene. However, the smaller core (49-180) antigen gene stimulated lower levels of killing of core-negative "self" PBL targets (GOR- and p450-positive) (p = 0.002). These AAV/core: DC-stimulated CTL displayed higher IFN-gamma expression, higher CD8:CD4 ratios, and lower CD56:CD8 ratios than controls. The rAAV-loading derived CD8+ T cells had more CD69+ cells and the CD4+ T populations had fewer CD25+ cells than controls. We conclude that the core (49-180) gene is an effect antigen, but has the advantage of stimulating less self-recognition. Thus, core (49-180) may be useful for further translational immunotherapy studies against HCV.
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Células Dendríticas/fisiología , Hepacivirus/inmunología , Hepatitis C/terapia , Linfocitos T Citotóxicos/inmunología , Proteínas del Núcleo Viral/inmunología , Autoinmunidad , Dependovirus/genética , Citometría de Flujo , Humanos , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Transducción Genética , Proteínas del Núcleo Viral/genéticaRESUMEN
OBJECTIVE: A difference in survival rates between black and white patients with cancer of the corpus uteri is well established. This study was conducted to determine whether the overexpression of HER2/neu oncogene is associated with poor outcome in uterine serous papillary endometrial cancer, which is a highly aggressive variant of endometrial cancer, and whether a racial difference in the frequency of HER2/neu overexpression may contribute to the disparity in endometrial cancer survival. STUDY DESIGN: Immunohistochemical evaluation was used to examine HER2/neu expression in paraffin blocks from 27 women with stage IA to IV uterine serous papillary endometrial cancer. Univariable analysis was performed and followed by multivariable analysis with Cox's proportional hazard model to evaluate whether HER2/neu expression was associated with poor outcome in uterine serous papillary endometrial cancer. RESULTS: Black patients tended to be younger (P = .02) and have higher HER2/neu expression than white patients (trend P = .02). Seven of 10 black patients (70%) showed heavy (3+) expression, compared with 4 of 17 white patients (24%; P = .04). The association of heavy HER2/neu expression with race persisted after age was controlled through stratification (P = .05). Earlier deaths from uterine serous papillary endometrial cancer were seen among heavy HER2/neu expressers (P = .002), black patients (P = .04), and patients < or = 65 years old (P = .04). However, multivariate Cox regression showed that short survival was associated significantly with heavy HER2/neu expression (P = .02) but not with age (P = .07) or race (P = .35), which indicates that HER2/neu expression accounted for much of the race disparity in survival in this patient population. CONCLUSION: Overexpression of HER2/neu in uterine serous papillary endometrial cancer is an independent variable that is associated with poor outcome, occurs more frequently in black women, and may contribute to racial disparity in survival. HER2/neu expression may guide clinical treatment of patients with uterine serous papillary endometrial cancer and may have implications for the implementation of novel treatment strategies.
Asunto(s)
Carcinoma Papilar/mortalidad , Cistadenocarcinoma Seroso/mortalidad , Regulación Neoplásica de la Expresión Génica , Genes erbB-2/genética , Neoplasias Uterinas/mortalidad , Anciano , Biomarcadores de Tumor/análisis , Población Negra , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Pronóstico , Modelos de Riesgos Proporcionales , Tasa de Supervivencia , Población BlancaRESUMEN
With the goal of identifying genes with a differential pattern of expression between invasive cervical carcinomas (CVX) and normal cervical keratinocytes (NCK), we used oligonucleotide microarrays to interrogate the expression of 14,500 known genes in 11 primary HPV16 and HPV18-infected stage IB-IIA cervical cancers and four primary normal cervical keratinocyte cultures. Hierarchical cluster analysis of gene expression data identified 240 and 265 genes that exhibited greater than twofold up-regulation and down-regulation, respectively, in primary CVX when compared to NCK. Cyclin-dependent kinase inhibitor 2A (CDKN2A/p16), mesoderm-specific transcript, forkhead box M1, v-myb myeloblastosis viral oncogene homolog (avian)-like2 (v-Myb), minichromosome maintenance proteins 2, 4, and 5, cyclin B1, prostaglandin E synthase (PTGES), topoisomerase II alpha (TOP2A), ubiquitin-conjugating enzyme E2C, CD97 antigen, E2F transcription factor 1, and dUTP pyrophosphatase were among the most highly overexpressed genes in CVX when compared to NCK. Down-regulated genes in CVX included transforming growth factor beta 1, transforming growth factor alpha, CFLAR, serine proteinase inhibitors (SERPING1 and SERPINF1), cadherin 13, protease inhibitor 3, keratin 16, and tissue factor pathway inhibitor-2 (TFPI-2). Differential expression of some of these genes including CDKN2A/p16, v-Myb, PTGES, and TOP2A was validated by quantitative real-time PCR. Flow cytometry on primary CVX and NCK and immunohistochemical staining of formalin fixed paraffin-embedded tumor specimens from which primary CVX cultures were derived as well as from a separate set of invasive cervical cancers confirmed differential expression of the CDKN2A/p16 and PTGES markers on CVX versus NCK. These results identify several genes that are coordinately disregulated in cervical cancer, likely representing common signaling pathways triggered by HPV transformation. Moreover, these data obtained with highly purified primary tumor cultures highlight novel molecular features of human cervical cancer and provide a foundation for the development of new type-specific diagnostic and therapeutic strategies for this disease.
Asunto(s)
Biomarcadores de Tumor/genética , Perfilación de la Expresión Génica , Papillomaviridae/metabolismo , Neoplasias del Cuello Uterino/diagnóstico , Células Cultivadas , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Papillomaviridae/clasificación , Papillomaviridae/genética , ARN Mensajero/genética , Neoplasias del Cuello Uterino/tratamiento farmacológico , Neoplasias del Cuello Uterino/genéticaRESUMEN
Human papillomavirus (HPV) DNA is preferentially found in spontaneous abortions, specifically residing in trophoblasts, and transfected HPV-16 DNA replicates and produces progeny in 3A trophoblasts in culture. In this study 3A trophoblasts were shown to display both HPV receptors and infection by HPV-31b and HPV-6 virus resulted in de novo (increasing) HPV DNA replication in these cells (inhibited by neutralizing anti-HPV31b antibodies). Reverse transcription-polymerase chain reaction analysis revealed that E1;E4, E6, and L1 were significantly expressed at days 5 (early) and 10 (late), respectively, and in situ immunocytochemistry verified L1 protein expression. Perhaps most important, HPV 31b virus infection caused both a decrease in 3A trophoblast cell numbers in a dose-dependent manner and a low trophoblast-endometrial cell adhesion (both inhibited by neutralizing anti-HPV-31 antibodies). These data further support the hypothesis that HPVs are fully active in trophoblasts and may cause some spontaneous abortions.