Persistence and Intra-Host Genetic Evolution of Zika Virus Infection in Symptomatic Adults: A Special View in the Male Reproductive System.

We followed the presence of Zika virus (ZIKV) in four healthy adults (two men and two women), for periods ranging from 78 to 298 days post symptom onset. The patients were evaluated regarding the presence of the virus in different body fluids (blood, saliva, urine and semen), development of immune responses (including antibodies, cytokines and chemokines), and virus genetic variation within samples collected from semen and urine during the infection course. The analysis was focused primarily on the two male patients who shed the virus for up to 158 days after the initial symptoms. ZIKV particles were detected in the spermatozoa cytoplasm and flagella, in immature sperm cells and could also be isolated from semen in cell culture, confirming that the virus is able to preserve integrity and infectivity during replication in the male reproductive system (MRS). Despite the damage caused by ZIKV infection within the MRS, our data showed that ZIKV infection did not result in infertility at least in one of the male patients. This patient was able to conceive a child after the infection. We also detected alterations in the male genital cytokine milieu, which could play an important role in the replication and transmission of the virus which could considerably increase the risk of ZIKV sexual spread. In addition, full genome ZIKV sequences were obtained from several samples (mainly semen), which allowed us to monitor the evolution of the virus within a patient during the infection course. We observed genetic changes over time in consensus sequences and lower frequency intra-host single nucleotide variants (iSNV), that suggested independent compartmentalization of ZIKV populations in the reproductive and urinary systems. Altogether, the present observations confirm the risks associated with the long-term replication and shedding of ZIKV in the MRS and help to elucidate patterns of intra-host genetic evolution during long term replication of the virus.

Persistence and intra-host genetic evolution of zika virus infection in symptomatic adults: A special view in the male reproductive system

We followed the presence of Zika virus (ZIKV) in four healthy adults (two men and two women), for periods ranging from 78 to 298 days post symptom onset. The patients were evaluated regarding the presence of the virus in different body fluids (blood, saliva, urine and semen), development of immune responses (including antibodies, cytokines and chemokines), and virus genetic variation within samples collected from semen and urine during the infection course. The analysis was focused primarily on the two male patients who shed the virus for up to 158 days after the initial symptoms. ZIKV particles were detected in the spermatozoa cytoplasm and flagella, in immature sperm cells and could also be isolated from semen in cell culture, confirming that the virus is able to preserve integrity and infectivity during replication in the male reproductive system (MRS). Despite the damage caused by ZIKV infection within the MRS, our data showed that ZIKV infection did not result in infertility at least in one of the male patients. This patient was able to conceive a child after the infection. We also detected alterations in the male genital cytokine milieu, which could play an important role in the replication and transmission of the virus which could considerably increase the risk of ZIKV sexual spread. In addition, full genome ZIKV sequences were obtained from several samples (mainly semen), which allowed us to monitor the evolution of the virus within a patient during the infection course. We observed genetic changes over time in consensus sequences and lower frequency intra-host single nucleotide variants (iSNV), that suggested independent compartmentalization of ZIKV populations in the reproductive and urinary systems. Altogether, the present observations confirm the risks associated with the long-term replication and shedding of ZIKV in the MRS and help to elucidate patterns of intra-host genetic evolution during long term replication of the virus

Persistence and intra-host genetic evolution of Zika virus infection in symptomatic adults: a special view in the male reproductive system

We followed the presence of Zika virus (ZIKV) in four healthy adults (two men and two women), for periods ranging from 78 to 298 days post symptom onset. The patients were evaluated regarding the presence of the virus in different body fluids (blood, saliva, urine and semen), development of immune responses (including antibodies, cytokines and chemokines), and virus genetic variation within samples collected from semen and urine during the infection course. The analysis was focused primarily on the two male patients who shed the virus for up to 158 days after the initial symptoms. ZIKV particles were detected in the spermatozoa cytoplasm and flagella, in immature sperm cells and could also be isolated from semen in cell culture, confirming that the virus is able to preserve integrity and infectivity during replication in the male reproductive system (MRS). Despite the damage caused by ZIKV infection within the MRS, our data showed that ZIKV infection did not result in infertility at least in one of the male patients. This patient was able to conceive a child after the infection. We also detected alterations in the male genital cytokine milieu, which could play an important role in the replication and transmission of the virus which could considerably increase the risk of ZIKV sexual spread. In addition, full genome ZIKV sequences were obtained from several samples (mainly semen), which allowed us to monitor the evolution of the virus within a patient during the infection course. We observed genetic changes over time in consensus sequences and lower frequency intra-host single nucleotide variants (iSNV), that suggested independent compartmentalization of ZIKV populations in the reproductive and urinary systems. Altogether, the present observations confirm the risks associated with the long-term replication and shedding of ZIKV in the MRS and help to elucidate patterns of intra-host genetic evolution during long term replication of the virus.

Persistence and intra-host genetic evolution of Zika virus infection in symptomatic adults: a special view in the male reproductive system

We followed the presence of Zika virus (ZIKV) in four healthy adults (two men and two women), for periods ranging from 78 to 298 days post symptom onset. The patients were evaluated regarding the presence of the virus in different body fluids (blood, saliva, urine and semen), development of immune responses (including antibodies, cytokines and chemokines), and virus genetic variation within samples collected from semen and urine during the infection course. The analysis was focused primarily on the two male patients who shed the virus for up to 158 days after the initial symptoms. ZIKV particles were detected in the spermatozoa cytoplasm and flagella, in immature sperm cells and could also be isolated from semen in cell culture, confirming that the virus is able to preserve integrity and infectivity during replication in the male reproductive system (MRS). Despite the damage caused by ZIKV infection within the MRS, our data showed that ZIKV infection did not result in infertility at least in one of the male patients. This patient was able to conceive a child after the infection. We also detected alterations in the male genital cytokine milieu, which could play an important role in the replication and transmission of the virus which could considerably increase the risk of ZIKV sexual spread. In addition, full genome ZIKV sequences were obtained from several samples (mainly semen), which allowed us to monitor the evolution of the virus within a patient during the infection course. We observed genetic changes over time in consensus sequences and lower frequency intra-host single nucleotide variants (iSNV), that suggested independent compartmentalization of ZIKV populations in the reproductive and urinary systems. Altogether, the present observations confirm the risks associated with the long-term replication and shedding of ZIKV in the MRS and help to elucidate patterns of intra-host genetic evolution during long term replication of the virus.

Macrophage migration inhibitory factor induces phosphorylation of Mdm2 mediated by phosphatidylinositol 3-kinase/Akt kinase: Role of this pathway in decidual cell survival.

The phosphatidylinositol 3-kinase (PI3K)/Akt pathway has an anti-apoptotic effect through several downstream targets, which includes activation of the transformed mouse 3T3 cell double-minute 2 (Mdm2) protein, its translocation to the nucleus and degradation of the tumor suppressor p53. We show that Mif, the Macrophage Migration Inhibitory Factor, an important cytokine at the maternal fetal interface in several species, triggers phosphorylation of Mdm2 protein in a PI3K/Akt-dependent manner, thereby preventing apoptosis in cultured mouse decidual cells. Inhibition of Akt and PI3K suppresses the pathway. Mif treatment also changes the nuclear translocation of p53 and interferes with the apoptotic fate of these cells when challenged with reactive oxygen species. In conclusion, an important mechanism has been found underlying decidual cell survival through Akt signaling pathway activated by Mif, suggesting a role for this cytokine in decidual homeostasis and in the integrity of the maternal-fetal barrier that is essential for successful gestation.

Decorin and biglycan immunolocalization in non-villous structures of healthy and pathological human placentas.

AIMS: Decorin and biglycan are members of the small leucine-rich proteoglycan family, and constituents of both the extracellular matrix (ECM) and the cell surface. They are recognized as important factors in the control of proliferation, migration and invasion in vivo and in vitro. In this study, the localization patterns of decorin and biglycan were determined in healthy placentas and in highly invasive placental pathologies. METHODS AND RESULTS: The study included immunolocalization of decorin and biglycan in samples of first-trimester and term placentas, placenta accreta, invasive mole, and choriocarcinoma. Extravillous cytotrophoblast (EVT) cells were positive for both proteoglycans in all pathologies and in first-trimester placentas, although not in term placentas. Biglycan was immunolocalized in the ECM of all healthy and pathological placentas, whereas decorin was observed only in term placenta ECM. CONCLUSIONS: The expression of both proteoglycans was cell-specific and gestation time-dependent in healthy placentas, and was associated with invasive EVT cells in pathological placentas. In view of the biological properties of these molecules, it is possible that the biglycan pattern found here is intrinsically implicated in the invasive activity of EVT cells in both healthy and disordered placentas.

Ectoplacental cone induces resistance to apoptosis in high doses of interferon (IFN)-γ-treated decidual cells.

PROBLEM In this study, we explored the relationship between decidual cells (DC) and interferon (IFN)-γ, in the presence or absence of ectoplacental cone (EC) using a coculture system. METHOD OF STUDY Decidual cells and EC were isolated from pregnant mice on gestation day 7.5. DCs were cultured for 48 hr and then treated with fresh EC. After characterization, they were treated with IFN-γ, and cell death was evaluated. RESULTS Interferon-γ drastically increased decidual apoptosis, which was partially reverted by the addition of EC to the IFN-γ-treated decidual culture. Moreover, the addition of EC to non-treated DC cultures was also capable of attenuating death rates. CONCLUSION Resistance to apoptosis may be induced in DC by the EC. This suggests that EC may participate in the inhibition of IFN-γ-dependent apoptosis and, therefore, play important role for DC survival in a cytokine-enriched placental environment.

Collagen fibril organization in the pregnant endometrium of decorin-deficient mice.

In the pregnant mouse endometrium, collagen fibrillogenesis is characterized by the presence of very thick collagen fibrils which are topographically located exclusively within the decidualized stroma. This dynamic biological process is in part regulated by the small leucine-rich proteoglycans decorin and biglycan. In the present study we utilized wild-type (Dcn(+/+)) and decorin-deficient (Dcn(-/-)) time-pregnant mice to investigate the evolution of non-decidualized and decidualized collagen matrix in the uterine wall of these animals. Ultrastructural and morphometric analyses revealed that the organization of collagen fibrils in the pregnant endometrium of both non-decidualized and decidualized stroma showed a great variability of shape and size, regardless of the genotype. However, the decidualized endometrium from Dcn(-/-) mice contained fibrils with larger diameter and more irregular contours as compared to the wild-type littermates. In the Dcn(-/-) animals, the proportion of thin (10-50 nm) fibrils was also higher as compared to Dcn(+/+) animals. On day 7 of pregnancy, biglycan was similarly localized in the decidualized endometrium in both genotypes. Lumican immunostaining was intense both in decidualized and non-decidualized stroma from Dcn(-/-) animals. The present results support previous findings suggesting that decorin participates in uterine collagen fibrillogenesis. In addition, we suggest that the absence of decorin disturbs the process of lateral assembly of thin fibrils, resulting in very thick collagen fibrils with irregular profiles. Our data further suggest that decorin, biglycan and lumican might play an interactive role in collagen fibrillogenesis in the mouse endometrium, a process modulated according to the stage of pregnancy.

An ultrastructural study of interstitial implantation in captive-bred, short-tailed fruit bats, Carollia perspicillata: trophoblastic adhesion and penetration of the uterine epithelium.

Trophoblastic adhesion to, and penetration of, the uterine epithelium during implantation have been examined in captive-bred Carollia perspicillata at the light and electron microscopic levels. Initial adhesion is localized to marginal ridges immediately over the apical intercellular junctions of the epithelial cells. Penetration then involves the intrusion of trophoblastic processes between the epithelial cells and the formation of junctional complexes between the two cell types. As larger areas of adhesion develop, they still occur most often near the intercellular boundaries between the more flattened epithelial cells or on their lateral sides. This suggests that many (if not all) of these broad areas of adhesion to the trophoblast had actually formed along what had originally been part of the lateral (rather than the apical) surfaces of the epithelial cells. Portions of the apical cell surfaces further removed from their intercellular boundaries usually were not adherent to the trophoblast and still had microvilli. Upon reaching the basal lamina of the uterine epithelium, invasion of the endometrium is temporarily retarded, and trophoblast cells migrate instead between the basal lamina and the epithelial cells. This occurs extensively along both the luminal and glandular epithelia around the implantation site, but not significantly along their apical surfaces. This again indicates that adhesive interactions between the trophoblast and those apical surfaces are at most very limited in Carollia. The epithelial cells appear to be viable until separated from their basal laminae by the trophoblast. They are then phagocytized by the trophoblast. During initial penetration of the uterine luminal epithelium, the trophoblast is still entirely of the cellular variety. Syncytiotrophoblast does not begin to appear until later, when the trophoblast first comes into contact with endometrial capillaries.

Cirurgia de revascularizaçäo do miocárdio em pacientes com cardiomiopatia isquêmica e disfunçäo ventricular acentuada: resultados imediatos e de longo prazo / Myocardial revascularization surgery in patients with ischemic cardiopathy and severe left ventricular dysfunction: short and long-term results

Objetivo: Estudar aspectos ainda näo esclarecidos relacionados a parâmetros clínicos pré-operatórios de prognóstico, risco cirúrgico, benefício funcional e sobrevida a longo prazo. Métodos: Foram analisados 71 pacientes com diagnóstico de cardiomiopatia isquêmica, com fraçäo de ejeçäo ventricular esquerda <30 por cento e perfusäo miocárdica pela cintilografia TL-201 antes e após a revascularizaçäo do miocárdio no período hospitalar e após a alta, com seguimento médio de 48 meses. Resultados: A mortalidade imediata foi de 2,8 por cento e a sobrevida tardia em seguimento de 5 anos foi de 62,8 por cento. Analisando todos os pacientes em relaçäo à curva de sobrevida, näo se verificou correlaçäo da presença ou ausência da onda Q no ECG, com a presença de isquemia na cintilografia TI-201, com o grau de fraçäo de ejeçäo do ventrículo esquerdo e com a intensidade de insuficiência cardíaca (ICC) ou de angina. Comparando-se os pacientes sobreviventes com os que faleceram, verificou-se diferença estatisticamente significante em relaçäo à maior presença de ICC classe funcional IV e de bloqueio de ramo esquerdo nos pacientes que faleceram. Conclusäo: As presenças de insuficiência cardíaca classe funcional IV e de bloqueio de ramo esquerdo relacionaram-se a pior prognóstico. Devido ao mecanismo de morte ser multifatorial e a miocardiopatia isquêmica apresentar padräo miocárdico e arterial heterogêneos, é dificil estabelecer índices pré-operatórios de prognóstico para a revascularizaçäo do miocárdio. Os resultados cirúrgicos obtidos a curto e longo prazos demonstram a segurança do procedimento cirúrgico e o seu benefício em aumentar a sobrevida e a qualidade de vida dos pacientes com cardiomiopatia isquêmica.

Isquemia silenciosa após infarto do miocárdio näo complicado: estudo em pacientes assintomáticos / Silent Myocardial Ischemia After Uncomplicated Myocardial Infarction

Objetivo - Verificar o valor prognóstico da isquemiasilenciosa após o infarto do miocárdio (IAM) não complicado. Métodos - Quarenta pacientes, assintomáticos após o 1° episódio de IAM não complicado, foram estudados através da eletrocardiografia dinâmica (48 h) e de esforço, na 2ª e 8ª semanas de evolução. Trinta e nove doentes foram submetidos ao estudo hemodinâmico e cinecoronariográfico e uma paciente à necrópsia. A eletrocardiografia identificou 11 (27,5%) indivíduos com isquemia silenciosa (grupo A); os demais constituiram o grupo B. Resultados - Os pacientes do grupos A e B não diferiram quanto a caracteres clínicos, localização eletrocardiográfica do infarto e função ventricular; os doentes do grupo A apresentaram doença arterial coronária significantemente mais extensa que os do grupo B; ao final de dois anos, os eventos coronários predominaram significantemente no grupo A (36,3%) em relação ao grupo B (3,4%). A análise pelas curvas de Kaplan-Meier mostrou maior probabilidade cumulativa de não ocorrência de eventos no grupo B em relação ao grupo A. Conclusão - A isquemia silenciosa parace ter valor prognóstico nos pacientes após o IAM não complicado, assim como nas outras modalidades de insuficiência coronária

Purpose - To verify the prognostic value of silent myocardial ischemia (SMI) after an uncomplicated myocardial infarction (MI). Methods - Forty asymptomatic patients were studied after afirst uncomplicated MI. They were submitted to 48 hour ambulatory eletrocardiographic monitoring and exercise-testing, during the 2nd and 8th weeks after the acute event. Thirty-nine patients were submitted to cardiac catheterization and coronary arteriography; one patient was submitted to necropsy. The eletrocardiographic study identified 11 (27,5%) individuals with SMI (group A); the other 29 patients were considered group B. Results - Groups A and B were similar in relation to clinical characteristics, infarct site and ventricular function. Group A had significantly more extensive coronary artery disease when comparad to group B. After a two-year follow-tip, patients from group A had significantly more coronary events (36,3%) when comparad to group B (3,4%). Kaplan-Meier analysis demonstrated a significantly higher cumulative probability of not experiencing a new coronary evens for the group B patients. Conclusion - SMI may have a prognostic value after uncomplicated MI, as in other clinical manifestations of coronary artery disease

Nitroglicerina transdérmica em portadores de angina estável / Nitroglycerin patches in patients with stable angina

Objetivo - Avaliar os efeitos clínicos da nitroglicerina transdérmica em portadores de angina estável. Métodos - Quinhentos e trinta e cinco especialistas avaliaram mil quinhentos e trinta e nove pacientes com angina estável, idade média de 61,0 ñ 10,3 anos, sendo 891 (57,97%) do sexo masculino. O estudo multicêntrico prospectivo consistiu na aplicaçäo inicial de 5 mg e, após duas semanas, caso persistisse quadro de angina ou infradesnivelamanto de ST, a dose era aumentada para 10 mg de nitroglicerina transdérmica, a cada 12-14 horas, por 12 semanas. Foram realizadas avaliaçöes clínicas no período inicial e nas semanas 2, 4, 8 e 12, e eletrocardiograma e eletrocardiograma de esforço (ECGE), no início e na 12ª semana. Resultados - Observou-se reduçäo estatisticamente significante (p < 0,005) no número de crises anginosas, no consumo de nitratos sublingual, na pressäo arterial e na porcentagem de pacientes com ECGE isquêmico. A frequência cardíaca e a dose de nitroglicerina transdérmica utilizada näo tiveram alteraçöes significativas. A tolerabilidade desta via de administraçäo foi considerada muito boa e boa em 92,2% dos pacientes. Conclusäo - A utilizaçäo da nitroglicerina transdérmica mostrou-se eficaz no tratamento da angina estável, com excelente tolerabilidade pelo paciente

Purpose - To evaluate the nitroglycerin patches efficacy and tolerability in patients with stable angina pectoris. Methods - One thousand and five hundred and thirty nine patients with stable angina pectoris, mean age 61.0 + 10.3, 891 men and 648 women were prospectively evaluated by five hundred and thirty five specialists after 5 mg or, posteriorly, if clinical necessary, 10 mg of nitroglycerin patches during 12 weeks. Clinical evaluation, electrocardiogram (ECG) and treadmill exercise were obtained on study entry and at weeks 2, 4, 8 and 12 for clinical evaluation, and at week 12 for ECG and treatmill exercise. Results - A significative reduction was observed in the number of angina crisis, sublingual nitrates consumption, arterial blood pressure and on the percentage of positive treadmill exercise tests. The heart rate and nitroglycerin patches dose did not show statistical differences. The compliance of transdermal administration was excellent. Conclusion - The nitroglycerin patches administration was effective for stable angina pectoris with excellent patient's compliance

Spermiogenesis in Odontophrynus cultripes (Amphibia, Anura, Leptodactylidae): Ultrastructural and cytochemical studies of proteins using E-PTA.

Spermiogenesis in the South American leptodactylid frog Odontophrynus cultripes was analyzed ultrastructurally. The spermatids undergo morphological modification while still enclosed in microtubule-rich processes of Sertoli cells. Electron-dense plates resembling junctional structures appear in regions at which the spermatids lie in close contact with the surface of Sertoli cell processes. Spermatid differentiation can be divided into five distinct stages based mainly on chromatin condensation. In the late stages, the densely compacted chromatin loses reactivity to ethanolic phosphotungstic acid (E-PTA). Helical arrangements of microtubules appear in the cytoplasm that surrounds the spermatid nucleus after the second stage. The acrosomal vesicle differentiates into a cone-shaped acrosome that caps the anterior region of the nucleus. The connecting piece, located in the flagellum implantation zone, has transverse striations, and is continuous with the axial rod. The tail is formed by a 9 + 2 axoneme, an undulating membrane, and an axial rod that is rich in basic proteins as demonstrated by E-PTA staining.

Angina de esforço em paciente adulto com origem anômala de artéria coronária esquerda / Effort angina in patient with anomalous origin of left main coronary artery

Origem anômala da artéria coronária esquerda na artéria pulmonar em paciente do sexo feminino, com 50 anos, cujo quadro clínico inicial foi angina de esforço há oito meses. Säo descritos o quadro clínico, a fisiopatologia e o diagnóstico

Anomalous origin of the left main coronary artery from the pulmonary artery in a 50 year-old patient whose initial symptom was effort angina during the last 8 months. The clinical features and treatment are also discussed.