HLA-G alleles and their impacts on placental HSV-1 infection in women from southern Brazil.

The Human Leukocyte Antigen G (HLA-G) is an immunoregulatory molecule with a critical role in pregnancy success. HLA-G alleles are associated with differential susceptibility to multiple conditions, including gestational problems, infectious diseases, and viral persistence. Of note, both herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) can impair HLA-G expression, interfering with HLA-G-associated immunoregulation. On the other hand, the impacts of HLA-G alleles on susceptibility to Herpesviridae infection is a neglected issue. Therefore, this study evaluated HLA-G allele frequencies and their associations with placental Herpesviridae infection in women from southern Brazil. Placenta samples were collected soon after delivery, and detection of viral DNA of HSV-1, HSV-2 and human cytomegalovirus (HCMV) was performed by polymerase chain reaction (PCR). A fragment of HLA-G (exons 2-4) was amplified by PCR, sequenced, and analyzed to allele determination. One hundred and seventy women had their alleles determined. Overall, 25 HLA-G alleles were found, distributed into 56 different genotypes. The most frequent alleles were G* 01:01:01 and G* 01:01:02, found in 37.9 % and 16.5 % of samples, respectively. Among the 170 women, 89 (52.4 %) tested positive for Herpesviridae DNA in the placenta, 55 (32.3 %) tested negative, 3 (1.8 %) were negative for HSV-1 and HSV-2 (with absent HCMV data), and 23 (13.5 %) were undetermined. The G* 01:01:01 allele was significantly associated with an increased risk of placental HSV-1 infection (p = 0.0151; OR=1.837; IC=1.108-3.045). This study describes new information concerning placental HLA-G alleles in women from southern Brazil and helps explain how genetic background can modify susceptibility to placental infections.

Impact of hepatitis C virus eradication with direct-acting antivirals on glycidic metabolism.

Objective: To compare the glucose metabolism of patients with chronic hepatitis C virus infection treated with direct-acting antivirals (DAAs) in pretreatment and sustained viral response (SVR) periods. Materials and methods: This was an intervention pre-post study of 273 patients with chronic hepatitis C virus infection treated with DAAs from March 2018 to December 2019. Glycidic metabolism was evaluated through homeostasis model assessment (HOMA) - insulin resistance (IR) and HOMA-ß indices and assessments of insulinemia and HbA1c levels. These parameters were analyzed with a T test by paired comparison of the means of the variables and Wilcoxon's test paired for the median; in the variables with an abnormal distribution, the Z score was generated for the mean in both the pretreatment and SVR periods. Statistical significance was considered at p ≤ 0.05. Results: Among 273 participants, 125 (45.8%) had prediabetes, and 50 (18.3%) had diabetes. In SVR, there was a significant increase in platelets, albumin, alkaline phosphatase, cholesterol and triglycerides and a significant decrease in aspartate aminotransferase, alanine aminotransferase, gamma GT and bilirubin. The HOMA-IR and HOMA-ß indices increased in SVR from 1.95 to 2.29 (p = 0.087) and 71.20 to 82.60 (p = 0.001), respectively. Insulinemia increased from 7.60 µU/mL to 8.90 µU/mL (p = 0.011). HbA1c decreased from 5.6 to 5.4 (p < 0.001). Among patients with prediabetes and those with diabetes, the reduction in HbA1c values was significant (p = 0.006 and p = 0.026, respectively). Conclusion: SVR significantly impacts and leads to improvement in glucose metabolism in patients with chronic liver disease induced by hepatitis C virus.

Impact of TLR7 and TLR9 polymorphisms on susceptibility to placental infections and pregnancy complications.

This study evaluated the impact of the TLR7 Gln11Leu (rs179008) and TLR9 -1237 T/C (rs5743836) single nucleotide polymorphisms (SNPs) on susceptibility to placental infections and pregnancy complications in 455 Brazilian women. Demographic, socioeconomic, gynecological, and clinical characteristics of the women were collected. Placental tissues were sampled from pregnant women and human and viral DNA was extracted. Human alphaherpesvirus 1 (Herpes simplex virus type 1, HSV-1), Human alphaherpesvirus 2 (Herpes simplex virus type 2, HSV-2) and Human betaherpesvirus 5 (Human cytomegalovirus, HCMV) were detected by nested PCR. TLR9 and TLR7 SNPs were genotyped by PCR amplification of bi-directional specific alleles (Bi-PASA) and restriction fragment length polymorphism (RFLP), respectively. Infections at the time of birth were detected in 45.71 % of women. The presence of the TT genotype (recessive model) of the TLR7 SNP was associated with increased susceptibility to HSV-1 infection (O.R. = 2.23, p = 0.05). The presence of the C allele of the TLR9 SNP, in heterozygosis or homozygosis (dominant model), decreased the infection risk by HCMV (O.R. = 0.31, p-mod<0.05). The TT genotype (recessive model) of the TLR7 SNP was significantly associated (p < 0.05) with increased occurrence of pre-treated hypertension. The codominant model of the TLR9 SNP was significantly associated (p < 0.05) with reduced risk of hospitalization during pregnancy. In combination, the AA/CT (TLR7-TLR9) genotypes significantly decreased the risk of placental infection by HSV-1 and/or HSV-2 (O.R. = 0.47, p = 0.02), the susceptibility to all infectious agents considered in combination (O.R. = 0.4, p = 0.00), and the need of hospitalization (O.R. = 0.48, p = 0.02). In conclusion, TLR7 and TLR9 SNPs are potential modulating factors for the risk of placental infections and pregnancy complications.

High Incidence of Herpes Simplex Virus-1 in Cord Blood and Placenta Infection of Women in Southern Brazil.

OBJECTIVE: Estimate the prevalence of human herpesvirus type 1 HSV-1 DNA in placental samples, its incidence in umbilical cord blood of newborns and the associated risk factors. METHODS: Placental biopsies and umbilical cord blood were analyzed, totaling 480 samples, from asymptomatic parturients and their newborns at a University Hospital. Nested polymerase chain reaction (PCR) and gene sequencing were used to identify the virus; odds ratio (OR) and relative risk (RR) were performed to compare risk factors associated with this condition. RESULTS: The prevalence of HSV-1 DNA in placental samples was 37.5%, and the incidence in cord blood was 27.5%. Hematogenous transplacental route was identified in 61.4% from HSV-1+ samples of umbilical cord blood paired with the placental tissue. No evidence of the virus was observed in the remaining 38.6% of placental tissues, suggesting an ascendant infection from the genital tract, without replication in the placental tissue, resulting in intra-amniotic infection and vertical transmission, seen by the virus in the cord blood. The lack of condom use increased the risk of finding HSV-1 in the placenta and umbilical cord blood. CONCLUSION: The occurrence of HSV-1 DNA in the placenta and in cord blood found suggests vertical transmission from asymptomatic pregnant women to the fetus.

OBJETIVO: Estimar a prevalência do DNA do vírus herpes humano 1 (HSV-1) em amostras de placenta, sua incidência no sangue do cordão umbilical de recém-nascidos e fatores de risco associados. MéTODOS: Biópsias de placenta e de sangue de cordão umbilical foram analisadas, totalizando 480 amostras de parturientes assintomáticas e seus recém-nascidos em um hospital universitário. Reação de cadeia de polimerase (RCP) nested e sequenciamento gênico foram usados para identificar o vírus; odds ratio (OR) e risco relativo (RR) foram realizados para comparar os fatores de risco associados à essa condição. RESULTADOS: A prevalência do DNA do HSV-1 em amostras de placenta foi de 37,5%, e a incidência no sangue do cordão foi de 27,5%. A via transplacentária hematogênica foi identificada em 61,4% das amostras de HSV-1 + do sangue do cordão umbilical, pareadas com o tecido placentário. Nenhuma evidência do vírus foi observada nos restantes 38,6% dos tecidos placentários, sugerindo uma infecção ascendente do trato genital. A falta de uso do preservativo aumentou o risco de encontrar o HSV-1 na placenta e no sangue do cordão umbilical. CONCLUSãO: A ocorrência de DNA do HSV-1 na placenta e no sangue do cordão umbilical sugere uma transmissão vertical de gestantes assintomáticas para o feto.

Herpes simplex virus type 2 IgG antibodies in sera of umbilical cord as a proxy for placental infection in asymptomatic pregnant women.

PROBLEM: Herpes simplex virus type 2 (HSV-2) infection is one of the most prevalent diseases worldwide and is mainly sexually transmitted. When infecting pregnant women, HSV-2 is able to infect the placenta, can reach the fetus, and may affect the fetal development. We sought to determine the prevalence of HSV-2 infection and reactivation in asymptomatic pregnant women, the correlation between IgG in the maternal circulation and cord blood, and the correlation between circulating IgG, placental, and newborn infection (blood cord). METHOD OF STUDY: Serum samples and placental tissues from pregnant women and umbilical cord blood samples from their newborns were collected. Anti-HSV-2 antibodies were identified by ELISA, and HSV-2 DNA was detected by nested PCR. RESULTS: The seropositivity of IgG in pregnant women was 29.7% and IgM was detected in 1 woman (0.5%). In the umbilical cord of newborns, 33.1% were IgG-positive and IgM was detected in 2 samples (1.5%). A positive correlation between HSV-2 IgG titers in serum from pregnant women and cord blood samples was found (r = .36, P = .001). A difference between the positive and negative placental groups (maternal side) was found in titers of IgG in sera of umbilical cord, which were significantly higher in the positive placental group (P = .004). CONCLUSION: We describe for the first time that newborns from mothers with HSV-2 placental infection have higher IgG titers in sera of umbilical cord, suggesting IgGs antibodies can be indicative of placental viral infection in asymptomatic women.

Seroepidemiology of Paracoccidioides brasiliensis infection in horses from Rio Grande do Sul, Brazil.

Paracoccidioides brasiliensis is the etiological agent of the major systemic mycosis in Brazil, called paracoccidioidomycosis. Although the Rio Grande do Sul is considered an endemic area of the disease, there are few studies on the ecology of P. brasiliensis in the state. Therefore, this study aimed to evaluate the infection of P. brasiliensis in horses from the mesoregion of Southwest Riograndense, using these animals as sentinels. Serological techniques, such as double immunodiffusion in agar gel (AGID) and indirect ELISA, were performed to detect the anti-gp43 P. brasiliensis antibody in horses from five different farms in the region of Bagé, RS, Brazil. Serology was performed in 200 Pure Blood English horses up to two years of age that were born and raised exclusively at the farms. Of these horses, 12% had anti-gp43 antibodies according to the ELISA results, with rates ranging from 0 to 30% according to the farm of origin (p < 0.001). Based on the immunodiffusion results, all equine serum samples were negative. These results indicate the presence of the fungus P. brasiliensis in the middle region of the southwestern state of Rio Grande do Sul, Brazil.

[Frequency of Human Papillomavirus in the placenta, in the colostrum and in the umbilical cord blood]. / Frequência do Papilomavírus Humano na placenta, no colostro e no sangue do cordão umbilical.

PURPOSE: To determine the frequency of Human Papillomavirus (HPV) in the placenta, in the colostrum and in the umbilical cord blood of parturient women and their newborns assisted at the Clinic of Gynecology and Obstetrics of the University Hospital of Rio Grande (RS), Brazil. METHODS: Biopsies were collected from 150 placentas on the maternal side, 150 on the fetal side, 138 samples of umbilical cord blood and 118 of the colostrum. The placenta biopsies were collected from the central and peripheral portions. DNA was extracted according to the manufacturer's protocol and to a reference found in the literature. HPV was detected by the nested polymerase chain reaction (PCR-Nested) using primers MY09/11 and GP5/GP6. Genotyping was performed by direct sequencing. The participants responded to a self-applied questionnaire with demographic and clinical data, in order to characterize the sample. RESULTS: HPV was detected in 4% (6/150) of cases on the mother's side of the placentas, in 3.3% (5/150) on the fetal side, in 2.2% (3/138) in umbilical cord blood and in 0.84% (1/118) in colostrum samples. The vertical transmission rate was 50%. HPV-6 was the low-risk genotype found (60%) and the high-risk genotypes were HPV-16 and HPV-18 (20% each). CONCLUSIONS: These results suggest that HPV can infect the placenta, the colostrum and the umbilical cord blood.

Seroepidemiology of Paracoccidioides brasiliensis infection in horses from Rio Grande do Sul, Brazil

Paracoccidioides brasiliensis is the etiological agent of the major systemic mycosis in Brazil, called paracoccidioidomycosis. Although the Rio Grande do Sul is considered an endemic area of the disease, there are few studies on the ecology of P. brasiliensis in the state. Therefore, this study aimed to evaluate the infection of P. brasiliensis in horses from the mesoregion of Southwest Riograndense, using these animals as sentinels. Serological techniques, such as double immunodiffusion in agar gel (AGID) and indirect ELISA, were performed to detect the anti-gp43 P. brasiliensis antibody in horses from five different farms in the region of Bagé, RS, Brazil. Serology was performed in 200 Pure Blood English horses up to two years of age that were born and raised exclusively at the farms. Of these horses, 12% had anti-gp43 antibodies according to the ELISA results, with rates ranging from 0 to 30% according to the farm of origin (p < 0.001). Based on the immunodiffusion results, all equine serum samples were negative. These results indicate the presence of the fungus P. brasiliensis in the middle region of the southwestern state of Rio Grande do Sul, Brazil.

Herpes simplex virus: prevalence in placental tissue and incidence in neonatal cord blood samples.

Herpes simplex virus (HSV) infection is one of the most prevalent infectious diseases worldwide, with HSV-2 being primarily associated with genital infections. HSV-2 is believed to account for the majority of cases of neonatal herpes, which may cause diverse of complications in infected newborns. The present study sought to estimate the prevalence of HSV-2 in placental tissue samples and the incidence of HSV-2 in the umbilical cord blood of newborn infants. Placental tissue samples from 201 women (maternal-side and fetal-side = 402 specimens) and 184 neonatal cord blood samples, all collected at the obstetric ward of a University hospital were studied. HSV-2 was detected by means of nested PCR. The prevalence of HSV-2 in placental samples was 9.0% (n = 18), and the incidence of neonatal HSV-2 infection was 1.1% (n = 2). All HSV-2-positive patients were asymptomatic at the time of delivery and none reported genital herpes. Women with a time between rupture of membranes and delivery of ≥360 min had an approximately fourfold risk of HSV-2 infection in the placental tissue (95% CI 0.93-5.66, P = 0.01). These results suggest that HSV-2 is present in the placenta of asymptomatic women and that a risk of transmission to the neonate exists. New strategies must be implemented for the management of asymptomatic patients who are capable of transmitting the virus to the newborn.

Evaluation of the initial and chronic phases of toxocariasis after consumption of liver treated by freezing or cooling.

Human toxocariasis is a neglected parasitic zoonosis of worldwide distribution. The consumption of raw or undercooked meat and offal from paratenic hosts of the Toxocara canis nematode can cause infection in humans, but there have been a lack of studies examining specific prophylactic measures to combat this mode of transmission. The aim of this study was to evaluate the establishment of infection by T. canis larvae at the initial and chronic phases of visceral toxocariasis after the consumption of mouse liver subjected to cold treatment. This study was divided into two stages using groups (G) of five donor mice inoculated with 2,000 eggs of T. canis. Two days post-inoculation, the livers of donor mice in G1 and G2 were kept at -20 °C and between 0 and 4 °C, respectively, for 10 days. In the first stage of the study, the livers of mice from G1, G2, and G3 (control) were subjected to a tissue digestion technique and found to be positive for infection. In the second stage, which evaluated infection in mice that had consumed livers from donor mice, receiver mice of G4 and G7 were fed with livers of donor mice from G1 (freezing), receiver mice of G5 and G8 were fed with livers of donor mice from G2 (cooling), and receiver mice of G6 and G9 with livers from G3 (control). Then, the tissue digestion technique was performed for recovering larvae from organs and carcasses of mice, at 2 days (G4, G5, and G6) and 60 days after liver consumption (G7, G8, and G9). It was observed that freezing inhibited the viability of 100 % of the larvae, while cooling promoted 87.7 and 95.7 % reductions in the intensity of infection at 2 and 60 days after liver consumption, respectively. Under the studied conditions, cold treatment shows great potential to help control this parasitosis, both in the initial and chronic phases of toxocariasis.