Genetic modifications of cytokine genes and Toxoplasma gondii infections in pregnant women.

PURPOSE: Toxoplasma gondii causes one of the most common intrauterine infections worldwide, thus being a severe threat during pregnancy. IL1, IL6, IL10, IL12, and TNF-α cytokines were reported to be involved in immune responses to infections with T. gondii. The research was aimed to reveal relationships between genetic changes within the polymorphisms of these cytokine genes and the incidence of T. gondii infection among pregnant women, as well as congenital transmission of the parasite to the foetuses of their infected mothers. METHODS: The primary study was performed in 148 Polish pregnant women, including 74 T. gondii-infected patients and 74 age-matched uninfected individuals; and further analysis - among the additional 142 pregnant women. Genotypes within IL1A -889 C>T, IL1B +3954 C>T, IL6 -174 G>C, IL10 -1082 G>A, IL12B -1188 A>C and TNFA -308 G>A single nucleotide polymorphisms (SNPs) were determined, using self-designed nested PCR-RFLP assays. Randomly selected PCR products, representing distinct genotypes in the analyzed polymorphisms, were confirmed by sequencing, using the Sanger method. A statistical analysis was carried out of relationships between genetic alterations within studied SNPs and the occurrence of T. gondii infection, using the following tools: cross-tabulation, Pearson's Chi-square test and the logistic regression model to estimate genetic models of inheritance. A power analysis of statistically significant outcomes was performed by Cramér's V test. RESULTS: A multiple-SNP analysis showed TC haplotype for IL1A and IL1B SNPs to be significantly associated with a decreased risk of the parasitic infection (OR 0.41, P≤0.050). The association remained important after power analysis (Cramér's V = 0.39, χ2 = 7.73, P≤0.050), and the additional analysis with larger groups of patients (OR 0.47, P≤0.050). Moreover, the CCCAGA complex variants were for all the studied polymorphisms at an increased risk of T. gondii infection (OR 8.14, P≤0.050), although this strong relationship was not significant in the further analysis (Cramér's V = 0.76, χ2 = 26.81, P = 0.310). Regarding the susceptibility to congenital transmission of T. gondii from mothers to their foetuses among the infected pregnant women, the presence of GA heterozygotic status within IL10 polymorphism significantly increased the risk of parasitic transmission (OR 5.73 in the codominant model and OR 5.18 in the overdominant model; P≤0.050). The correlation stayed important in the power analysis (Cramér's V = 0.29, χ2 = 6.03, P≤0.050), although it was non-significant in larger groups of patients. Important relationships specific for the first study cohort remained non-significant in the second group of studied pregnant women. CONCLUSIONS: Within the analyzed cohort of Polish pregnant women, the genetic modifications from SNPs of genes, encoding both the proinflammatory IL1α, IL1ß, IL6, IL12 and TNF-α, and anti-inflammatory IL10 cytokines, may have been associated with susceptibility to T. gondii infection. It is the first study on the contribution of cytokine genes polymorphisms to the occurrence of T. gondii infection during pregnancy. Further studies for other populations of pregnant women would be justified to reveal a detailed role of the analyzed polymorphisms for the occurrence of T. gondii infections during pregnancy.

What is the impact of gestational diabetes mellitus on frequency of structural chromosome aberrations in pregnant women and their offspring?

Gestational diabetes mellitus (GDM) is defined as carbohydrate intolerance which results in hyperglycemia first diagnosed during pregnancy. It is associated with an increased levels of oxidative stress due to overproduction of reactive oxygen species (ROS). Overproduction of ROS induces protein oxidation, lipid peroxidation and different types of DNA damage. The objective of this study was to determine the frequencies of structural chromosome aberrations (CA) in peripheral blood of pregnant women (mothers) with GDM and in cord blood of their newborns. Peripheral blood lymphocytes were collected from 35 GDM mothers and cord blood lymphocytes from their 35 newborns. The control group included 30 pregnant mothers without diabetes mellitus (DM) and their 30 newborns. CA were evaluated with in vitro chromosome aberration assays. We observed a moderate increase of the mean numbers of structural CA between GDM mothers and their newborns, GDM mothers and mothers without DM, GDM mothers' offspring and the offspring of mothers without DM, mothers without DM and their newborns, but this effect did not reach statistical significance (p>0.1).

Genetic alterations within TLR genes in development of Toxoplasma gondii infection among Polish pregnant women.

PURPOSE: The research was conducted to evaluate the role of genotypes, haplotypes and multiple-SNP variants in the range of TLR2, TLR4 and TLR9 single nucleotide polymorphisms (SNPs) in the development of Toxoplasma gondii infection among Polish pregnant women. MATERIAL AND METHODS: The study was performed for 116 Polish pregnant women, including 51 patients infected with T. gondii, and 65 age-matched control pregnant individuals. Genotypes in TLR2 2258 G>A, TLR4 896 A>G, TLR4 1196 C>T and TLR9 2848 G>A SNPs were estimated by self-designed, nested PCR-RFLP assays. Randomly selected PCR products, representative for distinct genotypes in the studied polymorphisms, were confirmed by sequencing. All the genotypes were calculated for Hardy-Weinberg (H-W) equilibrium and TLR4 variants were tested for linkage disequilibrium. Relationships were assessed between alleles, genotypes, haplotypes or multiple-SNP variants in TLR polymorphisms and the occurrence of T. gondii infection in pregnant women, using a logistic regression model. RESULTS: All the analyzed genotypes preserved the H-W equilibrium among the studied groups of patients (P>0.050). Similar distribution of distinct alleles and individual genotypes in TLR SNPs, as well as of haplotypes in TLR4 polymorphisms, were observed in T. gondii infected and control uninfected pregnant women. However, the GACG multiple-SNP variant, within the range of all the four studied polymorphisms, was correlated with a decreased risk of the parasitic infection (OR 0.52, 95% CI 0.28-0.97; P≤0.050). CONCLUSIONS: The polymorphisms, located within TLR2, TLR4 and TLR9 genes, may be involved together in occurrence of T. gondii infection among Polish pregnant women.

Both a frameshift and a missense mutation of the STRA6 gene observed in an infant with the Matthew-Wood syndrome.

BACKGROUND: The Matthew-Wood syndrome is associated with mutations of the STRA6 gene. It combines a pulmonary agenesis/hypoplasia; microphthalmia/anophthalmia; congenital cardiac, digestive, and urogenital malformations; and diaphragmatic defects. CASE: A 23-year-old nulliparous woman was referred to our center after a fetal ultrasound examination at 26 weeks of pregnancy revealed an abnormal head shape, a heart malformation, multiple cysts in both kidneys, and dilated ureters. A male baby (46, XY; 3600g; Apgar score 1) was delivered at 38 weeks of gestation and died 1 hr later due to respiratory failure. The diagnosis of Matthew-Wood syndrome was suspected given the association of bilateral anophthalmia, agenesis of the left lung, and heart and kidney defects. It was confirmed by the identification of two deleterious mutations of the STRA6 gene. RESULTS: The child was a compound heterozygote for two previously reported mutations, a paternally inherited missense mutation (c.878C>T [p.Pro293Leu] and a maternally inherited frameshift mutation (c.50_52delACTinsCC [p. Asp17Alafs*55]), producing a premature stop codon. CONCLUSION: The diagnosis of Matthew-Wood syndrome should be considered in all fetuses with microphthalmia/anophthalmia. It requires an extensive ultrasound/MRI examination of the lung, heart, and diaphragm. Birth Defects Research 109:251-253, 2017. © 2017 Wiley Periodicals, Inc.

Toll-like receptors genes polymorphisms and the occurrence of HCMV infection among pregnant women.

BACKGROUND: Human cytomegalovirus (HCMV) is the most common cause of intrauterine infections worldwide. The toll-like receptors (TLRs) have been reported as important factors in immune response against HCMV. Particularly, TLR2, TLR4 and TLR9 have been shown to be involved in antiviral immunity. Evaluation of the role of single nucleotide polymorphisms (SNPs), located within TLR2, TLR4 and TLR9 genes, in the development of human cytomegalovirus (HCMV) infection in pregnant women and their fetuses and neonates, was performed. METHODS: The study was performed for 131 pregnant women, including 66 patients infected with HCMV during pregnancy, and 65 age-matched control pregnant individuals. The patients were selected to the study, based on serological status of anti-HCMV IgG and IgM antibodies and on the presence of viral DNA in their body fluids. Genotypes in TLR2 2258 A > G, TLR4 896 G > A and 1196 C > T and TLR9 2848 G > A SNPs were determined by self-designed nested PCR-RFLP assays. Randomly selected PCR products, representative for distinct genotypes in TLR SNPs, were confirmed by sequencing. A relationship between the genotypes, alleles, haplotypes and multiple variants in the studied polymorphisms, and the occurrence of HCMV infection in pregnant women and their offsprings, was determined, using a logistic regression model. RESULTS: Genotypes in all the analyzed polymorphisms preserved the Hardy-Weinberg equilibrium in pregnant women, both infected and uninfected with HCMV (P > 0.050). GG homozygotic and GA heterozygotic status in TLR9 2848 G > A SNP decreased significantly the occurrence of HCMV infection (OR 0.44 95% CI 0.21-0.94 in the dominant model, P ≤ 0.050). The G allele in TLR9 SNP was significantly more frequent among the uninfected pregnant women than among the infected ones (χ2 = 4.14, P ≤ 0.050). Considering other polymorphisms, similar frequencies of distinct genotypes, haplotypes and multiple-SNP variants were observed between the studied groups of patients. CONCLUSIONS: TLR9 2848 G > A SNP may be associated with HCMV infection in pregnant women.

Association of SNPs from IL1A, IL1B, and IL6 Genes with Human Cytomegalovirus Infection Among Pregnant Women.

The study was aimed to estimate the role and prevalence rates of genotypes, haplotypes, and alleles, located within the single-nucleotide polymorphisms (SNPs) of interleukin (IL) 1A, IL1B, and IL6 genes, in the occurrence and development of human cytomegalovirus (HCMV) infection among pregnant women. A research was conducted in 129 pregnant women, out of whom, 65 were HCMV infected and 64 were age-matched control uninfected individuals. HCMV DNA was quantitated for UL55 gene by the real-time Q PCR in the body fluids. The genotypic statuses within the SNPs were determined by nested PCR-RFLP assays and confirmed, by sequencing for randomly selected representative PCR products. A relationship between the genotypes and alleles, as well as haplotypes and multiple variants in the studied polymorphisms, and the occurrence of HCMV infection in pregnant women, was determined using a logistic regression model. TT genotype within IL1A polymorphism significantly decreased the risk of HCMV infection (OR 0.32, 95% CI 0.09-1.05; p ≤ 0.050). Considering IL6 SNP, the prevalence rate of GC genotype was significantly decreased among the HCMV infected, compared to the uninfected control individuals (OR 0.45, 95% CI 0.21-0.99; p ≤ 0.050). Moreover, CC homozygotic status in IL6 SNP, found in pregnant women, significantly decreased the risk of congenital infection with HCMV in their offsprings (OR 0.12; p ≤ 0.050). In multiple SNP analysis, TC haplotype within the IL1 polymorphisms significantly decreased the risk of the infection in pregnant women (OR 0.38 95% CI 0.15-0.96; p ≤ 0.050). In addition, TTG complex variants for all the studied polymorphisms and TG variants for IL1B and IL6 SNPs were significantly more prevalent among the infected offsprings with symptomatic congenital cytomegaly than among the asymptomatic cases (p ≤ 0.050). In conclusion, the analyzed IL1A -889 C>T, IL1B +3954 C>T, and IL6 -174 G>C polymorphisms may be associated with the occurrence and development of HCMV infection among studied patients.

The role of single nucleotide polymorphisms, contained in proinflammatory cytokine genes, in the development of congenital infection with human cytomegalovirus in fetuses and neonates.

PURPOSE: The research project targeted the distribution of genotypes, alleles and haplotypes in single nucleotide polymorphisms (SNPs) within the interleukin (IL) 1A, IL1B, IL6, IL12B and TNFA genes, in fetuses and neonates, congenitally infected with human cytomegalovirus (HCMV), and among uninfected controls. METHODS: The study included 20 fetuses and neonates with congenital HCMV infection and 31 control individuals. The genotypes in SNPs of the studied cytokine genes were identified by a self-designed nested PCR-RFLP assays. Selected genotypes, representing distinct variants in analyzed polymorphisms, were confirmed by sequencing. The relationship between the genetic status of the studied polymorphisms and congenital infection development was estimated, using a logistic regression model. RESULTS: The CT haplotype, composed of C allele determined in IL1A -889 C > T and T allele in IL1B +3954 C > T SNP, increased the risk of congenital HCMV infection, as well as the onset of disease related symptoms (P ≤ 0.0001). Considering disease outcome, the risk of development of symptoms, was increased among the CT heterozygotes in IL1A -889 C > T polymorphism (OR 2.86, 95% CI 0.24-33.90; P = 0.045). Moreover, multiple-SNP variants CCGAG in the range of all the SNPs, analyzed in the study, increased the risk of congenital infection with HCMV (OR 7.94, 95% CI 1.38-45.69; P = 0.026). CONCLUSIONS: Polymorphisms within the proinflammatory cytokine genes may contribute to the development of congenital infection with HCMV.

TLR2 2258 G>A single nucleotide polymorphism and the risk of congenital infection with human cytomegalovirus.

BACKGROUND: Human cytomegalovirus (HCMV) is responsible for the most common intrauterine infections, which may be acquired congenitally from infected pregnant woman to fetus. The research was aimed to estimate the role of three single nucleotide polymorphisms (SNPs) located in TLR2 gene, and the common contribution of TLR2, and previously studied TLR4 and TLR9 SNPs, to the occurrence of congenital HCMV infection in fetuses and newborns. METHODS: The study was performed in 20 Polish fetuses and newborns, congenitally infected with HCMV, and in 31 uninfected controls, as well as with participation of pregnant women, the mothers of 16 infected and 14 uninfected offsprings. Genotypes in TLR2 SNPs were determined, using self-designed nested PCR-RFLP assays, and confirmed by sequencing. The genotypes were tested for Hardy-Weinberg (H-W) equilibrium, and for their relationship with the development of congenital cytomegaly, using a logistic regression model. The common influence of TLR2, TLR4 and TLR9 SNPs on the occurrence of congenital disease was estimated by multiple-SNP analysis. RESULTS: Distribution of the genotypes and alleles in TLR2 1350 T>C and 2029 C>T SNPs was similar between the studied groups of fetuses and neonates. In case of 2258 G>A polymorphism, the GA heterozygotic status was significantly more frequent in the infected cases than among the uninfected individuals (25.0% vs. 3.2%, respectively), and increased the risk of HCMV infection (OR 10.00, 95% CI 1.07-93.44; P ≤ 0.050). Similarly, the A allele within 2258 G>A polymorphism was significantly more frequent among the infected offsprings than in the uninfected ones (12.5% vs. 1.6%; P ≤ 0.050). Complex AA variants for both TLR2 2258 and TLR9 2848 G>A polymorphisms, were estimated to be at increased risk of congenital HCMV infection (OR 11.58, 95% CI 1.19-112.59; P ≤ 0.050). Additionally, significant relationships were observed between the occurrence of complex AA or GA variants for both TLR2 and TLR9 SNPs and the increased viral loads, determined in fetal amniotic fluids and in maternal blood or urine specimens (P ≤ 0.050). CONCLUSIONS: Among various TLR2, TLR4 and TLR9 polymorphisms, TLR2 2258 G>A SNP seems to be an important factor associated with increased risk of congenital HCMV infection in Polish fetuses and neonates.

Association of TLR3 L412F Polymorphism with Cytomegalovirus Infection in Children.

Intracellular Toll-like receptor 3 (TLR3) recognizes viral double-stranded RNA (dsRNA) and activates antiviral immune responses through the production of type I interferons (IFNs) and inflammatory cytokines. This receptor binds to dsRNA molecules produced during human cytomegalovirus (HCMV) replication. TLR7 senses viral single-stranded RNA (ssRNA) in endosomes, and it can interact with endogenous RNAs. We determined the genotype distribution of single-nucleotide polymorphisms (SNPs) within the TLR3 and TLR7 genes in children with HCMV infection and the relationship between TLR polymorphisms and viral infection. We genotyped 59 children with symptomatic HCMV infection and 78 healthy individuals for SNPs in the TLR3 (rs3775290, c.1377C>T, F459F; rs3775291, c.1234C>T, L412F; rs3775296, c.-7C>A) and TLR7 (rs179008, c.32A>T, Q11L; rs5741880, c.3+1716G>T) genes. SNP genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and capillary electrophoresis. The HCMV DNA load was quantified by real-time PCR. We found an increased frequency of the heterozygous genotype TLR3 L412F in children with HCMV infection compared with uninfected cases. In individuals with a mutation present in at least one allele of the L412F SNP, an increased risk of HCMV disease was found, and this result remained highly significant after Bonferroni's correction for multiple testing (Pc < 0.001). The heterozygous genotype of this SNP was associated with the increased risk of HCMV disease in an adjusted model that included the HCMV DNA copy number in whole blood and urine (P < 0.001 and P = 0.008, respectively). Moreover, those with a heterozygous genotype of rs3775296 showed an increased relative risk of HCMV infection (P = 0.042), but this association did not reach statistical significance after correction for multiple testing. In contrast, the rs3775290 SNP of TLR3 and TLR7 SNPs were not related to viral infection. A moderate linkage disequilibrium (LD) was observed between the SNPs rs3775291 and rs3775296 (r2 = 0.514). We suggest that the L412F polymorphism in the TLR3 gene could be a genetic risk factor for the development of HCMV disease.

New criteria for gestational diabetes mellitus - do they impact the outcome?

OBJECTIVE: The aim of the study was to compare the perinatal outcome of pregnancies in mothers who were diagnosed with gestational diabetes mellitus (GDM) with previous versus current Polish Gynecological Society (PTG) criteria. METHODS: 475 patients were divided into three groups. In group A, the patients only met the previous PTG criteria for a GDM diagnosis, i.e., those with a blood glucose level of 140-152 mg/dl 2 hours after administration, a fasting glucose level <92 mg/dl, and a blood glucose level <180 mg/dl 1 hour after administration. Group B included patients complying with both the previous and current PTG criteria for a GDM diagnosis. Group C included patients who only met the current PTG criteria for a GDM diagnosis, i.e., those with a fasting blood glucose level of 92-99 mg/dl, a blood glucose level <180 mg/dl 1 hour and <140 mg/dl 2 hours after administration, respectively. RESULTS: Women from group C were characterized by the highest fasting glycaemia in the first trimester of pregnancy (93.0 mg/dL vs. 88.0 mg/dL vs. 83.5 mg/dL, p=0.012) and during the OGTT (p=0.001). Gestational diabetes was diagnosed significantly earlier in patients from group C (23 vs. 26 vs. 26 weeks, p=0.005). The patients from group A significantly less frequently required insulin therapy for proper glycemic control (p=0.035). Women from group A were characterized by lower pre-pregnancy BMI (p=0.001). CONCLUSIONS: Current PTG criteria for diagnosing GDM according to the IADPSG allow for identification of women who often require insulin therapy to achieve proper glycemic control.

Evaluation of the amoxicillin concentrations in amniotic fluid, placenta, umbilical cord blood and maternal serum two hours after oral administration.

OBJECTIVES: Amoxicillin is a broad-spectrum beta-lactam antibiotic. Due to its low toxicity, it is commonly used in obstetrics. The objective of this study was to assess amoxicillin concentrations in amniotic fluid, umbilical blood, placenta and maternal serum two hours following oral administration among pregnant women at term and to assess obstetric and non-obstetric factors that might affect amoxicillin's penetration of these tissues. MATERIALS AND METHODS: A total of 30 full-term pregnant women who qualified for elective Caesarean delivery were included in the study. Amoxicillin at a dose of 500 mg was administered prior to surgery. Amoxicillin levels were determined by diffusion microbial assay. RESULTS: The maternal serum, placental, umbilical blood and amniotic fluid levels of amoxicillin two hours after oral administration were 2.18±1.30 µg/g, 1.00±0.71 µg/g, 1.00±0.73 µg/g, and 0.67±0.59 µg/g, respectively (Table 2). Maternal serum levels of amoxicillin were significantly higher compared to other tissues (p<0.05). CONCLUSION: If the target tissues for the use of antibiotic drugs in pregnant patients are the fetus and/or the placenta, the drug should be administered in a higher-than-standard dose than that used to treat infections in non-pregnant patients. Considering that there is a maximum absorbable dose following oral administration, intravenous administration should be considered to prevent failure of antibiotic treatment. A higher dose of amoxicillin should be considered in obese mothers.

TLR9 -1486T/C and 2848C/T SNPs Are Associated with Human Cytomegalovirus Infection in Infants.

Toll-like receptor 9 (TLR9) recognizes non-methylated viral CpG-containing DNA and serves as a pattern recognition receptor that signals the presence of human cytomegalovirus (HCMV). Here, we present the genotype distribution of single-nucleotide polymorphisms (SNPs) of the TLR9 gene in infants and the relationship between TLR9 polymorphisms and HCMV infection. Four polymorphisms (-1237T/C, rs5743836; -1486T/C, rs187084; 1174G/A, rs352139; and 2848C/T, rs352140) in the TLR9 gene were genotyped in 72 infants with symptomatic HCMV infection and 70 healthy individuals. SNP genotyping was performed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Digested fragments were separated and identified by capillary electrophoresis. The HCMV DNA copy number was measured by a quantitative real-time PCR assay. We found an increased frequency of heterozygous genotypes TLR9 -1486T/C and 2848C/T in infants with HCMV infection compared with uninfected cases. Heterozygous variants of these two SNPs increased the risk of HCMV disease in children (P = 0.044 and P = 0.029, respectively). In infants with a mutation present in at least one allele of -1486T/C and 2848C/T SNPs, a trend towards increased risk of cytomegaly was confirmed after Bonferroni's correction for multiple testing (Pc = 0.063). The rs352139 GG genotype showed a significantly reduced relative risk for HCMV infection (Pc = 0.006). In contrast, the -1237T/C SNP was not related to viral infection. We found no evidence for linkage disequilibrium with the four examined TLR9 SNPs. The findings suggest that the TLR9 -1486T/C and 2848C/T polymorphisms could be a genetic risk factor for the development of HCMV disease.

Emergency peripartum hysterectomy - a challenge or an obstetrical defeat?

OBJECTIVES: Peripartum hysterectomy remains an obstetric nightmare. Most obstetricians consider it a defeat. The aim of our study was to assess the prevalence, indications, procedures and complications of emergency peripartum hysterectomy (EPH) in the 2nd Department of Obstetrics & Gynecology, Medical University of Warsaw during a 7 year period (2007-2013). METHODS: A retrospective evaluation of 21,144 deliveries was performed. We analyzed all cases of EPH, including the maternal characteristics, obstetrical history, course of pregnancy and delivery, type of surgery and complications. RESULTS: Nineteen peripartum hysterectomies were performed between January 1, 2007 and October 30, 2013 (0.9/1000), including 16 EPH (0.76/1000). The rate of EPH was between 0.66 and 1.0 per 1000 deliveries. The majority of the patients were multiparous (79.0%), and EPH was performed after at least one cesarean section (75.0%). Fifteen women had a singleton pregnancy and one woman had a triplet pregnancy. The mean gestational age was 34.2 weeks. The delivery mode was cesarean section in 93.8% of the cases. The most common reason for peripartum hemorrhage and the indication for EPH was abnormal placentation (75.0%). All patients underwent a total hysterectomy, including 43.8% during the same operation and 50.0% during a reoperation. There was no maternal death. The serious maternal complication rates were relatively low in our study and included one case of cardiac arrest that required cardiopulmonary resuscitation and one case of sepsis with pulmonary embolism. CONCLUSIONS: EPH is typically performed as a result of massive hemorrhage associated with abnormal placentation, and it should be treated as a challenging, life-saving procedure.

Evaluation of the amoxicillin concentrations in amniotic fluid, placenta, umbilical cord blood and maternal serum two hours after intravenous administration.

OBJECTIVES: The aims of this study were to evaluate amoxicillin concentrations in amniotic fluid, placenta, umbilical cord blood and maternal blood two hours after intravenous administration to assess obstetric and non-obstetric factors that could have influences on the penetration of the antibiotic into the examined tissues and to analyze the sensitivity to amoxicillin of the most common pathogens isolated from the genital tract. METHODS: A total of 35 full-term pregnant women who qualified for elective Caesarean delivery were included in the study. Amoxicillin at a dose of 1000 mg was administered prior to surgery. Amoxicillin levels were determined by diffusion microbial assay. RESULTS: The drug concentration was highest in umbilical cord blood compared with amniotic fluid, maternal blood and placenta (4.20±1.06 µg/g versus 3.96±0.79 µg/g, 3.22±0.64 µg/g and 2.81±0.64 µg/g, respectively). Obstetric and non-obstetric factors had no influence on the amoxicillin concentration. The most common bacteria isolated from the genital tracts of pregnant women (Streptococcus agalactiae, Enterococcus faecalis, Escherichia coli) were sensitive to amoxicillin. The MIC for the sensitive strain of Streptococcus agalactiae was seen in the majority of tissues of all of the patients; however, the MICs for E. faecalis and E. coli were not observed in any compartment. CONCLUSIONS: Amoxicillin proved to have good penetration into the fetal tissues and placenta after intravenous administration. The most common bacteria isolated from the genital tracts of pregnant women were sensitive to amoxicillin. Pregnancy complications were not found to have an influence on the amoxicillin concentrations in the examined tissues.

Cytological evaluation of the nasal mucosa in neonates exposed to tobacco smoke during fetal life.

OBJECTIVES: The objective of this study is to assess the cytological picture of the nasal mucosa of neonates born to mothers who are active smokers, passive smokers and non-smokers. METHODS: A prospective study was conducted in a group of 86 neonates born between 23 and 41 weeks of gestation. The assignation of neonates to one of the three aforementioned groups was based on a questionnaire concerning exposure to tobacco smoke, and on the concentration of cotinine in maternal urine. A cytological examination was performed using exfoliative cytology with a semi-quantitative evaluation of the cells present in the specimens. Hematological summation equipment was used to assess the number of neutrophils, eosinophils, columnar, goblet, basal and squamous cells out of 500 cells counted. The number of specific cells was expressed as a percentage and a cytogram was created. RESULTS: The most common type of cytogram contained neutrophils, columnar cells, and squamous cells. No significant differences were observed between the subgroups. Similarly, there was no correlation between the median of each type of cell and the cotinine concentration in the mothers' urine. CONCLUSION: Active and passive smoking during pregnancy do not influence the cytological picture of the nasal mucosa of neonates.

Continuous Glucose Monitoring in Type 1 Diabetes Pregnancy Shows that Fetal Heart Rate Correlates with Maternal Glycemia.

BACKGROUND: Much evidence has shown that pregnancies in women with preexisting diabetes are affected by an increased risk of maternal and fetal adverse outcomes, probably linked to poor glycemic control. Despite great progress in medical care, the rate of stillbirths remains much higher in diabetes patients than in the general population. Recent technological advances in the field of glucose monitoring and noninvasive fetal heart rate monitoring made it possible to observe the fetal-maternal dependencies in a continuous manner. SUBJECTS AND METHODS: Fourteen type 1 diabetes patients were involved into the study and fitted with a blinded continuous glucose monitoring (CGM) recorder. Fetal electrocardiogram data were recorded using the Monica AN24™ device (Monica Healthcare Ltd., Nottingham, United Kingdom), the recordings of which were matched with CGM data. Statistical analysis was performed using a generalized mixed-effect logistic regression to account for individual factors. RESULTS: The mean number of paired data points per patient was 254±106, representing an observation period of 21.2±8.8 h. Mean glycemia equaled 5.64±0.68 mmol/L, and mean fetal heart rate was 135±6 beats/min. Higher glycemia correlated with fetal heart rate (R=0.32; P<0.0001) and was associated with higher odds of the fetus developing small accelerations (odds ratio=1.05; 95% confidence interval, 1.00-1.10; P=0.04). CONCLUSIONS: Elevated maternal glycemia of mothers with diabetes is associated with accelerations of fetal heart rate.

TLR9 2848 GA heterozygotic status possibly predisposes fetuses and newborns to congenital infection with human cytomegalovirus.

BACKGROUND: Some single nucleotide polymorphisms (SNP), located in Toll-like receptor (TLR) genes, were reported to be associated with human cytomegalovirus (HCMV) infections. The study was aimed to assess the correlation of SNPs at TLR4 and TLR9 genes with the occurrence of congenital cytomegaly, based on available samples. METHODS: Reported case-control study included both HCMV infected and non-infected fetuses and newborns. The specimens were classified to the molecular analyses, based on serological features of the recent infection and HCMV DNAemia in body fluids. TLR SNPs were studied, using multiplex nested PCR-RFLP assay, and determined genotypes were confirmed by sequencing. Hardy-Weinberg equilibrium was assessed for the identified genotypes. The linkage disequilibrium was also estimated for TLR4 SNPs. A relationship between the status of TLR genotypes and congenital cytomegaly development was estimated, using a logistic regression model. RESULTS: Hardy Weinberg equilibrium was observed for almost all SNPs, both infected and non-infected patients, with exception of TLR4 896 A>G polymorphism in the control group (P≤0.050). TLR4 896 A>G and 1196 C>T SNPs were found in linkage disequilibrium in both study groups (P≤0.050). The CC genotype at TLR4 1196 SNP and the GA variant at TLR9 2848 G>A SNP were significantly associated with HCMV infection (P≤0.050). The risk of congenital cytomegaly was higher in heterozygotes at TLR9 SNP than in the carriers of other genotypic variants at the reported locus (OR 4.81; P≤0.050). The GC haplotype at TLR4 SNPs and GCA variants at TLR4 and TLR9 SNPs were significantly associated with HCMV infection (P≤0.0001). The ACA variants were more frequent among fetuses and neonates with symptomatic, rather than asymptomatic cytomegaly (P≤0.0001). CONCLUSIONS: TLR4 and TLR9 polymorphisms may contribute to the development of congenital infection with HCMV in fetuses and neonates. The TLR9 2848 GA heterozygotic status possibly predisposes to HCMV infection, increasing the risk of congenital cytomegaly development.

Prenatal sonographic diagnosis of premature constriction of the fetal ductus arteriosus after maternal self-medication with benzydamine hydrochloride: report of 3 cases and review of the literature.

We report 3 cases of prenatal diagnosis of premature constriction of the ductus arteriosus after maternal benzydamine hydrochloride therapy (3-mg lozenges) in third-trimester pregnancies. In each case, fetal echocardiography revealed a dilated, hypocontractile right ventricle with severe tricuspid regurgitation and constriction of the ductus arteriosus. Although the effect of indomethacin and other nonsteroidal anti-inflammatory drugs on prenatal ductal constriction is well known, readily available over-the-counter nonsteroidal anti-inflammatory drugs such as benzydamine can have an equally deleterious effect and are best avoided in the third trimester of pregnancy.

DNA repair mechanisms and human cytomegalovirus (HCMV) infection.

Herpesvirus infections, such as those induced by human cytomegalovirus (HCMV), induce specific DNA damages. DNA damages can lead to cell mutation, death, apoptosis and immune system activation. Various types of DNA damage are repaired through multiple repair pathways, such as base excision, nucleotide excision, homologous recombination and nonhomologous end joining. Changes in the activity of DNA repair proteins during viral infection can cause disturbances in the DNA repair system and change its mechanisms. This report reviews results from studies, assaying a DNA repair system in HCMV infection.

Pregnancy and Delivery in Women with Uterine Malformations.

BACKGROUND: Uterine defects are the most common malformations of the female reproductive system. They can lead to many obstetric complications, e.g. preterm delivery, intrauterine growth restriction, oligohydramnios and operational delivery. OBJECTIVES: Our aim was to analyze the impact of different types of uterine defects on pregnancy outcomes. MATERIAL AND METHODS: The study involved 94 pregnant women with different types of uterine defects hospitalized at the Department of Fetal--Maternal Medicine and Gynecology, RIPMMH in Lódz, between 1994 and 2012. The patients were divided into 5 groups on the basis of diagnosed defects: arcuate (n=6), bicornuate (n=50), duplex (n=29), septate (n=5) and unicornuate uterus (n=4). In order to avoid correlated data in statistical analysis, our research did not consider the total number of pregnancies and births but the number of patients. The first pregnancy of each patient, if completed after 22-week gestation, was studied and analyzed. RESULTS: Preterm delivery was the most common complication in pregnancy (55 women, 58.5%). The caesarean section was performed in 73 (78%) women. IUGR was diagnosed in 16% of cases. Placental abruption occurred in 13 (14%) and cervical insufficiency in 10 cases (11%), respectively. Prenatal diagnostic showed abnormalities in 12 fetuses (13%). The Apgar score from 0 to 4 points was assigned to 9 newborns (9.6%), 5-7 to 20 children (21.3%) and 8-10 points to 75 cases (69.1%). Normal birth weight (>2500 g) was determined in 51 newborns (54.3%). CONCLUSIONS: Women with uterine defects are subject to an increased risk of complications in pregnancy and delivery, including premature births, low birth weights, births by cesarean section.