Contribution of IL6 -174 G>C and IL1B +3954 C>T polymorphisms to congenital infection with Toxoplasma gondii.

The purpose of this investigation was the determination of the distribution of genotypes and alleles, residing within interleukin 6 (IL6) and interleukin 1 (IL1) polymorphisms, among fetuses and neonates, congenitally infected with Toxoplasma gondii, and among uninfected control cases. The study included 22 fetuses and newborns infected with T. gondii and 49 control cases. Screening for IgG and IgM antibodies against the parasite and IgG avidity was performed by enzyme-linked fluorescent assay (ELFA) tests. Quantitation of T. gondii DNA in amniotic fluids was assayed by the real-time Q PCR technique for the parasitic B1 gene. Genotypes at IL6 and IL1 single nucleotide polymorphisms (SNPs) were determined by a self-designed, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Representative genotypes at the studied loci were confirmed by sequencing. All the genotypes were estimated for Hardy-Weinberg equilibrium and IL1 genotypes were tested for linkage disequilibrium. Genotypes and haplotypes at the studied SNPs were investigated for their possible association with the occurrence of congenital T. gondii infection, using a logistic regression model. GC heterozygotes at the IL6 -174 G>C SNP were significantly associated with toxoplasmosis and increased the risk of T. gondii infection [odds ratio (OR) 4.24, 95 % confidence interval (CI) 1.24-14.50 in the codominant model, p ≤ 0.050]. In case of IL1 SNPs, similar prevalence rates were observed between T. gondii-infected and -uninfected offspring. Regarding allelic variability, the C alleles at both IL6 and IL1B SNPs were significantly more frequent in the infected than in the uninfected cases (p ≤ 0.050). It is concluded that IL6 -174 G>C and IL1B +3954 C>T SNPs might be involved in the development of congenital T. gondii infection.

Possible role of TLR4 and TLR9 SNPs in protection against congenital toxoplasmosis.

The purpose of this investigation was the determination of the distribution of genotypes at single nucleotide polymorphisms (SNPs) of the toll-like receptor 4 (TLR4) and the toll-like receptor 9 (TLR9) in fetuses and newborns congenitally infected with Toxoplasma gondii and the identification of genetic changes predisposing to infection development. The study involved 20 fetuses and newborns with congenital toxoplasmosis and 50 uninfected controls. The levels of IgG and IgM antibodies against T. gondii, as well as IgG avidity, were estimated by enzyme-linked fluorescent assay (ELFA) tests. T. gondii DNA loads in amniotic fluids were assayed by the real-time (RT) quantitative polymerase chain reaction (Q PCR) technique for parasitic B1 gene. TLR4 and TLR9 SNPs were identified using a self-designed multiplex nested PCR-restriction fragment length polymorphism (RFLP) assay. Randomly selected genotypes at SNPs were confirmed by sequencing. All the genotypes were tested for Hardy-Weinberg equilibrium and TLR4 genotypes were analyzed for linkage disequilibrium. A correlation was studied between the genotypes or haplotypes and the development of congenital toxoplasmosis using a logistic regression model. Single SNP analysis showed no statistically significant differences in the distribution of distinct genotypes at the analyzed TLR4 and TLR9 SNPs between T. gondii-infected fetuses and newborns and the controls. Taking into account the prevalence of alleles residing within polymorphic sites, similar prevalence rates were observed in both of the studied groups. The multiple SNP analysis indicated GTG variants at the TLR4 and TLR9 SNPs to be significantly less frequent in offspring with congenital toxoplasmosis than in uninfected offspring (p ≤ 0.0001). TLR4 and TLR9 SNPs seem to be involved in protection against congenital toxoplasmosis.

Mixed infections with distinct cytomegalovirus glycoprotein B genotypes in Polish pregnant women, fetuses, and newborns.

The purpose of this investigation was to describe a distribution of cytomegalovirus (CMV) single and multiple genotypes among infected pregnant women, their fetuses, and newborns coming from Central Poland, as well as congenital cytomegaly outcome. The study involved 278 CMV-seropositive pregnant women, of whom 192 were tested for viral DNAemia. Human cytomegalovirus (HCMV) genotyping was performed for 18 of 34 pregnant women carrying the viral DNA and for 12 of their 15 offspring with confirmed HCMV infections. Anti-HCMV antibodies levels were assessed by chemiluminescence immunoassay (CLIA) and enzyme-linked fluorescence assay (ELFA) tests. Viral DNA loads and genotypes were determined by real-time polymerase chain reaction (PCR) assays for the UL55 gene. In the pregnant women, we identified HCMV gB1, gB2, gB3, and gB4 genotypes. Single gB2, gB3, or gB4 genotypes were observed in 14 (77.8 %) women, while multiple gB1-gB2 or gB2-gB3 genotypes were observed in four (22.2 %). Maternal HCMV genotypes determined the genotypes identified in their fetuses and newborns (p ≤ 0.050). Half of them were infected with single HCMV gB1, gB2, or gB3 genotypes and the other half with multiple gB1-gB2 or gB2-gB3 genotypes. Single and multiple genotypes were observed in both asymptomatic and symptomatic congenital cytomegaly, although no gB3 genotype was identified among asymptomatic cases. In Central Poland, infections with single and multiple HCMV strains occur in pregnant women, as well as in their fetuses and neonates, with both asymptomatic and symptomatic infections. HCMV infections identified in mothers seem to be associated with the viral genotypes in their children.

Impact of socioeconomic risk factors on the seroprevalence of cytomegalovirus infections in a cohort of pregnant Polish women between 2010 and 2011.

The purpose of this investigation was to perform an evaluation of the prevalence and socioeconomic risk factors for human cytomegalovirus (HCMV) infections in a cohort of Polish pregnant women between 2010 and 2011. HCMV-specific IgG and IgM antibody levels were assayed with enzyme-linked immunosorbent assay (ELISA) tests in serum samples collected from 1,250 pregnant women attending outpatient obstetric clinics and hospitalized at two hospitals in Lodz. The seroprevalence of anti-HCMV IgG and IgM antibodies was 62.4 and 2.2 %, respectively, and differed significantly between age-stratified groups (p ≤ 0.05). The highest IgG prevalence was observed in women above 36 years of age (76.2 %) and IgM in adolescent women aged 16-20 years (6.0 %). Of the various socioeconomic factors, age above 36 years, basic and professional education, and offspring were significantly associated with HCMV IgG prevalence rates (PRs; 1.89, 1.80, and 1.56, respectively). Financial status, occupational risk related to contact with children, and transfusions were not related to the prevalence of IgG antibodies. The IgM prevalence was not associated with any of the analyzed risk factors. A slightly higher prevalence was observed in women who were transfused in the past, but the relationship was not significant. The current data have revealed a decrease in HCMV IgG seroprevalence in our region during recent years (62.4 vs. 76.7 %). Basic and professional education, as well as bringing up offspring, were determined as significant risk factors for HCMV infections in Polish pregnant women [risk ratio (RR) 1.20 and 1.17, respectively], suggesting that the primary and secondary prophylaxis of cytomegaly is necessary during pregnancy, even if screening is not mandatory.

Decrease in 3-nitrotyrosine in the amniotic fluid of women with cytomegalovirus infection.

The aim of this study was to assess oxidative stress in pregnant women infected with cytomegalovirus on the basis of 3-nitrotyrosine levels in amniotic fluid (AF). The 3-nitrotyrosine (3-NT) level in AF was measured using the competitive ELISA method. The study groups were as follows: group I consisted of women with IgM and/or IgA; group II were women with only IgG anti-CMV antibodies and group III were seronegative women, used as the control group. Age, gestational age and socioeconomic status were also assessed. The average level of 3-nitrotyrosine in group II and the control group was similar: 53.14 nM 3-NT and 49.78 nM 3-NT, respectively. However, in group I, the lowest level 3-NT in AF was observed - 39.17 nM 3-NT and statistical analysis showed significant differences in levels of 3-NT between group I and the control group (p < 0.01). We conclude that significantly lower levels of 3-nitrotyrosine in pregnant women with CMV infection may indicate an increase in the antioxidant defence mechanisms in these patients.

Age-associated prevalence of Toxoplasma gondii in 8281 pregnant women in Poland between 2004 and 2012.

This study aimed to describe Toxoplasma gondii prevalence in Polish pregnant women and the incidence rates of congenital infections in their neonates observed between 2004 and 2012. Serological tests for T. gondii-specific IgG and IgM antibodies were performed on serum samples of 8281 pregnant women treated at the Polish Mother's Memorial Hospital Research Institute in Lodz. The yearly seroconversion rate for T. gondii IgG antibodies was estimated using a mathematical model to determine the dependency between age and prevalence. Mean prevalence of IgG antibodies between 2004 and 2012 in pregnant women was 40·6% [95% confidence interval (CI) 39·6-41·7] and increased with age with a yearly seroconversion rate of 0·8% (95% CI 0·6-1·0, P<0·001). Assuming a T. gondii materno-fetal transmission rate of 30% gave an estimate of 1·80/1000 neonates as congenitally infected. The increased mean age (28·7 vs 26·7 years, P<0·001) of pregnant women was probably the most important factor in abolishing the effect of falling prevalence rates.

Do the placental barrier, parasite genotype and Toll-like receptor polymorphisms contribute to the course of primary infection with various Toxoplasma gondii genotypes in pregnant women?

Toxoplasma gondii has a highly clonal genetic structure classified into three major genetic types, I, II, and III, plus additional recombinant and atypical strains. In humans, type I and atypical strains usually associate with severe toxoplasmosis. Type II strains, predominantly identified in European countries and the United States, correlate with a differential course of toxoplasmosis. During pregnancy, the important protective role of the placenta against maternal-fetal T. gondii transmission has been reported. T. gondii preferentially colonizes extravillous trophoblasts as compared to syncytiotrophoblasts. The latter compartment was suggested to act as the real barrier to the fetal dissemination of T. gondii. Alterations in immune response to particular T. gondii strains were observed. Higher transcription levels of IP-10, IL-1ß, IL-6, IL-10, IL-12 cytokines, and NF-κB translocation to the nucleus were more often documented for type II strains than type I strains. Since the induction of IL-12 during type II infection was Myd88-dependent, the involvement of Toll-like receptors (TLRs) in the immunity against these strains was suggested. Differential expression of TLRs depends on placental cell types and gestational age. The expression of TLR2 and TLR4 in the first trimester of pregnancy was reported only for villous cytotrophoblasts and extravillous trophoblasts, but not for syncytiotrophoblasts. The involvement of single-nucleotide polymorphisms (SNPs) in the TLR genes in infectious pathogenicity, including toxoplasmic retinochoroiditis, points at a possible involvement of TLR alterations in immunity against T. gondii. We conclude that studies on TLR contributions in the maternal-fetal transmission of particular parasite strains and congenital toxoplasmosis are warranted.

SNPs in toll-like receptor (TLR) genes as new genetic alterations associated with congenital toxoplasmosis?

Nearly 40 % of pregnant women are infected with Toxoplasma gondii. Primary infections in pregnant women result, in approximately 30-50 % of patients, in transmission of T. gondii through the placenta to the fetus and then in congenital infections with severe, sometimes fatal course. Studies still do not provide sufficient data on the genetic bases of the immunity in fetuses, newborns, and infants with congenital toxoplasmosis. Previous research showed the contribution of toll-like receptors (TLRs) to non-specific immunity against T. gondii invasion, observed in T. gondii-infected animals, especially mice. So far, the activity of TLRs in defense against T. gondii infections was observed particularly for TLR2, TLR4, and TLR9 molecules. Differential TLR activity associates with both cell types, including a variety of placental cells and stage of pregnancy. Several single-nucleotide polymorphisms (SNPs) residing in three genes encoding these receptors were reported as significant genetic modifications of TLRs associated with different pregnancy disorders. Despite those data, genetic alterations of TLRs which have contributed to innate immune response against T. gondii infections are still not precisely described. In this article, we present reasons for the research of the plausible role of SNPs residing in TLR2, TLR4, and TLR9 genes in congenital toxoplasmosis development.

Cytotoxic potential of vasoconstrictor experimental gingival retraction agents: in vitro study on primary human gingival fibroblasts.

The aim of this in vitro study was to evaluate the cytotoxic effects of the vasoconstrictor experimental gingival retraction agents (VEGRAs) in a dynamic setting. The strong cytotoxic effects of the astringent-based conventional gingival retraction agents (ACGRAs) on human gingival fibroblasts (HGFs) in vitro was our motivation to evaluate the biocompatibility of the vasoconstrictor-based experimental gingival retraction agents (VEGRAs) for the selected minimally invasive chemical agent. These agents were used to create three self-made retraction gels. Human gingival fibroblasts (HGFs) were treated with two groups of retraction agents: 1) three α- and ß-adrenergic agents (VEGRA-αß-s) based on 0.1%, 0.01% and 0.05% HCl-epinephrine, and 2) seven α-adrenergic agents (VEGRA-α-s), including two commercially available 0.05% HCl-tetrahydrozoline solutions, one 0.05% HCl-oxymetazoline solution, 10% HCl-phenylephrine solution, and three new self-made experimental 0.05% HCl-tetrahydro zoline-based gels. The methyl thiazolyl tetrazolium (MTT) colorimetric assay was performed to determine the oxidoreductive mitochondrial function after 3, 5, 10 min and 24 h of incubation. The cytotoxic effect, measured by cell viability lower than the 50% threshold, was not observed at any time period, even 24 h after application of 0.05% HCl-tetrahydrozolinebased self-manufactured retraction gels. High cell viability values of human gingival fibroblasts after the treatment with the three self-made 0.05% HCl-tetrahydrozoline- based gels may serve as a basis for further studies aimed at selecting the best retraction agents biocompatible with gingival margin tissues.

Distribution of UL144, US28 and UL55 genotypes in Polish newborns with congenital cytomegalovirus infections.

Human cytomegalovirus (HCMV) is the most common congenital infection. HCMV strains display genetic variability in different regions. Distribution of HCMV genotypes in the population of congenitally infected newborns from Central Poland and viral load in newborns' blood is described and discussed. HCMV isolates were analysed by sequencing at three sites on the genome: the UL144 tumour necrosis factor-alpha (TNFα)-like receptor gene, the US28 beta-chemokine receptor gene and the UL55 envelope glycoprotein B (gB) gene. The newborns' blood was examined for HCMV DNA with a nested (UL144, UL55) or heminested (US28) polymerase chain reaction, and the genotypes were determined by sequence analysis. HCMV DNA was detectable in 25 out of 55 examined newborns born by HCMV-infected mothers (45.5%). The blood viral load in mother-infant pairs was determined. Most of the newborns had identical virus genotype, gB2 (96%), UL144 B1 (88%) and US28 A2 (84%). These genotypes were detected in all newborns with asymptomatic congenital infection. The occurrence of UL144 B1 or US28 A2 genotypes in the babies examined was significant in comparison to other genotypes (p=0.0002 and p=0.040 respectively). There was no association between specific gB subtypes in all patients groups (p=0.463). There was no correlation between HCMV genotypes and the outcome.

Dynamic oxidoreductive potential of astringent retraction agents.

The aim of this study was to evaluate the dynamics of the cytotoxicity of gingival margin retraction astringents based on aluminium chloride, aluminium sulphate, and ferric sulphate (solutions and gels) in human fibroblasts isolated from gingiva. The cytocompatibility of ten astringent-based chemical retraction agents: Gingiva Liquid, Alustin, Racestypine, Orbat sensitive, Astringedent®, Alustat, Hemostat, Racécord, Gel cord and ViscoStat®, in dilutions of 1 : 10 and 1 : 20, with human gingival fibroblasts was investigated. The MTT assay was performed to determine oxidoreductive mitochondrial function after 3, 5, 10 min and 24 h of incubation. Cell viability was determined according to the chemical group, concentration, exposure time, and the clinical form of the gingival retraction agents. Ferric sulphate- based agents were the most cytotoxic, followed by aluminium chloride and aluminium sulphate. The form of the astrigents influenced cell viability. The evaluated astringents may have cytotoxic potential for gingival margin tissues under clinical conditions.

The AAPC case, with an early onset of colorectal cancer.

INTRODUCTION: Attenuated adenomatous polyposis coli (AAPC) is a variant of the familial adenomatous polyposis (FAP) characterized by the occurrence of sparse polyps in the colon, stomach, and duodenum with a late onset of colorectal cancer. The AAPC syndrome is associated with mutations at the 5' region of the APC gene. Until recently, the fragment encompassing codons 157 and 170 was considered as boundary for the described cases of AAPC and FAP syndromes. MATERIALS AND METHODS: This study describes a case of the AAPC syndrome caused by a CCTT deletion at codon 173, with polyps diagnosed at the age of 17. The father and grandfather of the proband died of colorectal cancer (CRC), which developed from untreated polyps, at the age 35 and 40, respectively. RESULTS AND DISCUSSIONS: In the case of the proband's father, the untreated polyps led to death after 12 years. The proband revealed a low number of polyps and an extra colon feature characteristic of AAPC, but the polyps onset and the death of CRC of two family members, who refused colectomy, was very early and characteristic for FAP. An atypical course of AAPC must be taken into consideration both in genetic counseling and in qualifying the patients with AAPC for the surgical treatment.

Prevalence and estimated incidence of Toxoplasma infection among pregnant women in Poland: a decreasing trend in the younger population.

This study investigated the prevalence of specific Toxoplasma gondii IgG in pregnancy, the incidence of congenital toxoplasmosis and the prevalence trend of T. gondii infection among pregnant Polish women between 1998 and 2003. The study population comprised 4916 women who were admitted to the Polish Mother's Memorial Hospital Research Institute in Lódz. Their sera were tested for specific IgG and IgM antibodies to T. gondii, and the incidence of T. gondii infection was calculated from the increase in prevalence rates of IgG antibodies in various age groups. Specific IgG antibody was found in 41.3% (95% CI 39.9-42.7) of pregnant women, and the prevalence of IgG increased with age. The linear trend was significant (p <0.001), with an annual seroconversion rate of 0.7% (95% CI 0.004-0.010). The risk of primary infection was estimated to be 0.5% for 9 months, i.e., an incidence of 5/1000 pregnancies. Assuming a 30% maternofetal transmission rate, 1.5/1000 neonates were infected in utero. Seroprevalence during the 6-year study period decreased from 45.4% in 1998 to 39.4% in 2003, with a yearly decline in prevalence of 1.0% (p 0.02). The most important contributory factor to this decline was the group of women aged 19-29 years, among whom seroprevalence decreased significantly (p 0.007). Specific IgM was found in 244 (4.9%) women.

Low frequency of the CHEK2*1100delC mutation among breast cancer probands from three regions of Poland.

The 1100delC germline mutation of the CHEK2 gene appears to contribute significantly to the overall breast cancer incidence in some West and North European countries, but seems to be much less frequent among breast cancer patients from other regions of Europe. In the present study we found, respectively, 3/487, 1/296 and 0/279 carriers of this mutation among breast cancer patients from the East-Central, South-East and West-Central regions of Poland. Two carriers of the 1100delC mutation were found among 120 patients with bilateral breast cancer, but only one had a previous family incidence of breast cancer. We found no carriers among 182 patients with unilateral breast cancer with family history of this tumor and among 64 patients with breast cancer and a second primary tumor at an other site. We conclude that the 1100delC mutation of the CHEK2 gene contributes little to the overall breast cancer burden in Poland, including familial cases of this malignancy. Further studies are still needed to evaluate the contribution of this mutation to the development of bilateral breast tumors.

[Anti - T. gondii antibody concentration in sera of pregnant women in the sample of Lódz population]. / Stezenie przeciwcial anty -T. gondii w surowicy kobiet ciezarnych w próbie populacji regionu lódzkiego w roku 1998.

The aims of the study were: to estimate the prevalence of Toxoplasma gondii in pregnant women in the Lódz region; to assess the rate of incidence of suspected recent infection based on the results of the standard test for specific anty -T. gondii IgG and IgM antibodies. The study covered 1920 pregnant women served in 1998 by Toxoplasma Reference Laboratory in Lódz. T. gondii specific IgG and IgM were quantitated by an enzym-linked immunoabsorbent assay (ELISA, Organon). We have demonstrated high prevalence of T. gondii among pregnant women in the Lódz region. In the analysed sample of Polish population the T. gondii-specific IgG with negative IgM occurred in 43.4% (n = 837). In 1.42% of pregnant women IgG and IgM specific antibodies were detected.

[Multifocal fungal infections in pregnant women]. / Zarazenia wieloogniskowe grzybami u kobiet ciezarnych.

The aim of the study was to estimate the prevalence of fungi in monofocal and multifocal infections in pregnant women. 251 pregnant women were examined for presence of fungi in vagina, oral cavity and rectum. Fungal strains were recovered in 259 out of 753 samples (49.2% of all pregnant women). Presence of fungi in vagina was associated with the invasion of oral cavity and/or rectum in 98% of all women. Monofocal vulvovaginal mycosis was found only in 1.9% of the cases. Bifocal infections depended on the ontocenoses: vagina - oral cavity (32.1%); vagina--rectum (7.5%); oral cavity--rectum (13.2%). Trifocal infections (vagina - oral cavity--rectum) occurred in 58.5% of all examined women.

Search for the pathogenic fungi in amniotic fluid.

Intra-amniotic infection with Candida is rare but considers an etiological factor for preterm rupture of membranes (PROM). The aim of the study was the investigation of the mycotic invasion of the amniotic cavity in pregnant women with PROM and with intact fetal memranes. 170 women were included in two groups: I (n = 150)--women with intact membranes; II (n = 20)--women with PROM. Samples of amniotic fluid were cultured on Sabouraud's medium. Fungal strains were isolated also from vagina, oral cavity and rectum of women with PROM. Intraamniotic infections were absent both in women with PROM and with intact membranes. Candida strains were isolated from vagina, oral cavity, and rectum in the group of women with PROM. Coexisting of multifocal Candida infections in pregnant women with PROM indicates that in those cases mycologic diagnosics should be recomended.

[Twin pregnancy complicated by intrauterine death of one twin]. / Ciaza blizniacza powiklana obumarciem wewnatrzmacicznym jednego plodu.

In twin pregnancies single intrauterine death of one fetus is associated with significant morbidity and mortality of the surviving infant. The aims of our retrospective study were to review conditions of twin pregnancies complicated with SIUD in Polish Mother's Memorial Hospital in Lódz between 1989-1999 and to assess the fetal outcome when conservative management had undergone. In this study we reviewed 30 twin gestations involving the intrauterine death of one fetus. The incidence of preterm delivery among pregnancies with fetal death was 83.3%; Caesarean section was the method of delivery in 53.3% cases. Monochorionic placentation was found in 60%. Conservative management until there is no risk for the fetus is apt.