HIF-1α regulated pathomechanism of low birth weight through angiogenesis factors in placental Plasmodium vivax infection.

Background: Malaria in pregnancy leads to placental malaria. The primary pathogenesis of the complex fetal implications in placental malaria is tissue hypoxia due to sequestrations of Plasmodium falciparum-infected erythrocytes in the placenta. However, the pathomechanism of placental Plasmodium vivax infection has not been thoroughly investigated. Hypoxia-inducible factor-1α (HIF-1α) is a key transcriptional mediator of the response to hypoxic conditions, which interacts with the change and imbalances of many chemical mediators, including angiogenic factors, leading to fetal growth abnormality. Methods: This study was conducted cross-sectionally in Maumere, Sikka Regency, East Nusa Tenggara Province, previously known as one of the malaria endemic areas with a high incidence of low birth weight (LBW) cases. This study collected peripheral and umbilical blood samples and placental tissues from mothers who delivered their babies with LBW at the TC Hiller Regional Hospital. All of the blood samples were examined for parasites by microscopic and PCR techniques, while the plasma levels of VEGF, PlGF, VEGFR-1, VEGFR-2, and HIF-1α were determined using ELISA. The sequestration of infected erythrocytes and hemozoin was determined from placental histological slides, and the expression of placenta angiogenic factors was observed using the immunofluorescent technique. Results: In this study, 33 cases had complete data to be analyzed. Of them, 19 samples were diagnosed as vivax malaria and none of falciparum malaria. There were significant differences in Δ 10th percentile growth curve of baby's body weights and also all angiogenic factors in placental tissues {VEGF, PlGF, and VEGFR-1, VEGFR-2, and HIF-1α} between those infected and not infected cases (p<0.05), but not for VEGF and VEGFR-2 in the plasma. Conclusion: This study indicated that Plasmodium vivax sequestration may promote LBW through alterations and imbalances in angiogenic factors led by HIF-1α.

Dynamics of PfEMP1 Antibody Profile From Birth to 12 Months of Age in Beninese Infants.

BACKGROUND: Plasmodium falciparum-infected erythrocytes bind to specific endothelial cell receptors via members of the PfEMP1 family exported onto the erythrocyte surface. These interactions are mediated by different types of cysteine-rich interdomain region (CIDR) domains found in the N-terminal region of all PfEMP1. CIDRα1 domains bind endothelial protein C receptor (EPCR), CIDRα2-6 domains bind CD36, whereas the receptor specificity of CIDRß/γ/δ domains is unknown. METHODS: In this study, we investigated the level of immunoglobulin (Ig)G targeting the different types of PfEMP1 CIDR during the first year of life. We used plasma collected longitudinally from children of pregnant women who had been followed closely through pregnancy. RESULTS: Antibodies to CIDRα1 domains were more frequent in cord blood compared with antibodies to CIDRα2-6 domains. Higher IgG levels to EPCR-binding CIDRα1 variants positively correlated with the timing of first infections. Antibodies to all PfEMP1 types declined at similar rates to the point of disappearance over the first 6 months of life. At 12 months, children had acquired antibody to all types of CIDR domains, mostly in children with documented P falciparum infections. CONCLUSIONS: These observations agree with the notion that the timing and phenotype of first P falciparum infections in life are influenced by the immune status of the mother.

Blood cytokine, chemokine and growth factor profiling in a cohort of pregnant women from tropical countries.

The immune status of women changes during and after pregnancy, differs between blood compartments at delivery and is affected by environmental factors particularly in tropical areas endemic for multiple infections. We quantified the plasma concentration of a set of thirty-one TH1, TH2, TH17 and regulatory cytokines, pro-inflammatory and anti-inflammatory cytokines and chemokines, and growth factors (altogether biomarkers), in a cohort of 540 pregnant women from five malaria-endemic tropical countries. Samples were collected at recruitment (first antenatal visit), delivery (periphery, cord and placenta) and postpartum, allowing a longitudinal analysis. We found the lowest concentration of biomarkers at recruitment and the highest at postpartum, with few exceptions. Among them, IL-6, HGF and TGF-ß had the highest levels at delivery, and even higher concentrations in the placenta compared to peripheral blood. Placental concentrations were generally higher than peripheral, except for eotaxin that was lower. We also compared plasma biomarker concentrations between the tropical cohort and a control group from Spain at delivery, presenting overall higher biomarker levels the tropical cohort, particularly pro-inflammatory cytokines and growth factors. Only IL-6 presented lower levels in the tropical group. Moreover, a principal component analysis of biomarker concentrations at delivery showed that women from Spain grouped more homogenously, and that IL-6 and IL-8 clustered together in the tropical cohort but not in the Spanish one. Plasma cytokine concentrations correlated with Plasmodium antibody levels at postpartum but not during pregnancy. This basal profiling of immune mediators over gestation and in different compartments at delivery is important to subsequently understand response to infections and clinical outcomes in mothers and infants in tropical areas.

Update of pre- and postnatal iron supplementation in malaria endemic settings.

This review focuses on pre- and post-natal iron supplementation in malaria endemic settings. Although iron supplementation can reduce iron deficiency, malaria infection may counteract this effect by the increase of hepcidin, and iron supplementation may further worsen malaria infection by providing additional iron for the parasites. However, most iron supplementation intervention studies in pregnant women with malaria have not shown a negative impact, although malaria treatment with iron supplementation may be beneficial in terms of improving birth outcomes. In infants and young children in malaria endemic settings, the adverse effects of iron supplementation has been well documented and malaria prevention and treatment with iron supplementation is recommended. Besides fostering the growth of malaria parasites, iron may also promote potential pathogens in the gut and cause an inflammatory response in young children. Overall, iron supplementation is beneficial for treating iron deficiency, but needs to be considered in the context of malaria prevention and treatment in pregnant women, infants and young children for safety and effectiveness.

Investigation of the Applicability of a Rapid Diagnosis Test in the Diagnosis of Cystic Echinococcosis

OBJECTIVE: Echinococcus granulosus, the etiological agent of cystic echinococcosis (CE) in humans and livestock, is a widely distributed zoonotic pathogen tapeworm. The infection is transmitted to humans by the ingestion of E. granulosus eggs released in the feces of definitive hosts such as dogs. The larval stage of the parasite develops a slowly enlarging cyst in the visceral organs, particularly in the liver and/or lung. The aim of the present study was to evaluate the diagnostic value of an immunochromatographic test (ICT) for CE. METHODS: A total of 50 sera from surgically and/or pathologically confirmed patients with CE were included in the study as the study group; the control group comprised patients who tested negative for enzyme-linked immunosorbent assay (ELISA). Sera were selected from the collection at Adnan Menderes University, Faculty of Medicine, Parasitology Laboratory, by simple random sampling. The collection included sera obtained between 2010 and 2014; antibody titers of each serum sample were determined using in-house ELISA, before storage at -20°C. The presence of E. granulosus antibody in the sera was determined using a commercially available ICT (VIRAPID® HYDATIDOSIS) kit method. RESULTS: In the study group (E. granulosus-confirmed cases), two (4%) of the 50 sera were negative and 48 (96%) were positive with ICT. In the control group (ELISA-negative), all were negative with ICT. CONCLUSION: The rapid diagnostic test has been evaluated as a practical, easy-to-use method for detecting CE, and it can be used as a screening test in routine diagnosis and research.

Serum Levels of Inflammatory Cytokines in Helminth Infested Pregnant Women and Cord Blood of their Babies in Relation to Pregnancy Outcome.

Pregnancy places a very high demand on physical, physiological and immunological responses of females,especially when aggravated by parasitic infestation. There is strong evidence that maternal infestations with helminth haveprofound effects on immunity to helminths and other pathogens. This case-control study involved 245 pregnant women aged18-40 years (>30 weeks of gestation) recruited from three secondary level hospitals in Ibadan, Nigeria. Morning stoolsamples collected from pregnant women were examine for intestinal helminths using formol-ether concentration method. Atotal of 38 participants comprising 17 Helminth Positive (HP) and 21 Helminth Negative (HN) pregnant women werepurposely selected for the study. Sera from these women (38) and their babies' cord (38) were analysed for immune factors[interleukins 6 and 8 (IL-6, IL-8), tumor necrosis factor-alpha (TNF-α) and immunoglobulin E (IgE)] were analyzed usingELISA. Anthropometric indices [weight and height in mothers and babies and Chest Circumference (CC) in babies] weremeasured using standard methods. Data were subjected to descriptive statistics and analysed using Student t-test and Pearsoncorrelation at α0.05. Only Ascaris lumbricoides was found in the 17 (6.9%) infested pregnant women. The mean levels of IL-6 (57.8 ± 32.8 vs 52.8±39.6 pg/mL), IL-8 (24.3±3.5 vs 22.0±7.1 pg/mL) and IgE (333.3±96.6 vs 242.3±96.8 IU/mL) weresimilar in HP when compared with HN. In cord sera, IL-8 level was significantly higher in babies of HP (23.7±3.9 pg/mL)compared with babies of HN (20.1±5.9 pg/mL). The levels of IL-6, TNF-α and nutritional indices in HP had significantpositive correlation with corresponding levels in babies of HP mothers. Only CC was significantly lowered in babies of HPcompared with HN mothers. Other anthropometric indices were not significantly different. Therefore, this present studysuggests that helminth infestation may lead to strong Th2 immune responses as is reflected by the cytokine levels of mothersand babies as well as anthropometric measurements of babies of infested mothers. The outcomes of this study provide basisto deworm pregnant women during pregnancy.

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.

Role of IL-1ß, IL-6 and TNF-α cytokines and TNF-α promoter variability in Plasmodium vivax infection during pregnancy in endemic population of Jharkhand, India.

BACKGROUND: The combinatorial effects of Plasmodium infection, perturbation of inflammatory responses and the dichotomic role of TNF promoter polymorphism has potential clinical and physiological relevance during pregnancy. OBJECTIVE AND METHODS: This coordinated orchestration instigated us to investigate the circulating level of inflammatory cytokines (IL-1ß, TNF-α and IL-6) employing ELISA in a stratified group of samples and the plausible genetic association of TNF-α -308 G/A using PCR-RFLP/sequencing during Plasmodium vivax infection in pregnancy. RESULTS: We observed significantly elevated concentrations of IL-1ß were observed, followed by IL-6 and TNF-α in women with malaria (WWM) and in malaria in pregnancy (MIP). Further, elevated IL-1ß, followed by TNF-α and IL-6 were detected in the non-infected pregnancy group. The differential dynamics of inflammatory cytokine concentration during each trimester of pregnancy with and without P. vivax infection were detected. For the first time, a high level of IL-6 was observed in the first trimester of MIP and high IL-1ß in healthy pregnancies. In the second trimester, however, we observed a high level of IL-1ß in the MIP group compared to a sustained high level of IL-1ß in the healthy pregnancy group. In the third trimester, high IL-1ß was sustained in the MIP group and healthy pregnancies acquired a high TNF-α level. The genotypic distribution for the TNF-α promoter -308 G/A position was observed to be nonsignificant and mildly associated during MIP (OR = 1.4) and in WWM (OR = 1.2). Moreover, based on genotypic distribution, we observed a well-correlated and significantly elevated TNF-α concentration in the mutant homozygote genotype (AA; p = 0.001) followed by heterozygotes (GA; p = 0.0001) and ancestral genotypes (GG; p = 0.0001) in both MIP and WWM subjects. CONCLUSION: The observation of elevated IL-1ß and IL-6 in MIP and TNF-α in WWM may be regarded as a prognostic inflammatory marker of infection and pregnancy. Most particularly, the TNF-α concentration and its polymorphic variability in the promoter region may indicate genetic susceptibility and mildly influence the risk for P. vivax infection during pregnancy and in women with malaria.

Changes in some pregnancy biomarkers of Yankasa ewes experimentally infected with Trypanosoma evansi.

The study was designed to determine the effect of Trypanosoma evansi infection on some pregnancy biomarkers of Yankasa ewes (YE). Twenty pregnant YE were assigned into 3 groups (A, B and C) comprising 7 ewes each in groups A and B, while group C comprise 6 YE. Groups A and B were each inoculated with blood containing approximately 1.0 × 10(6) of T. evansi through the jugular vein on days 59 and 110 of pregnancy, representing second and third trimesters, respectively, while group C served as the uninfected control. Progesterone (P4) and pregnancy specific protein-B (PSPB) of YE in group A were significantly (p < 0.05) high at weeks 4 and 12 post infection (pi) respectively, while there was no significant (p > 0.05) difference in P4 and PSPB of YE in groups B. Estrone sulfate (E1S) significantly (p < 0.05) decrease for YE in group A at weeks 2 and 11 pi. However, it was not significantly (p > 0.05) different in group B. Cortisol concentration of YE in group A was significantly (p < 0.05) decreased at week 12 pi. Conversely, the cortisol concentration of YE in group B significantly (p < 0.05) increased at week 3 pi. There was no significant (p > 0.05) association among the pregnancy biomarkers of YE in groups A and B throughout the study, except between progesterone and cortisol in group B, which were significantly associated (r = 0.77, p < 0.05). It was therefore concluded that T. evansi infection affects pregnancy biomarkers more at mid pregnancy than at late pregnancy.

Evaluation of a recombinant rhoptry protein 2 enzyme-linked immunoassay for the diagnosis of toxoplasmosis acquired during pregnancy

The aim of this study was to evaluate an enzyme-linked immunoassay with recombinant rhoptry protein 2 (ELISA-rROP2) for its ability to detectToxoplasma gondii ROP2-specific IgG in samples from pregnant women. The study included 236 samples that were divided into groups according to serological screening profiles for toxoplasmosis: unexposed (n = 65), probable acute infection (n = 48), possible acute infection (n = 58) and exposed to the parasite (n = 65). When an indirect immunofluorescence assay forT. gondii-specific IgG was considered as a reference test, the ELISA-rROP2 had a sensitivity of 61.8%, specificity of 62.8%, predictive positive value of 76.6% and predictive negative value of 45.4% (p = 0.0002). The ELISA-rROP2 reacted with 62.5% of the samples from pregnant women with probable acute infection and 40% of the samples from pregnant women with previous exposure (p = 0.0180). Seropositivity was observed in 50/57 (87.7%) pregnant women with possible infection. The results underscored that T. gondii rROP2 is recognised by specific IgG antibodies in both the acute and chronic phases of toxoplasmosis acquired during pregnancy. However, the sensitivity of the ELISA-rROP2 was higher in the pregnant women with probable and possible acute infections and IgM reactivity.

Impact of placental Plasmodium falciparum malaria on the profile of some oxidative stress biomarkers in women living in Yaoundé, Cameroon.

BACKGROUND: Impact of the pathophysiology of Plasmodium falciparum placental malaria (PM) on the profile of some oxidative stress biomarkers and their relationship with poor pregnancy outcomes in women remain unknown. METHODS: Between 2013 and 2014, peripheral blood and placenta tissue from 120 Cameroonian women at delivery were assessed for maternal haemoglobin and, parasitaemia respectively. Parasite accumulation in the placenta was investigated histologically. The levels of oxidative stress biomarkers Malondialdehyde (MDA), Nitric Oxide (NO), Superoxide dismutase (SOD), Catalase (CAT) and Gluthatione (GSH) in the supernatant of teased placenta tissues were determined by Colorimetric enzymatic assays. RESULTS: Parasitaemia was inversely related to haemoglobin levels and birth weight (P <0.001 and 0.012, respectively). The level of lipid peroxide product (MDA) was significantly higher in the malaria infected (P = 0.0047) and anaemic (P = 0.024) women compared to their non-infected and non-anaemic counterparts, respectively. A similar trend was observed with SOD levels, though not significant. The levels of MDA also correlated positively with parasitaemia (P = 0.0024) but negatively with haemoglobin levels (P = 0.002). There was no association between parasitaemia, haemoglobin level and the other oxidative stress biomarkers. From histological studies, levels of MDA associated positively and significantly with placenta malaria infection and the presence of malaria pigments. The levels of SOD, NO and CAT increased with decreasing leukocyte accumulation in the intervillous space. Baby birth weight increased significantly with SOD and CAT levels, but decreased with levels of GSH. CONCLUSIONS: Placental P. falciparum infection may cause oxidative stress of the placenta tissue with MDA as a potential biomarker of PM, which alongside GSH could lead to poor pregnancy outcomes (anaemia and low birth weight). This finding contributes to the understanding of the pathophysiology of P. falciparum placental malaria in women.

Insights Into Circulating Cytokine Dynamics During Pregnancy in HIV-Infected Beninese Exposed to Plasmodium falciparum Malaria.

We investigated the circulating plasma levels of Th1- (Interleukin-2 [IL-2], tumor necrosis factor-α [TNF-α], interferon-gamma [IFN-γ]) and Th2-type (IL-4, IL-5, IL-10) cytokines in human immunodeficiency virus (HIV)-infected pregnant women living in a malaria-endemic area. We analyzed samples from 200 pregnant women included in the prevention of pregnancy-associated malaria in HIV-infected women: cotrimoxazole prophylaxis versus mefloquine (PACOME) clinical trial who were followed until delivery. Cytokine concentrations were measured by flow cytometry-based multiplex bead array. Significantly elevated levels of IL-10 and lower levels of TNF-α were observed at delivery compared with inclusion (P = 0.005). At inclusion, the presence of circulating IFN-γ, a higher CD4(+) T cell count and having initiated intermittent preventive treatment of malaria with sulfadoxine pyrimethamine (SP-IPTp) were all associated with a lower likelihood of Plasmodium falciparum infection. At delivery, the inverse relationship between the presence of infection and circulating IFN-γ persisted, although there was a positive association between the likelihood of infection and the presence of circulating TNF-α. Initiation of antiretroviral therapy was associated with elevated IL-5 production. Consistent with our own and others' observations in HIV seronegative subjects, this study shows circulating IL-10 to be a marker of infection with P. falciparum during pregnancy even in HIV-infected women, although plasma IFN-γ may be a marker of anti-malarial protection in such women.

Prevalência de toxoplasmose em gestantes atendidas em dois centros de referência em uma cidade do Nordeste, Brasil / Prevalence of toxoplasmosis in pregnant women in two reference centers in a city in Northeast Brazil

OBJETIVO: Determinar a prevalência de toxoplasmose e identificar os principais fatores associados à sororeatividade em gestantes atendidas em dois centros de referência em uma cidade do Nordeste do Brasil. MÉTODOS: Foi realizado um estudo transversal com 561 gestantes atendidas em dois centros de referência para pré-natal de alto risco em uma cidade do Nordeste do Brasil. Todas foram entrevistadas por meio de um questionário epidemiológico e foram coletadas amostras de sangue em que foram realizadas sorologia anti-Toxoplasma gondii para IgG e IgM (ELISA), teste de avidez da IgG e reação em cadeia da polimerase (PCR). A análise estatística foi realizada com o programa SPSS version 18.0 Windows, usando odds ratio e intervalo de confiança de 95%, considerando-se o nível de significância de 5%. RESULTADOS: Constatou-se sororeatividade para toxoplasmose em 437 (77,0%), susceptibilidade em 124 (22,1%) e 5 (0,9%) gestantes com infecção ativa. Não encontramos associação significativa entre sororeatividade para toxoplasmose e idade, procedência, renda, escolaridade, situação da rede de esgotos, número de gestações e idade gestacional. As variáveis com associação significativa (p≤0,05) para sororeatividade foram: multigestas (p=0,03) e convívio com cães soltos na rua (p=0,001). CONCLUSÕES: O estudo permitiu identificar uma alta sororeatividade para toxoplasmose entre as pacientes atendidas no pré-natal, assim como os fatores associados à sororeatividade, devendo ser reforçadas orientações apropriadas sobre medidas de prevenção primária e monitoramento sorológico trimestral das gestantes nesse município e outras regiões do Nordeste do Brasil. .

PURPOSE: To determine the prevalence of toxoplasmosis and to identify the main factors associated with seroreactivity in pregnant women cared for at two reference centers in a city in Northeast Brazil. METHODS: A cross-sectional study was conducted on 561 pregnant women at two high-risk prenatal reference centers in a city in Northeast Brazil. All women were interviewed using an epidemiological questionnaire and had their blood samples collected for the following serological tests: anti-Toxoplasma gondii IgG and IgM (ELISA), IgG avidity test, and polymerase chain reaction (PCR). Statistical analysis was carried out using SPSS version 18.0 for Windows, calculating odds ratio, confidence interval of 95% and with the level of significance set at 5%. RESULTS: Seroreactivity for toxoplasmosis was detected in 437 women (77.0%), susceptibility in 124 (22.1%) and active infection in 5 (0.9%). There was no significant association between seroreactivity for toxoplasmosis and age, location, income, education, availability of sewage, number of pregnancies or gestational age. The variables significantly associated (p≤0.05) with seroreactivity were multiparity (p=0.03) and living with stray dogs (p=0.01). CONCLUSIONS: This study identified high seroreactivity for toxoplasmosis among patients seen during prenatal care, as well as factors associated with seroreactivity. Appropriate guidelines about primary preventive measures should be emphasized and quarterly serological monitoring is recommended for pregnant women in this city and elsewhere in the Northeast of Brazil. .

Effects of crossbreeding on endocrine patterns determined in pregnant beef/dairy cows naturally infected with Neospora caninum.

Neospora caninum is an intracellular coccidian parasite causing abortion worldwide in dairy cattle. Studies have shown that N caninum infection modifies endocrine patterns and that beef cows or cows with a crossbreed pregnancy, especially for a greater maternal-paternal genetic distance, show a reduced risk of abortion when naturally infected compared with pure-breed Holstein Friesian (HF) pregnancies. This study examined the effects of crossbreeding on plasma progesterone (P4), pregnancy-associated glycoprotein (PAG)-1 and -2, and prolactin patterns produced during gestation in N caninum-infected beef/dairy cows. We analyzed 74 pregnancies in nonaborting cows carrying a single fetus established in 26 lactating HF cows bred with Holstein bulls (H-H), 13 HF cows bred with Limousin bulls (H-L), 12 HF cows used as recipients of transferred in vivo-produced frozen-thawed Rubia Gallega (beef breed; RG) embryos (H-RG-ET), and 23 RG suckling cows bred with RG bulls (RG-RG). Of the cows, 29 (39%) were seropositive for N caninum. Blood samples for hormone and placental protein determinations were collected on Days 30, 60, 90, 120, 150, and 180 of gestation. Procedures of repeated measures analysis of variance throughout gestation revealed higher PAG-1 levels in the H-L and H-RG-ET groups compared with H-H and RG-RG. Higher prolactin levels were detected in RG-RG than in the remaining groups. N caninum seropositivity and fetal genotype had a significant effect on PAG-2 concentrations, such that highest levels of PAG-2 occurred in RG-RG seropositive cows and lowest in H-H seropositive cows, whereas Neospora-seropositive cows showed lower P4 concentrations than their seronegative partners. In conclusion, chronic N caninum infection modifies endocrine patterns of PAG-2 and P4 during pregnancy, whereas PAG-1 and prolactin concentrations are affected by breed and fetal genotype, irrespective of Neospora infection status.

IFN-gamma and TNF associated with severe falciparum malaria infection in Saudi pregnant women.

BACKGROUND: Tumour necrosis factor (TNF) and interferon gamma (IFN-γ), encoded by TNF-836 C/A (rs 1800630) and IFN-γ -1616 C/T (rs2069705) genes, are key immunological mediators that are believed to both play protective and pathological roles in malaria. The aim of this study was to investigate the relationship between TNF-836 C/A and IFN-γ-1616 C/T polymorphism and susceptibility to severe malaria in pregnant women. METHODS: A prospective cohort (cross-sectional) study was conducted in pregnant women attending the out-patient clinic in King Fahad Specialist Hospital in Jazan (KFSHJ), with a clinical diagnosis of malaria. A total of one hundred and eighty six pregnant women were genotyped for single nucleotide polymorphism (SNP) for TNF and IFN-γ using Taqman® MGB Probes. Serum cytokine concentrations were measured by sandwich ELISA method. RESULTS: A hospital case-control study of severe malaria in a Saudi population identified strong associations with individual single-nucleotide polymorphisms in the TNF and IFN-γ genes, and defined TNF-836 C and IFN-γ-1616 T genotypes and alleles which were statistically significantly associated with severe malaria infection. Furthermore, TNF-836 CC and IFN-γ-1616 TT genotypes were associated with higher serum concentration of TNF and IFN-γ, respectively, and with susceptibility to severe malaria. CONCLUSIONS: This data provides a starting point for functional and genetic analysis of the TNF and IFN-γ genomic region in malaria infection affecting Saudi populations.

The association between malaria and iron status or supplementation in pregnancy: a systematic review and meta-analysis.

INTRODUCTION: Malaria prevention and iron supplementation are associated with improved maternal and infant outcomes. However, evidence from studies in children suggests iron may adversely modify the risk of malaria. We reviewed the evidence in pregnancy of the association between malaria and markers of iron status, iron supplementation or parenteral treatment. METHODS AND FINDINGS: We searched MEDLINE, EMBASE, the Cochrane Central Register of Controlled Trials, the Global Health Library, and the Malaria in Pregnancy library to identify studies that investigated the association between iron status, iron treatment or supplementation during pregnancy and malaria. Thirty one studies contributed to the analysis; 3 experimental and 28 observational studies. Iron supplementation was not associated with an increased risk of P. falciparum malaria during pregnancy or delivery in Africa (summary Relative Risk = 0.89, 95% Confidence Interval (CI) 0.66-1.20, I(2) = 78.8%, 5 studies). One study in Asia reported an increased risk of P. vivax within 30 days of iron supplementation (e.g. adjusted Hazard Ratio = 1.75, 95% CI 1.14-2.70 for 1-15 days), but not after 60 days. Iron deficiency (based on ferritin and C-reactive protein) was associated with lower odds for malaria infection (summary Odds Ratio = 0.35, 0.24-0.51, I(2) = 59.2%, 5 studies). With the exception of the acute phase protein ferritin, biomarkers of iron deficiency were generally not associated with malaria infection. CONCLUSIONS: Iron supplementation was associated with a temporal increase in P vivax, but not with an increased risk of P. falciparum; however, data are insufficient to rule out the potential for an increased risk of P. falciparum. Iron deficiency was associated with a decreased malaria risk in pregnancy only when measured with ferritin. Until there is more evidence, it is prudent to provide iron in combination with malaria prevention during pregnancy.

IgG western blot for confirmatory diagnosis of equivocal cases of toxoplasmosis by EIA-IgG and fluorescent antibody test.

The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.

Prevalence of congenital toxoplasmosis among a series of Turkish women / Prevalencia de toxoplasmosis congénita en una serie de mujeres en Turquía

Background: Toxoplasma gondii infection during pregnancy causes congenital malformations. Pregnant women should be screened for this infection since it is preventable and treatable. Aim: To study the sero prevalence of Toxoplasma gondii infection among pregnant women living in lzmir, Turkey. Material and Methods: A blood sample was obtained from 4651 women aged between 15 and 45years, during their first trimester of pregnancy. IgM and IgG antibodies against Toxoplasma gondii were measured using an ELISA assay. Among women with both IgG and IgM antibodies positive, an IgG avidity test was performed, using a VIDAS kit. Results: IgG antibodies were positive in 1871 (39.9%) participants. Of these, 48 (2.5%) also had positive IgM antibodies. In 41 ofthese 48 women, the IgG avidity test was performed and only one woman had a low avidity. This woman was treated with Spiramycin. Her offspring had an intrauterine growth retardation and oligohydramnios. A chorioretinitis was diagnosed in the offspring of other woman with both antibodies positive. Conclusions: In this series, the prevalence of congenital toxoplasmosis was low. However, women with positive antibodies against Toxoplasma Gondii should be further studied and followed during their pregnancy.

Antecedentes: La infección por Toxoplasma gondii durante el embarazo causa malformaciones congénitas. Se debe efectuar serologíapara esta infección en mujeres embarazadas ya que es prevenible y tratable. Objetivo: Estudiar la seroprevalencia de infección por Toxoplasma gondii en mujeres embarazadas que viven en Esmirna, Turquía. Material y Métodos: Se obtuvo una muestra de sangre en 4.651 mujeres cuyas edades fluctuaban entre 15 y 45, años, durante su primer trimestre de embarazo. Los anticuerpos IgM e IgG en contra de Toxoplasma gondii se midieron por ELISA. En mujeres que tenían anticuerpos IgG e IgM positivos, un ensayo de avidez de IgG se efectuó utilizando el kit VIDAS. Resultados: Los anticuerpos IgG fueron positivos en 1.871 participantes (39,9%). De estas, 48 (2,5%) también tenían anticuerpos IgM positivos. En 41 de estas 48 mujeres, se efectuó el test de avidez y sólo una tenía una baja avidez. Esta mujer se trató con espiramicina y su producto de concepción tuvo un retardo de crecimiento intrauterino y un oligohidroamnios. Una corioretinitis se diagnosticó en el producto de concepción de otra mujer con ambos anticuerpos positivos. Conclusiones: La seroprevalencia de toxoplasmosis congénita en esta serie de pacientes fue baja, sin embargo, las mujeres con anticuerpos positivos deben ser tratadas y seguidas.

Antibody to P. falciparum in pregnancy varies with intermittent preventive treatment regime and bed net use.

BACKGROUND: Antibodies towards placental-binding P. falciparum are thought to protect against pregnancy malaria; however, environmental factors may affect antibody development. METHODS AND FINDINGS: Using plasma from pregnant Malawian women, we measured IgG against placental-binding P. falciparum parasites by flow cytometry, and related results to intermittent preventive treatment (IPTp) regime, and bed net use. Bed net use was associated with decreased antibody levels at mid-pregnancy but not at 1 month post partum (1 mpp). At 1 mpp a more intensive IPTp regime was associated with decreased antibody levels in primigravidae, but not multigravidae. CONCLUSIONS/SIGNIFICANCE: Results suggest bed nets and IPTp regime influence acquisition of pregnancy-specific P. falciparum immunity.

Soroepidemiologia da toxoplasmose em gestantes a partir da implantação do Programa de Vigilância da Toxoplasmose Adquirida e Congênita em municípios da região oeste do Paraná / Seroepidemiology of toxoplasmosis in pregnant women since the implementation of the Surveillance Program of Toxoplasmosis Acquired in Pregnancy and Congenital in the western region of Paraná, Brazil

OBJETIVO: Avaliar a suscetibilidade das gestantes à toxoplasmose em serviço público de saúde de dois municípios da região oeste do Paraná. MÉTODOS: Foram avaliadas 422 gestantes por meio da pesquisa sorológica de anticorpos IgG e IgM anti-Toxoplasma gondii (ELISA e MEIA). As soronegativas repetiram a sorologia no segundo e terceiro trimestre de gestação. Em um dos municípios, também foi realizada a triagem neonatal em 27 recém-nascidos para detecção de IgM anti-Toxoplasma gondii pelo teste de fluorometria. Todas as gestantes responderam a um questionário epidemiológico, para análise dos fatores associados ao risco da infecção pelo Toxoplasma gondii. Para análise estatística, foram consideradas a variável dependente da presença de IgG anti-Toxoplasma gondii e as variáveis independentes contidas no questionário epidemiológico. RESULTADOS: A prevalência de anticorpos IgG anti-Toxoplasma gondii nas gestantes foi de 59,8 e 60,6 por cento. Em um dos municípios, as variáveis associadas à presença de anticorpos IgG foram baixo nível de escolaridade e mais de uma gestação. Não houve associação com os outros fatores investigados, como a ingestão de carnes cruas ou mal cozidas, vegetais crus, salames coloniais, manipulação de terra ou areia, horta em casa e gatos em casa. No outro município, não foi observada associação estatística com nenhuma das variáveis estudadas. Não foi confirmado nenhum caso de infecção aguda nem de soroconversão em ambos os municípios. Nenhuns dos recém-nascidos avaliados apresentou positividade para toxoplasmose. CONCLUSÃO: A toxoplasmose é comum nas gestantes atendidas pelo serviço público de saúde da região estudada e há 40 por cento delas suscetíveis à infecção. Esse dado reforça a necessidade de manter o programa implantado nesses municípios.

PURPOSE: To evaluate the susceptibility to toxoplasmosis in pregnant women in the public health service from two cities in the western region of Paraná, Brazil. METHODS: Four thousand twenty-two pregnant women were evaluated for anti-Toxoplasma gondii IgG and IgM by ELISA and MEIA. Seronegative pregnant women repeated the serology in the second and third trimester of pregnancy. Neonatal screening of 27 newborns was also performed in one of the cities to detect IgM anti- Toxoplasma gondii by fluorometry. All pregnant women answered an epidemiological questionnaire to analyze the factors associated with the risk of infection by Toxoplasma gondii. For statistical analysis, the presence of IgG anti-Toxoplasma gondii was considered as the dependent variable and the variables contained in the epidemiological questionnaire as the independent ones. RESULTS: The prevalence of anti-Toxoplasma gondii IgG in pregnant women was 59.8 and 60.6 percent. In one of the cities, the variables associated with the presence of IgG antibodies were low educational level and more than one pregnancy. There was no association with other factors studied such as consumption of raw or undercooked meat, consumption of raw vegetables, consumption of colonial salami, handling soil or sand, the presence of a home vegetable garden and cats in the household. In the other city there was no statistical association with the variables studied. No case of acute infection and no seroconversion were confirmed in either city. None of the infants evaluated were positive for toxoplasmosis. CONCLUSION: Toxoplasmosis is common in pregnant women attended by the public health service in the region studied and 40 percent of them are susceptible to the infection. These data reinforce the need to keep the screening program in these cities.