CCR5 contributes to adverse outcomes during malaria in pregnancy.

CCR5 is a chemokine receptor that mediates cell recruitment to sites of inflammation. It has been previously reported that the expression of CCR5 is increased in the placentas of women with malaria, a disease characterized by causing deliveries with low birth weight among other complications. CCR5 has been associated with pathology of protozoan infections during pregnancy but its role during malaria in pregnancy has not been elucidated. In the present work, we assessed the pregnancy outcome, placental structure, and levels of inflammatory markers of pregnant C57BL/6 and CCR5-/- mice infected or not with Plasmodium berghei NK65, with the purpose of determine the role of CCR5 in pregnancy associated malaria complications. We demonstrated that the expression of CCR5 mRNA increases in late pregnancy placentas of C57BL/6 when compared to uninfected controls. Infected pregnant C57BL/6 mice showed preterm birth, decreased fetal weight, placental inefficiency, and reduced placental vascular space. On the other hand, CCR5 deficiency led to increased levels of maternal parasitemia, reduced fetal weight and placental inefficiency compared to C57BL/6 mice. However, the infection did not cause additional changes in these parameters or in the incidence of preterm delivery in infected CCR5-/- mice in relation to C57BL/6 mice, showing that CCR5 may contribute to the adverse effects caused by infection during pregnancy. This improvement in pregnancy outcome, observed in infected CCR5-/- mice, was accompanied by lower placental levels of the inflammatory markers, such as TNF and NAG. Furthermore, it was observed that the placentas of CCR5-/- animals showed structural differences in relation to C57BL/6 mice, which could improve the efficiency of maternal-fetal exchanges, reflecting on fetal weight. Taken together, these results indicate that CCR5 expression contributes to the adverse outcomes caused by malaria in late pregnancy.

Peptides DLL37-1 and LL37-1, an alternative to inhibit biofilm formation in clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis.

Staphylococcus aureus and Staphylococcus epidermidis have microbial surface components recognizing adhesive matrix molecules (MSCRAMM) adhesion proteins that enhance their biofilm formation ability, as well as virulence factors that influence morbidity and mortality in hospital settings. In this work, four peptides analogous of the peptide LL-37 that were evaluated to inhibit biofilm formation and its action potential on the expression of MSCRAMM proteins in clinical isolates through different tests, such as crystal violet, PCR and qPCR. In total, 96.8% of S. aureus were strong in biofilm formation in contrast to 48.4% of S. epidermidis. sdrG and sdrF genes were present in 100% of S. epidermidis strains and in all isolates. In S. aureus, specific genes that code for MSCRAMM proteins were detected: clfA (89%), clFB, sdrC and fnBA (94%). The peptides did not show hemolytic or cytotoxic activity. In this study, it was evidenced that of the peptides DLL37-1 at a 5 µM concentration was an efficacious antimicrobial agent and depicted greater biofilm inhibition in both bacterial species. Exhibiting a significant inhibition rate in S. aureus, this peptide caused a negative regulation in the expression of the genes clfA and sdrC, showed greater biological activity.

Inhibition of Heme Oxygenase-1 by Zinc Protoporphyrin IX Improves Adverse Pregnancy Outcomes in Malaria During Early Gestation.

The enzyme heme oxygenase-1 (HO-1) has cytoprotective effects by catalyzing the degradation of heme to produce carbon monoxide, iron and biliverdin. Furthermore, HO-1 activity has been associated with successful pregnancy. On the other hand, in the context of certain inflammatory conditions, HO-1 can induce iron overload and cell death. To investigate the role of HO-1 in gestational malaria, pregnant BALB/c mice were infected with Plasmodium berghei ANKA in early, mid and late gestation. We found that malaria affected the pregnancy outcome in the three periods evaluated. However, only poor pregnancy outcomes in early pregnancy were related to HO-1 upregulation, iron overload, lipid peroxidation and necrosis of the decidua, which were prevented by HO-1 inhibition. In conclusion, HO-1 expression must be finely tuned in gestational malaria to avoid the deleterious effect of increased enzyme activity.

Detección molecular de sífilis gestacional y congénita / Molecular detection of gestational and congenital syphilis

Objetivo: estandarizar una qPCR para la determinación de T. pallidum en muestras de suero de pacientes con sífilis gestacional y congénita. Materiales y métodos: se optimizó una qPCR con sonda para la amplificación del gen TpN47 en muestras de suero, se evaluó la sensibilidad, especificidad y eficiencia analítica de la técnica. Se comparó con pruebas serológicas (VDRL y TPPA) y se calculó índice de concordancia Kappa. Resultados: la qPCR mostró un límite de detección de 0.113 femtogramos, especificidad analítica del 100% y fidelidad de 104%. Se evidenció correlación optima en la prueba, sugerida por un r2 de 0.99 y un valor p <0,0001 de la qPCR. Se observó acuerdo entre las pruebas serológicas y moleculares. Conclusiones: se desarrolló una herramienta molecular prometedora con buena sensibilidad, óptima especificidad analítica y gran potencial diagnóstico para la detección y hallazgo de T. pallidum subsp. pallidum, a través de la amplificación del gen TpN47 en muestras clínicas de pacientes con diagnóstico presuntivo de sífilis gestacional y congénita.

Objective: to evaluate a qPCR to detect T.pallidum in serum samples from patients with gestational and congenital syphilis. Methodology: qPCR with probe was optimized for the amplification of the TpN47 gene in serum samples, the sensitivity, specificity and analytical efficiency of the technique were evaluated. It was compared with the serological tests (VDRL and TPPA) and the Kappa concordance index was calculated. Results: the qPCR showed a detection limit of 0.113 femtograms, an analytical specificity of 100% and an accuracy of 104%. Optimal correlation was evidenced in the test, suggested by an r2 of 0.99 and a p value <0.0001 of the qPCR. An agreement was observed between serological and molecular tests. Conclusion: a promising molecular tool was developed with good sensitivity, excellent analytical specificity and great diagnostic potential for the detection and finding of T. pallidum subsp. pallidum, through the amplification of the TpN47 gene in serum from patients with gestational and congenital syphilis.