Impact of placental malaria on maternal, placental and fetal cord responses and its role in pregnancy outcomes in women from Blue Nile State, Sudan.

BACKGROUND: The sequestration of Plasmodium falciparum infected cells in the placenta results in placental malaria (PM). It activates the mother's immune cells and induces secretion of inflammatory cytokines, which might influence pregnancy outcomes. This study aims to investigate the cytokines (levels IL-4, IL-6, IL-10, IL-17A, and INF γ) in maternal peripheral, placental, and umbilical cord blood in response to PM and the extent to which this may influence maternal haemoglobin levels and birth weight. METHODS: A total of 185 consenting Sudanese women from Blue Nile State were enrolled at delivery time in a cross-sectional study conducted between Jan 2012-Dec 2015. Malaria infection in the collected maternal peripheral, placental, umbilical cord samples was determined microscopically, and ELISA was used to measure the plasma levels IL-4, IL-6, IL-10, IL-17A, and INF γ in the collected positive and negative malaria samples. RESULTS: Elevated levels of IL-4 and IL-10 and reduced levels of IL-6 were detected in the malaria positive samples in comparison to the negative ones in the three types of the samples investigated. Maternal, IL-4 and IL-10 were significantly higher in the samples collected from the PM infected group compared to the non-infected control (P < 0.001). While the absence of PM was significantly associated with the IL-6 and maternal IFN-γ levels, maternal IL-17A, placental and umbilical cord IFN-γ levels showed no significant difference (P = 0.214, P = 0.065, P = 0.536, respectively) due to infection. Haemoglobin level and birth weight were increased in the group with high levels of IL-6 and IL-17A, but not in the group with IL-4 and IL-10 levels. While significantly negative correlation was found between IFN-γ levels and birth weight for all three types of samples, only maternal peripheral IFN-γ level was significantly positively correlated with maternal haemoglobin (r = 0.171, P = 0.020). CONCLUSION: These results suggest that PM induces mother's immune response and impairs her cytokine profile, which might alter maternal haemoglobin levels and the baby's birth weight.

Congenital Malaria in Newborns Delivered to Mothers with Malaria-Infected Placenta in Blue Nile State, Sudan.

Diagnosis of congenital malaria is complicated by the low density of the parasite circulating in the cord blood and/or the peripheral blood of the newborns. Molecular techniques are significantly more sensitive than blood smears in detecting low-level parasitemia. This study investigated the prevalence of congenital malaria by the use of the real-time polymerase chain reaction (real-time PCR) in 102 babies born to mothers with microscopically confirmed infected placenta from Blue Nile state, Sudan. At delivery time, placental, maternal peripheral and cord blood samples in addition to samples collected from the newborns' peripheral blood were examined for malaria infection using Giemsa-stained thick smear and parasite DNA detection by real-time PCR. The overall prevalence of congenital malaria includes the total babies with cord blood parasitaemia and peripheral blood parasitaemia was 18.6 and 56.8% using microscopy and real-time PCR, respectively. Even though all the neonates were aparasitaemic by microscopy, 19 (18.6%) of the babies had congenital malaria detected by real-time PCR, 15 (25.9%) of the babies with congenital malaria were born to mothers with both placental and peripheral blood malaria infections detected using the two techniques. Congenital malaria was significantly associated with cord blood malaria infections, maternal age and maternal haemoglobin level (p < 0.001). This first study investigating congenital malaria in Blue Nile state, Sudan shows that malaria-infected placenta resulted in infant and cord blood infections.

Placental malaria and its effect on pregnancy outcomes in Sudanese women from Blue Nile State.

BACKGROUND: Malaria infection during pregnancy can result in placental malaria and is associated with adverse pregnancy outcomes particularly among primigravidae. The aim of this study was to assess the prevalence and risk factors for placental malaria and its effect on pregnancy outcomes in Blue Nile state, Sudan. METHODS: A cross-sectional hospital-based study was conducted consecutively during January 2012-December 2015 in three main hospitals in Blue Nile State, Sudan. At delivery, peripheral and placental blood samples were collected from consenting women. Finger prick blood was used for preparation of peripheral smears and for haemoglobin measurement. Smears were stained with Giemsa and examined microscopically for malaria parasites. Pregnancy outcomes in association to placental malaria were investigated. RESULTS: A total of 1149 mothers and their newborns were recruited. The mean (SD) of the age was 23.3 (5.2) years. Detection of malaria parasites was confirmed in 37.8% of the peripheral blood films and 59.3% of the placental films with Plasmodium falciparum as the only species detected. In multivariate analysis, younger age ≤23.2 years old (AOR = 3.2, 95% CI 1.9-5.5; P < 0.001), primiparae (AOR = 3.9, CI 2.1-7.6; P < 0.001), secundiparae (AOR = 2.8, 95% CI 1.5-5.1; P < 0.001, no antenatal care (ANC) visits (AOR = 11.9, 95% CI 7.8-18.1; P < 0.001) and not using bed nets (AOR = 3.5, 95% CI 1.7-6.8; P < 0.001) were risk factors for placental malaria. Education and residence were not associated with placental malaria infection. Placental malaria was significantly associated with maternal anaemia (AOR = 41.6, 95% CI 23.3-74.4; P < 0.001) and low birth weight (LBW) (AOR = 25.2, 95% CI 15.1-41.3; P < 0.001). CONCLUSION: During the study, there was a high prevalence of placental malaria in Blue Nile State-Sudan, as the enhanced control activities were not practiced, leading to adverse pregnancy outcomes, such as maternal anaemia and LBW.

Drug repurposing for SARS-CoV-2 main protease: Molecular docking and molecular dynamics investigations.

The current novel corona virus illness (COVID-19) is a developing viral disease that was discovered in 2019. There is currently no viable therapeutic strategy for this illness management. Because traditional medication development and discovery has lagged behind the threat of emerging and re-emerging illnesses like Ebola, MERS-CoV, and, more recently, SARS-CoV-2. Drug developers began to consider drug repurposing (or repositioning) as a viable option to the more traditional drug development method. The goal of drug repurposing is to uncover new uses for an approved or investigational medicine that aren't related to its original use. The main benefits of this strategy are that there is less developmental risk and that it takes less time because the safety and pharmacologic requirements are met. The main protease (Mpro) of corona viruses is one of the well-studied and appealing therapeutic targets. As a result, the current research examines the molecular docking of Mpro (PDB ID: 5R81) conjugated repurposed drugs. 12,432 approved drugs were collected from ChEMBL and drugbank libraries, and docked separately into the receptor grid created on 5R81, using the three phases of molecular docking including high throughput virtual screening (HTVS), standard precision (SP), and extra precision (XP). Based on docking scores and MM-GBSA binding free energy calculation, top three drugs (kanamycin, sulfinalol and carvedilol) were chosen for further analyses for molecular dynamic simulations.

The diagnostic performance of rapid diagnostic tests and microscopy for malaria diagnosis in eastern Sudan using a nested polymerase chain reaction assay as a reference standard.

BACKGROUND: Accurate diagnosis of malaria infection is essential for successful control and management of the disease. Both microscopy and rapid diagnostic tests (RDTs) are recommended for malaria diagnosis, however, RDTs are more commonly used. The aim of the current study was to assess the performance of microscopy and RDTs in the diagnosis of Plasmodium falciparum infection using a nested polymerase chain reaction (PCR) assay as the gold standard. METHODS: A cross-sectional study was carried out in Kassala Hospital, eastern Sudan. A total of 341 febrile participants of all ages were recruited. Blood specimens were collected and malaria testing was performed using an RDT (SD Bioline Malaria Ag Pf), microscopy and nested PCR. The sensitivity, specificity, positive and negative predictive values (PPV and NPV, respectively) of microscopy and the RDT were investigated. RESULTS: The prevalence of P. falciparum malaria infections in this study was 22.9%, 24.3% and 26.7% by PCR, microscopy and RDT, respectively. Compared with microscopy, the RDT had slightly higher sensitivity (80.7% vs 74.3%; p=0.442), equivalent specificity (89.3% vs 90.4%), a similar PPV (69.2% vs 69.8%) and a higher NPV (94.0% vs 92.2%). CONCLUSIONS: The diagnostic performance of the RDT was better than that of microscopy in the diagnosis of P. falciparum malaria when nested PCR was used as the gold standard.

Impact of Submicroscopic Plasmodium falciparum Parasitaemia on Maternal Anaemia and Low Birth Weight in Blue Nile State, Sudan.

The aim of the present study was to investigate the prevalence of submicroscopic infections and to assess its impact on maternal anaemia and low birth weight. A cross-sectional study was carried out with 1149 consented pregnant women who delivered at 3 main hospitals in the Blue Nile State, between January 2012 and December 2015. From a matched maternal peripheral, placental maternal side, and cord blood sample, blood films and dried spots were prepared for microscopic examination and nested polymerase chain reaction (n-PCR), respectively. 107 out of 447 negative blood films were found to have submicroscopic infection detected using n-PCR in any of the three compartments. Placental samples had a significantly higher prevalence (142) of submicroscopic infections compared with the peripheral (6.5%) and cord (8.1%) samples. The mean (SD) of the maternal haemoglobin (Hb) was significantly lower in cases with submicroscopic parasitaemia (10.9 (0.8) vs. 12.1 (0.7) g/dl, P < 0.001) compared with those who had no submicroscopic parasitaemia. Submicroscopic malaria infection was associated with anaemia (OR 19.7, (95% CI, 10.3-37.8)). Thirty-eight babies born to women with submicroscopic infections were low birth weight (LBW) and was significantly lower in submicroscopic parasitaemia (2.663 (0.235) vs. 2.926 (0.341) kg, P < 0.001). Submicroscopic malaria infection was associated with LBW (OR = 2.7, (95% CI, 1.2-5.6)). There is a high incidence of submicroscopic infections in any of the three compartments regardless of age or parity. Submicroscopic infection is a risk of maternal anaemia and low birth weight in women in this area of high seasonal malaria transmission.

Evolution of Plasmodium falciparum drug resistance genes following artemisinin combination therapy in Sudan.

BACKGROUND: Malaria control efforts in Sudan rely heavily on case management. In 2004, health authorities adopted artemisinin-based combination therapies (ACTs) for the treatment of uncomplicated malaria. However, some recent surveys have reported ACT failure and a prevalent irrational malaria treatment practice. Here we examine whether the widespread use of ACT and failure to adhere to national guidelines have led to the evolution of drug resistance genes. METHODS: We genotyped known drug resistance markers (Pfcrt, Pfmdr-1, Pfdhfr, Pfdhps, Pfk13 propeller) and their flanking microsatellites among Plasmodium falciparum isolates obtained between 2009 and 2016 in different geographical regions in Sudan. Data were then compared with published findings pre-ACT (1992-2003). RESULTS: A high prevalence of Pfcrt76T, Pfmdr-1-86Y, Pfdhfr51I, Pfdhfr108N, Pfdhps37G was observed in all regions, while no Pfk13 mutations were detected. Compared with pre-ACT data, Pfcrt-76T and Pfmdr-1-86Y have decayed, while Pfdhfr-51I, Pfdhfr-108N and Pfdhps-437G strengthened. Haplotypes Pfcrt-CVIET, Pfmdr-1-NFSND/YFSND, Pfdhfr-ICNI and Pfdhps-SGKAA predominated in all sites. Microsatellites flanking drug resistance genes showed lower diversity than neutral ones, signifying high ACT pressure/selection. CONCLUSIONS: Evaluation of P. falciparum drug resistance genes in Sudan matches the drug deployment pattern. Regular monitoring of these genes, coupled with clinical response, should be considered to combat the spread of ACT resistance.

Innovative serum-free medium for in vitro cultivation of promastigote forms of Leishmania species.

We described a comparatively simple medium formula (CML) using common, available and reasonably priced ingredients that could be used in place of medium that requires calf serum enhancement for cultivation of Leishmania promastigote forms. This medium equivalently supported the growth of parasites at rates comparable with those obtained with serum supplemented RPMI-1640 medium. Leishmania promastigotes reproduced in CML exhibited moderate to high infectivity capacities when tested against J774 macrophage cell line. No significant difference was noted between Leishmania strains cultivated in the newly modified medium and those grown in RPMI-1640 medium in their cells infectivity and replication potentials. The use of new CML can easily take the place of other biphasic or liquid media because of its easy preparation and instantaneous use, reasonable price, availability of ingredients, and its long shelf life, which is 30-45 days. The fact that this medium is similar to other culture media as far as durability and quantity of produced parasites might give it an advantage over the other currently used media.

Submicroscopic and multiple plasmodium falciparum infections in pregnant Sudanese women.

BACKGROUND: Control of malaria during pregnancy remains a major public health challenge in developing countries. Microscopic parasite detection represents a pivotal step in malaria control, while modern molecular techniques are deemed to improve detection rates markedly. AIMS: This study aimed to investigate the frequency of submicroscopic and multiple Plasmodium falciparum (P. falciparum) infections during pregnancy, using the P. falciparum merozoite surface protein1 (MSP-1) gene as a polymorphic marker. MATERIALS AND METHODS: The study was a cross-sectional, analytical study that was conducted at Omdurman Maternity Hospital, Sudan, between July 2003 and December 2004. Following informed consent, 836 pregnant women between the ages of 16-47 years with different gestational ages were enrolled in the study. Thin and thick blood films were stained with Giemsa and examined by experienced microscopists. Parasite DNA was extracted using Chelex method. Nested polymerase chain reaction (PCR) assays specific for P. falciparum were carried out to detect infections below the threshold of microscopy and to genotype different strains in the samples using merozoite surface protein-1. RESULTS: More than a quarter of the study participants (219/836; 26.2%) were smear-positive for malaria infection. The results of the PCR-based assays showed that 41.8 % (257/617) of the smear-negative women were PCR positive and therefore had submicroscopic infections. The mean number of genetically different P. falciparum parasites detected was 2.7 (range 1-9). The multiplicity of infection identified by at least two alleles of MSP-1 was significantly higher among paucigravidae (45.6%) compared to multigravidae (28.9%), with mean number of alleles of 2.4 and 1.9, respectively (p=0.009). This likely indicates the gradual acquisition of immunity. CONCLUSION: Conventional microscopy underestimates the actual extent of malaria infections during pregnancy in endemic regions. Multiplicity of infection may be an important factor in the gradual acquisition of strain-specific immunity.

Leishmania donovani: an in vitro study of antimony-resistant amphotericin B-sensitive isolates.

Drug sensitivity of clinically antimony-unresponsive Leishmania donovani isolates from Eastern Sudan was evaluated in an in vitro culture system against sodium stibogluconate (Pentostam) and Amphotericin B. Eight isolates, six from antimony-resistant and two from clinically responsive patients were included in the study. Parasites were tested as promastigotes and four of them were selected to be tested as amastigotes using a murine macrophage-like cell line. The results indicated that the conventional promastigotes and amastigotes-screening assays did not correlate with the clinical picture of patients. In vivo unresponsiveness does not necessarily mean primary parasite resistance. Amphotericin B could be a suitable second line drug in patients unresponsive to pentostam and without concomitant diseases, if close hospital monitoring is available. Promastigotes sensitivity testing concentrations are virtually incomparable with the in vivo clinically curable doses and the amastigotes/macrophage test concentrations.