Background & objectives: Malaria continues to be a significant public health problem in tropical countries including India; however, there are limited tools to predict occurrence of severe disease due to malaria. This study was designed to evaluate the role of Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2), Vascular endothelial growth factor (VEGF) and Asymmetric Dimethylarginine (ADMA)as disease biomarkers in uncomplicated malaria (UM) and severe malaria (SM). Methods: This is a prospective observational study carried out at All India Institute of Medical Sciences (AIIMS), tertiary referral hospital in New Delhi, India. The study population included patients diagnosed with malaria (Plasmodium falciparum or Plasmodium vivax) either by rapid diagnostic kit test or positive peripheral smear and age more than 12 years. Forty-nine patients (25 with SM, 24 with UM) and 22 controls were recruited. In addition to routine investigations, serum concentrations of Ang-1, Ang-2, VEGF and ADMA were measured using ELISA technique. Results: We observed Ang-1 serum levels to be significantly lower in patients with severe malaria (7775 pg/ml) compared to uncomplicated malaria (17629 pg/ml) and healthy controls (43472 pg/ml) [p <0.001]. Ang-2 levels were significantly higher in severe malaria (11100 pg/ml) compared to uncomplicated malaria (7315 pg/ml) and healthy controls (3679 pg/ml) (p <0.001). The ratio of Ang-2/Ang-1 was significantly higher in patients with severe malaria. VEGF serum levels was significantly lower in severe malaria (130.36 pg/ml) compared to uncomplicated malaria (317.3 pg/ml). The Ang-1, Ang-2 and VEGF levels were able to differentiate severe malaria from uncomplicated malaria caused by P. vivax but not with P. falciparum. Interpretation & conclusion: We conclude that Ang-1, Ang-2 and VEGF are markers of disease severity in vivax malaria.

Background & objectives: Lymphatic filariasis (LF) is a neglected parasitic disease of tropics and subtropics including India. Although the disease mostly affects adults, there are evidences that LF infection is often acquired during childhood. This study assessed the filarial epidemiology as well as awareness about the disease among school children in the rural areas of Bankura district, West Bengal, India. Methods: Night blood samples of 1193 school children were examined to detect microfilariae. They were examined clinically by a physician for filarial manifestations and inquired about their awareness on the disease. Results: Microfilaria rate and disease rate among school children of the study area were assessed as 3.69% and 8.97% respectively. Only 12.74% of the school children had proper knowledge about filariasis and 72.09% had filarial patients living nearby. Among those affected children, 53.79% never received proper medical attention and 66.21% were found to be anxious about the fate of their filarial problems. School going males were found to be more affected than the females. Anxiety about the effect of the disease in girls was higher due to societal and medical ignorance among women that might develop psychological concerns. Interpretation & conclusion: This study suggests that, in the present study area, about 12.15% population acquired filarial infection at the stage of schooling. But effort for developing awareness about filariasis among the school children is scanty, which is presumably one of the major causes of high endemicity. To rectify the situation, knowledge about mosquito and mosquito borne diseases, including LF, should be properly incorporated in school curriculum. Control measures like anti-filarial drug administration and morbidity-management can be implemented in schools by the local governments, as it should be.

An outbreak of Japanese encephalitis (JE) was reported in Solan district of Himachal Pradesh, India in May 2018 wherein a total of eight JE cases were reported by the state health department, of which seven cases were confirmed by ELISA. An entomological survey was carried out to investigate the presence of vector mosquitoes in the affected area. Field visits were undertaken in eight villages of two blocks in Solan district. Larval collections were made from the major aquatic sites and emerging mosquito species were identified. Seepage water ditches were the main source of JE vector mosquitoes Culex tritaeniorhynchus and Cx. bitaeniorhynchus. During hand catch collections of adult mosquitoes, three vector species of JE,Culex tritaeniorhynchus (Man Hour Density 1–14), Cx. vishnui (MHD 2) and Cx. bitaeniorhynchus (MHD 1–2) were collected. A few specimens of Cx. tritaeniorhynchus (6 no.) were also collected in light trap collections. Since this was the first reported outbreak of JE from Himachal Pradesh, India, studies on sero-surveillance in addition to bionomics of JE vectors are required for better understanding of epidemiology of JE in Himachal Pradesh. Moreover, there is a need to study the role of climate change especially rising temperature in the context of JE in Himachal Pradesh.

Background & objectives: Scrub typhus has a wide spectrum of clinical presentations from acute febrile illness to sepsis with multi-organ failure with poor prognosis. The aim was to study the clinical presentation of scrub typhus and application of SOFA and qSOFA scores of sepsis-3 criteria to determine sepsis and consequent in-hospital outcomes. Methods: A prospective study was carried out in adults (≥18 years), with scrub typhus diagnosed by IgM ELISA. Sepsis-3 criteria were used to identify patients with sepsis and plan subsequent management. Statistical analysis was done using software SPSS 16.0 and p value of less than 0.05 was taken as significant. Results: Of the 78 scrub typhus patients, 38 (48.71%) presented with sepsis. Mean age in the sepsis group was significantly lower than the non-sepsis group (35.41±11.14 versus 43.56±14.31 years, p<0.05). Mortality was 71.05% in the sepsis group compared to nil in the non-sepsis group. On regression analysis, the presence of eschar (OR=5.2, 95% CI=1.15-23.544, P= 0.032), Acute Respiratory Distress Syndrome (ARDS) (OR=5.33, 95% CI= 1.13-25.16, P = 0.034) and a GCS <10 (OR=9.29, 95% CI=1.04-82.96, P=0.046) were significant predictors of mortality in patients of scrub typhus with sepsis. Interpretation & conclusion: Scrub typhus is more common in young age, with significantly younger patients developing sepsis. Presence of eschar, ARDS and GCS<10 were risk factors for mortality in scrub typhus patients with sepsis.

Background & objectives: Molecular methods for malaria vector species and parasite identification have received great attention in recent years. Accurate and precise identification of the target species has direct medical and practical implications, such as in malaria diagnosis and vector dynamics study. Translation of molecular techniques will help in evaluation of epidemiological and entomological profile of malaria even in highly inaccessible areas where there is lack of an expert microscopist or entomologist. Methods: In the present study, we have developed a simple yet accurate molecular tool for malaria diagnosis as well as for malaria vector studies. We have standardized, simplified and improvised the DNA isolation (using Chelex; a cationic exchanger), its storage and multiplex PCR for parasite detection from dried blood spot (DBS) filter paper as well as malaria vector identification and infection status study. Results: The chelex-PCR based molecular method was highly sensitive (sensitivity >90%) and specific (specificity >80%) for parasite detection as well as vector species identification. This method has proven readily adaptable for use in the clinical diagnostic/research laboratory for epidemiological investigation and vector dynamics study that can challenge the conventional gold standard approach such as microscopy/ morphological methods not only in response to accuracy but also in relation to cost, time and technical expertise. Interpretation & conclusion: Transfer of this molecular technology from laboratory to field condition is highly essential for its availability to the common public rather than being restricted to only academic research. This can be achieved by implementation of the technology in terms of conducting mass training and awareness programs in various resource-limited endemic zones for the purpose of malaria elimination.

Background and objectives: The mitochondrial electron transport chain (mtETC) of Plasmodium falciparum is an important drug target. Identification and functional validation of putative mitochondrial proteins of the mtETC is critical for drug development. Many of the regulatory subunits and assembly factors of cytochrome c oxidase readily identifiable in humans and yeast are missing in P. falciparum. Here, we describe our efforts to identify and validate the function of putative Pfsurf1, a key assembly factor of complex IV of the mtETC. Methods: Multiple sequence alignment of SURF 1/Shy 1 was carried out in Clustal X 2.1. Phylogenetic tree was constructed using “Draw tree” option in Clustal X, and was analyzed using interactive Tree of Life software. To identify the conserved sequences, domain search was done using Jalview version 2.8.2 (BLOSUM 62 scoring). The haploid Saccharomyces cerevisiae strain (BY4741) containing the null allele shy1 (Orf: YGR112w) (shy1::Kan) was complemented with putative Pfsurf1 to study its ability to rescue the growth defect. Results: Similarity searches of PfSURF1-like protein in the Pfam shows statistically significant E = 4.7e-10 match to SURF1 family. Sequence alignment of PfSURF1 with other SURF1-like proteins reveals the conservation of transmembrane domains, α-helices and β-pleated sheets. Phylogenetic analysis clusters putative PfSURF1 with apicomplexan SURF1-like proteins. Yeast complementation studies show that Pfsurf1 can partially rescue the yeast shy1 mutant, YGR112w. Interpretation & conclusion: Bioinformatics and complementation studies in yeast show that P. falciparum’s SURF1 is the functional ortholog of human SURF1 and yeast Shy1.

Background & objectives: Dengue is considered one of the greatest public health challenges in the world, especially, for tropical and subtropical countries. Brazil highlights with considerable number of registers associated with this arbovirus, with emphasis on the state of Minas Gerais and its municipalities in regions such as the midwest. We aimed to evaluate the economic and epidemiological impact of dengue in the midwest region of minas Gerais, from SUS perspective. Methods: Our study evaluated the epidemiological impact of dengue fever in the midwest region of Minas Gerais using data applied to Divinópolis city from SUS perspective between 2000 to 2015 considering SIH/SUS database (hospitalization registers) and 2007 to 2017 using the SINAN database (notification cases). We analyzed the number of notification registers and hospital services, as well as their costs, from a SUS perspective associated with this infection. Results: There were 26,516 notifications associated with dengue, with 21,953 cases being confirmed by SEMUSA, with a median incidence of 211 cases/100,000 inhabitants of dengue notifications between 2007 and 2017. The southeast and southwest regions in the municipality are highlighted with high number of cases. Additionally, 160 hospitalizations were recorded with 33.79% in individuals between 15 and 34 years old and expenses of USD 53,737.21 for SUS between 2000 and 2015. Interpretation & conclusion: Divinópolis is the reference (social and economic) municipality of the midwest region in Minas Gerais state. Our study is the first conducted involving a long period of follow up applied to dengue context in this locality, especially, including the costs associated with hospitalization services. We hope to contribute to the discussions regarding the coping strategies of the disease, considering the impact in its different regions, highlighting the need for continued efforts and initiatives aimed at combating the vector of this and other arboviruses such as Zika and chikungunya.

Background & objectives: Cutaneous leishmaniasis (CL) is a zoonotic and anthropogenic protozoal disease. We aimedto develop a new therapeutic approach for the treatment of CL. Methods: BALB/c mice have infected L. major amastigotes from their footpads. Twenty-one days later after injection, the animals were divided into three control and three experimental groups. The intralesional injection of graphene oxide and photothermal application (GO+PA) were applied to the first experimental group (Group 1); graphene oxide modified with a macrophage-specific antibody and photothermal application (MSA+GO+PA) were applied to the second experimental group (Group 2), and the photothermal application (PA) was applied to the third experimental group (Group 3). Miltefosine was administered orally to the first control group (Group 4); the second control group that is not treated was assigned as the positive control (Group 5) and the third control group was assigned as the negative control (Group 6). Lesions were examined (erythema and edema) after the 5^th day and 10^th of the treatment, clinically. On the 10^th day of the treatment, the level of TNF-α, IL-1, IL-6, and IFN-ɤ were detected histopathologically and immunohistochemically. Results: In the 5^th day of the treatment it was observed that 50% of the animals were completely treated with Group 2, and in the 10^th day, the ration raised to 75%. Interpretation & conclusion: We showed a novel application to treat CL by using MSA modified GO and PA within 10 days. According to our study outcomes, this application could be a new treatment approach for CL cure.

Acute acalculous cholecystitis is usually seen in association with systemic medical illness, or after surgery, trauma or burn and is considered as a more severe disease than acute calculous cholecystitis. We recently had the opportunity of observing a patient who was admitted in a surgical emergency with clinical features of acute cholecystitis and was found to have Orientia tsutsugamushi infection after a strong clinical suspicion of acute tropical fever illness.