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Backgrounds: Leucas cephalotes is a common ethnomedicinal plant widely used by traditional healers for the treatment of Malaria and other types of fever. Oleanolic acid and its derivatives have been reported for various types of pharmacological activities, such as anti-inflammatory, antioxidant, anticancer, hepatoprotective, anti-HIV and anti-HCV activity., Methods: L.cephalotes plant extracts were prepared by supercritical fluid extraction (SFE) method and oleanolic acid was isolated by preparatory thin-layer chromatography. The compound was identified and characterize by using ultraviolet-visible spectroscopy (UV-VIS), Fourier transform infra-Red spectroscopy (FT-IR) and high-performance thin-layer chromatography (HPTLC). The structure of the compound was elucidated by proton nuclear magnetic resonance ( 1 HNMR) and carbon nuclear magnetic resonance ( 1 CNMR) and the purity checked by differential scanning calorimetry (DSC). The MTT assay was used to determine the toxicity of plant extract and oleanolic acid using a microplate reader at 595 nm. The anti-dengue activity of plant extract and oleanolic acid was tested in vitro and in silico using real-time RT-PCR., Results: The optimum yield of the extract was obtained at 40 °C temperature and 15Mpa pressure. The maximum non-toxic dose (MNTD) of plant extract and oleanolic acid were found as 46.87 μg/ml and 93.75 μg/ml, respectively in C6/36 cell lines. UV spectrophotometer curve of the isolated compound was overlapped with standard oleanolic acid at 232 nm. Superimposed FT-IR structure of the isolated compound was indicated the same spectra at 3433, 2939, 2871, 1690, 1500,1463, 1387, 1250, 1209, 1137 and 656 position as per marker compound. HPTLC analysis showed the retention factor of L. cephalotes extract was 0.19 + 0.06 as similar to the standard oleanolic acid chromatogram. The NMR structure of the isolated compound was identified as similar to the marker oleanolic acid structure. DSC analysis revealed the purity of isolated oleanolic acid was 98.27% with a melting point of 311.16 °C. Real-time RT PCR results revealed that L. cephalotes supercritical extract and isolated oleanolic acid showed 100 and 99.17% inhibition against the dengue - 2 virus when treated with MNTD value of plant extract (46.87 μg/ml) and the test compound (93.75 μg/ml), respectively. The molecular study demonstrated the binding energy of oleanolic acid with NS1and NS5 (non-structural protein) were - 9.42 & -8.32Kcal/mol, respectively., Conclusions: The SFE extract L. cephalotes and its active compound, oleanolic acid inhibiting the activity of dengue-2 serotype in the in vitro and in silico assays. Thus, the L.cephalotes plant could be an excellent source for drug design for the treatment of dengue infection., (© 2021. The Author(s).)

Ethnopharmacological Relevance: About 2.5 billion peoples are at risk of dengue virus and the majority of people, use traditional plant-based medicines to combat dengue. The whole plant of Andrographis paniculata used traditionally over past decades for health promotion. Andrographolide isolated from Andrographis paniculata is used as natural remedy for the treatment of various diseases in different parts of the world. Andrographolide has been reported to have antiviral activity against hepatitis B virus, hepatitis C virus, herpes simplex virus, influenza virus, chikungunya virus, dengue virus 2 and 4., Aim of the Study: The aim of the present study to isolate the andrographolide from the A. paniculata by supercritical fluid extraction technique and to characterize the isolated compound along with it anti-dengue activity against DENV-2 in vitro and in silico methods., Materials and Methods: Supercritical extraction condition for A. paniculata was standardised to isolate andrographolide compound at definite temperature and pressure on the basis of previous study. The andrographolide was identified by using Ultraviolet-Visible Spectroscopy (UV-VIS), Fourier-Transform Infrared Spectroscopy (FT-IR) and High Performance Thin Layer Chromatography (HPTLC) and Proton Nuclear Magnetic Resonance ( 1 HNMR). The maximum non-toxic dose of isolated andrographolide was detected by MTT assay using a micro plate reader at 595 nm. One hundred (100) copies/ml of the DENV-2 virus was used for antiviral assay in C6/36 cells lines and inhibition of virus due to andrographolide was determined by real-time PCR assay. The purity of isolated andrographolide was determined by Differential Scanning Calorimetry (DSC). The dengue NS5 receptor protein was docked with andrographolide and evaluated on the basis of the total energy and binding affinity score by Auto Dock (V4.2.6) software., Results: Andrographolide, a diterpene lactone was isolated from the A. paniculata supercritical extract at 40 °C temperature and 15 Mpa pressure. UV spectrophotometer analysis revealed that the curve of andrographolide plant extract was overlapped with reference compound at 228 nm and the similar bands were detected from FT-IR spectroscopy analysis at 3315, 2917, 2849, 1673, 1462 and 1454 cm - 1 in isolated and standard andrographolide. HPTLC analysis shows the retention factor (R f values. The purity of isolated andrographolide was 99.76%. The maximum non-toxic dose of isolated andrographolide was found as 15.62 μg/ml on the C6/36 cell line calculated by using MTT assay. The andrographolide showed the 97.23% anti-dengue activity against the dengue-2 virus in C6/36 cell lines. Results of molecular docking showed that the interaction between andrographolide and NS5 of dengue protein with the maximum binding energy as -7.35 kcal/mol.f values. The purity of isolated andrographolide was 99.76%. The maximum non-toxic dose of isolated andrographolide was found as 15.62 μg/ml on the C6/36 cell line calculated by using MTT assay. The andrographolide showed the 97.23% anti-dengue activity against the dengue-2 virus in C6/36 cell lines. Results of molecular docking showed that the interaction between andrographolide and NS5 of dengue protein with the maximum binding energy as -7.35 kcal/mol., Conclusions: It is concluded that isolated andrographolide from the A. paniculata possess anti-dengue activity against dengue-2 virus as revealed from in vitro and in silico method. Due to lack of the vaccine and anti-viral agents, andrographolide extracted from A. paniculata play a major role to inhibit the dengue replication. Hence, it could be a source for drug design and help to reduce the dengue infection., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Dengue fever management is guided by WHO guideline, the recent one being 2009; however, compliance to the guideline is difficult to assess and in India there is no data on it. The present study, a longitudinal pre-post interventional quality improvement study, was done to determine the compliance to the guideline on dengue patients before and after resident physicians' training during two peak seasons and their impact on survival., Methods: This study was conducted in a tertiary health care centre in North India over 18 months. Data of hospitalized patients who admitted with dengue fever diagnosis in a peak season was collected in the form of quality indicators as described by the WHO-2009 guideline on dengue. Resident physicians were then given appropriate training about the guideline during the off season. Data of new dengue patients in next peak season after resident training was collected and compared with the baseline by standard statistical tests., Results: The post-intervention compliances of all components increased (total mean score by giving one point to each of the quality indicators reached 7.9 from 6.4). The compliance to individual indicator also increased: the admission criteria (baseline, 44% to post-intervention, 52%, p = 0.37), classification criteria (91.7% to 96%, p = 0.33), correct staging/triage (42.9% to 86%, p <0.001), vitals monitoring (85.7% to 92%, p = 0.28), correct usage of bolus fluids (34.3% to 69.5%, p <0.001), crystalloid as choice of fluid (100% in both groups), proper fluid titration (26.2% to 56%, p <0.001), hematocrit monitoring (95.2% to 98%, p = 0.42), platelet transfusion when indicated (65.5% to 58%, p = 0.39), antibiotic use when required (61.5% to 80%, p = 0.03), and discharge criteria (100% in both groups). The mortality decreased from 7.1% (baseline) to zero (post-intervention). The median duration of hospital stay also reduced by 1 day., Conclusion: The study affirms that the compliance to WHO guideline on dengue management in India can be further improved by regular physician training on the guideline. Simultaneously, this educational intervention not only improves patient outcomes but also direct proper resource utilization especially platelet transfusion and antibiotic use. Furthermore, every hospital/institute should have an internal quality improvement program like this to improve the management of dengue patients. Future studies are needed to understand various barriers to 100% implementation of the guideline., (© Journal of the Association of Physicians of India 2011.)

A 50-year-old male presented to us with features of diabetic ketoacidosis which was managed with adequate hydration and insulin therapy. His routine laboratory investigation revealed transaminitis, acute kidney injury and pancytopenia. Further evaluation for hematological and biochemical derangements uncovered positive dengue test (NS1 antigen and polymerase chain reaction assay). Patient distinctively reported no history of fever and remained afebrile during the course of illness. We report this case to highlight the possibility of afebrile dengue in endemic areas., (© Journal of the Association of Physicians of India 2011.)

Thrombocytopenia is common in patients with dengue virus (DENV) infections. With a focus on understanding the possible mechanism of thrombocytopenia in DENV infections we described a direct correlation between activation and depletion of platelets in patients. Our data showed a sharp decrease in platelet counts at day 4 of fever in patients. The high DENV genome copies in platelets correlated directly with the elevated platelet activation along with increased binding of complement factor C3 and IgG on their surface at day 4. Recovery in platelet count was observed on day 10 through day 6 and 8 with simultaneous decrease in platelet activation markers. Further, our in vitro data supported the above observations describing a concentration-dependent increase in platelet activation by DENV serotype-2. The high copy number of DENV2 genome in the platelet pellet correlated directly with platelet activation, microparticle generation and clot formation. Furthermore the DENV2-activated platelets were phagocytosed in large numbers by the monocytes. The DENV2-mediated lysis and clearance of platelets were abrogated in presence of platelet activation inhibitor, prostacyclin. These observations collectively suggest that platelet activation status is an important determinant of thrombocytopenia in dengue infections. A careful strategy of inactivation of platelets may rescue them from rapid destruction during DENV infections., Competing Interests: The authors declare no competing financial interests.

Infection with dengue virus (DENV) is the most rapidly spreading mosquito-borne viral disease in the world. The clinical spectrum of dengue, caused by any of the four serotypes of DENV, ranges from mild self-limiting dengue fever to severe dengue, in the form dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Increased rates of hospitalization due to severe dengue, during outbreaks, result in massive economic losses and strained health services. In the absence of specific antiviral therapy, control of transmission of DENV by vector management is the sole method available for decreasing dengue-associated morbidity. Since vector control strategies alone have not been able to satisfactorily achieve reduction in viral transmission, the implementation of a safe, efficacious and cost-effective dengue vaccine as a supplementary measure is a high public health priority. However, the unique and complex immunopathology of dengue has complicated vaccine development. Dengue vaccines have also been challenged by critical issues like lack of animal models for the disease and absence of suitable markers of protective immunity. Although no licensed dengue vaccine is yet available, several vaccine candidates are under phases of development, including live attenuated virus vaccines, live chimeric virus vaccines, inactivated virus vaccines, subunit vaccines, DNA vaccines and viral-vectored vaccines. Although some vaccine candidates have progressed from animal trials to phase II and III in humans, a number of issues regarding implementation of dengue vaccine in countries like India still need to be addressed. Despite the current limitations, collaborative effects of regulatory bodies like World Health Organization with vaccine manufacturers and policy makers, to facilitate vaccine development and standardize field trials can make a safe and efficacious dengue vaccine a reality in near future.

Dengue virus (DV) is the most widespread arbovirus, being endemic in over 100 countries, and is estimated to cause 50 million infections annually. Viral factors, such as the genetic composition of the virus strain can play a role in determining the virus virulence and subsequent clinical disease severity. Virus vector competence plays an integral role in virus transmission and is a critical factor in determining the severity and impact of DV outbreaks. Host genetic variations in immune-related genes, including the human leukocyte antigen, have also been shown to correlate with clinical disease and thus may play a role in regulating disease severity. The host's immune system, however, appears to be the primary factor in DV pathogenesis with the delicate interplay of innate and acquired immunity playing a crucial role. Although current research of DV pathogenesis has been limited by the lack of an appropriate animal model, the development of DV therapeutics has been a primary focus of research groups around the world. In the past decade advances in both the development of vaccines and anti-virals have increased in dramatically. This review summarises the current understanding of viral, vector and host factors which contribute to dengue virus pathogenesis and how this knowledge is critically important in the development of pharmaceutical interventions., (Copyright © 2012. Published by Elsevier Inc.)

A case of mixed infection due to leptospira, dengue and hepatitis E is reported. Similar clinical presentation and simultaneous transmission of these diseases during rainy season can cause substantial misdiagnosis. Serological tests play a mainstay in diagnosis. Early detection by appropriate serological tests and institution of therapy is crucial and life saving.

Aedes aegypti mosquitoes are common vectors for dengue virus and chikungunya virus. In areas where both viruses cocirculate, they can be transmitted together. During a dengue outbreak in Delhi in 2006, 17 of 69 serum samples were positive for chikungunya virus by reverse transcription-PCR; 6 samples were positive for both viruses.

Background: A major DHF outbreak occurred in Delhi in 1996. Following this another outbreak was reported in the year 2003. In the years 2004 and 2005, though no outbreak was reported, a definitely higher number of samples were received in the virology laboratory of A.I.I.M.S. from suspected cases of dengue infection. This study was designed to compare the serological and virological profiles of confirmed dengue cases in the years 2003, 2004 and 2005., Results: Out of 1820 serum samples received from suspected cases in all three years, 811 (44.56%) were confirmed as dengue infection serologically. Out of these confirmed dengue cases maximum cases, in all three years, were seen in the age group 21-30 years. There was an increase in the number of samples received in the post monsoon period (September to November) with a peak in the second and third week of October. More samples were received from DHF cases in the year 2005 than 2004 and 2003. All four dengue serotypes were seen co-circulating in the year 2003, followed by complete predominance of dengue serotype 3 in 2005., Conclusion: Epidemiology of dengue is changing rapidly in Delhi. Dengue infections are seen every year thus making it an endemic disease. After co-circulation of all serotypes in 2003, now dengue serotype 3 is emerging as the predominant serotype.

Background and Objectives: Neurological manifestations of dengue fever are rarely reported during acute illness and clinical presentation commonly observed is of acute encephalitis or one of the post-infectious immune mediated manifestations. We describe a case of dengue fever having mild encephalopathy and papilledema at presentation., Case Report: Twenty-year-old female presented with fever, headache and vomiting. On examination she did not have classical signs of dengue fever and was found to have bilateral papilledema on fundus examination. Detailed work-up did not reveal any other cause of papilledema. Diagnosis of dengue fever was established by blood IgM antibody test on day 7 of illness. Retrospective analysis of CSF (drawn on day 5 of illness) by RT-PCR assay showed a characteristic band of dengue-3 virus. Papilledema was transient and subsided following symptomatic treatment. The patient recovered from acute illness and follow-up was unremarkable., Conclusion: Especially in dengue endemic areas, in the patients having acute febrile illness with subtle signs and symptoms suggestive of CNS involvement, dengue virus infection should also be ruled out early in the clinical course.

Dengue, an important human arboviral infection, is endemic in many parts of India. Outbreaks are now reported quite frequently from different parts of the country. We report here findings of serological investigation of serum samples received during a recent outbreak of dengue infection in Delhi from September to November 2003. Acute phase serum samples from suspected cases of dengue infection (including dengue fever, dengue haemorrhagic fever and dengue shock syndrome) of duration 5 days, were tested for dengue specific IgM antibodies by enzyme linked immuno sorbant assay (ELISA). Of the 874 serum samples tested, 456 (52.3% ) were positive for dengue specific IgM antibodies. The maximum number of positive cases was reported in October, peaking in second and third week. Patients in the age group of 21-30 yr accounted for 34.2 per cent of positive cases. This outbreak was less severe as compared to the previous 1996 outbreak and showed a shift towards an older age group.

Dengue fever (DF) and dengue hemorrhagic fever (DHF) are major public health problems in India. During the period following an epidemic, a study was carried out using virological and serological tests for confirmation of suspected cases of dengue virus infection in fever cases presenting to the All India Institute of Medical Sciences. Serum samples of suspected DF/DHF cases were processed from January to December 1997. In 37 samples from patients with fever of less than 5-day duration, received on ice, virus isolation was attempted in C6/36 clone of Aedes albopictus cell line, followed by indirect fluorescent antibody staining with monoclonal antibodies to dengue viruses 1 to 4. One hundred and forty-three serum samples from patients with more than 5 days fever were tested for dengue specific IgM antibody by either MAC-ELISA or a rapid immunochromatographic assay. Dengue virus type 1 was demonstrated by culture in 8 (21.6%) of 37 serum samples and IgM antibody could be detected in 42 (29.4%) of the 143 serum samples by the serological methods. The peak of dengue virus infection was seen from September to November 1997.

An outbreak of dengue hemorrhagic fever/dengue shock syndrome (DHS/DSS) occurred in 1996 in India in and near Delhi. The cause was confirmed as dengue virus type 2, by virus cultivation and indirect immunofluorescence with type-specific monoclonal antibodies. This is the largest such outbreak reported from India, indicating a serious resurgence of dengue virus infection. [ABSTRACT FROM AUTHOR]

Background: Dengue epidemic has become a serious emerging infectious diseases and a major public health problem in India. Some of the Dengue Fever patients present with mild symptoms and some present with severe manifestations involvement of vital organs responsible for high morbidity and mortality. Exact pathogenetic mechanism of severity is still not clearly understood. It is hypothesized that high level of serum TNF-alpha, IL-6 and soluble Thrombomodulin are associated with severe Dengue Fever, bleeding and shock. Methods & Materials: It was a prospective observational case control study conducted at tertiary care hospital. All cases with either +ve for IgM or NS1 were enrolled for the study. Patients with shock were enrolled as cases and patients without shock were enrolled as control. We estimated serum TNF-alpha, IL-6 and sTM among shock and non-shock patients at the time of 1st visit and followed up till death, discharge or 14 days. Results: Thirty Dengue patients with shock were enrolled as cases and 50 patients without shock were enrolled as control. Mean value of sTM was 15.8±6.4 ng/ml among shock and 6.4±4.4 ng/ml among non-shock and their difference was highly statistically significant (p<0.001). About 21 cases (70%) from shock and 3 cases (6%) from noshock were found to have high sTM level (>15 ng/ml) and their association was also found to have statistically significant (<0.001). About 12 cases (39.9%) from shock and 10 cases (20%) from non-shock were found to have elevated TNF-alpha (>100 pg/ml). About 11 cases (37.7%) from shock and 15 cases (30%) from non-shock were found to have elevated IL-6 (>100 pg/ml). However serum level of TNF-alpha and IL-6 were not found to be significant. We found 22% incidence (11) of shock among Dengue Fever and found to have significant rise of sTM (13±4.67) and IL-6 (136±76.4) at the time of 1st visit and they all had normal blood pressure but subsequently developed shock. Conclusion: In our study we found high serum soluble Thrombomoduline level (sTM) and IL-6 were both predictor and risk factor for the development of severe Dengue Fever and shock. [ABSTRACT FROM AUTHOR]