Hypericum japonicum: a Double-Headed Sword to Combat Vector Control and Cancer.

Mosquito control with naturally derived herbal insecticides has gained much momentum, with the increased insecticide resistance of vectors and the multiple infectious diseases spread by them. Yet, recent studies also suggest that mosquitoes could probably transmit some cancerous cells or cancer-causing viruses from one individual to another between their blood meals. The current research thus focused on the screening and characterization of novel plants with both mosquitocidal and anticancerous properties. Accordingly, different solvent extracts of Hypericum japonicum, a key plant in Chinese medicine, were screened for its larvicidal efficacy using the fourth instar larvae of Aedes aegypti (major vector of Dengue and chikungunya). Methanolic extracts of the plant showed effective larvicidal property with LC50 7.37 ppm and LC9011.59 ppm values. The anticancerous property of the plant extract was also evaluated by in vitro cytotoxicity assay against Daltons Lymphoma Ascites (DLA) cells. The results indicated that H. japonicum plant extracts at very low concentrations of LC500.95 ppm and LC901.85 ppm were potent cytotoxic agents. To the best of our knowledge, this is the first and the foremost report of Hypericum japonicum as a potent mosquitocidal and anticancerous agent. Identification and characterization of such plant-derived bioactive plants thus could serve as a double-headed sword against the spread of infectious diseases and cancer.

Enhanced sensing of dengue virus DNA detection using O2 plasma treated-silicon nanowire based electrical biosensor.

Dengue Virus (DENV) has become one of the most serious arthropod-borne viral diseases, causing death globally. The existing methods for DENV detection suffer from the late stage treatment due to antibodies-based detection which is feasible only after five days following the onset of the illness. Here, we demonstrated the highly effective molecular electronic based detection utilizing silicon nanowire (SiNW) integrated with standard complementary metal-oxide-semiconductor (CMOS) process as a sensing device for detecting deoxyribonucleic acid (DNA) related to DENV in an early stage diagnosis. To transform the fabricated devices as a functional sensing element, three-step procedure consist of SiNW surface modification, DNA immobilization and DNA hybridization were employed. The detection principle works by detecting the changes in current of SiNW which bridge the source and drain terminal to sense the immobilization of probe DNA and their hybridization with target DNA. The oxygen (O2) plasma was proposed as an effective strategy for increasing the binding amounts of target DNA by modified the SiNW surface. It was found that the detection limit of the optimized O2 plasma treated-SiNW device could be reduced to 1.985 × 10-14 M with a linear detection range of the sequence-specific DNA from 1.0 × 10-9 M to 1.0 × 10-13 M. In addition, the developed biosensor device was able to discriminate between complementary, single mismatch and non-complementary DNA sequences. This highly sensitive assay was then applied to the detection of reverse transcription-polymerase chain reaction (RT-PCR) product of DENV-DNA, making it as a potential method for disease diagnosis through electrical biosensor.

A Multi-centric, Double-blind, Placebo-controlled, Randomized, Prospective Study to Evaluate the Efficacy and Safety of Carica papaya Leaf Extract, as Empirical Therapy for Thrombocytopenia associated with Dengue Fever.

INTRODUCTION: Dengue is a rapidly expanding global health problem. Approximately 2.5 billion people live in dengue-risk regions with about 100 million new cases each year worldwide. The cumulative dengue diseases burden has attained an unprecedented proportion in recent times with sharp increase in the size of human population at risk. The management of dengue virus infection is essentially supportive and symptomatic and no specific treatment is available for increasing the fallen platelets, which have a significant role in causing the mortality of dengue patient.This study was conducted to evaluate the platelet increasing efficacy of Carica papaya leaf extract (CPLE) in patients with dengue fever (DF). AIM: The administration of Carica papaya leaf extract should significantly increase the platelet count in cases of thrombocytopenia associated with dengue, preventing the patient to go in DHF or DSS conditions. MATERIAL AND METHODS: A Multi-centric, Double blind, Placebo controlled, Randomized, prospective study was conducted in 300 patients across 5 centres', to evaluate the Efficacy and Safety of Carica Papaya Leaf Extract, as empirical therapy for thrombocytopenia associated with dengue fever. The subjects were randomized into two groups, as control and intervention group. Both the groups were managed by the standard management guidelines for dengue except steroid administration. In addition to this, the intervention group received CPLE tablet three times daily for five days. All of them were followed daily with platelet monitoring. This study has been registered in the clinical trial registry-India (CTRI Registration number: CTRI/2015/05/005806). RESULTS: The results indicate that CPLE had significant increase(p< 0.01) in the platelet count over the therapy duration, in dengue fever patients, confirming CPLE accelerates the increase in platelet count compared to the control group. There were few adverse events related to GI disturbance like nausea and vomiting which were similar in both groups. CONCLUSIONS: Thus this study concluded that Carica papaya leaf extract (CPLE) does significantly increase the platelet count in patients with thrombocytopenia associated with dengue with fewer side effects and good tolerability.

Manifestações clínicas na dengue: diagnóstico laboratorial / Clinical manifestations in dengue: laboratory diagnosis

A dengue é uma doença infecciosa de evolução aguda, transmitida por vírus (RNA vírus). Infecta o homem através da picada do inseto fêmea Aedes aegypti. Seus sinais e sintomas são variáveis, com formas oligossintomáticas, formas clássicas (febris) e formas graves hemorrágicas, podendo até apresentar síndrome cardiovascular hipovolêmica. O diagnóstico envolve critérios clínico-laboratoriais. O diagnóstico sorológico tem fundamental importância na classificação de infecção primária ou secundária, já que a dengue hemorrágica surge com maior frequência nas infecções secundárias. O isolamento do vírus é geralmente realizado para fins de pesquisa ou epidemiológicos. As epidemias ocorrem principalmente no verão, durante ou após períodos chuvosos.

The dengue is an infectious disease of acute evolution transmitted by virus (RNA virus), infecting humans through the bite of the Aedes aegypti female insect. Presenting signs and symptoms variables with oligosymptomatic forms, classical forms (fever) and severe hemorrhagic form (DHF), this can lead to cardiovascular hypovolemic syndrome. The diagnoses of dengue disease involves clinical and laboratory criteria. Serological diagnosis has fundamental importance in the classification of primary or secondary infection, since DHF appears most often in secondary infections.Virus isolation is usually carried out for research or epidemiological studies. Epidemics occur mainly in the summer, during or after rainy periods.

Clinical implications and treatment of dengue.

Dengue is a common pathogenic disease often proving fatal, more commonly affecting the tropics. Aedes mosquito is the vector for this disease, and outbreaks of dengue often cause mass damage to life. The current review is an effort to present an insight into the causes, etiology, symptoms, transmission, diagnosis, major organs affected, mitigation and line of treatment of this disease with special emphasis on drugs of natural origin. The disease has a potential to spread as an endemic, often claiming several lives and thus requires concerted efforts to work out better treatment options. Traditional medicine offers an alternative solution and could be explored as a safer treatment option. Development of a successful vaccine and immunization technique largely remains a challenge and a better antiviral approach needs to be worked out to complement the supportive therapy. No single synthetic molecule has found to be wholly effective enough to offer curative control and the line of treatment mostly utilizes a combination of fluid replacement and antipyretics-analgesics like molecules to provide symptomatic relief.

Serotype-specific identification of Dengue virus by silicon nanowire array biosensor.

In this work, we demonstrated a silicon nanowire (SiNW) biosensing platform capable of simultaneously identifying different Dengue serotypes on a single sensing chip. Four peptide nucleic acids (PNAs), specific to each Dengue serotypes (DENV-1 to DENV-4), were spotted on different areas of the SiNW array surface, and the covalently immobilized PNA probes were then interacted with different Dengue serotypes target to establish the specificity of detection. Detection scheme is based on the changes in resistances due to accumulation of negative charges contributed by the hybridized DNA target. The results show that resistance changes only occur in regions where the Dengue target hybridizes with its complementary probe. What is more, a mixture of two different Dengue serotypes obtained from a one-step duplex RT-PCR was applied to the multiplex SiNW surface to validate SiNW capability to identify multiple Dengue serotypes on a single sensing platform. Through this study, we have established the multiplex SiNW biosensor as a promising device to detect multiple Dengue infections with high specificity.

Mentha x piperita L. y su potencial en el control del vector del dengue (Aedes aegypti L.). Revisión / Mentha x piperita L. and its potential in controlling the dengue vector (Aedes aegypti L.). Review

El dengue es una de las enfermedades tropicales más fatales, afectando a 50-100 millones de personas al año. Endémico en más de cien países y transmitido a los humanos por la picadura subcutánea del mosquito Aedes aegypti L., puede desencadenar los síntomas de una gripe normal, episodios de fiebre hemorrágica y shock. Las medidas de erradicación del vector adoptadas por las autoridades sanitarias mundiales, como la eliminación del hábitat y uso de repelentes y/o insecticidas) no logran frenar la propagación del dengue, ya extendido por las áreas no endémicas, como Portugal. Se sabe que el mosquito es vulnerable a algunos productos vegetales y varios estudios los han considerado como alternativas potenciales a los productos químicos sintéticos, a la que el mosquito se ha vuelto resistente. Algunas de esas plantas pertenecen a la flora nativa, espontánea y subespontánea, estando accesibles a un gran número de personas. Es el caso de la Mentha x piperita., elegida como ejemplo por las propiedades insecticidas y repelentes de su aceite esencial (AU)

Dengue is one of the most fatal tropical diseases which affect 50-100 billion people/ year. It is endemic in more than 100 countries and transmitted to humans by the subcutaneous bite of the Aedes aegypti L., it can trigger different symptoms like normal flu, hemorrhagic fever and shock. The eradication of the vector adopted by global health authorities, like the elimination of its habitat, the use of repellents and/or insecticides, fail to curb the spread of the disease, which are expanding into non-endemic areas (Portugal). Vector is vulnerable to some herbal products and scientific studies have considered them as a potential alternative to the synthetic ones in use, to which the mosquito may become resistant. Some of these herbs belong to native, spontaneous and subspontaneous flora, resulting accessible to the population. Mentha x piperita L., was chosen as example due to insecticide and repellent properties of it essential oil (AU)

Dengue virus detection using impedance measured across nanoporous alumina membrane.

The prevalence of dengue around the world makes it critical to develop a simple diagnostic device that can be easily handled by end users and provides fast results. In this paper, we described the use of a small and thin piece of alumina membrane, 60 µm thick and 13 mm in diameter as the sensing platform for the detection of dengue infection. The electrochemical setup is simplified by using the membrane as both the working and the counter electrode. This is achieved by coating both sides of the membrane with a submicron layer of platinum. Electrochemical impedance spectroscopy was utilized for the characterization of the immunosensor as well as the acquisition of data. The change in the pore resistance of the membrane displayed a good correlation with the concentration of the dengue 2 and dengue 3 viruses in plaque forming unit (PFU mL⁻¹), giving detection limit of 0.230 PFU mL⁻¹ and 0.710 PFU mL⁻¹ respectively. This thin piece of membrane sensor, coupled with the simple electrochemical setup, fast detection time of 40 min and high sensitivity, showed potential to be developed into a disposable point-of-care diagnostic tool for clinical uses.

Electrochemical impedance spectroscopy characterization of nanoporous alumina dengue virus biosensor.

The Faradaic electrochemical impedance technique is employed to characterize the impedance change of a nanoporous alumina biosensor in response towards the specific binding of dengue serotype 2 (Denv2) viral particles to its serotype 2-specific immunoglobulin G antibody within the thin alumina layer. The optimal equivalent circuit model that matches the impedimetric responses of the sensor describes three distinct regions: the electrolyte solution (R(s)), the porous alumina channels (including biomaterials) (Q(1), R(1)) and the conductive electrode substrate layer (Q(2), R(2)). Both channel resistance R(1) and capacitance Q(1) change in response to the increase of the Denv2 virus concentration. A linear relationship between R(1) and Denv2 concentration from 1 to 900 plaque forming unit per mL (pfu mL(-1)) can be derived using Langmuir-Freundlich isotherm model. At 1pfu mL(-1) Denv2 concentration, R(1) can be distinguished from that of the cell culture control sample. Moreover, Q(1) doubles when Denv2 is added but remains unchanged in the presence of two other non-specific viruses - West Nile virus and Chikungunya virus indicates biosensor specificity can be quantitatively measured using channel capacitance.

Development of an electrochemical membrane-based nanobiosensor for ultrasensitive detection of dengue virus.

A sensitive membrane-based electrochemical nanobiosensor is developed for the detection of dengue type 2 virus (DENV-2) using nanoporous alumina-modified platinum electrode. Its sensing mechanism relies on the monitoring of electrode's Faradaic current response toward redox probe, ferrocenemethanol, which is sensitive toward the formation of immune complexes within the alumina nanochannels. Anti-DENV-2 monoclonal antibody (clone 3H5, isotype IgG) is used as the biorecognition element in this work. The stepwise additions of antibody, bovine serum albumin (BSA) and DENV-2 are characterized by differential pulse voltammetry (DPV). A low detection limit of 1 pfu mL(-1) with linear range from 1 to 10(3) pfu mL(-1) (R(2)=0.98) can be achieved by the nanobiosensor. The nanobiosensor is selective toward DENV-2 with insignificant cross reaction with non-specific viruses, Chikungunya virus, West Nile virus and dengue type 3 virus (DENV-3). Relative standard deviation (RSD) for triplicate analysis of 5.9% indicates an acceptable level of reproducibility. The first direct quantitation of DENV-2 concentration in whole mosquito vector is demonstrated using this electrochemical nanobiosensor.

Development of dengue virus plaques under serum-free overlay medium.

An improved plaque assay for dengue virus was developed utilizing baby hamster kidney (BHK-21) cells initially grown in shaker culture. Different media preparations were tested for uniform and fast formation of BHK-21 cell sheets. Several overlay formulas were tested to develop a rapid plaque assay in 6- and 24-well plastic plates. The best results were obtained utilizing Eagle minimal essential medium (pH 7.2 to 7.4) supplemented with 1 mg of NaHCO3 per ml and 5% newborn calf serum for the formation of cell monolayers after 8 to 24 h of incubation at 37 degrees C. Serum-free Eagle minimal essential medium supplemented with 1% methylcellulose and buffered with 10 mM N-2-hydroxyethyl piperazine-N'-2-ethanesulfonic acid (pH 7.4 to 7.6) was used as an overlay medium. This system allowed for plaque formation after 3 days of incubation of dengue type 2 virus and after 4 days for dengue type 1 and 4 viruses.