Viremia and clinical manifestations in acute febrile patients of Chikungunya infection during the 2016 CHIKV outbreak in Delhi, India.

Background: Chikungunya virus (CHIKV) is an infectious agent that caused several outbreaks among different countries and affected approximately 1.3 million Indian populations. It is transmitted by Aedes mosquito-either A. albopictus or A. aegypti. Generally, the clinical manifestations of CHIKV infection involve high-grade fever, joint pain, skin rashes, headache, and myalgia. The present study aims to investigate the relationship between the CHIKV virus load and clinical symptoms of the CHIKV infection so that better patient management can be done in the background of the CHIKV outbreak as there is no licensed anti-viral drug and approved vaccines available against CHIKV. Methods: CHIKV RTPCR positive samples (n = 18) (Acute febrile patients having D.O.F ≤ 7 days) were taken for the quantification of CHIKV viremia by Real-Time PCR. Clinical features of the febrile patients were recorded during the collection of blood samples. Results: The log mean virus load of 18 RT-PCR-positive samples was 1.3 × 106 copies/mL (1.21 × 103-2.33 × 108 copies/mL). Among the observed clinical features, the log mean virus load (CHIKV) of the patients without skin rash is higher than in the patients with skin rash (6.61 vs 5.5, P = 0.0435). Conclusion: The conclusion of the study was that the patients with skin rashes had lower viral load and those without skin rashes had higher viral load.

Mutation of conserved histidine residues of dengue virus envelope protein impairs viral like particle maturation and secretion.

Dengue virus (DENV) envelope protein plays crucial role in virus entry and maturation of virus during infection. Maturation of DENV occurs in the trans Golgi network at slightly acidic pH which is close to pKa of histidine. When exposed to the acidic environment of the late secretory pathway, dengue virus particles go through a significant conformational change, whereby interactions of structural proteins envelope (E) and prM proteins are reorganised and enable furin protease to cleave prM resulting in mature virus. In order to study the role of histidine of E protein in DENV maturation, we mutated 7 conserved histidine residues of envelope protein and assessed the percent of budding using viral like particle (VLP) system. Histidine mutants; H144A, H244A, H261A and H282A severely disrupted VLP formation without any significant change in expression in cell and its oligomerization ability. Treatment with acidotropic amine reversed the defect for all 4 mutants suggesting that these histidines could be involved in maturation and release. Over expression of capsid protein slightly enhanced VLP release of H244A and H261A. Similarly, furin over expression increased VLP release of these mutants. Co-immunoprecipitation studies revealed that prM and E interaction is lost for H244A, H261A and H282A mutants at acidic pH but not at neutral pH indicating that they could be involved in histidine switch during maturation at acidic pH. Detailed analysis of the mutants could provide novel insights on the interplay of envelop protein during maturation and aid in target for drug development.

Artemisinin resistance in P. falciparum: probing the interacting partners of Kelch13 protein in parasite.

OBJECTIVES: Artemisinin (ART) resistance in Plasmodium is threatening the artemisinin combination therapies-the first line of defence against malaria. ART resistance has been established to be mediated by the Plasmodium Kelch13 (PfK13) protein. For the crucial role of PfK13 in multiple pathways of the Plasmodium life cycle and ART resistance, it is imperative that we investigate its interacting partners. METHODS: We recombinantly expressed PfK13-p (Bric a brac/Poxvirus and zinc finger and propeller domains), generating anti-PfK13-p antibodies to perform co-immunoprecipitation assays and probed PfK13 interacting partners. Surface plasmon resonance and pull-down assays were performed to establish physical interactions of representative proteins with PfK13-p. RESULTS: The co-immunoprecipitation assays identified 17 proteins with distinct functions in the parasite life cycle- protein folding, cellular metabolism, and protein binding and invasion. In addition to the overlap with previously identified proteins, our study identified 10 unique proteins. Fructose-biphosphate aldolase and heat shock protein 70 demonstrated strong biophysical interaction with PfK13-p, with KD values of 6.6 µM and 7.6 µM, respectively. Additionally, Plasmodium merozoite surface protein 1 formed a complex with PfK13-p, which is evident from the pull-down assay. CONCLUSION: This study adds to our knowledge of the PfK13 protein in mediating ART resistance by identifying new PfK13 interacting partners. Three representative proteins-fructose-biphosphate aldolase, heat shock protein 70, and merozoite surface protein 1-demonstrated clear evidence of biophysical interactions with PfK13-p. However, elucidation of the functional relevance of these physical interactions are crucial in context of PfK13 role in ART resistance.

Novel development of Lecanicillium lecanii-based granules as a platform against malarial vector Anopheles culicifacies.

Mosquitoes are infectious vectors for a wide range of pathogens and parasites thereby transmitting several diseases including malaria, dengue, Zika, Japanese encephalitis and chikungunya which pose a major public health concern. Mostly synthetic insecticides are usually applied as a primary control strategy to manage vector-borne diseases. However excessive and non-judicious usage of such chemically derived insecticides has led to serious environmental and health issues owing to their biomagnification ability and increased toxicity towards non-target organisms. In this context, many such bioactive compounds originating from entomopathogenic microbes serve as an alternative strategy and environmentally benign tool for vector control. In the present paper, the entomopathogenic fungus, Lecanicillium lecanii (LL) was processed to make the granules. Developed 4% LL granules have been characterized using the technique of Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). The developed formulation was also subjected to an accelerated temperature study at 40 °C and was found to be stable for 3 months. Further, GCMS of the L. lecanii was also performed to screen the potential biomolecules present. The developed formulation was found to be lethal against Anopheles culicifacies with an LC50 value of 11.836 µg/mL. The findings from SEM and histopathology also substantiated the mortality effects. Further, the SEM EDX (energy dispersive X-ray) studies revealed that the treated larvae have lower nitrogen content which is correlated to a lower level of chitin whereas the control ones has higher chitin content and healthy membranes. The developed LL granule formulation exhibited high toxicity against Anopheles mosquitoes. The granule formulations can be used as an effective biocontrol strategy against malaria-causing mosquitoes.

Efficient generation and characterization of chimeric dengue viral-like particles.

Viral-like particles (VLPs) because of their non-infectious and high immunogenic properties have important applications in diagnostics, drug delivery, and vaccine production. They also serve as an attractive model system to study virus assembly and fusion processes. Unlike other flaviviruses, Dengue virus (DENV) is not very efficient in the production of VLPs on the expression of DENV structural proteins. On the other hand, the stem region and transmembrane region (TM) of G protein of Vesicular Stomatitis virus (VSV) alone are sufficient for budding. Here we generated chimeric VLPs replacing regions of stem and transmembrane domain (STEM) or only transmembrane domain (TM) of E protein of DENV-2 with corresponding regions of VSV G protein. Both chimeric proteins secreted VLPs at higher levels than the wild type (2-4 folds) without any significant change in the expression in the cell. Chimeric VLPs could be recognized by a conformational monoclonal antibody, 4G2. They were also found to interact with dengue-infected patient sera effectively thus implying that their antigenic determinants are preserved. In addition, they were able to bind to its putative receptor, heparin with similar affinity as the parent counterpart thus retaining its functional property. However, cell-cell fusion revealed that there is no significant increase in the fusion ability of chimeras as compared to the parent clone, whereas VSV G protein displayed high cell-cell fusion activity. Overall, this study suggests that chimeric dengue VLPs can be taken forward for their likely potential as vaccine production and serodiagnosis.

Molecular Tools for Early Detection of Invasive Malaria Vector Anopheles stephensi Mosquitoes.

Reports of the expansion of the Asia malaria vector Anopheles stephensi mosquito into new geographic areas are increasing, which poses a threat to the elimination of urban malaria. Efficient surveillance of this vector in affected areas and early detection in new geographic areas is key to containing and controlling this species. To overcome the practical difficulties associated with the morphological identification of immature stages and adults of An. stephensi mosquitoes, we developed a species-specific PCR and a real-time PCR targeting a unique segment of the second internal transcribed spacer lacking homology to any other organism. Both PCRs can be used to identify An. stephensi mosquitoes individually or in pooled samples of mixed species, including when present in extremely low proportions (1:500). This study also reports a method for selective amplification and sequencing of partial ribosomal DNA from An. stephensi mosquitoes for their confirmation in pooled samples of mixed species.

Tandem duplication of a genomic region encoding glutathione S-transferase epsilon-2 and -4 genes in DDT-resistant Anopheles stephensi strain from India.

The glutathione S-transferases (GST) genes are a multigene family of enzymes involved in the metabolism of endogenous and xenobiotic compounds by catalysing the conjugation of the reduced form of glutathione to the substrate. The epsilon class of GST (GSTe), unique to arthropods, is known to be involved in the detoxification process of several classes of insecticides, and GSTe2 in particular is known to have DDT dehydrochlorinase activity. This communication reports a tandem duplication of a genomic region encoding GSTe2 and GSTe4 genes in a laboratory-colonized DDT-resistant Anopheles stephensi. We identified duplication breakpoints and the organization of gene duplication through Sanger sequencing performed on long-PCR products. Manual annotation of sequences revealed a tandemly-arrayed duplication of a 3.62 kb segment of GST epsilon gene clusters comprised of five genes: a partial GSTe1, GSTe2, GSTe2-pseudogene, GSTe4 and partial GSTe5, interconnected by a conserved 2.42 kb DNA insert segment major part of which is homologous to a genomic region located on a different chromosome. The tandemly duplicated array contained a total of two GSTe2 and three GSTe4 functional paralog genes. Read-depth coverage and split-read analysis of Illumina-based whole-genome sequence reads confirmed the presence of duplication in the corresponding region of the genome. The increased gene dose in mosquitoes as a result of the GSTe gene-duplication may be an adaptive process to increase levels of detoxifying enzymes to counter insecticide pressure.

Evaluation of intron-1 of odorant-binding protein-1 of Anopheles stephensi as a marker for the identification of biological forms or putative sibling species.

BACKGROUND: Anopheles stephensi, an invasive malaria vector, has been reported to have three biological forms identifiable mainly based on the number of ridges present on the egg's floats. Recently, the first intron of the odorant-binding protein-1 (AsteObp1) has been introduced as a molecular marker for the identification of these forms, and based on this marker, the presence of three putative sibling species (designated as species A, B and C) has been proposed. However, there is no data on the association of proposed markers with biological form or putative species on field populations. METHODS: Field collected and laboratory-reared An. stephensi were characterized for biological forms based on the number of ridges on the egg's float. DNA sequencing of the partial AsteObp1 gene of An. stephensi individuals were performed by Sanger's method, either directly or after cloning with a plasmid vector. Additionally, AsteObp1 sequences of various laboratory lines of An. stephensi were retrieved from a public sequence database. RESULTS: AsteObp1 intron-1 in Indian An. stephensi populations are highly polymorphic with the presence of more than 13 haplotypes exhibiting nucleotides as well as length-polymorphism (90-to-121 bp). No specific haplotype or a group of closely related haplotypes of intron-1 was found associated with any biological form identified morphologically. High heterozygosity for this marker with a low inbreeding coefficient in field and laboratory populations indicates that this marker is not suitable for the delimitation of putative sibling species, at least in Indian populations. CONCLUSIONS: AsteObp1 cannot serve as a marker for identifying biological forms of An. stephensi or putative sibling species in Indian populations.

Utilization of the castor seed cake (biowaste) for mosquito vector control.

The present work is related to the utilization of castor (Ricinus communis) seed cake, biowaste produced during the oil extraction of castor seeds, as efficient mosquitocidal composition against Aedes aegypti and Anopheles culicifacies. The efficacy of coil formulations was evaluated in the Peet Grady chamber and resulted in 90% and 100% knocked down and mortality against A. aegypti and A. Culicifacies, respectively. Further heavy metals' (Cr, Pb, Co, As, Cd, Cu, Mn, and Zn) analysis of the coil was performed using Inductively Coupled Plasma mass spectrometry and was compared with commercially available mosquito repellent coil. Heavy metal analysis revealed that commercial repellent coil had a higher content of heavy metals than the castor seed cake coil. Finding of the present research study indicates that castor seed cake coil has the potential to be used in mosquito vector control. Castor seed cake coil formulation will also open up avenues in future for sustainable utilization of the biowaste.

A new knockdown resistance (kdr) mutation, F1534L, in the voltage-gated sodium channel of Aedes aegypti, co-occurring with F1534C, S989P and V1016G.

BACKGROUND: Aedes aegypti is a primary vector of dengue, chikungunya and Zika infections in India. In the absence of specific drugs or safe and effective vaccines for these infections, their control relies mainly on vector control measures. The emergence of insecticide resistance in vectors, especially against pyrethroids, is a serious threat to the insecticide-based vector control programme. This study reports the presence of multiple knockdown resistance (kdr) mutations present in an Ae. aegypti population from Bengaluru (India), including a new mutation F1534L. METHODS: Aedes aegypti collected from Bengaluru were subjected to insecticide susceptibility tests with DDT, deltamethrin and permethrin. The DNA sequencing of partial domain II, III and IV of the voltage-gated sodium channel (VGSC) was performed to screen kdr mutations present in the population and PCR-based assays were developed for their detection. Genotyping of kdr mutations was done using PCR-based assays, allelic frequencies were determined, and tests of genetic association of kdr mutations with the insecticide resistance phenotype were performed. RESULTS: The Ae. aegypti population was resistant to DDT, deltamethrin and permethrin. The DNA sequencing of the VGSC revealed the presence of four kdr mutations, i.e. S989P and V1016G in domain II and two alternative kdr mutations F1534C and F1534L in domain III. Allele-specific PCR assays (ASPCR) were developed for the detection of kdr mutations S989P and V1016G and an existing PCR-RFLP based strategy was modified for the genotyping of all three known kdr mutations in domain III (F1534L, F1534C and T1520I). Genotyping of Ae. aegypti samples revealed a moderate frequency of S989P/V1016G (18.27%) and F1534L (17.48%), a relatively high frequency of F1534C (50.61%) and absence of T1520I in the population. Mutations S989P and V1016G were in complete linkage disequilibrium in this population while they were in linkage equilibrium with kdr mutations F1534C and F1534L. The alleles F1534C and F1534L are genetically associated with permethrin resistance. CONCLUSIONS: A new kdr mutation, F1534L, was found in an Ae. aegypti population from Bengaluru (India), co-occurring with the other three mutations S989P, V1016G and F1534C. The findings of a new mutation have implications for insecticide resistance management.

Assessment of neuro-optometric rehabilitation using the Developmental Eye Movement (DEM) test in adults with acquired brain injury / Valoración de la rehabilitación neuro-optométrica mediante la prueba DEM (Developmental Eye Movement) en adultos con daño cerebral adquirido

Purpose: This pilot study sought to determine the efficacy of using the Developmental Eye Movement (DEM) test in the adult, acquired brain injury (ABI) population to quantify clinically the effects of controlled, laboratory-performed, oculomotor-based vision therapy/vision rehabilitation. Methods: Nine adult subjects with mild traumatic brain injury (mTBI) and five with stroke were assessed before and after an eight-week, computer-based, versional oculomotor (fixation, saccades, pursuit, and simulated reading) training program (9.6 h total). The protocol incorporated a cross-over, interventional design with and without the addition of auditory feedback regarding two-dimensional eye position. The clinical outcome measure was the Developmental Eye Movement (DEM) test score (ratio, errors) taken before, midway, and immediately following training. Results: For the DEM ratio parameter, improvements were found in 80-89% of the subjects. For the DEM error parameter, improvements were found in 100% of the subjects. Incorporation of the auditory feedback component revealed a trend toward enhanced performance. The findings were similar for both DEM parameters, as well as for incorporation of the auditory feedback, in both diagnostic groups. Discussion: The results of the present study demonstrated considerable improvements in the DEM test scores following the oculomotor-based training, thus reflecting more time-optimal and accurate saccadic tracking after the training. The DEM test should be considered as another clinical test of global saccadic tracking performance in the ABI population

Objetivo: Este estudio piloto trató de determinar la eficacia del uso de la prueba DEM (Developmental Eye Movement) en la población adulta con daño cerebral adquirido (DCA) para cuantificar clínicamente los efectos de la rehabilitación/terapia visual controlada, realizada en laboratorio, y de carácter oculomotor. Métodos: Se valoraron nueve sujetos adultos con daño cerebral traumático leve (mTBI) y cinco con ictus, con anterioridad y posterioridad a un programa de entrenamiento de ocho semanas, informático oculomotor y versional (movimientos de fijación, sacádicos, de persecución y lectura simulada, de 9,6 horas en total). El protocolo incorporó un diseño cruzado e intervencional, con y sin adición de retroalimentación auditiva en relación a la posición bi-dimensional de los ojos. La medida del resultado clínico se llevó a cabo mediante la puntuación de la prueba DEM (Developmental Eye Movement) (ratio, errores) realizada con anterioridad, en el punto medio, e inmediatamente tras el entrenamiento. Resultados: Para el parámetro de ratio DEM, se encontraron mejoras en el 80-89% de los sujetos. Para el parámetro de error DEM, se encontraron mejoras en el 100% de los sujetos. La incorporación del componente de retroalimentación auditiva reveló una tendencia hacia la mejora del rendimiento. Los hallazgos fueron similares para ambos parámetros DEM, así como para la incorporación de la retroalimentación auditiva, en ambos grupos diagnósticos. Discusión: Los resultados del presente estudio demostraron mejoras considerables de las puntuaciones de la prueba DEM tras la terapia oculomotora, lo que refleja una mayor optimización y precisión del seguimiento sacádico en el tiempo tras la terapia. Deberá considerarse la prueba DEM como una prueba clínica para valorar el rendimiento del seguimiento sacádico global en la población de DCA

Assessment of neuro-optometric rehabilitation using the Developmental Eye Movement (DEM) test in adults with acquired brain injury.

PURPOSE: This pilot study sought to determine the efficacy of using the Developmental Eye Movement (DEM) test in the adult, acquired brain injury (ABI) population to quantify clinically the effects of controlled, laboratory-performed, oculomotor-based vision therapy/vision rehabilitation. METHODS: Nine adult subjects with mild traumatic brain injury (mTBI) and five with stroke were assessed before and after an eight-week, computer-based, versional oculomotor (fixation, saccades, pursuit, and simulated reading) training program (9.6h total). The protocol incorporated a cross-over, interventional design with and without the addition of auditory feedback regarding two-dimensional eye position. The clinical outcome measure was the Developmental Eye Movement (DEM) test score (ratio, errors) taken before, midway, and immediately following training. RESULTS: For the DEM ratio parameter, improvements were found in 80-89% of the subjects. For the DEM error parameter, improvements were found in 100% of the subjects. Incorporation of the auditory feedback component revealed a trend toward enhanced performance. The findings were similar for both DEM parameters, as well as for incorporation of the auditory feedback, in both diagnostic groups. DISCUSSION: The results of the present study demonstrated considerable improvements in the DEM test scores following the oculomotor-based training, thus reflecting more time-optimal and accurate saccadic tracking after the training. The DEM test should be considered as another clinical test of global saccadic tracking performance in the ABI population.

Larvicidal activity of Ricinus communis extract against mosquitoes.

BACKGROUND & OBJECTIVES: Vector control strategies play significant role in reducing the transmission of malaria, dengue and other vector-borne diseases. The control of vector population using synthetic insecticides has resulted in development of insecticide resistance and negative effects on humans and environment. The present investigation evaluated the larvicidal potential of methanol, dichloromethane and hexane extracts of leaves and seeds of Ricinus communis (castor) plant against the early IV instar larvae of the dengue vector, Aedes aegypti, and malaria vector, Anopheles culicifacies. METHODS: Plant extracts were screened for their efficacy against Ae. aegypti and An. culicifacies using WHO standard larval susceptibility test method. Dose response bioassay was performed to get lethal concentrations. Further, gas chromatography-mass spectroscopy (GC-MS) analysis was carried out to identify the bioactive chemical constituents of the extracts of R. communis. Toxicity of the extracts towards non-target organism, Poecilia reticulata was also evaluated. RESULTS: The leaf and seed extracts of R. communis showed significant mortality against the larvae of Ae. aegypti and An. culicifacies at concentrations of 31.25, 62.5, 125, 250, 500 ppm; and 2, 4, 8, 16, 32, 64 ppm, respectively. At 24 h of the exposure period, the larvicidal activities were highest for the methanol extract of seeds with LC50 15.52 and 9.37 ppm and LC90 45.24 and 31.1 ppm for Ae. aegypti and An. culicifacies, respectively. The methanol extract of seeds and leaves was found to be safe towards non-target organism, P. reticulata. The GC-MS profile showed that seed extracts were having higher concentration of stigmasterol (7.5%), ß-sitosterol (11.48%), methyl linoleate (2.5%), vitamin E (11.93%), and ricinoleic acid (34%) than the leaf extracts. INTERPRETATION & CONCLUSION: The seed extract of R. communis has better larvicidal activity than the leaf extract and can be used as an effective larvicide against mosquitoes. The non-toxicity of the extracts towards P. reticulata further suggests that these plant extracts could be used along with predatory fishes in integrated vector control approaches.

Health Impact Assessment of Indira Sagar Project: a paramount to studies on Water Development Projects.

BACKGROUND: Very limited studies on Health Impact Assessment (HIA) of Water Development Projects (WDP) in relation to mosquito-borne diseases have been carried out in India. The current study focuses on using HIA as a tool for finding impact of Indira Sagar Project, Madhya Pradesh on human health in relation to mosquito borne diseases, and emphasizing its incorporation as an integral part of any WDP. METHODS: Screening, scoping, assessment, recommendation, reporting, and evaluation were carried out in selected study areas. Entomological, epidemiological, socio-economic and knowledge, attitudes and practices data related to malaria transmission in three dam components: Submergence (SUB), Command (CMD) and Resettlement and Rehabilitation (RR) colonies were generated for the period of January 2013-December 2014. Statistical analysis was attempted to compare data among dam components and to identify risk factors. Component-specific mitigation measures were suggested based on observations. RESULTS: Anopheles culicifacies was the dominating species in all three dam components and its man-hour density in CMD areas was higher compared to SUB and RR. The odds of finding a positive malaria case was much higher in CMD compared to SUB (OR 1.24, CI 95% 0.71-2.43) and RR (OR 5.48, CI 95% 0.73-40.63). Respondents of CMD stated more previous episodes of malaria (81.8%) compared to RR (61.4%) and SUB (55.7%). The canonical discriminant analysis concluded that distance from reservoir/Indira Sagar canal had the highest discriminating ability of malaria cases in different components followed by treatment-seeking behaviour and malaria history. The analysis identified these risk factors with 70% accuracy. CONCLUSION: Engineering manipulations may be carried out in CMD areas to control seepage and RR colonies should be established beyond 3 km from reservoir/Indira Sagar canal considering the flight range of A. culicifacies. Strengthening of surveillance with early detection and complete treatment was recommended for CMD areas. To avoid future transmission in other areas and projects HIA should be carried out at planning stage for planning better control activities.

Role of An. culicifacies as a vector of malaria in changing ecological scenario of Northeastern states of India.

BACKGROUND & OBJECTIVES: Malaria has become endemic and subject of concern in most part of the India especially Northeastern states of India. Surveys before 2000 revealed that Anopheles minimus was major vector responsible for transmission of malaria in this region followed by An. dirus and An. fluviatilis. However, recent studies indicate replacement of An. minimus vector by An. culicifacies due to different ecological changes and change in landuse pattern etc. The objective of present study was to explore the vectorial role of An. culicifacies in transmission of malaria in four malaria endemic states, viz. Assam, Meghalaya, Manipur and Sikkim of India. METHODS: The seven surveys were conducted in 176 selected villages belonging to eight districts of the four states in both pre-monsoon (March-April) and post-monsoon (September-October) seasons from 2010 to 2013. However, in 2011 surveys could not be carried out due to public inconvenience in pre-monsoon season. For vectorial role of all vector species collected, ELISA and PCR were assayed. RESULTS: A total of 19,173 specimens belonging to 30 anopheline species were collected, out of which 4315 belonged to four established vector species. In total, 4183 specimens were processed through ELISA, out of which 236 specimens were found positive for circumsporozoite (CS) protein. Further, infectivity was confirmed by PCR in 35 samples, of which 12 samples were found positive for Plasmodium falciparum and three for P. vivax. Out of 12 Plasmodium falciparum positive samples, nine samples were positive for An. culicifacies, two for An. fluviatilis and one for An. minimus. While out of three Plasmodium vivax positive samples, two samples were positive for An. dirus and one sample was positive for An. culicifacies. INTERPRETATION & CONCLUSION: Anopheles culicifacies replaced the An. minimus, the vector of malaria in Northeastern states of India, as it was found to be highly infected with malaria parasite as compared to An. minimus by ELISA and PCR analysis, and thus playing a major role in malaria transmission in this region. The ecological changes like deforestation, development of irrigation channels and change in landuse pattern, have helped in evolution of An. culicifacies in the study area. Therefore, modified vector control strategies are required on urgent basis.

Pyrethroid-resistance and presence of two knockdown resistance (kdr) mutations, F1534C and a novel mutation T1520I, in Indian Aedes aegypti.

BACKGROUND: Control of Aedes aegypti, the mosquito vector of dengue, chikungunya and yellow fever, is a challenging task. Pyrethroid insecticides have emerged as a preferred choice for vector control but are threatened by the emergence of resistance. The present study reports a focus of pyrethroid resistance and presence of two kdr mutations--F1534C and a novel mutation T1520I, in Ae. aegypti from Delhi, India. METHODOLOGY/PRINCIPAL FINDINGS: Insecticide susceptibility status of adult-female Ae. aegypti against DDT (4%), deltamethrin (0.05%) and permethrin (0.75%) was determined using WHO's standard insecticide susceptibility kit, which revealed resistance to DDT, deltamethrin and permethrin with corrected mortalities of 35%, 72% and 76% respectively. Mosquitoes were screened for the presence of kdr mutations including those reported earlier (I1011V/M, V1016G/I, F1534C, D1794Y and S989P), which revealed the presence of F1534C and a novel mutation T1520I. Highly specific PCR-RFLP assays were developed for genotyping of these two mutations. Genotyping using allele specific PCR and new PCR-RFLP assays revealed a high frequency of F1534C (0.41-0.79) and low frequency of novel mutation T1520I (0.13). The latter was observed to be tightly linked with F1534C and possibly serve as a compensatory mutation. A positive association of F1534C mutation with DDT and deltamethrin resistance in Ae. aegypti was established. However, F1534C-kdr did not show significant protection against permethrin. CONCLUSIONS/SIGNIFICANCE: The Aedes aegypti population of Delhi is resistant to DDT, deltamethrin and permethrin. Two kdr mutations, F1534C and a novel mutation T1520I, were identified in this population. This is the first report of kdr mutations being present in the Indian Ae. aegypti population. Highly specific PCR-RFLP assays were developed for discrimination of alleles at both kdr loci. A positive association of F1534C mutation with DDT and deltamethrin resistance was confirmed.

Oculomotor neurorehabilitation for reading in mild traumatic brain injury (mTBI): an integrative approach.

BACKGROUND: Considering the extensive neural network of the oculomotor subsystems, traumatic brain injury (TBI) could affect oculomotor control and related reading dysfunction. OBJECTIVE: To evaluate comprehensively the effect of oculomotor-based vision rehabilitation (OBVR) in individuals with mTBI. METHODS: Twelve subjects with mTBI participated in a cross-over, interventional study involving oculomotor training (OMT) and sham training (ST). Each training was performed for 6 weeks, 2 sessions a week. During each training session, all three oculomotor subsystems (vergence/accommodation/version) were trained in a randomized order across sessions. All laboratory and clinical parameters were determined before and after OMT and ST. In addition, nearvision-related symptoms using the Convergence Insufficiency Symptom Survey (CISS) scale and subjective visual attention using the Visual Search and Attention Test (VSAT) were assessed. RESULTS: Following the OMT, over 80% of the abnormal parameters significantly improved. Reading rate, along with the amplitudes of vergence and accommodation, improved markedly. Saccadic eye movements demonstrated enhanced rhythmicity and accuracy. The improved reading-related oculomotor behavior was reflected in reduced symptoms and increased visual attention. None of the parameters changed with ST. CONCLUSIONS: OBVR had a strong positive effect on oculomotor control, reading rate, and overall reading ability. This oculomotor learning effect suggests considerable residual neuroplasticity following mTBI.

Field evaluation of Icon®Life, a long-lasting insecticidal net (LLIN) against Anopheles culicifacies and transmission of malaria in District Gautam Budh Nagar (Uttar Pradesh), India.

BACKGROUND & OBJECTIVES: In the present study, Icon®Life net, a long-lasting polyethylene net, 100 denier and bursting strength of minimum 280 kpa incorporated with deltamethrin @ 65 mg/m2 was evaluated for its efficacy in reducing the density of malaria vector Anopheles culicifacies and impact on malaria prevalence in a malaria endemic area of District Gautam Budh Nagar, India. METHODS: Wash resistance of Icon®Life LLIN was determined up to 20 serial washings using An. culicifacies in cone bioassays under field conditions. Efficacy of Icon®Life LLIN was determined in the field in three sets of villages in District Gautam Budh Nagar (Uttar Pradesh), India, selected randomly for the intervention with Icon®Life LLIN, untreated nets and a control without any intervention for the period of August 2008-July 2009. Entomological and malariometric indices in all the three villages were compared during pre- and post-intervention periods for one year against An. culicifacies. A survey was also conducted in the village provided with Icon®Life LLIN to assessing the perception of community regarding acceptance of these nets by the community. RESULTS: In cone bioassays on Icon®Life LLIN with An. culicifacies, >95% knockdown within 1 h and 100% mortality after 24 h exposure were reported even after 20× serial washings under field conditions. Results of the field study revealed reduced entry rate, resting density and parity rate of An. culicifacies in the village with Icon®Life LLIN when compared to no net and untreated net villages. Number of malaria cases reported were less in the Icon®Life LLIN used villages when compared to other two villages. The community compliance and acceptance was high and no adverse health events were reported by the households using these nets. CONCLUSIONS: Icon®Life LLIN is an effective intervention for the control of An. culicifacies transmitted malaria in India. Long-term studies are indicated for the duration of effectiveness and to ascertain the epidemiological impact of the use of Icon®Life nets.

Visual impairments in the first year after traumatic brain injury.

BACKGROUND: This article reviews literature regarding individuals with traumatic brain injury who have vision related impairments up to one year, post-injury. Such impairments may impact rehabilitation of activities of daily living and mobility since vision is integral in much of what one does on a daily basis. METHODS: Search of Medline, Ovid, and PubMed was performed using terms including: traumatic brain injury, visual deficits after brain injury, vision and traumatic brain injury, and ADLs after brain injury. RESULTS: Eighteen studies were analyzed and reviewed. A range of visual and visual-motor impairments are seen across the severity of traumatic brain injury. Visual impairment negatively impacts independence in mobility and activities of daily living. Common sensorimotor visual symptoms reported by those with traumatic brain injury include blurred vision, reading problems, double vision or eyestrain, dizziness or disequilibrium in visually-crowded environments, visual field defects, light sensitivity, and color blindness. CONCLUSIONS: This review should alert the reader to common visual complaints and defects seen after traumatic brain injury. It is important to screen persons who have suffered traumatic brain injury for sensorimotor vision deficits early on in recovery so that these issues may be addressed and recovery of function and independence in the community are not delayed.