Current status of insecticide resistance and its underlying mechanisms in Aedes aegypti (L.) in Punjab, Pakistan

Aedes aegypti (Linnaeus) plays an important role as a vector of different deadly diseases particularly dengue fever. Insecticides are used as a primary tool to control Ae. aegypti. However, due to the excessive use of insecticides on agricultural, public health, and industrial levels, mosquitoes have developed resistance. In this study, the current susceptibility status of Ae. aegypti mosquitoes against different insecticides (Temephos, DDT, dieldrin, Malathion, Bendiocarb, Permethrin, Cypermethrin, and Lambda-cyhalothrin) was evaluated in district Lahore and district Muzaffargarh of Punjab, Pakistan. For this purpose, WHO bioassays and biochemical assays were performed on Ae. aegypti population from Lahore (APLa) and Aedes population from Muzaffargarh (APMg). Results of APLa and APMg showed high levels of resistance against the larvicide Temephos. Resistance against all adulticides was also observed in APLa and APMg (% mortality < 98%). The biochemical assays indicated statistically significant elevated levels of detoxification enzymes in APLa and APMg. APLa showed slightly higher levels as compared to APMg. Mosquitoes were also screened for the presence of kdr mutations. The results revealed no mutation in domain II while the presence of mutation F1534C in domain III was found in both field populations. The results showed the presence of moderate to high grade resistance against all insecticides in Ae. aegypti in district Lahore and district Muzaffargarh of Punjab, Pakistan

Genetic diversity and Kdr mutations of natural Aedes (Stegomyia) aegypti (Diptera: Culicidae) populations of Brazil

ABSTRACT Aedes (Stegomyia) aegypti is an important vector of dengue, yellow fever, chikungunya and Zika virus. It is well known that resistance monitoring and genetic diversity data help designing the vector control programs. This study aimed to evaluate resistance to pyrethroids (PYs) through the frequency of kdr mutations Val1016IIe and F1534C, and the genetic variation of the mitochondrial gene ND4 in six natural populations of A. aegypti from Paraná - Brazil. Adults were obtained from eggs collected from Alvorada do Sul, Marilena, Maringá, Nova Londrina, Paranavaí and São Carlos do Ivaí. From these adults, 345 were used to identify the 1016 and 1534 sites, and 120 were used to perform the ND4 gene analysis. The studied populations from Paraná showed PYs resistance, low gene flow and genetic diversity. Additionally, a relationship was observed among the haplotypes of populations from the Amazon and Southeastern Brazil, Peru, Mexico, and North America.

Insecticide resistance levels and associated mechanisms in three Aedes aegypti populations from Venezuela

BACKGROUND The massive use of insecticides in public health has exerted selective pressure resulting in the development of resistance in Aedes aegypti to different insecticides in Venezuela. Between 2010 and 2020, the only insecticides available for vector control were the organophosphates (Ops) fenitrothion and temephos which were focally applied. OBJECTIVES To determine the state of insecticide resistance and to identify the possible biochemical and molecular mechanisms involved in three populations of Ae. aegypti from Venezuela. METHODS CDC bottle bioassays were conducted on Ae. aegypti collected between October 2019 and February 2020 in two hyperendemic localities for dengue in Aragua State and in a malaria endemic area in Bolívar State. Insecticide resistance mechanisms were studied using biochemical assays and polymerase chain reaction (PCR) to detect kdr mutations. FINDINGS Bioassays showed contrasting results among populations; Las Brisas was resistant to malathion, permethrin and deltamethrin, Urbanización 19 de Abril was resistant to permethrin and Nacupay to malathion. All populations showed significantly higher activity of mixed function oxidases and glutathione-S-transferases (GSTs) in comparison with the susceptible strain. The kdr mutations V410L, F1534C, and V1016I were detected in all populations, with F1534C at higher frequencies. MAIN CONCLUSION Insecticide resistance persists in three Ae. aegypti populations from Venezuela even in the relative absence of insecticide application.

Estudo de associação entre polimorfismos genéticos e níveis plasmáticos do VEGF, FLT1, KDR e o risco para depressão / Study of association between genetic polymorphisms and plasma levels of VEGF, FLT1, KDR and the risk for depression

Introdução: a depressão é um transtorno mental comum, grave e incapacitante que afeta mais de 350 milhões de pessoas em todo o mundo. A depressão é caracterizada principalmente por sintomas como tristeza, perda de interesse, diminuição da energia, perda de confiança e autoestima, culpa inadequada, distúrbios do sono e do apetite, pensamentos de morte e suicídio. Além disso, essa patologia também tem um forte impacto na qualidade de vida dos indivíduos afetados e de suas famílias. Sabe-se que fatores genéticos interagem com as condições socioambientais de modo a influenciar a predisposição das pessoas ao adoecimento. Estudos identificaram polimorfismos de nucleotídeos simples (SNPs) que podem ser marcadores genéticos apropriados para prever inflamação sistêmica, por exemplo, e a atual tese teve como foco o efeito de SNPs na via do fator de crescimento endotelial vascular (VEGF). Esta proteína é uma potente molécula angiogênica e está envolvida na neurogênese do hipocampo, uma das principais estruturas límbicas afetadas em pessoas com depressão. O VEGF está implicado em uma das principais teorias que tentam explicar a fisiopatologia deste transtorno mental grave, a teoria neurotrófica, a qual diz que a diminuição ou desregulação da sinalização de neurotrofinas pode contribuir para a manifestação do transtorno depressivo (TD). Objetivo: avaliar se polimorfismos do VEGF e seus receptores, KDR e FLT1, estão associados à depressão e à gravidade dos sintomas, à ideação e tentativas de suicídio, independentemente tanto de um tratamento otimizado quanto da presença de estresse precoce (do inglês, early-life stress, ELS), também verificar se há efeito destes polimorfismos nas concentrações plasmáticas de proteínas expressas pelos seus respectivos genes e observar se existe correlação entre VEGF e seus inibidores, VEGF e s100ß. Metodologia: participaram do presente estudo 160 pacientes com depressão e 114 controles saudáveis. Foram aplicados durante entrevista questionários que avaliaram o perfil clínico dos pacientes como o MINI-International Neuropsychiatric Interview, GRID-HAMD21, CTQ, BSI e foi registrado o número de tentativas de suicídio. Os controles passaram por uma entrevista para serem avaliados quanto aos critérios de inclusão e exclusão do grupo. A genotipagem dos participantes foi realizada através da técnica de Real Time PCR e as mensurações de proteínas por meio do ensaio ensaio imunoenzimático (ELISA). Resultados: indivíduos com depressão, homozigotos AA do polimorfismo rs699947, apresentaram maiores concentrações plasmáticas de VEGF (P-valor= 0.006) e se associaram a um maior número de tentativas de suicídio na análise direta (P-valor= 0.041) e na análise corrigida foi observada uma tendência para a confirmação deste resultado (P-valor= 0.076). O genótipo homozigoto GG do polimorfismo rs7993418 do FLT1 se associou à severidade de sintomas (P-valor= 0.040), bem como uma tendência de associação com um aumento nas tentativas de suicídio e uma maior pontuação na escala que avaliou ideação suicida. Entre os pacientes quanto maior foram as concentrações plasmáticas de VEGF, maior foram as de KDR, FLT1 e s100ß. Conclusão: os resultados sugerem que os polimorfismos da via VEGF estão associados ao número de tentativas de suicídio e severidade dos sintomas depressivos

Introduction: Depression is a common, serious, and disabling mental disorder that affects more than 350 million people worldwide. Depression is mainly characterized by symptoms such as sadness, loss of interest, decreased energy, loss of confidence and self-esteem, inadequate guilt, sleep and appetite disturbances, thoughts of death and suicide. Furthermore, this pathology also has a strong impact on the quality of life of those affected and their families. It is known that genetic factors interact with social and environmental conditions to influence people's predisposition to illness. Studies have identified single nucleotide polymorphisms (SNPs) that may be appropriate genetic markers to predict systemic inflammation, for example, and the current thesis focused on the effect of SNPs on the vascular endothelial growth factor (VEGF) pathway. This protein is a potent angiogenic molecule and is involved in hippocampal neurogenesis, one of the main limbic structures affected in people with depression. VEGF is implicated in one of the main theories that try to explain the pathophysiology of this severe mental disorder, the neurotrophic theory, which says that the decrease or dysregulation of neurotrophin signaling can contribute to the manifestation of depressive disorder (DT). Objective: to assess whether polymorphisms of VEGF and its receptors, KDR and FLT1, are associated with depression and severity of symptoms, suicide ideation and attempts, regardless of both optimal treatment and the presence of early-life stress (ELS) in these associations. also check whether there is an effect of these polymorphisms on the plasma concentrations of proteins expressed by their respective genes and observe whether there is a correlation between VEGF and its inhibitors, VEGF and s100ß. Methodology: 160 patients with depression and 114 healthy controls participated in this study. Questionnaires that assessed the clinical profile of patients, such as the MINI-International Neuropsychiatric Interview, GRID-HAMD21, CTQ, BSI, were applied during interviews, and the number of suicide attempts was recorded. The controls underwent an interview to be evaluated regarding the inclusion and exclusion criteria. The genotyping of the participants was performed using the Real Time PCR technique and protein measurements were performed using the enzyme-linked immunosorbent assay (ELISA). Results: individuals with depression, homozygous AA of the rs699947 polymorphism, had higher plasma concentrations of VEGF (P-value = 0.006) and a greater number of suicide attempts in the direct analysis (P-value = 0.041) and in the corrected analysis a trend towards confirmation of this result was observed (P-value = 0.076). The GG genotype of the FLT1 polymorphism rs7993418 was associated with symptom severity (P-value = 0.040), as well as with a trend for association with increase in suicide attempts and a higher score on the scale that evaluated suicidal ideation. The bigger the plasma concentrations of VEGF, the higher were those of KDR, FLT1 and s100ß. Conclusion: the results indicate that VEGF pathway polymorphisms are associated with the number of suicides and severity of depressive symptoms

Evaluación de primers reportados para detección de KDR en especies de Anopheles colectadas en los departamentos de Caaguazú y Alto Paraná en Paraguay / Evaluation of primers reported for KDR detection in Anopheles species collected in the departments of Caaguazú and Alto Paraná in Paraguay

Las mutaciones KDR en el gen del canal del sodio (VGSC) han sido ya detectadas en al menos 13 especies de mosquitos Anopheles en su mayoría especies de África, pero aún resta por determinar los cebadores específicos para la detección en especies de Latinoamérica. En nuestro país la especie Anopheles darlingi es el vector principal de la malaria, y el A. albitarsis, el vector secundario. Se emplearon muestras de mosquitos Anoheles de las especies A. strodei, A. albitarsis, A. fluminensis, A. evansae, A. nuneztovari, A. nyssorhynchela lutzi y A. oswaldoi capturadas en los departamentos de Caaguazú y Alto Paraná en Paraguay. Para la amplificación y secuenciación se usaron cebadores reportados para el gen VGSC de A. albimanus en Guatemala, que resultaron ser específicos solo para la especie A. strodei. La secuencia revela el codón TTA que codifica para una Leucina como la secuencia TTG, reportada para la versión susceptible en la posición L1014. El fragmento amplificado es de aproximadamente 225 pares de bases. A nuestro entender, esta es la primera caracterización del gen VGSC en mosquitos Anopheles del Paraguay y para la especie A. strodei

KDR mutations in the sodium channel gene (VGSC) have already been detected in at least 13 species of Anopheles mosquitoes, mostly African species, but the molecular techniques for detection in Latin American species have yet to be determined. In our country, Anopheles darlingi species is the main vector of Malaria, and A. albitarsis, the secondary vector. We used samples of Anoheles from the species A. strodei, A. albitarsis, A. fluminensis, A. evansae, A. nuneztovari, A. nyssorhynchela lutzi and A. oswaldoi collected at the departments of Caaguazú and Alto Paraná in Paraguay. For the amplification and sequentiation, primers reported for the VGSC gen of A. strodei in Guatemala were used and were specific only for A. strode in this case. The sequence revealed the TTA codon that codifies for a leucine as the TTG sequence, reported for the susceptible version at position L1014. The amplified fragment is approximately 225 base pairs. To our knowledge, this is the first characterization of the VGSC gene in Anopheles mosquitoes in Paraguay and for the species A. strodei

Study on kdr allele mutation of Culex pipiens pallens in Zichuan District, Zibo City, Shandong Province / 中国血吸虫病防治杂志

Objective To study the seasonal dynamics of Culex pipiens pallens and the distribution of knockdown resistance (kdr) gene related sodium channel gene polymorphism in Zichuan District, Zibo City, Shandong Province. Methods Cx. pipiens pallens mosquitoes were collected in Zichuan District during the peak period of mosquito vector activity from 2017 to 2018. The DNA from Cx. pipiens pallens was extracted, and the genotypes and frequencies of kdr allele mutation were detected by polymerase chain reaction. Results Totally 830 mosquitoes belonging to six species, including Cx. pipiens pallens, Armigeres subalbatus, Aedes albopictus, Ae. vexans, Anopheles sinensis, and Cx. tritaeniorhchus were collected in this study. The number of Cx. pipiens pallens accounted for 83.13% in total, with the density of 12.32 per lamp per night. The annual density monitoring curve of Cx. pipiens pallens showed a bimodal trend, and the peaks were observed in June and September respectively. In this study, five kdr alleles were detected at the 1 014 locus of kdr gene, with TTA (75.71%), TTT (10.00%), CTA (5.71%), TCA (4.29%), and TTC (4.29%). Two nonsynonymous nucleotide mutations were detected at site 1 014 of kdr gene, namely leucine (L1014) mutated to phenylalanine (L1014F) and serine (L1014S). The kdr gene mutation frequency (%) of Cx. pipiens pallens in Luochun Town and Taihe Town was 10.53% and 40.63%, respectively, and the difference was statistically significant ( χ2 = 8.559, P = 0.003). Conclusions Cx. pipiens pallens is the dominant mosquito species in Zichuan District. In addition, two novel mutations, CTA and TTC, are identified in the voltage-dependent sodium channel gene of Cx. pipiens pallens. The kdr genotype of Cx. pipiens pallens in Zichuan area was polymorphic.

Study on kdr allele mutation of Culex pipiens pallens in Zichuan District, Zibo City, Shandong Province / 中国血吸虫病防治杂志

Objective To study the seasonal dynamics of Culex pipiens pallens and the distribution of knockdown resistance (kdr) gene related sodium channel gene polymorphism in Zichuan District, Zibo City, Shandong Province. Methods Cx. pipiens pallens mosquitoes were collected in Zichuan District during the peak period of mosquito vector activity from 2017 to 2018. The DNA from Cx. pipiens pallens was extracted, and the genotypes and frequencies of kdr allele mutation were detected by polymerase chain reaction. Results Totally 830 mosquitoes belonging to six species, including Cx. pipiens pallens, Armigeres subalbatus, Aedes albopictus, Ae. vexans, Anopheles sinensis, and Cx. tritaeniorhchus were collected in this study. The number of Cx. pipiens pallens accounted for 83.13% in total, with the density of 12.32 per lamp per night. The annual density monitoring curve of Cx. pipiens pallens showed a bimodal trend, and the peaks were observed in June and September respectively. In this study, five kdr alleles were detected at the 1 014 locus of kdr gene, with TTA (75.71%), TTT (10.00%), CTA (5.71%), TCA (4.29%), and TTC (4.29%). Two nonsynonymous nucleotide mutations were detected at site 1 014 of kdr gene, namely leucine (L1014) mutated to phenylalanine (L1014F) and serine (L1014S). The kdr gene mutation frequency (%) of Cx. pipiens pallens in Luochun Town and Taihe Town was 10.53% and 40.63%, respectively, and the difference was statistically significant ( χ2 = 8.559, P = 0.003). Conclusions Cx. pipiens pallens is the dominant mosquito species in Zichuan District. In addition, two novel mutations, CTA and TTC, are identified in the voltage-dependent sodium channel gene of Cx. pipiens pallens. The kdr genotype of Cx. pipiens pallens in Zichuan area was polymorphic.

The role of KDR in intrauterine adhesions may involve the TGF-ß1/Smads signaling pathway

The aim of this study was to investigate the role of kinase-insert domain-containing receptor (KDR) in intrauterine adhesions (IUA) and its mechanism. The Case group consisted of 92 patients diagnosed with IUA, and the Control group included 86 patients with uterine septum who had normal endometrium verified with an uteroscope. In addition, 50 rats were randomly assigned into Control, Sham, Model, NC-siRNA, and KDR-siRNA groups. Rats in the Model, NC-siRNA, and KDR-siRNA groups were induced by uterine curettage and lipopolysaccharide (LPS) treatment to establish the IUA model. Then, immunohistochemistry was applied for detection of VEGF and KDR expression, HE staining was used for observation of the endometrial morphology and gland counting, Masson staining for measurement of the degree of endometrial fibrosis, and qRT-PCR and western blot for the expression of KDR, VEGF, MMP-9, as well as TGF-β1/Smads pathway-related proteins. Compared with the Control group, the mRNA and protein expressions of KDR were significantly higher in IUA endometrial tissues, and the expression of KDR was positively correlated to the severity of IUA. In addition, the injection of si-KDR increased the number of endometrial glands, reduced the area of fibrosis, inhibited mRNA and protein expression of KDR and VEGF, up-regulated the expression of MMP-9 and Smad7, and decreased the expression level of TGF-β1, p-Smad2, p-Smad3, and Smad4 in rats with IUA. Highly-expressed KDR was related to patients' severity of IUA, and silencing KDR may prevent the occurrence and development of IUA via TGF-β1/Smads signaling pathway and up-regulating the expression of MMP-9.

Evaluation of insecticide resistance in Aedes aegypti populations connected by roads and rivers: the case of Tocantins state in Brazil

BACKGROUND The longstanding application of insecticides for vector control without periodic monitoring of the population response to these chemicals can directly drive the selection of resistant populations of vector mosquitoes. Tocantins is the newest state of the Brazilian Federation. Despite a historically high number of dengue cases, studies and monitoring data concerning insecticide resistance in the state are lacking. OBJECTIVES To verify the resistance status of Aedes aegypti from 10 populations distributed throughout the state connected by rivers and roads. METHODS Between 50 and 150 ovitraps were installed in house gardens within each municipality. Collection points were established based on the importance of the towns and on geographic aspects. Dose response bioassays were performed in accordance with World Health Organization guidelines. Molecular assays were conducted to detect kdr mutations, which are related to pyrethroid resistance. FINDINGS Of the 3,200 ovitrap paddles analysed, 25.8% contained eggs, with a total of 55,687 eggs collected. With the exception of Caseara, all evaluated populations were considered to be resistant to temephos. The data showed different levels of resistance to deltamethrin among the samples. Caseara and Guaraí showed the lowest RR95 values. On average, the NaVR1 kdr allele was most frequent (40.3%), followed by NaVS (38.1%), and NaVR2 (21.6%). Palmas, the capital of the state, had the highest frequency of kdr alleles (87.5%). MAIN CONCLUSIONS With the exception of Palmas, the towns with the highest indexes (ovitrap positivity, number and density of eggs), as well with high levels of resistance and kdr alleles were located along the BR-153 road, indicating that the flow of people and cargo can contribute to the dispersion of the vector and potentially resistance. This study contributes substantially to knowledge regarding the insecticide resistance profile of Tocantins mosquito populations; the data generated via the study could facilitate the judicious use of insecticides by vector control programs.

Association between genetic variation of kinase insert domain receptor and prognosis in colorectal cancer patients received 5-FU based adjuvant chemotherapy / 中国肿瘤生物治疗杂志

@# Objective: To investigate the association between genetic variation of kinase insert domain receptor (KDR) and the prognosis in colorectal cancer (CRC) patients received 5-FU based adjuvant chemotherapy. Methods: The clinical data of 176 CRC patients, who underwent surgical treatment at the Department of Anus and Intestine Surgery, People’s Hospital of Zhengzhou during January 2012 and December 2017, were retrospectively analyzed, and 93 cases of tumor tissues were collected for this study. The genotype of KDR polymorphism locus was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). qPCR was used to detect the expression of KDR mRNAin colorectal cancer tissues. The correlation between the polymorphism genotypes and other variables was analyzed by logistic regression model. The expression of different genotypes of KDR was analyzed by nonparametric test. The relationship between KDR genotype and prognosis of patients was analyzed by Kaplan-Meier survival analysis, and the other variables were adjusted by Cox risk scale model. Results: Of the polymorphisms analyzed, only rs2071559 was of clinical significance. The distribution frequency of KDR rs2071559 in 176 CRC patients was as follows: TT genotype in 95 cases (53.98%), TC genotype in 70 cases (39.77%) and CC genotype in 11 cases (6.25%); the minor allele frequency was 0.26; and the distribution of three genotypes was in accordance with Hardy-Weinberg's Equilibrium (P=0.690). The median disease free survival (mDFS) of patients carrying C allele and wild type TT genotype was 4.4 and 3.2 years, respectively (P<0.05); The median overall survival (mOS) of patients with TC/CC genotype and TT genotype was 5.2 and 4.0 years, respectively (P<0.05). After COX model modification, the effect of TC/CC genotype on mOS was still statistically significant (OR=0.55, P<0.05). The mRNA expression of KDR in cancer tissues of the patients with TC/CC genotypes were significantly lower than those of the wild type TT genotype (P<0.01). Conclusion: The polymorphism of KDR rs2071559 is associated with clinical outcomes in patients with colorectal cancer. KDR rs2071559 may affect the prognosis of colorectal cancer patients by affecting the mRNAexpression of KDR.

First investigation of deltamethrin pyrethroid susceptibility and resistance status of Anopheles labranchiae (Falleroni, 1926), potential malaria vector in Tunisia

Objective To evaluate the deltamethrin pyrethroid insecticides against Anopheles labranchiae, potential malaria vector in Tunisia. Methods Six field populations of Anopheles labranchiae mosquitoes were collected from six localities in Northern and Central Tunisia between October and November 2016. Different bioassays were performed to estimate the level of resistance in each collected population. Two synergists were used to estimate the involvement of detoxification enzymes in insecticide resistance. Results All studied strains were resistant and the RR

First investigation of deltamethrin pyrethroid susceptibility and resistance status of Anopheles labranchiae (Falleroni, 1926), potential malaria vector in Tunisia

Objective:To evaluate the deltamethrin pyrethroid insecticides against Anopheles labranchiae,potential malaria vector in Tunisia.Methods:Six field populations of Anopheles labranchiae mosquitoes were collected from six localities in Northern and Central Tunisia between October and November 2016.Different bioassays were performed to estimate the level of resistance in each collected population.Two synergists were used to estimate the involvement of detoxification enzymes in insecticide resistance.Results:All studied strains were resistant and the RRs0 ranged from 12.5 in sample #1 to 72.5 in sample #6.Synergist tests using piperonyl butoxide indicated the involvement of monoxygenases enzymes in the recorded resistance.In contrast,the increase of deltamethrin mortality was not significant in presence of S,S,sributyl phosphorothioate (0.8 ＜ SR ＜ 1.2),suggesting no role of esterases (and/or GST) in the resistance phenotype.The correlation recorded between mortality due to DDT and the LC50 of deltamethrin insecticide indicated an insensitive sodium channel affected by Kdr mutation (Spearman rank correlation,r =-0.59,P ＜ 0.01).Conclusions:These results should be considered in the current mosquitoes control programs in Tunisia.The use of pesticides and insecticides by both agricultural and public health departments in Tunisia should be more rational to reduce the development of resistance in populations.Different insecticide applications should be implemented alternately.

Polymorphisms in VEGF and KDR genes in the development of endometriosis: a systematic review / Polimorfismos nos genes VEGF e KDR no desenvolvimento da endometriose: revisão sistemática

Abstract Objectives: to review studies that used case-control design to verify the association of polymorphisms in VEGF and KDR genes in the development of endometriosis. Methods: the systematic review selected articles published until September 1, 2015 from PubMed, MEDLINE, BVS, SciELO databases, considering the following key words: endometriosis and ("polymorphism" or "SNP" or "genetic polymorphism") and ("VEGF" OR "Vascular endothelial growth factor" or "VEGFR-2" or "Vascular endothelial growth factor-2" or "KDR" or "Kinase Insert Domain Receptor"). Results: 106 articles were identified, only 11 were eligible. Discrepant results were observed regarding polymorphisms in VEGF gene in the development of endometriosis, which can be explained by methodological differences, sample size, eligible control type, using the unadjusted risk estimates and the heterogeneity of the studied population. Only one study investigated polymorphisms in KDR gene in the development of endometriosis, however it was ineligible for this review. Conclusions: to avoid discrepancy in the results, we suggest that the ideal control group should be formed by fertile women and free of gynecological diseases. Multicentric studies with adequate design, involving different population besides the combined analysis on polymorphisms in VEGF and KDR genes are still necessary to contribute in the understanding of this disease, which are social, clinical and economical problems.

Resumo Objetivos: revisar os trabalhos que utilizaram o delineamento caso-controle para verificar a associação de polimorfismos nos genes VEGF e KDR no desenvolvimento da endometriose. Métodos: revisão sistemática que pesquisou nas bases de dados PubMed, MEDLINE, BVS, SciELO os artigos publicados até o dia 1 de setembro de 2015, considerando os descritores: endometriosis and ("polymorphism" or "SNP" or "geneticpolymorphism") and ("VEGF" or "Vascular endothelial growth factor" or "VEGFR-2" or "Vascular endothelial growth factor-2" or "KDR" or "Kinase Insert Domain Receptor"). Resultados: identificou-se 106 artigos, sendo 11 considerados elegíveis. Observou-se resultados discrepantes quanto aos polimorfismos no gene VEGF no desenvolvimento da endometriose, podendo ser explicados pelas diferenças metodológicas, pelo tamanho amostral, pelo tipo de controle elegível, pela utilização da estimativa de risco não ajustada e pela heterogeneidade das populações estudadas. Apenas um estudo investigou polimorfismos no gene KDR no desenvolvimento da endometriose, contudo não foi elegível nesta revisão. Conclusões: para evitar as discrepâncias dos resultados observados sugerimos que o grupo controle ideal deva ser formado por mulheres férteis e livres de doenças ginecológicas. Estudos multicêntricos com delineamento adequado, envolvendo diferentes populações, além da análise combinada de polimorfismos nos genes VEGF e KDR ainda são necessários para contribuir no entendimento desta doença que é um problema social, clínico e econômico.

The impact of insecticides management linked with resistance expression in Anopheles spp. populations / O impacto do manejo de inseticidas relacionado com a expressão de resistência em populações de Anopheles spp

Abstract The resistance of some species of Anopheles to chemical insecticides is spreading quickly throughout the world and has hindered the actions of prevention and control of malaria. The main mechanism responsible for resistance in these insects appears to be the target site known as knock-down resistance (kdr), which causes mutations in the sodium channel. Even so, many countries have made significant progress in the prevention of malaria, focusing largely on vector control through long-lasting insecticide nets (LLINs), indoor residual spraying and (IRS) of insecticides. The objective of this review is to contribute with information on the more applied insecticides for the control of the main vectors of malaria, its effects, and the different mechanisms of resistance. Currently it is necessary to look for others alternatives, e.g. biological control and products derived from plants and fungi, by using other organisms as a possible regulator of the populations of malaria vectors in critical outbreaks.

Resumo A resistência de algumas espécies de Anopheles a inseticidas químicos está se espalhando rapidamente por todo o mundo e tem dificultado as ações de prevenção e controle da malária. O principal mecanismo responsável pela resistência nestes insetos parece ser o sítio-alvo conhecido como resistência knock-down resistance (kdr), que causa mutações no canal de sódio. Mesmo assim, muitos países fizeram progressos significativos na prevenção da malária, concentrando-se em grande parte no controle do vetor através redes inseticidas de longa duração (RILD), e de pulverização residual interna (PRI) de inseticidas. O objetivo desta revisão é contribuir com informações sobre os inseticidas mais aplicados para o controle dos principais vetores da malária, seus efeitos, e os diferentes mecanismos de resistências. Atualmente é necessário olhar para outras alternativas, como por exemplo, controle biológico e produtos derivados de plantas e fungos, pela utilização de outros organismos como um possível regulador de populações de vetores da malária em surtos críticos.

Mecanismos de resistencia a la permetrina en dos poblaciones de Aedes aegypti del occidente de Venezuela / Mechanisms of Permethrin resistance in two populations of Aedes aegypti in westernVenezuela

Se determinó la resistencia a la permetrina en Aedes aegypti de los estados Trujillo y Zulia mediante bioensayos de botella. Los insectos derribados durante la hora de exposición fueron registrados y usados para calcular la Concentración Knock-down cincuenta (CK50) y los muertos a las 24 horas para la Concentración Letal cincuenta(CL50). La resistencia al derribo y post-recuperación fueron determinadas calculando del Factor de Resistencia FRCK50 y FRCL50, comparando los valores de CK50 y CL50de las poblaciones de Ae. aegypti de campo con los de la cepa susceptible New Orleans obtenidos mediante análisis de regresión log-probit. Mecanismos metabólicos y no metabólicos asociados a la resistencia, fueron evaluados midiendo los niveles de las enzimas alfa-esterasas, beta-esterasas, oxidasas de función múltiple y glutatión-S-transferasas mediante la técnica de microplacas y determinando la frecuencia alélica I1016 por PCR alelo específico. Ambas poblaciones mostraron baja resistencia al derribo (FRCK50 < 5) y moderada resistencia post-recuperación (FRCL50 entre 5 y 10). Sobre-expresión de alfa-esterasas fue observada en la población Loma Linda la cual se correlacionó significativamente con la CL50. En la población Pampanito la frecuencia del alelo I1016 fue de 0,1 y en Loma Linda de 0,17, observándose homocigotos mutantes solo en esta última población. Se evidencia la presencia de mecanismos metabólicos y no metabólicos asociados a la resistencia al derribo y post-recuperación a la permetrina en las poblaciones bajo estudio, lo cual debe ser considerado antes de la aplicación de piretroides para el control de Ae. aegypti en la zona de estudio.

Permethrin resistance was determined in Aedes aegypti populations from Trujillo and Zulia states using the bottle bioassay method. Insect knock-down rates during 1h of exposure were recorded and used to calculate the 50% knock-down concentration (KC50) and the mortality after 24 h (LC50). Knock-down and post-recovery resistance were determined by calculating the resistance factors, FRKC50 and FRLC50 . This was done by comparing the KC50 and LC50 values (obtained by regression analysis log-probit) of the field populations with a susceptible New Orleans strain. Metabolic and non-metabolic mechanisms associated with resistance were assessed by measuring the levels of the following enzymes: alpha-esterases, beta-esterases, mixed function oxidases (MFOs) and glutathione-S-transferases (GSTs) using the microplate technique. We also determined the alellic frequency of I1016 by allele specific PCR. Both populations showed a low knock-down resistance (FRKC50 < 5) and moderate post-recovery resistance (FRLC50 between 5 and 10). Overexpression of alpha-esterases was observed in the Loma Linda population and was significantly correlated with the LC50 . The frequency of the I1016 allele was 0.1 for the Pampanito population and 0.17 for the Loma Linda population, and in the latter we also observed homozygous mutants. The existence of metabolic and non-metabolic mechanisms associated with knockdown resistance and post-recovery to permethrin in the populations studied was demonstrated. This should be taken into account before introducing these insecticides to control populations of Ae. aegypti in the region.

Multiple insecticide resistances and kdr mutation of Culex pipiens pallens in north-central Anhui Province / 中国血吸虫病防治杂志

Objective To understand the status of resistance to multiple insecticides and the frequencies of kdr mutations in Culex pipiens pallens from north?central Anhui Province. Methods From July to September,2014,the C. pipiens pallens mosquito larvae were collected in Huaibei,Bengbu and Chuzhou cities of the north?central Anhui Province and reared to adults. The female adult mosquitoes at 3-5 days post emergence were tested for susceptibility to the four insecticides,namely 0.05%deltamethrin,5%malathion,0.1%bendiocarb and 4%DDT,by using the standard WHO resistance tube bioassay. The detec?tion of the point mutations of the kdr gene at codon 1014 was conducted by PCR and DNA sequencing in the deltamethrin?resis?tant and?susceptible mosquitoes. Results High levels of resistance to all the four insecticides were found in all the three tested populations,although mosquito mortality varied among populations and test insecticides. Among the test insecticides,DDT showed lowest mortality with no significant difference(F=1.027,P>0.05)in all test populations,whereas significantly differ?ent mortalities were observed among populations for the remained three insecticides tested(deltamethrin,malathion,and ben?diocarb)(F = 23.823,33.955,128.841;all P < 0.01). Two types of non?synonymous kdr mutation at codon position 1014 (L1014F and L1014S)were observed. A positive correlation between L1014F mutation frequencies and deltamethrin resistance levels were detected in the three mosquito populations(r2=0.718,P<0.01). Conclusions The observed high levels of resis?tance to multiple?insecticides coupled with the occurrence of medium to high kdr frequencies in populations of C. pipiens pallens could profoundly affect the mosquito vector control programme in China. The local health departments need to strengthen vector dynamic monitoring and implement rational resistance management strategies.

Resistance to pyrethroid insecticides and analysis of knockdown resistance (kdr) gene mutations in Aedes albopictus from Haikou City / 第二军医大学学报

Objective To investigate the resistance to deltamethrin and permethrin in Aedes albopictus in Haikou City, and to detect the knockdown resistance (kdr) mutants of A. albopictus, elucidating the association of resistant phenotype with kdr mutation. Methods The A. albopictus samples were collected from Xinfu Island, Longtang Town, Shishan Town, Baishamen Park, and the flower market in Haikou City. Their susceptibility to deltamethrin and permethrin was tested by larval bioassay. The resistance level was determined by the R/S ratio. The kdr gene was amplified and analyzed, and the relationship between the resistant phenotype and kdr mutation was determined by Chi-square test. Results The A. albopictus samples from the flower market and Baishamen Park showed resistance to deltamethrin and permethrin, of which the flower market population had an R/S ratio of up to 436.36, indicating a high level of resistance. The A. albopictus samples from Longtang Town and Shishan Town were sensitive to permethrin, but had low (R/S ratio, 9.09) and moderate (18.18) degrees of resistance to deltamethrin, respectively. The Xinfu Island population was sensitive to both deltamethrin and permethrin. The codon 1 534 kdr was detected point mutations among 317 A. albopictus samples, with two mutant alleles, TGC/C and TCC/S. The five genotypes were wild-type TTC/TTC (frequency, 62.15%), wild/mutant heterozygous TTC/TGC (0.63%) and TTC/TCC (16.09%), homozygous mutant TCC/TCC (20.82%), and mutant heterozygous TGC/TCC (0.32%). Chi-square test revealed significant differences in the frequency of kdr mutant genotype between A. albopictu sensitive to the insecticides (31.97%) and those resistant to the insecticides (42.94%), P <0.05. Conclusion Some A. albopictus populations in Haikou City have developed resistance to both deltamethrin and permethrin. The resistant phenotype and kdr mutant genotype are closely associated. This study has also discovered for the first time the novel TCC/S point mutation of kdr.

Expression of vascular endothelial cell growth factor and its receptors in vitreous of patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

To investigate the expression of vascular endothelial cell growth factor (VEGF) and its receptors in vitreous of patients with diabetic retinopathy ( DR), and to discuss its role in the development of DR. ●METHODS: Selected 13 patients (16 eyes) with DR and 15 healthy people (15 eyes), the expression of VEGF and its receptors (fms-like tyrosine kinase-1, Flt-1 and kinase insert domain containing receptor, KDR) were evaluated by immunohistochemistry in vitreous. The levels of VEGF, the Flt-1 and KDR in vitreous of patients with DR were examined with enzyme linked immunosorbent assay (ELlSA). ● RESULTS: lmmunohistochemical staining showed that the expression of VEGF, Flt-1 and KDR in vitreous vessel membrane of patients with DR was increased significantly. And the levels of VEGF, Flt - 1 and KDR in vitreous of patients with DR were obviously higher than that in the control group (P ● CONCLUSlON: VEGF, Flt - 1 and KDR were widely expressed in vitreous of patients with DR, and were positively related to micro-angiogenesis of DR patients. lt proved that VEGF and its receptors played important roles in the occurrence and development of DR.

Evidence of increasing L1014F kdr mutation frequency in Anopheles gambiae s.l. pyrethroid resistant following a nationwide distribution of LLINs by the Beninese National Malaria Control Programme

<p><b>OBJECTIVE</b>To determine the susceptibility status to pyrethroid in Anopheles gambiae s.l. (An. gambiae), the distribution of kdr "Leu-Phe" mutation in malaria vectors in Benin and to compare the current frequency of kdr "Leu-Phe" mutation to the previous frequency after long-lasting insecticide treated nets implementation.</p><p><b>METHODS</b>Larvae and pupae of An. gambiae s.l. mosquitoes were collected from the breeding sites in Littoral, Zou, Borgou and Alibori provinces. CDC susceptibility tests were conducted on unfed females mosquitoes aged 2-5 d old. An. gambiae mosquitoes were identified to species using PCR techniques. Molecular assays were also carried out to identify kdr mutations in individual mosquitoes.</p><p><b>RESULTS</b>The results showed that An. gambiae Malanville and Suru-lere populations were resistant to deltamethrin. Regarding An. gambiae Parakou and Bohicon populations, they were resistant to permethrin. PCR revealed 100% of mosquitoes tested were An. gambiae s.s. The L1014F kdr mutation was found in An. gambiae s.s. Malanville and Parakou at various allelic frequencies. The increase of kdr allelic frequency was positively correlated with CDC bioassays data.</p><p><b>CONCLUSIONS</b>: Pyrethroid resistance is widespread in malaria vector in Benin and kdr mutation is the main resistance mechanism involved. More attention may be paid for the future success of malaria control programmes based on LLINs with pyrethroids in the country.</p>

Evidence of increasing L1014F kdr mutation frequency in Anopheles gambiae s.l. pyrethroid resistant following a nationwide distribution of LLINs by the Beninese National Malaria Control Programme

Objective:To determine the susceptibility status to pyrethroid in Anopheles gambiae s.l. (An. gambiae), the distribution of kdr“Leu-Phe”mutation in malaria vectors in Benin and to compare the current frequency of kdr“Leu-Phe”mutation to the previous frequency after long-lasting insecticide treated nets implementation. Methods: Larvae and pupae of An. gambiae s.l. mosquitoes were collected from the breeding sites in Littoral, Zou, Borgou and Alibori provinces. CDC susceptibility tests were conducted on unfed females mosquitoes aged 2-5 d old. An. gambiae mosquitoes were identified to species using PCR techniques. Molecular assays were also carried out to identify kdr mutations in individual mosquitoes. Results: The results showed that An. gambiae Malanville and Suru-lere populations were resistant to deltamethrin. Regarding An. gambiae Parakou and Bohicon populations, they were resistant to permethrin. PCR revealed 100%of mosquitoes tested were An. gambiae s.s. The L1014F kdr mutation was found in An. gambiae s.s. Malanville and Parakou at various allelic frequencies. The increase of kdr allelic frequency was positively correlated with CDC bioassays data. Conclusions: Pyrethroid resistance is widespread in malaria vector in Benin and kdr mutation is the main resistance mechanism involved. More attention may be paid for the future success of malaria control programmes based on LLINs with pyrethroids in the country.