Zika virus outbreak and the case for building effective and sustainable rapid diagnostics laboratory capacity globally

New and re-emerging pathogens with epidemic potential have threatened global health security for the past century.1 As with the recent Ebola Virus Disease (EVD) epidemic, the Zika Virus (ZIKV) outbreak has yet again surprised and overwhelmed the international health community with an unexpected event for which it might have been better prepared

Implicación de NF-κB y p53 en la expresión de receptores de muerte-TRAIL y apoptosis por procianidinas en células metastásicas humanas SW620

Introduction. The nuclear factor-kappaB (NF-κB) has been shown to upregulate pro-apoptotic mediators such as TRAIL-DR4/-DR5 receptors and the p53 transcription factor depending on the type of stimulus and the cell type involved. Previously, apple procyanidins (Pcy) have been shown to upregulate the expression of TRAIL-DR4/-DR5 and thereby overcoming the resistance of human colon cancer-derived metastatic SW620 cells to TRAIL.Objectives. NF-κB and p53 were investigated for their involvement in the Pcy-triggered apoptosis of human derived-metastatic colon cancer (SW620) cells.Materials and methods. Cell death, p53, TRAIL-DR4/-DR5 proteins were analyzed by flow cytometry. DR4/DR5 mRNA was analyzed by RT-PCR in real time. Activated p50/p65 and p53 forms were studied by ELISA and immunoblotting.Results. Pcy-triggered cell death was prevented by specific inhibitors of NF-κB and of p53: amino-4-(4-phenoxy-phenylethylamino) quinazoline (QNZ) and pifithrin α (Pα), respectively. QNZ and Pα inhibited the Pcy-dependent activation of TRAIL-DR4/-DR5 death receptors. However, the upregulation of TRAIL-DR4 by Pcy was significantly decreased only when NF-κB and p53 inhibitors were used in combination; this effect was not observed with a single inhibitor. This effect was not observed for TRAIL-DR5 and suggested that the expression of each TRAIL-death receptor may be regulated differently.Conclusions. These data suggested that NF-κB and p53 are partially required in Pcy-triggered apoptosis of SW620 cells by up-regulating the expression of TRAIL-DR4/-DR5. In addition, the ratio between TRAIL-DR4/-DR5 may be a determining factor in the activation of TRAIL-death receptor mediated apoptosis.  Introducción. Se ha demostrado que el factor nuclear-κB y p53 aumentan los mediadores proapoptósicos como los receptores de muerte TRAIL-DR4/-DR5, según el estímulo y el tipo celular. Previamente demostramos que las procianidinas de manzana aumentaban la expresión de TRAIL-DR4/-DR5, superando la resistencia a TRAIL característica en células humanas metastásicas SW620 derivadas del cáncer de colon.Objetivo. Investigar si NF-κB y p53 están involucrados en la apoptosis inducida por procianidinas en las células SW620.Materiales y métodos. La muerte celular y las proteínas p53, TRAIL-DR4/-DR5 se analizaron por citometría de flujo. Los ARN mensajeros (ARNm) de DR4/DR5 se analizaron por RT-PCR. Las formas activadas de p50/p65 y p53 se estudiaron por ELISA e inmunodetección.Resultados. La muerte celular activada por procianidinas fue prevenida por inhibidores específicos de NF-κB y de p53: amino-4-(4-fenoxi-feniletilamino)-quinazolina y pifitrina α, respectivamente. La quinazolina y la pifitrina α inhibieron la activación dependiente de procianidinas de TRAIL-DR4/DR5. Sin embargo, el aumento en la expresión de TRAIL-DR4 disminuyó significativamente sólo cuando la quinazolina y la pifitrina α se usaron simultáneamente; este efecto no se observó con cada uno por separado. No se observaron para TRAIL-DR5 estos efectos, lo cual sugiere que la expresión de cada receptor de muerte TRAIL puede estar regulada en forma diferente. Conclusiones. Estos datos sugieren que NF-κB y p53 se requieren parcialmente en la apoptosis de células SW620 inducida por procianidinas mediante el aumento en TRAIL-DR4/-DR5. La proporción de DR4/DR5 podría ser un factor determinante en la activación de la apoptosis por vía de TRAIL-DR4/-DR5

Activación diferencial de la apoptosis vía Fas (CD95) por procianidinas de manzana en células humanas de cáncer de colon y sus derivadas metastásicas

Introduction: We investigated the effects of apple procyanidins (Pcy), oligomers of catechins and epicatechins on Fas receptor expression and function in human colon adenocarcinoma cells (SW480) and in their derived metastatic cells (SW620). Methods: Pcy were characterized by reverse-phase HPLC. Cell death, Fas proteins, DNA fragmentation, and mitochondrial membrane potential were analyzed by flow cytometry. Fas mRNA was analyzed by RT-PCR in real time. Results: Pcy up-regulated the expression of the Fas receptor at the cell surface of both cell lines but activated Fas gene transcription only in SW620 cells. In SW480 cells, Pcy combined with Fas agonist CH-11 enhanced Fas-mediated apoptosis involving the loss of mitochondrial membrane potential and DNA fragmentation, which were abrogated by the antagonist antibody of Fas receptor, the anti-Fas ZB4. On the contrary, in SW620 cells, CH-11 was not able to enhance Pcy-triggered apoptosis indicating that Fas receptor-mediated apoptosis was not activated in these cells despite an up-regulation of Fas receptor gene expression. However, it was observed in SW620 cells that Pcy activated the Fas receptor-mediated apoptotic pathway after a specific blockage of TRAIL-death DR4/DR5 receptors. Conclusions: The present data showed that Pcy were able to activate the Fas receptor apoptotic pathway in SW480 cells and favored a cross-talk between TRAIL and Fas receptors in SW620 cells because specific blocking of TRAIL death receptors favored activation of the Fas receptor-mediated apoptosis. These important data may allow the emergence of new therapeutic protocols targeting death receptors against resistant metastatic cells. Introducción: Se estudiaron los efectos de procianidinas (Pcy) de manzana, oligómeros de catequinas y epicatequinas en la expresión y función del receptor Fas en células humanas de cáncer de colon (SW480) y sus derivadas metastásicas (SW620). Métodos: Las Pcy se caracterizaron por cromatografía líquida de alta presión (HPLC) en fase-reversa. Se analizaron por citometría de flujo la muerte celular, la proteína Fas, la fragmentación del ADN y el potencial de la membrana mitocondrial. Se analizaron los transcriptos de Fas por RT-PCR en tiempo real.Resultados: Las Pcy aumentaron la expresión del receptor Fas en la superficie celular de ambas líneas celulares pero la transcripción del gen Fas fue activado transcripcionalmente sólo en las células SW620. En las células SW480, las Pcy combinadas con el agonista de Fas CH-11 potenció la apoptosis mediada por Fas involucrando la pérdida del potencial mitocondrial de membrana y la fragmentación del ADN los cuales fueron evadidos por el anticuerpo antagonista del receptor Fas anti-ZB4. Por el contrario, en las células SW620, CH-11 no fue capaz de potenciar la apoptosis activada por Pcy indicando que la apoptosis mediada por el receptor Fas no fue activada en estas células a pesar del aumento en la expresión de Fas por regulación a nivel transcripcional. Sin embargo, se observó en las células SW620 que las Pcy activaron la vía apoptótica mediada por el receptor Fas después de un bloqueo específico de los receptores de muerte TRAIL DR4/DR5. Conclusiones: Estos datos muestran que las Pcy fueron capaces de activar la apoptosis a través del receptor Fas en las células SW480 y favorecieron una intercomunicación entre los receptores TRAIL y Fas en las células SW620 debido a que el bloqueo específico de los receptores de muerte TRAIL favoreció la activación de la apoptosis mediada por el receptor Fas. Estos datos podrían permitir el surgimiento de nuevos protocolos terapéuticos dirigidos contra receptores de muerte en células metastásicas resistentes

A comparative analysis of the goal orientation and test anxiety of high school students with and without private tutors

The purpose of the study was to find out whether the goal orientation and test anxiety of high school students engaged in shadow education will be different from those whose only source of learning is that of mainstream education. A total of 387 high school students participated in this comparative research. They completed the Patterns of Adaptive Learning Scales and the Test Anxiety Inventory and results show that high school students without exposure to shadow education are more mastery-oriented while those with those with private tutors are more performance-oriented. In terms of test anxiety, it appears that those engaged in shadow education are more anxious about the testing process as compared to those without private tutors. Specifically, significant differences between the two groups were observed in their mastery orientation, level of emotionality and total test anxiety

Brazilian version of the Segmental Assessment of Trunk Control (SATCo)

The Segmental Assessment of Trunk Control (SATCo) test was translated and adapted to Brazilian Portuguese. Two English language proficient professionals independently translated the original scale for the Brazilian Portuguese (T1 and T2). Following, the translated version of consensus was generated (TU). Two translators performed two versions in English (BT1 and BT2) of the TU version. A new process of consensus between translators and researchers resulted in an English version (BTfinal) that was compared with the original version, aiming to detect possible semantic differences. The version of the instrument in Brazilian Portuguese (TU), called Segmental Assessment of Trunk Control, was revised by the experts’ committee, composed of three physical therapists for content checking and the second version of agreement was generated (Tfinal). Tfinal was submitted to one of the original scale authors to check the understanding of the Brazilian Portuguese version. After this step, 20 physical therapists applied the scale in children with Cerebral Palsy. Part of the physical therapists indicates the need for complementary information in the description of instructions and score.Se tradujo y se adaptó al portugués de Brasil la prueba Segmental Assessment of Trunk Control (SATCo). La escala original fue traducida de forma independiente por dos expertos en lengua inglesa al portugués de Brasil (T1 y T2). Después se produjo una versión traducida en acuerdo (TU). De esta versión, fueron realizadas dos versiones en lengua inglesa por dos traductores (RT1 y RT2). Se formó un nuevo consenso entre traductores e investigadores del cual generó una versión en lengua inglesa (RTfinal), en que se comparó al original para encontrar diferencias semánticas. La versión del instrumento en portugués brasileño (TU) se llamó Avaliação Segmentar do Controle de Tronco (Evaluación Segmentaria de Control del Tronco) y fue corregida por un conjunto de expertos, constituido de tres fisioterapeutas, para chequear el contenido, siendo que al final generó una segunda versión por consenso (Tfinal). Esta segunda versión la mandaron a una de las autoras de la escala original para chequear la comprensión de la versión en portugués de Brasil. Tras esta etapa, veinte fisioterapeutas aplicaron esta escala a niños con parálisis cerebral. Una parte de los fisioterapeutas señalan que es necesario complementar informaciones sobre la descripción de las instrucciones y de los puntajes.Traduziu-se e adaptou-se para o português do Brasil o teste Segmental Assessment of Trunk Control (SATCo). Dois profissionais proficientes na língua inglesa traduziram, independentemente, a escala original para o português do Brasil (T1 e T2). Em seguida, gerou-se a versão traduzida de consenso (TU). Dois tradutores realizaram duas versões em inglês (RT1 e RT2) da versão TU. Um novo processo de consenso entre tradutores e pesquisadores resultou em uma versão em inglês (RTfinal), que foi comparada com a versão original, com vistas a possíveis diferenças semânticas. A versão do instrumento em português do Brasil (TU), denominada “Avaliação Segmentar do Controle de Tronco”, foi revisada pela comissão de especialistas, composta por três fisioterapeutas, para verificação do conteúdo e gerou a segunda versão de concordância (Tfinal). Tfinal foi encaminhada a uma das autoras da escala original para verificar o entendimento da versão em português do Brasil. Após essa etapa, 20 fisioterapeutas aplicaram a escala em crianças com paralisia cerebral. Parte dos fisioterapeutas indica a necessidade de complementação de informação na descrição das instruções e na descrição da pontuação

Lifespan extension and the doctrine of double effect

Recent developments in biogerontology—the study of the biology of ageing—suggest that it may eventually be possible to intervene in the human ageing process. This, in turn, offers the prospect of significantly postponing the onset of age-related diseases. The biogerontological project, however, has met with strong resistance, especially by deontologists. They consider the act of intervening in the ageing process impermissible on the grounds that it would (most probably) bring about an extended maximum lifespan—a state of affairs that they deem intrinsically bad. In a bid to convince their deontological opponents of the permissibility of this act, proponents of biogerontology invoke an argument which is grounded in the doctrine of double effect. Surprisingly, their argument, which we refer to as the ‘double effect argument’, has gone unnoticed. This article exposes and critically evaluates this ‘double effect argument’. To this end, we first review a series of excerpts from the ethical debate on biogerontology in order to substantiate the presence of double effect reasoning. Next, we attempt to determine the role that the ‘double effect argument’ is meant to fulfil within this debate. Finally, we assess whether the act of intervening in ageing actually can be justified using double effect reasoning

Plasmodium malariae contributes to high levels of malaria transmission in a forest–savannah transition area in Cameroon

Background: Malaria control efforts are highly skewed towards Plasmodium falciparum while overlooking other Plasmodium species such as P. malariae. A better understanding of the role of Plasmodium species other than P. falciparum is needed to strengthen malaria elimination initiatives. The aim of the present study was to elucidate the contribution of P. malariae to malaria transmission in Cameroon. Methods: The study was conducted in the Ngatti Health District, a forest–savannah transition area in the Adamawa Region, Cameroon. A total of 497 individuals aged from 1 to 85 years were diagnosed with malaria in November 2020 using a rapid diagnostic test (RDT) and microscopy. Adult mosquitoes were collected between September 2019 and March 2020 by indoor aspiration and identified morphologically and molecularly. The infection status of Plasmodium spp. was also determined by quantitative PCR, and dried blood spots were collected from 156 participants with the aim to detect different Plasmodium species by nested PCR. Results: The overall Plasmodium prevalence was 50.3%, 51.8% and 64.7%, as detected by microscopy, the RDT and PCR, respectively. Based on the PCR results, P. falciparum was the most prevalent species (43%); followed by co-infections P. falciparum/P. malariae (17%), P. falciparum/P. ovale (1.3%), P. falciparum/P. ovale/P. malariae (1.3%); and then by P. malariae mono-infection (2.5%). The same trend was observed using microscopy, with 35% of participants infected with P. falciparum, 11% co-infected with P. falciparum/P. malariae and 4% infected with P. malariae. The prevalence and parasite density of malaria infection varied significantly with age group (P < 0.05), with the highest prevalence rate observed in children aged 6–10 years (P = 0.0001) while the density of Plasmodium infection increased significantly in children aged < 5 years compared to the other age groups (P = 10−3). Among the 757 Anopheles mosquitoes collected, 737 (97.35%) were An. funestus sensu stricto, 15 (1.9%) were An. gambiae and 5 (0.6%) were An. hancocki. The Plasmodium species recorded at the head/thorax level were P. falciparum and P. malariae, with a sporozoite infection rate of 8.4%; the highest sporozoite infection rate was recorded at Mibellon village (13.6%). Conclusion: The results of this study reveal the significant contribution of P. malariae, in addition to P. falciparum, to the high malaria transmission rate in this region. These findings highlight the need to deploy initiatives to also tackle this Plasmodium species to eliminate malaria in the region

Elevated Plasmodium sporozoite infection and multiple insecticide resistance in the principal malaria vectors Anopheles funestus and Anopheles gambiae in a forested locality close to the Yaoundé airport, Cameroon

Background: Reducing the burden of malaria requires better understanding of vector populations, particularly in forested regions where the incidence remains elevated. Here, we characterized malaria vectors in a locality near the Yaoundé international airport, Cameroon, including species composition, abundance, Plasmodium infection rate, insecticide resistance profiles and underlying resistance mechanisms. Methods: Blood-fed adult mosquitoes resting indoors were aspirated from houses in April 2019 at Elende, a village located 2 km from the Yaoundé-Nsimalen airport. Female mosquitoes were forced to lay eggs to generate F1 adult progeny. Bioassays were performed to assess resistance profile to insecticides. The threshold of insecticide susceptibility was defined above 98% mortality rate and mortality rates below 90% were indicative of confirmed insecticide resistance. Furthermore, the molecular basis of resistance and Plasmodium infection rates were investigated. Results: Anopheles funestus s.s. was most abundant species in Elende (85%) followed by Anopheles gambiae s.s. (15%) with both having a similar sporozoite rate. Both species exhibited high levels of resistance to pyrethroids (<40% mortality). An. gambiae s.s. was also resistant to DDT (9.9% mortality) and bendiocarb (54% mortality) while susceptible to organophosphate. An. funestus s.s. was resistant to dieldrin (1% mortality), DDT (86% mortality) but susceptible to carbamates and organophosphates. The L119F-GSTe2 resistance allele (8%) and G119S ace-1 resistance allele (15%) were detected in An. funestus s.s. and An. gambiae s.s., respectively. Furthermore, the high pyrethroid/DDT resistances in An. gambiae s.s. corresponded with an increase frequency of 1014F kdr allele (95%). Transcriptional profiling of candidate cytochrome P450 genes reveals the over-expression of CYP6P5, CYP6P9a and CYP6P9b. Conclusion: The resistance to multiple insecticide classes observed in these vector populations alongside the high Plasmodium sporozoite rate highlights the challenges that vector control programs encounter in sustaining the regular benefits of contemporary insecticide-based control interventions in forested areas

A2 gene of Old World cutaneous Leishmania is a single highly conserved functional gene

BACKGROUND: Leishmaniases are among the most proteiform parasitic infections in humans ranging from unapparent to cutaneous, mucocutaneous or visceral diseases. The various clinical issues depend on complex and still poorly understood mechanisms where both host and parasite factors are interacting. Among the candidate factors of parasite virulence are the A2 genes, a family of multiple genes that are developmentally expressed in species of the Leishmania donovani group responsible for visceral diseases (VL). By contrast, in L. major determining cutaneous infections (CL) we showed that A2 genes are present in a truncated form only. Furthermore, the A2 genomic sequences of L. major were considered subsequently to represent non-expressed pseudogenes [1]. Consequently, it was suggested that the structural and functional properties of A2 genes could play a role in the differential tropism of CL and VL leishmanias. On this basis, it was of importance to determine whether the observed structural/functional particularities of the L. major A2 genes were shared by other CL Leishmania, therefore representing a proper characteristic of CL A2 genes as opposed to those of VL isolates. METHODS: In the present study we amplified by PCR and sequenced the A2 genes from genomic DNA and from clonal libraries of the four Old World CL species comparatively to a clonal population of L. infantum VL parasites. Using RT-PCR we also amplified and sequenced A2 mRNA transcripts from L. major. RESULTS: A unique A2 sequence was identified in Old World cutaneous Leishmania by sequencing. The shared sequence was highly conserved among the various CL strains and species analysed, showing a single polymorphism C/G at position 58. The CL A2 gene was found to be functionally transcribed at both parasite stages. CONCLUSION: The present study shows that cutaneous strains of leishmania share a conserved functional A2 gene. As opposed to the multiple A2 genes described in VL isolates, the CL A2 gene is unique, lacking most of the nucleotide repeats that constitute the variable region at the 5'end of the VL A2 sequences. As the variable region of the VL A2 gene has been shown to correspond to a portion of the protein which is highly immunogenic, the present results support the hypothesis of a possible role of the A2 gene in the differential tropism of CL and VL leishmania parasites

Entomological indicators of Plasmodium species transmission in Goma Tsé-Tsé and Madibou districts, in the Republic of Congo

Background: Malaria remains a major public health problem in the Republic of Congo, with Plasmodium falciparum being the deadliest species of Plasmodium in humans. Vector transmission of malaria is poorly studied in the country and no previous report compared rural and urban data. This study aimed to determine the Anopheles fauna and the entomological indices of malaria transmission in the rural and urban areas in the south of Brazzaville, and beyond. Methods: Indoor household mosquitoes capture using electric aspirator was performed in rural and urban areas during raining and dry seasons in 2021. The identification of Anopheles species was done using binocular magnifier and nested-PCR. TaqMan and nested-PCR were used to detect the Plasmodium species in the head/thorax and abdomens of Anopheles. Some entomological indices including the sporozoite infection rate, the entomological inoculation rate and the man biting rate were estimated. Results: A total of 699 Anopheles mosquitoes were collected: Anopheles gambiae sensu lato (s.l.) (90.7%), Anopheles funestus s.l. (6.9%), and Anopheles moucheti (2.4%). Three species of An. gambiae s.l. were identified including Anopheles gambiae sensu stricto (78.9%), Anopheles coluzzii (15.4%) and Anopheles arabiensis (5.7%). The overall sporozoite infection rate was 22.3% with a predominance of Plasmodium falciparum, followed by Plasmodium malariae and Plasmodium ovale. Anopheles aggressiveness rate was higher in households from rural area (1.1 bites/night) compared to that from urban area (0.8 ib/p/n). The overall entomological inoculation rate was 0.13 ib/p/n. This index was 0.17 ib/p/n and 0.092 ib/p/n in rural and in urban area, respectively, and was similar during the dry (0.18 ib/p/n) and rainy (0.14 ib/p/n) seasons. Conclusion: These findings highlight that malaria transmission remains high in rural and urban area in the south of Republic of Congo despite the ongoing control efforts, thereby indicating the need for more robust interventions