The four serotypes of dengue recognize the same putative receptors in Aedes aegypti midgut and Ae. albopictus cells

BACKGROUND: Dengue viruses (DENV) attach to the host cell surface and subsequently enter the cell by receptor-mediated endocytosis. Several primary and low affinity co-receptors for this flavivirus have been identified. However, the presence of these binding molecules on the cell surface does not necessarily render the cell susceptible to infection. Determination of which of them serve as bona fide receptors for this virus in the vector may be relevant to treating DENV infection and in designing control strategies. RESULTS: (1) Overlay protein binding assay showed two proteins with molecular masses of 80 and 67 kDa (R80 and R67). (2) Specific antibodies against these two proteins inhibited cell binding and infection. (3) Both proteins were bound by all four serotypes of dengue virus. (4) R80 and R67 were purified by affinity chromatography from Ae. aegypti mosquito midguts and from Ae albopictus C6/36 cells. (5) In addition, a protein with molecular mass of 57 kDa was purified by affinity chromatography from the midgut extracts. (6) R80 and R67 from radiolabeled surface membrane proteins of C6/36 cells were immunoprecipitated by antibodies against Ae. aegypti midgut. CONCLUSION: Our results strongly suggest that R67 and R80 are receptors for the four serotypes of dengue virus in the midgut cells of Ae. aegypti and in C6/36 Ae. albopictus cells

Specific genetic markers for detecting subtypes of dengue virus serotype-2 in isolates from the states of Oaxaca and Veracruz, Mexico

<p>Abstract</p> <p>Background</p> <p>Dengue (DEN) is an infectious disease caused by the DEN virus (DENV), which belongs to the <it>Flavivirus </it>genus in the family <it>Flaviviridae</it>. It has a (+) sense RNA genome and is mainly transmitted to humans by the vector mosquito <it>Aedes aegypti</it>. Dengue fever (DF) and dengue hemorrhagic fever (DHF) are caused by one of four closely related virus serotypes (DENV-1, DENV-2, DENV-3 and DENV-4). Epidemiological and evolutionary studies have indicated that host and viral factors are involved in determining disease outcome and have proved the importance of viral genotype in causing severe epidemics. Host immune status and mosquito vectorial capacity are also important influences on the severity of infection. Therefore, an understanding of the relationship between virus variants with altered amino acids and high pathogenicity will provide more information on the molecular epidemiology of DEN. Accordingly, knowledge of the DENV serotypes and genotypes circulating in the latest DEN outbreaks around the world, including Mexico, will contribute to understanding DEN infections.</p> <p>Results</p> <p>1. We obtained 88 isolates of DENV, 27 from Oaxaca and 61 from Veracruz. 2. Of these 88 isolates, 16 were serotype 1; 62 serotype 2; 7 serotype 3; and 2 serotype 4. One isolate had 2 serotypes (DENV-2 and -1). 3. Partial nucleotide sequences of the genes encoding C- prM (14 sequences), the NS3 helicase domain (7 sequences), the NS5 S-adenosyl methionine transferase domain (7 sequences) and the RNA-dependent RNA polymerase (RdRp) domain (18 sequences) were obtained. Phylogenetic analysis showed that DENV-2 isolates belonged to the Asian/American genotype. In addition, the Asian/American genotype was divided into two clusters, one containing the isolates from 2001 and the other the isolates from 2005–2006 with high bootstrap support of 94%.</p> <p>Conclusion</p> <p>DENV-2 was the predominant serotype in the DF and DHF outbreak from 2005 to 2006 in Oaxaca State as well as in the 2006 outbreak in Veracruz State, with the Asian/American genotype prevalent in both states. Interestingly, DENV-1 and DENV-2 were the only serotypes related to DHF cases. In contrast, DENV-3 and DENV-4 were poorly represented according to epidemiological data reported in Mexico. We found that isoleucine was replaced by valine at residue 106 of protein C in the isolates from these 2005–2006 outbreaks and in those from the 1997, 1998 and 2001 outbreaks in the Caribbean islands. We suggested that this amino acid change may be used as a signature for isolates arising in the Caribbean islands and pertaining to the Asian/American genotype. Other amino acid changes are specific for the Asian/American, Asian and American strains.</p

The distribution of potential West Nile virus vectors, Culex pipiens pipiens and Culex pipiens quinquefasciatus (Diptera: Culicidae), in Mexico City

<p>Abstract</p> <p>Background</p> <p><it>Culex </it>spp. mosquitoes are considered to be the most important vectors of West Nile virus (WNV) detected in at least 34 species of mosquitoes in the United States. In North America, <it>Culex pipiens pipiens, Culex pipiens quinquefasciatus</it>, and <it>Culex tarsalis </it>are all competent vectors of WNV, which is considered to be enzootic in the United States and has also been detected in equines and birds in many states of Mexico and in humans in Nuevo Leon. There is potential for WNV to be introduced into Mexico City by various means including infected mosquitoes on airplanes, migrating birds, ground transportation and infected humans. Little is known of the geographic distribution of <it>Culex pipiens </it>complex mosquitoes and hybrids in Mexico City. <it>Culex pipiens pipiens </it>preferentially feed on avian hosts; <it>Culex pipiens quinquefasciatus </it>have historically been considered to prefer mammalian hosts; and hybrids of these two species could theoretically serve as bridge vectors to transmit WNV from avian hosts to humans and other mammalian hosts. In order to address the potential of WNV being introduced into Mexico City, we have determined the identity and spatial distribution of <it>Culex pipiens </it>complex mosquitoes and their hybrids.</p> <p>Results</p> <p>Mosquito larvae collected from 103 sites throughout Mexico City during 2004-2005 were identified as <it>Culex, Culiseta </it>or <it>Ochlerotatus </it>by morphological analysis. Within the genus <it>Culex</it>, specimens were further identified as <it>Culex tarsalis </it>or as belonging to the <it>Culex pipiens </it>complex. Members of the <it>Culex pipiens </it>complex were separated by measuring the ratio of the dorsal and ventral arms (DV/D ratio) of the male genitalia and also by using diagnostic primers designed for the <it>Ace.2 </it>gene. <it>Culex pipiens quinquefasciatus </it>was the most abundant form collected.</p> <p>Conclusions</p> <p>Important WNV vectors species, <it>Cx. p. pipiens</it>, <it>Cx. p. quinquefasciatus </it>and <it>Cx. tarsalis</it>, are all present in Mexico City. Hybrids of <it>Cx. p. pipiens </it>and <it>Cx. p. quinquefasciatus </it>were also collected and identified. The presence and abundance of these WNV competent vectors is a cause for concern. Understanding the distribution of these vectors can help improve viral surveillance activities and mosquito control efforts in Mexico City.</p

The Role of miR-107 in Prostate Cancer: A Review and Experimental Evidence

Over the past two decades, several research groups have focused on the functioning of microRNAs (miRNAs), because many of them function as positive or negative endogenous regulators of processes that alter during the development of cancer. Prostate cancer is the second most commonly occurring cancer in men. New biomarkers are needed to support the diagnosis of prostate cancer. Although it is necessary to deepen the research on this molecule to explore its potential utility in the diagnosis, follow-up, and prognosis of cancer, our results support a role of miR-107 in the signaling cascades that allow cancer progression, and as shown here, in the progression of Prostate Cancer (PCa). These findings strongly suggest that miR-107 may be a potential circulating biomarker for the diagnosis and prognosis of prostate cancer

Rab11 and Actin Cytoskeleton Participate in Giardia lamblia Encystation, Guiding the Specific Vesicles to the Cyst Wall

The encystation process is crucial for survival and transmission of Giardia lamblia to new hosts. During this process, vesicular trafficking and the cytoskeleton play important roles. In eukaryotic cells, intracellular transport is regulated by proteins, including Rab-GTPases and SNAREs, which regulate vesicle formation along with recognition of and binding to the target membrane. Cytoskeletal structures are also involved in these processes. In this study, we demonstrate the participation of Rab11 in the transport of encystation-specific vesicles (ESVs). Additionally, we demonstrate that disruption of actin microfilaments affects ESVs transport. The modification of actin dynamics was also correlated with a reduction in rab11 and cwp1 expression. Furthermore, down-regulation of rab11 mRNA by a specific hammerhead ribozyme caused nonspecific localization of CWP1. We thus provide new information about the molecular machinery that regulates Giardia lamblia encystation. Given our findings, Rab11 and actin may be useful targets to block Giardia encystation

Multiple recombinants in two dengue virus, serotype-2 isolates from patients from Oaxaca, Mexico

Abstract Background Dengue (DEN) is a serious cause of mortality and morbidity in the world including Mexico, where the infection is endemic. One of the states with the highest rate of dengue cases is Oaxaca. The cause of DEN is a positive-sense RNA virus, the dengue virus (DENV) that evolves rapidly increasing its variability due to the absence of a repair mechanism that leads to approximately one mutational event per genome replication; which results in enhancement of viral adaptation, including the escape from host immune responses. Additionally, recombination may play a role in driving the evolution of DENV, which may potentially affect virulence and cause host tropism changes. Recombination in DENV has not been described in Mexican strains, neither has been described the relevance in virus evolution in an endemic state such as Oaxaca where the four serotypes of DENV are circulating. Results To study whether there are isolates from Oaxaca having recombination, we obtained the sequence of 6 different isolates of DENV-2 Asian/American genotype from the outbreak 2005-6, one clone of the C(91)-prM-E-NS1(2400) structural genes, and 10 clones of the E gene from the isolate MEX_OAX_1656_05. Evidence of recombination was found by using different methods along with two softwares: RDP3 and GARD. The Oaxaca MEX_OAX_1656_05 and MEX_OAX_1038_05 isolates sequenced in this study were recombinant viruses that incorporate the genome sequence from the Cosmopolitan genotype. Furthermore, the clone of the E gene namely MEX_OAX_165607_05 from this study was also recombinant, incorporating genome sequence from the American genotype. Conclusions This is the first report of recombination in DENV-2 in Mexico. Given such a recombinant activity new genomic combinations were produced, this could play a significant role in the DENV evolution and must be considered as a potentially important mechanism generating genetic variation in this virus with serious implications for the vaccines and drugs formulation as occurs for other viruses like poliovirus, influenza and HIV.</p

Estrategias genómicas y moleculares para el control de la babesiosis bovina

The control of bovine babesiosis around the world is restricted to chemotherapeutic treatment and tick population reduction by acaricide agents. There are no control strategies based on herd immunity studies, programs on integral control of ticks and tick-transmitted diseases, neither are commercially available safe vaccines against babesiosis. In order to develop these tools it is necessary the use of genomics, proteomics, and bioinformatics together with research on genes with a potential use as diagnostics or vaccines. Studies on the function of those genes and, the degree of conservation of variation are mandatory to determine their usefulness. First, it is necessary to identify genes with a potential use in the development of these tools and then, evaluate their variation or conservation between different parasite populations. Second, specific domains of those genes must be selected in order for them to be used as desired, whether they are on conserved or variable regions of those genes in different strains. Finally, the best evaluation method for those genes must be employed to develop adequate control methods. Although there is some research in the study of Babesia bovis genes, there is practically no information about B. bigemina. Therefore it is necessary to take advantage of the genomics and bioinformatics strategies to identify new genes as diagnostics and vaccine potential. The development of the Mexican livestock depends upon the implementation of these tools. KEY WORDS: Bovine babesiosis, diagnostics, bioinformatics.El control de la babesiosis bovina en muchas partes del mundo está restringido al tratamiento quimioterapéutico y al control de la población de garrapatas con agentes acaricidas. No hay programas de control basados en estudios de inmunidad de hato, control integral de la garrapata y las enfermedades que transmite, ni vacunas contra la babesiosis disponibles comercialmente. Para poder desarrollar estas herramientas es necesario utilizar tecnologías que incluyan conocimientos de genómica, proteómica y bioinformática, apoyadas en la investigación de genes con potencial diagnóstico o vacunal. El estudio de la función de los genes, y de la conservación o variabilidad son indispensables para determinar su utilidad. Es necesario, primero identificar los genes con potencial a incluirse en el desarrollo de estas herramientas, y después, evaluar su variabilidad o conservación en distintas poblaciones de parásitos. En segundo término, es necesario seleccionar regiones específicas de estos genes, que cumplan la función deseada, ya sean regiones conservadas o diferentes entre cepas. Finalmente, es necesario utilizar el método adecuado de evaluación de estos candidatos para el desarrollo de métodos de control adecuados. A pesar de que hay ciertos avances en el estudio de genes de B. bovis, hay prácticamente nula información respecto a B. bigemina. Es necesario aprovechar las nuevas estrategias genómicas y de bioinformática para identificar nuevos genes con potencial diagnóstico y de vacunación. El desarrollo de la ganadería mexicana está supeditado al establecimiento e implementación de estas herramientas

Expression of Polymorphic msp1β Genes during Acute Anaplasma marginale Rickettsemia

Immunization of cattle with native MSP1 induces protection against Anaplasma marginale . The native immunogen is composed of a single MSP1a protein and multiple, undefined MSP1b polypeptides. In addition to the originally sequenced gene, designated msp1β(F1) , we identified three complete msp1 β genes in the Florida strain: msp1β(F2) , msp1β(F3) , and msp1β(F4) . Each of these polymorphic genes encodes a structurally unique MSP1b protein, and unique transcripts can be identified during acute A. marginale rickettsemia. The structural polymorphism is clustered in discrete variable regions, and each MSP1b protein results from a unique mosaic of five variable regions. Although each of the MSP1b proteins in the Florida strain contains epitopes recognized by serum antibody induced by protective immunization with the native MSP1 complex, the variable regions also include epitopes expressed by some but not all of the MSP1b proteins. These data support testing recombinant vaccines composed of the multiple antigenically and structurally unique MSP1b proteins combined with MSP1a in order to mimic the efficacy of native MSP1 immunization