Dengue is the most prevalent arboviral disease of humans. The host and virus variables associated with dengue virus (DENV) transmission from symptomatic dengue cases (n = 208) to Aedes aegypti mosquitoes during 407 independent exposure events was defined. The 50% mosquito infectious dose for each of DENV-1-4 ranged from 6.29 to 7.52 log10 RNA copies/mL of plasma. Increasing day of illness, declining viremia, and rising antibody titers were independently associated with reduced risk of DENV transmission. High early DENV plasma viremia levels in patients were a marker of the duration of human infectiousness, and blood meals containing high concentrations of DENV were positively associated with the prevalence of infectious mosquitoes 14 d after blood feeding. Ambulatory dengue cases had lower viremia levels compared with hospitalized dengue cases but nonetheless at levels predicted to be infectious to mosquitoes. These data define serotype-specific viremia levels that vaccines or drugs must inhibit to prevent DENV transmission.engue is globally the most important mosquito-borne viral disease of humans, with a global burden of ∼100 million cases per annum (1, 2). Aedes aegypti mosquitoes are the primary mosquito vectors of dengue viruses (DENV), of which there are four virus types (DENV-1-4). Multiple factors influence the likelihood of infection and dissemination of DENV in Ae. aegypti and include the amplitude of daily temperature fluctuations (3), mean temperature (4), and the genotype of mosquito and virus (5), among others (6). The extrinsic incubation period (EIP), a critical determinant of vector competence (7,8), is widely accepted to be 7-14 d for DENV in Ae. aegypti, although a recent modeling analysis of historical DENV transmission data has suggested a wider range of 2-15 d at 30°C (9). A major caveat to many of these observations is that they stem from laboratory experiments with artificially generated virus-spiked blood meals and often in-bred colony mosquitoes.The temporal and virological variables associated with the transmission of DENV from a naturally infected human to a biting Ae. aegypti mosquito are poorly understood. Natural history studies of experimental DENV infection of small cohorts of human volunteers in the 1920s by Siler et al. (10,11), likely using DENV-4 (12), and subsequent studies by Simmons et al. (13), likely using DENV-1 (12), suggested that the window of time before the onset of clinical symptoms that DENV-1 or DENV-4 could be transmitted to Ae. aegypti mosquitoes was 6-18 h or 2 d, respectively (14). After fever onset, the duration of infectiousness was 4-5 d for DENV-1 and up to 2 d for DENV-4, with an EIP in the mosquito of 10 d or more. Consistent with this, mosquitobiting studies by Gubler et al. in the 1960s (15-18) collectively estimated that dengue cases were infectious for 4-5 d after illness onset (range, 2-12 d). The human viremia level required to infect Ae. aegypti mosquitoes is unknown, and therefore it is uncertain what percentage of symptomatic (or asymptomatic...

Background. Dengue is the most common arboviral infection of humans. There are currently no specific treatments for dengue. Balapiravir is a prodrug of a nucleoside analogue (called R1479) and an inhibitor of hepatitis C virus replication in vivo.Methods. We conducted in vitro experiments to determine the potency of balapiravir against dengue viruses and then an exploratory, dose-escalating, randomized placebo-controlled trial in adult male patients with dengue with <48 hours of fever.Results. The clinical and laboratory adverse event profile in patients receiving balapiravir at doses of 1500 mg (n = 10) or 3000 mg (n = 22) orally for 5 days was similar to that of patients receiving placebo (n = 32), indicating balapiravir was well tolerated. However, twice daily assessment of viremia and daily assessment of NS1 antigenemia indicated balapiravir did not measurably alter the kinetics of these virological markers, nor did it reduce the fever clearance time. The kinetics of plasma cytokine concentrations and the whole blood transcriptional profile were also not attenuated by balapiravir treatment.Conclusions. Although this trial, the first of its kind in dengue, does not support balapiravir as a candidate drug, it does establish a framework for antiviral treatment trials in dengue and provides the field with a clinically evaluated benchmark molecule.Clinical Trials Registration. NCT01096576.

Background: Understanding the genetic diversity in threatened species that occur in forest remnants is necessary to establish efficient strategies for the species conservation, restoration and management. Panax vietnamensis Ha et Grushv. is medicinally important, endemic and endangered species of Vietnam. However, genetic diversity and structure of population is unknown due to lack of efficient molecular markers.Results: In this study, we employed Illumina HiSeq TM 4000 sequencing to analyze the transcriptomes of P. vietnamensis (roots, leaves and stems). A total of 23,741,783 raw reads were obtained and assembled, from which, 89,271 unigenes with an average length of 598.3191 nt were generated. During functional annotation, 31,686 unigenes were annotated in Gene Ontology categories, Kyoto Encyclopedia of Genes and Genomes pathways, Swiss-Prot database, and Nucleotide Collection (NR/NT) database. In addition, 11,343 expressed sequence tag-simple sequence repeat (EST-SSRs) were detected. From 7,774 primer pairs, 101 were selected for polymorphism validation, in which, 20 primer pairs were successfully amplified to DNA fragments and significant amounts of polymorphism was observed within population. The nine polymorphic microsatellite loci were used to analyze genetic diversity and structure of the natural populations. The obtained results revealed that the shows high levels of genetic diversity in populations, the average observed and expected heterozygosity were H O = 0.422 and H E = 0.479. During the Bottleneck analysis using TPM and SMM models (p < 0.01) shows that targeted population is significantly heterozygote deficient. This suggests sign of bottleneck in all populations. Genetic differentiation among populations was moderate (F ST = 0.133) and indicating limited gene flow (Nm = 1.63). Analysis of molecular variance (AMOVA) showed 63.17% of variation within individuals and 12.45% among populations. These results showed a moderate genetic structure of P. vietnamensis. STRUCTURE analysis and the unweighted pair-group method with arithmetic means (UPGMA) tree revealed strong genetic structure and two genetic clusters related to geographical distances, as well. Conclusion: Our study will assist conservators in future conservation management, breeding, production and habitats restoration of the species.

Abstract Background: Understanding the genetic diversity in endangered species that occur in forest remnants is necessary to establish efficient strategies for the species conservation, restoration and management. Panax vietnamensis Ha et Grushv. is medicinally important, endemic and endangered species of Vietnam. However, genetic diversity and structure of population are unknown due to lack of efficient molecular markers. Results: In this study, we employed Illumina HiSeqTM 4000 sequencing to analyze the transcriptomes of P. vietnamensis (roots, leaves and stems). Raw reads total of 23,741,783 was obtained and then assembled, from which the generated unigenes were 89,271 (average length = 598.3191 nt). The 31,686 unigenes were annotated in different databases i.e. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, Nucleotide Collection (NR/NT) and Swiss-Prot for functional annotation. Further, 11,343 EST-SSRs were detected. From 7,774 primer pairs, 101 were selected for polymorphism validation, in which; 20 primer pairs were successfully amplified to DNA fragments and significant amounts of polymorphism was observed within population. The nine polymorphic microsatellite loci were used for population structure and diversity analyses. The obtained results revealed high levels of genetic diversity in populations, the average observed and expected heterozygosity were HO = 0.422 and HE = 0.479, respectively. During the Bottleneck analysis using TPM and SMM models (p < 0.01) shows that targeted population is significantly heterozygote deficient. This suggests sign of the bottleneck in all populations. Genetic differentiation between populations was moderate (FST = 0.133) and indicating slightly high level of gene flow (Nm = 1.63). Analysis of molecular variance (AMOVA) showed 63.17% of variation within individuals and 12.45% among populations. Our results shows two genetic clusters related to geographical distances. Conclusion: Our study will assist conservators in future conservation management, breeding, production and habitats restoration of the species.

Tacca chantrieri is a useful medicinal plant native to lowland tropical forests in northern Vietnam. However, it is threatened with extinction owing to habitat fragmentation and overexploitation. Genetic diversity in 5 natural populations of T. chantrieri was examined using 20 ISSR primers polymorphism. Our study showed that average PIC was 0.253 (0.018 - 0.460) and average PPB was 62.49 % (40- 85.71). 5 populations had modest genetic variability (Na = 1.384, Ne = 1.32, Ho = 0.182, He = 0.178 and I = 0.258) and the species level had a low value (Na = 1.892, Ne = 1.744, Ho = 0.293, He = 0.389 and I = 0.549). The lowest levels of genetic diversity were detected in Cuc Phuong population (Na = 1.135, Ne = 1.021, Ho = 0.091, He = 0.078 and I = 0.116) and the highest levels were detected in Ba Vi population (Na = 1.555, Ne = 1.436, Ho = 0.247, He = 0.242 and I = 0.350). A substantial positive connection between genetic distance and geographic distance was discovered using the Mantel test (R2 = 0.5086). The implications of the results of this study to conserve the genetic resources of T. chantrieri were discussed.

The medicinal plant resources in K9-Da Chong relic area (Ba Vi, Ha Noi) and surroundings are quite diversity. The survey results show that 328 species of medicinal plants belong to 249 genera of 95 families. Magnoliophyta was found the richest with 321 species (98.85%). The richest 12 families and genus with 137 (41.8%) species and 98 (39.35%), respectively are Euphorbiaceae, Asteraceae, Rubiaceae, Fabaceae, Rutaceae, Apocynaceae, Moraceae, Caesalpiniaceae, Amarantaceae, Verbenaceae, Lauraceae, Zingiberaceae. Ten life-forms of medicinnal species used at the research forest in K9-Da Chong relic area and surroundings with 328 species. The most of them are belong to herb 84 (25.85%), shrub 51 (15.79%) and others. The diversity of plant, depending on their used parts such as: leaves 60, bark 24, trunk 50 and whole plant 35 species, respectively. Several species of medicinal plants were identified, which are able to cure 10 disease groups. K9-Da Chong relic area and surroundings should perform synchronously conservation solutions, encouraging local communities to plant medicinal plants, especially species of rare medicinal plants.

With the development of molecular marker technology in the 1980s, specially their applications solving basic questions and ecological research. To explore the knowledge about molecular ecology, several reviews have been published in the sincelast three decades. The molecular markers have been widely used in study and selection of plants and animals. Molecular marker used in studies of genetic diversity, phylogeny, classification, gene tagging and identification; in germplasm and marker assisted selection. The presence of various types of molecular marker, and differences in their principles, methodologies, and applications require careful consideration in choosing one or more of such methods. Meanwhile, no molecular markeris available that fulfill all requirements needed by researchers. Depending on the nature of each study, one can choose among the variety of molecular marker techniques, each of which combines at least some desirable properties. However, all these reviews were meant for researchers with advanced knowledge of molecular genetics(gene identification and gene mapping), genetic diversity and structure of population, phylogenetic relationship and breeding (plant and animal). This review will provide better understanding in science and technology about recent developments in molecular markers, their applications and possibly devoted to early researchers with a little or no knowledge of molecular markers.

Background: Understanding the genetic diversity in threatened species that occur in forest remnants is necessary to establish efficient strategies for the species conservation, restoration and management. Panax vietnamensis Ha et Grushv. is medicinally important, endemic and endangered species of Vietnam. However, genetic diversity and structure of population is unknown due to lack of efficient molecular markers. Results: In this study, we employed Illumina HiSeq TM 4000 sequencing to analyze the transcriptomes of P. vietnamensis (roots, leaves and stems). A total of 23,741,783 raw reads were obtained and assembled, from which, 89,271 unigenes with an average length of 598.3191 nt were generated. During functional annotation, 31,686 unigenes were annotated in Gene Ontology categories, Kyoto Encyclopedia of Genes and Genomes pathways, Swiss-Prot database, and Nucleotide Collection (NR/NT) database. In addition, 11,343 expressed sequence tag-simple sequence repeat (EST-SSRs) were detected. From 7,774 primer pairs, 101 were selected for polymorphism validation, in which, 20 primer pairs were successfully amplified to DNA fragments and significant amounts of polymorphism was observed within population. The nine polymorphic microsatellite loci were used to analyze genetic diversity and structure of the natural populations. The obtained results revealed that the shows high levels of genetic diversity in populations, the average observed and expected heterozygosity were H O = 0.422 and H E = 0.479. During the Bottleneck analysis using TPM and SMM models (p < 0.01) shows that targeted population is significantly heterozygote deficient. This suggests sign of bottleneck in all populations. Genetic differentiation among populations was moderate (F ST = 0.133) and indicating limited gene flow (Nm = 1.63). Analysis of molecular variance (AMOVA) showed 63.17% of variation within individuals and 12.45% among populations. These results showed a moderate genetic structure of P. vietnamensis. STRUCTURE analysis and the unweighted pair-group method with arithmetic means (UPGMA) tree revealed strong genetic structure and two genetic clusters related to geographical distances, as well. Conclusion: Our study will assist conservators in future conservation management, breeding, production and habitats restoration of the species.