Chitosanase may enhance anti-fungal defense responses in transgenic tobacco

Chitosanase is an enzyme, similar to chitinase, capable of hydrolyzing the β-1,4-linkages between N-acetyl-D-glucosamine and D-glucosamine residues in partially acetylated chitosan polymers found in fungal cell walls. When attacked by pathogenic fungi, many plants exploit this hydrolytic action as a component of a larger post-attack defense response, but these enzymes may also play a role in the initial plant-pathogen interaction via the generation of elicitors resulting from the hydrolysis of fungal cell walls. To gain insight into these mechanisms, a Paenbacillus chitosanase was cloned, sequenced, and modified for plant expression. The modified gene was delivered to tobacco (Nicotiana tabacum L. cv. Xanthine) leaf disks via Agrobacterium tumenfaciensmediated transformation. Whole plants were regenerated from the transformed cells. The putative transformants were tested for transgene integration, transcription, and translation. Confirmed transformants were then screened for enhanced responses to a Rhizoctonia solani cellwall preparation by measuring time-course production of hydrogen peroxide, phenylalanine ammonia lyase, and peroxidase. These compounds play roles at different points in a pathogenesis-related signal transduction pathway and thus allow for an initial assessment of the global defense response. Preliminary data suggest that transgenic tobacco constitutively expressing a Paenbacillus chitosanase may activate pathogenesis-related defense responses more quickly than wild type tobacco

Chitosanase May Enhance Anti-Fungal Defense Responses in Transgenic Tobacco

Fungicides are expensive, dangerous, and can be harmful to the environment, but they are often necessary for profitable farming operations. New technologies may soon allow farmers to replace these chemicals with genetically engineered plants producing antifungal enzymes that degrade fungal cell walls. To explore this option, a Paenbacillus chitosanase gene was cloned, sequenced, and modified for plant expression. The modified gene was delivered to tobacco (Nicotiana tabacum L cv. Xanthine) leaf disks via Agrobacterium tumenfaciens-mediated transformation. The putative GMOs were tested for transgene integration, transcription, and translation. Confirmed transformants were then screened for enhanced responses to a Rhizoctonia solani cell wall preparation by measuring time-course production of hydrogen peroxide, phenyalanine ammonia lyase, and peroxidase. These compounds play roles at different points in a pathogensis-related signal transduction pathway and, thus, allow for an initial assessment of the global defense response. Preliminary data suggest that transgenic tobacco constitutively expressing a Paenbacillus chitosanase may activate pathogenesis-related defense responses more quickly than wild type tobacco

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Brief Report: Phase IIa Safety Study of a Vaginal Ring Containing Dapivirine in Adolescent Young Women.

BackgroundYoung women aged 15-24 years are disproportionately affected by the HIV epidemic. Two phase III trials of a vaginal ring containing 25-mg dapivirine demonstrated HIV-1 risk reduction in adult women older than 21 years but not in those aged 18-21 years. Lack of protection was correlated with low adherence.MethodsIn this phase-IIa, randomized, double-blind, placebo-controlled, US, multicenter trial of the dapivirine ring in sexually active females, aged 15-17 years, participants were randomized 3:1 to a dapivirine or placebo ring to be inserted monthly for 6 months (NCT02028338). Primary safety end points included grade 2 product related adverse events and any grade 3 and higher adverse events. Adherence to ring use was assessed by plasma dapivirine concentrations, residual levels in used rings, and self-report. A plasma dapivirine concentration of >95 pg/mL was used to define short-term adherence; a residual ring level of <23.5 mg was used to define long-term adherence. Acceptability was assessed through computer-assisted self-interviews.ResultsNinety-six participants were enrolled across 6 US sites. The median age was 16.0 years. There were no differences in safety outcomes between treatment arms. Adherence to the dapivirine ring was demonstrated by both plasma measurements (87%) and residual drug levels in rings (95%). Forty-two percent (95% confidence interval: 32 to 52) of participants reported that they never removed the ring. Participants noted no discomfort due to the ring at 87% of visits and "liking" the ring at 93% of visits.ConclusionThe dapivirine vaginal ring, a promising topical microbicide, was well tolerated and acceptable in young US adolescents

Exosomes and breast cancer drug resistance

Lymphatic filariasis (LF) is a vector-borne parasitic disease that is being targeted for elimination through mass drug administration (MDA). The co-distribution of Loa loa in Central Africa poses a significant barrier to the expansion of the MDA due to risk of severe adverse events (SAEs) associated with the drug ivermectin that is routinely used. National LF programmes are yet to significantly scale up in co-endemic areas and need a practical approach to make preliminary decisions based on the mapping status and potential treatment strategies. We reviewed relevant data available to WHO and in the literature for LF-L. loa endemic countries to develop a simple method to support the scale-up of MDA to eliminate LF. A basic model for national LF programmes to work from at the administrative or implementation unit (IU) level has been developed for LF - L. loa co-endemic countries. The model includes five practical steps, which comprise the development of a national filarial database and a simple classification system to help determine the mapping status and most appropriate treatment strategy. Steps are colour-coded and linked to a general decision tree, which is also presented. This IU-level model is simple to follow and will help LF elimination programmes develop an action plan and scale up the implementation of alternative treatment strategies in L. loa co-endemic areas. The model could be further developed to incorporate the additional complexity of IUs where an intervention is required to eliminate onchocerciasis, particularly in hypo-endemic areas where ivermectin has not been used