Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants.

Cowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium-mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we report a highly efficient transformation system using embryonic axis explants isolated from imbibed mature seeds. We found that removal of the shoot apical meristem from the explants stimulated direct multiple shoot organogenesis from the cotyledonary node tissue. The application of a previously reported ternary transformation vector system provided efficient Agrobacterium-mediated gene delivery, while the utilization of spcN as selectable marker enabled more robust transgenic selection, plant recovery and transgenic plant generation without escapes and chimera formation. Transgenic cowpea plantlets developed exclusively from the cotyledonary nodes at frequencies of 4% to 37% across a wide range of cowpea genotypes. CRISPR/Cas-mediated gene editing was successfully demonstrated. The transformation principles established here could also be applied to other legumes to increase transformation efficiencies.

Contact lens use and its compliance for care among healthcare workers in Pakistan.

BACKGROUND: Poor care and hygiene of contact lens (CL) results in eye problems and infections. Healthcare workers have an important role in advocating correct lens care. OBJECTIVES: To determine the practices of CL care and the adverse consequences of poor CL care among healthcare workers. SETTING AND DESIGN: A cross-sectional study in one public and three private sector hospitals in Karachi, Pakistan, in 2009-2010. MATERIALS AND METHODS: We questioned 500 healthcare workers of all ages and both sexes, who wore CL, about compliance with advice on care and any complications due to improper hygiene practices. Ethical approval was obtained. Chi-square tests were used to determine significance and p-value less than 0.05 was considered statistically significant. RESULTS: Of the total CL users, 385 (77%) were females. Most (75%) respondents wore CL to correct myopia, whereas 54% wore CL only occasionally. Surprisingly, only 24% knew the CL cleaning protocol. Lens solution was changed daily by 33% of users and after more than 2 weeks by 42%. Although 412 (82%) participants practised reasonable hand hygiene before inserting CL, 88 (18%) did not. Infection and eye dryness were statistically significantly (P < 0.01) associated with sex, hand-washing, and frequency of CL use. CONCLUSION: Noncompliance with the CL protocol was common among healthcare workers in our society. This behavior calls for targeted health education and awareness programs for healthcare workers.

Phase III trial of casopitant, a novel neurokinin-1 receptor antagonist, for the prevention of nausea and vomiting in patients receiving moderately emetogenic chemotherapy.

PURPOSE: The purpose of this phase III trial was to evaluate the efficacy and safety of regimens containing casopitant, a novel neurokinin-1 receptor antagonist, for the prevention of chemotherapy-induced nausea and vomiting during the first cycle in patients receiving moderately emetogenic chemotherapy (MEC). PATIENTS AND METHODS: Predominantly female patients (98%) diagnosed with breast cancer (96%) who were chemotherapy-naïve and scheduled to receive an anthracycline and cyclophosphamide (AC) -based regimen were enrolled onto this multinational, randomized, double-blind, parallel-group, placebo-controlled clinical trial. All patients received dexamethasone 8 mg intravenously (IV) on day 1 and oral ondansetron 8 mg twice daily on days 1 to 3. Patients were randomly assigned to a control arm (placebo), a single oral dose casopitant arm (150 mg orally [PO] on day 1), a 3-day oral casopitant arm (150 mg PO on day 1 plus 50 mg PO on days 2 to 3), or a 3-day IV/oral casopitant arm (90 mg IV on day 1 plus 50 mg PO on days 2 to 3). The primary end point was the proportion of patients achieving complete response (no vomiting/retching or rescue medications) in the first 120 hours after the initiation of MEC. RESULTS: A significantly greater proportion of patients in the single-dose oral casopitant arm, 3-day oral casopitant arm, and 3-day IV/oral casopitant arm achieved complete response (73%, 73%, and 74%, respectively) versus control (59%; P < .0001). The study did not demonstrate a reduced proportion of patients with nausea or significant nausea in those receiving casopitant. Adverse events were balanced among study arms. CONCLUSION: All casopitant regimens studied were more effective than the control regimen. Casopitant was generally well tolerated.