The MelA gene from Citrobacter freundii, which encodes a glycosyl hydrolase family 4 (GH4) α-galactosidase, has been cloned and expressed in Escherichia coli. The recombinant enzyme catalyzes the hydrolysis of phenyl α-galactosides via a redox elimination-addition mechanism involving oxidation of the hydroxyl group at C-3 and elimination of phenol across the C-1-C-2 bond to give an enzyme-bound glycal intermediate. For optimal activity, the MelA enzyme requires two cofactors, NAD(+) and Mn(2+), and the addition of a reducing agent, such as mercaptoethanol. To delineate the mechanism of action for this GH4 enzyme, we measured leaving group effects, and the derived β(lg) values on V and V/K are indistinguishable from zero (-0.01 ± 0.02 and 0.02 ± 0.04, respectively). Deuterium kinetic isotope effects (KIEs) were measured for the weakly activated substrate phenyl α-D-galactopyranoside in which isotopic substitution was incorporated at C-1, C-2, or C-3. KIEs of 1.06 ± 0.07, 0.91 ± 0.04, and 1.02 ± 0.06 were measured on V for the 1-(2)H, 2-(2)H, and 3-(2)H isotopic substrates, respectively. The corresponding values on V/K were 1.13 ± 0.07, 1.74 ± 0.06, and 1.74 ± 0.05, respectively. To determine if the KIEs report on a single step or on a virtual transition state, we measured KIEs using doubly deuterated substrates. The measured (D)V/K KIEs for MelA-catalyzed hydrolysis of phenyl α-D-galactopyranoside on the dideuterated substrates, (D)V/K((3-D)/(2-D,3-D)) and (D)V/K((2-D)/(2-D,3-D)), are 1.71 ± 0.12 and 1.71 ± 0.13, respectively. In addition, the corresponding values on V, (D)V((3-D)/(2-D,3-D)) and (D)V((2-D)/(2-D,3-D)), are 0.91 ± 0.06 and 1.01 ± 0.06, respectively. These observations are consistent with oxidation at C-3, which occurs via the transfer of a hydride to the on-board NAD(+), being concerted with proton removal at C-2 and the fact that this step is the first irreversible step for the MelA α-galactosidase-catalyzed reactions of aryl substrates. In addition, the rate-limiting step for V(max) must come after this irreversible step in the reaction mechanism.

Abstract.Integrated community case management (iCCM) aims to reduce child mortality in areas with poor access to health care. iCCM was implemented in 2009 in Kono district, Sierra Leone, a postconflict area with high under-five mortality rates (U5MRs). We evaluated iCCM’s impact and effects on child health using cluster surveys in 2010 (midterm) and 2013 (endline) to compare indicators on child mortality, coverage of appropriate treatment, timely access to care, quality of care, and recognition of community health workers (CHWs). The sample size was powered to detect a 28% decline in U5MR. Clusters were selected proportional to population size. All households were sampled to measure mortality and systematic random sampling was used to measure coverage in a subset of households. We used program data to evaluate utilization and access; 5,257 (2010) and 3,649 (2013) households were surveyed. U5MR did not change significantly (4.54 [95% confidence interval [CI]: 3.47–5.60] to 3.95 [95% CI: 3.06–4.83] deaths per 1,000 per month (P = 0.4)) though a relative change smaller than 28% could not be detected. CHWs were the first source of care for 52% (2010) and 50.9% (2013) of children. Coverage of appropriate treatment of fever by CHWs or peripheral health units increased from 45.5% [95% CI: 39.2–52.0] to 58.2% [95% CI: 50.5–65.5] (P = 0.01); changes for diarrhea and pneumonia were not significant. The continued reliance on the CHW as the first source of care and improved coverage for the appropriate treatment of fever support iCCM’s role in Kono district.

Introductory Summary: Barbados is a tourism dependent island state whose income is very sensitive to perturbations in the global economy and is thus seeking ways to diversify its service exports  Barbados has a two-tiered health system with a publicly funded health system operating alongside numerous private clinics  Private provision of health services in Barbados has grown between

Objective: Promoting healthy communities through the provision of accessible quality healthcare services is a common mission shared by schools of pharmacy, public health departments, and governmental agencies. The following study seeks to identify and detail the benefits of collaboration between these different groups. Methods: In total, 112 mobile clinics targeting Medicare beneficiaries were held in 20 cities across Northern/Central California from 2007 to 2016. Under the supervision of licensed pharmacists, trained student pharmacists provided vaccinations, health screenings, Medicare Part D plan optimization services, and Medication Therapy Management (MTM) to patients at each clinic site. Clinic support was extended by public health departments, governmental agency partners, and a health professional program. Results: Since clinic inception, 8,996 patients were provided services. In total, 19,441 health screenings and 3,643 vaccinations were collectively provided to clinic patients. We assisted 5,549 beneficiaries with their Part D benefit, resulting in an estimated aggregate out-of-pocket drug cost savings of $5.7 million. Comprehensive MTM services were provided to 4,717 patients during which 8,184 medication-related problem (MRP) were identified. In 15.3% of patients, the MRP was determined severe enough to warrant prescriber follow-up. In total, 42.9% of clinic patients were from racial/ethnic minority groups and 25.5% had incomes ≤150% of the Federal Poverty Level. Conclusion: Collaboration between a school of pharmacy, public health departments, and governmental organizations can effectively serve Medicare beneficiary populations and result in: 1) lower out-of-pocket drug costs, 2) minimization of medication-related problems, 3) increased vaccination uptake, and 4) increased utilization of health screenings. Conflict of Interest We declare no conflicts of interest or financial interests that the authors or members of their immediate families have in any product or service discussed in the manuscript, including grants (pending or received), employment, gifts, stock holdings or options, honoraria, consultancies, expert testimony, patents and royalties. Treatment of Human Subjects: IRB review/approval required and obtained   Type: Original Research