A solid-phase enzyme-linked assay for vitamin B 12

A new solid-phase enzyme-linked competitive binding assay for vitamin B 12 (cyanocobalamin) is described. The assay is based on the competition between analyte B 12 molecules and a glucose-6-phosphate dehydrogenase-vitamin B 12 conjugate for a limited number of R-protein binding sites immobilized on sepharose particles. After appropriate incubation and washing steps, the enzyme activity bound to the solid-phase is inversely related to the concentration of B 12 in the sample. Under optimized conditions, the method can detect B 12 in the range of 3×10 −10 −1×10 −8 M (using 100 μ l sample) with high selectivity over other biological molecules.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41626/1/604_2005_Article_BF01197285.pd

Swallowing difficulties with oral drugs among polypharmacy patients attending community pharmacies.

Background Swallowing difficulties are common and can affect patients' ability to take solid oral dosage forms, thus compromising medication adherence. Strategies developed by patients to overcome such difficulties while taking medicines have seldom been described. Objective To determine prevalence and characteristics of swallowing difficulties among primary care patients attending their community pharmacies; to explore strategies developed by patients to overcome their difficulties, and health professionals' awareness of these problems. Setting Prospective study with a semi-structured questionnaire in random community pharmacies located in two Swiss regions. Method In each pharmacy, an interviewer asked 16 questions to each consecutive patient (18 years and older) with a prescription for at least 3 different solid oral forms. Main outcome measure Quantification of number of patients with swallowing difficulties and detailed description of difficulties. Results Among 122 pharmacies, 59 (48 %) accepted to join the study and 410 patients were enrolled. Thirty-seven patients (9.0 %) reported ongoing swallowing difficulties, while 55 patients (13.4 %) reported past difficulties. For the majority of patients, difficulties occurred at each single dose (83.7 %), with a single medication (59.8 %) and lasted for less than 12 months (53.8 %). Number of tablets was not the main trigger. Swallowing difficulties impaired extremely daily life in 12 % of the patients. Intentional non adherence (23 % of patients) and altering the oral dose formulation were the most common and potentially harmful strategies used by patients to overcome their swallowing difficulties. According to the patients, pharmacists and physicians rarely inquired about their swallowing difficulties. Conclusion We report a fairly high prevalence of swallowing difficulties in polypharmacy patients attending their community pharmacies. Pharmacists have to interview patients on their swallowing difficulties in a more systematic way, support patients in finding solutions and refer them to their physician if necessary to ensure continuity in care

ING116070: A Study of the Pharmacokinetics and Antiviral Activity of Dolutegravir in Cerebrospinal Fluid in HIV-1-Infected, Antiretroviral Therapy-Naive Subjects

Background. Dolutegravir (DTG), a once-daily, human immunodeficiency virus type 1 (HIV-1) integrase inhibitor, was evaluated for distribution and antiviral activity in cerebrospinal fluid (CSF). Methods. ING116070 is an ongoing, single-arm, open-label, multicenter study in antiretroviral therapy–naive, HIV-1–infected adults. Subjects received DTG (50 mg) plus abacavir/lamivudine (600/300 mg) once daily. The CSF and plasma (total and unbound) DTG concentrations were measured at weeks 2 and 16. The HIV-1 RNA levels were measured in CSF at baseline and weeks 2 and 16 and in plasma at baseline and weeks 2, 4, 8, 12, and 16. Results. Thirteen white men enrolled in the study; 2 withdrew prematurely, 1 because of a non–drug-related serious adverse event (pharyngitis) and 1 because of lack of treatment efficacy. The median DTG concentrations in CSF were 18 ng/mL (range, 4–23 ng/mL) at week 2 and 13 ng/mL (4–18 ng/mL) at week 16. Ratios of DTG CSF to total plasma concentration were similar to the unbound fraction of DTG in plasma. Median changes from baseline in CSF (n = 11) and plasma (n = 12) HIV-1 RNA were −3.42 and −3.04 log(10) copies/mL, respectively. Nine of 11 subjects (82%) had plasma and CSF HIV-1 RNA levels <50 copies/mL and 10 of 11 (91%) had CSF HIV-1 RNA levels <2 copies/mL at week 16. Conclusions. The DTG concentrations in CSF were similar to unbound plasma concentrations and exceeded the in vitro 50% inhibitory concentration for wild-type HIV (0.2 ng/mL), suggesting that DTG achieves therapeutic concentrations in the central nervous system. The HIV-1 RNA reductions were similar in CSF and plasma. Clinical Trials Registration. NCT01499199