THE NECESSITY OF INTRODUCTION THE DRUG INSURANCE SYSTEM IN ARMENIA

The increase in the cost of the medicinal component of the treatment, the spread of chronic diseases, and the maintenance of socio-economic inequality in access to health services require the provision of adequate access to medicines. These issues create prerequisites for the improvement of the state health policy and, first, the drug supply system, which is an integral part of the treatment process. The financing of healthcare in Armenia is mainly formed from budget allocations and out of pocket expenditures of the population. Reducing the financial burden on the state and ensuring the rational use of drugs contributes to improving the health of the population. The implementation of a drug insurance scheme, which partially or fully cover the cost of drugs in RA, is one of the solutions for resolving the issue of access to medicines. This article studies the problems of financing healthcare system in Armenia and highlights the need of introduction a drug insurance system in Armeni

Elevated PGC-1α Activity Sustains Mitochondrial Biogenesis and Muscle Function without Extending Survival in a Mouse Model of Inherited ALS

SummaryThe transcriptional coactivator PGC-1α induces multiple effects on muscle, including increased mitochondrial mass and activity. Amyotrophic lateral sclerosis (ALS) is a progressive, fatal, adult-onset neurodegenerative disorder characterized by selective loss of motor neurons and skeletal muscle degeneration. An early event is thought to be denervation-induced muscle atrophy accompanied by alterations in mitochondrial activity and morphology within muscle. We now report that elevation of PGC-1α levels in muscles of mice that develop fatal paralysis from an ALS-causing SOD1 mutant elevates PGC-1α-dependent pathways throughout disease course. Mitochondrial biogenesis and activity are maintained through end-stage disease, accompanied by retention of muscle function, delayed muscle atrophy, and significantly improved muscle endurance even at late disease stages. However, survival was not extended. Therefore, muscle is not a primary target of mutant SOD1-mediated toxicity, but drugs increasing PGC-1α activity in muscle represent an attractive therapy for maintaining muscle function during progression of ALS

High Throughput Microplate Respiratory Measurements Using Minimal Quantities Of Isolated Mitochondria

Recently developed technologies have enabled multi-well measurement of O2 consumption, facilitating the rate of mitochondrial research, particularly regarding the mechanism of action of drugs and proteins that modulate metabolism. Among these technologies, the Seahorse XF24 Analyzer was designed for use with intact cells attached in a monolayer to a multi-well tissue culture plate. In order to have a high throughput assay system in which both energy demand and substrate availability can be tightly controlled, we have developed a protocol to expand the application of the XF24 Analyzer to include isolated mitochondria. Acquisition of optimal rates requires assay conditions that are unexpectedly distinct from those of conventional polarography. The optimized conditions, derived from experiments with isolated mouse liver mitochondria, allow multi-well assessment of rates of respiration and proton production by mitochondria attached to the bottom of the XF assay plate, and require extremely small quantities of material (1–10 µg of mitochondrial protein per well). Sequential measurement of basal, State 3, State 4, and uncoupler-stimulated respiration can be made in each well through additions of reagents from the injection ports. We describe optimization and validation of this technique using isolated mouse liver and rat heart mitochondria, and apply the approach to discover that inclusion of phosphatase inhibitors in the preparation of the heart mitochondria results in a specific decrease in rates of Complex I-dependent respiration. We believe this new technique will be particularly useful for drug screening and for generating previously unobtainable respiratory data on small mitochondrial samples

Probiotic Lactobacillus acidophilus Strain INMIA 9602 Er 317/402 Administration Reduces the Numbers of Candida albicans and Abundance of Enterobacteria in the Gut Microbiota of Familial Mediterranean Fever Patients

Intestinal microorganisms play a crucial role in health and disease. The disruption of host–microbiota homeostasis has been reported to occur not only during disease development but also as a result of medication. Familial Mediterranean fever (FMF) is an inflammatory genetic disease characterized by elevated systemic reactivity against the commensal gut microbiota and high levels of Candida albicans in the gut. This study’s major objective was to investigate the effects of commercial probiotic Narine on the relative abundance of gut bacteria (specifically, enterobacteria, lactobacilli, Staphylococcus aureus, and enterococci) of C. albicans carrier and non-carrier FMF patients in remission. Our main finding indicates that the probiotic reduces numbers of C. albicans and abundance of enterobacteria in male and female patients of C. albicans carriers and non-carriers. It has pivotal effect on Enterococcus faecalis: increase in male non-carriers and decrease in female ones regardless of C. albicans status. No effect was seen for Lactobacillus and S. aureus. Our data suggest that M694V/V726A pyrin inflammasome mutations leading to FMF disease may contribute to gender-specific differences in microbial community structure in FMF patients. The study’s secondary objective was to elucidate the gender-specific differences in the gut’s microbial community of FMF patients. The tendency was detected for higher counts of enterobacteria in female FMF subjects. However, the small number of patients of these groups preclude from conclusive statements, pointing at the need for additional investigations with appropriate for statistical analysis groups of subjects involved in the study

Using the Coupling and Electron Flow assays in tandem to elucidate mechanistic activity of test agents.

<p>Coupling (A, C) and electron flow experiments (B, D) were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021746#s2" target="_blank">Methods</a>. Initial conditions are as follows (with final concentrations listed): A–B, Controls (no additives) or 20 mM sodium azide or 4 µM antimycin-A; C–D, Controls (no additives) or 10 mM malonate or 2.5 µg/ml oligomycin or 2 µM rotenone. See text for further explanation of results.</p

Effects of Phosphatase Inhibitor (PPI) treatment on rat heart mitochondrial respiration.

<p>Mitochondria were treated with a cocktail of phosphatase inhibitors during the isolation procedure as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021746#s2" target="_blank">Methods</a>. Respiratory States 3, 4_o and 3_u were measured in the presence (+PPI) or absence (-PPI) of phosphatase inhibitor treatment in the presence of either succinate/rotenone (3 µg/well) or glutamate/malate (6 µg/well) as oxidizable substrates, and rates are expressed per µg mitochondrial protein.</p

Mix and Measure Cycle Times.

<p>*Measure times for State 3 respiration may be extended beyond 3 min to observe the transition from State 3 to State 4 due to exhaustion of ADP in the microchamber.</p

Optimization of isolated mitochondria XF assays.

<p>2A–B, Determination of optimal µg amount of mitochondria/well. 1.25–40 µg/well of mouse liver mitochondria were attached to a V7 polystyrene XF24 plate and the coupling experiment was performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021746#s2" target="_blank">Methods</a> in the presence of succinate/rotenone. Blue vertical lines denote injections of indicated compounds. 2A shows OCR for 1.25–40 µg samples. 2B shows the absolute O₂ tension (in mm Hg) in the microchamber for 1.25–40 µg samples. Note that samples at 10 µg and above show unstable State 3 rates for OCR and depletion of O₂ in the microchamber in panels A and B, respectively. Lettering within data points indicates the group identification number.</p