Pre-Clinical Tools for Predicting Drug Efficacy in Treatment of Tuberculosis

Combination therapy has, to some extent, been successful in limiting the emergence of drug-resistant tuberculosis. Drug combinations achieve this advantage by simultaneously acting on different targets and metabolic pathways. Additionally, drug combination therapies are shown to shorten the duration of therapy for tuberculosis. As new drugs are being developed, to overcome the challenge of finding new and effective drug combinations, systems biology commonly uses approaches that analyse mycobacterial cellular processes. These approaches identify the regulatory networks, metabolic pathways, and signaling programs associated with M. tuberculosis infection and survival. Different preclinical models that assess anti-tuberculosis drug activity are available, but the combination of models that is most predictive of clinical treatment efficacy remains unclear. In this structured literature review, we appraise the options to accelerate the TB drug development pipeline through the evaluation of preclinical testing assays of drug combinations

Burial of two closely related infants under a “dragon stone” from prehistoric Armenia

“Dragon stones” are prehistoric basalt stelae carved with animal imagery found in Armenia and surrounding regions. These monuments have a complex history of use and reuse across millennia, and the original date of creation is still a matter of debate. In this article, we present a unique dragon stone context excavated at the site of Lchashen, Armenia, where a three-and-a-half-meter high basalt stela with an image of a sacrificed bovid was found above a burial dating to the 16th century BC. The burial stands out among hundreds from this site as the only one in connection with a “dragon stone”, and one of very few containing the remains of newborn babies. Furthermore, our analyses of ancient DNA extracted from the well-preserved skeletal remains of two 0–2-month-old individuals showed them to be second-degree related females with identical mitochondrial sequences of the haplogroup U5a1a1 lineage, thus indicating that the infants are closely related. Additionally, we assessed that the buried individuals displayed genetic ancestry profiles similar to other Bronze Age individuals from the region

Evaluation of genetic mutations associated with Mycobacterium tuberculosis resistance to isoniazid, rifampicin (MTBDR plus), fluoroquinolone and injectable second-line drugs (MTBDRsl)

Aims and objectives: Tuberculosis (TB) is one of the major public health problems in Armenia. In 2009, TB incidence in Armenia was reported to be as high as 45.5 per 100,000 population, and TB mortality was 3.9 per 100,000 population. Only 35% of estimated new sputum smear-positive pulmonary TB patients are notified annually. From 1729 TB cases notified in 2010, only 329 patients were sputum smear-positive. The treatment success rate of new sputum smear-positive pulmonary TB patients was 76%, which is below the WHO target of 85%. Poor treatment outcome is partially explained by the high prevalence of drug resistant forms of TB. According to the 2007 drug resistance survey, MDR-TB among never-treated patients was 9.4% and 43.2% among previously treated cases with 4% of XDR cases. This represents an enormous public health challenge for Armenia; early identification and effective treatment of patients with MDR-TB is crucial to prevent further spread of the disease. Methods: A total of 583 specimens are sent to the NRL Armenia being either acid-fast bacterium positive or negative, but culture positive. Rifampin and isoniazid resistance was performed on 77 out of the 583 specimens of DR TB clinical isolates to analyze resistance for FQL, AM/CP and EMB. The DNA preparation method used was as described and recommended by the manufacturer (MTBDR plus version 1.0 and MTBDR sl version 2.0 Hain Lifescience, Nehren). For 99 (88.4%) and 160 (76.2%) strains, the mutations causing rifampicin and isoniazid resistance were located in the codon of rpoB S531L and katG S315T 1, respectively. Regarding isoniazid, 14 (6.6%) strains have a mutation in the inhA regulatory region (C15T) and 31 (14.7%) in both katG and InhA. Mutations detected in the FQ, EMB and rrs-resistant respectively are: 13 (16.9%) strains with the majority of mutations (10 [13%]) in the codon of 94 of the gyrA gen; 20 (26%) strains, with the majority of mutations (7 [9.1%]) in the codon of 306 of the EMB gen; and 13 (16.8%) strains with the majority of mutations (9 [11.6%]) in the codon of 1401 of the rrs gen. Comparing with phenotypically DST sensitivity and specificity for RMP was 97.6% (69 strains) and 100%, respectively. False resistance was detected for 2 strains containing missed bands of rpoB (W6, 7, 8) and 1 strain W7. Sensitivity and specificity for INH was 98.33%, 87.5% (69 strains). From 2 strains, 1 was false resistant (missing of WT 1, 2 bands) and 1 false sensitive results for INH. The sensitivity and specificity for AM/CM was 80%, 100%, and for the FQL 85.7%, 96.72%, respectively (69 strains). Conclusion: Rapid and accurate detection of resistance to first- and second-line anti-TB drugs is the key to successful therapy and interruption of the transmission chain of MDRTB strains. In summary, this data represent an important addition to the rare epidemiological data concerning resistance patterns of MTB in Armenia and showed that the application of the GenoType MTBDRplus and GenoType MTBDRsl assays might be useful additional tools to allow for a rapid and safe diagnosis of MDR and XDR TB. In mutations associated with katG, high dosages of isoniazid in future will be considered for treatment

Effect of diabetes mellitus on reproductive parameters in mice

The decrease in population fertility has become a major concern in many developed countries. Recent studies show that infertility is affecting an estimated 15% of all couples (World Health Organization, WHO, 2010). Male infertility is the primary or contributing cause in 60% of cases. Male infertility is caused by a number of factors, such as genetic background, various environmental factors and disease. Diabetes mellitus (DM), a serious health problem on its own, is also suspected to be a contributing factor to male infertility. The aim of this project was to analyze the cellular, molecular and genetic effects of diabetic environment on spermatogenesis and sperm quality and to determine the impact of DM on the in vivo reproduction, using the mouse model (Mus musculus) inbred FVB. Diabetes was induced using streptozotocin. We used our knowledge and tools (unique monoclonal antibodies developed by our group) to determine the status of reproductive organs, anogenital distance, and the quality of sperms. Genetic analysis was performed by a quantitative Reverse Transcription Polymerase Chain Reaction (qPCR). We tested selected genes which are expressed in testicular tissue and thus can influence process of spermatogenesis and consequently the sperm quality. Our preliminary data strongly suggest that DM impairs male fertility. We have found significant changes in the body and reproductive organ weight of mice with DM. We have identified qualitative and quantitative changes in the expression of proteins in epididymal fluid and sperms. We have also detected an increased number of apoptotic cells in sperm of diabetic mice compared to the control group. To our knowledge, there is no study assessing the correlation between DM and “unexplained infertility”. In view of this, it is essential to analyze the effects of DM on male fertility, sperm quality, and reproduction parameters

Possible role of spermatogenic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) in mammalian sperm

Sperm proteins are important for the structure and function of these specific, highly differentiated cells. Certain of these proteins play a role in sperm-egg recognition during primary or secondary binding at zona pellucida glycoprotein matrix. The aim of this study was to characterize the acrosomal sperm protein recognized by a monoclonal antibody (MoAb) Hs-8, prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperm, and to test the possible role of this protein in gamete interaction. MoAb Hs-8 specifically labelled a 45 kDa protein from the sperm extract in the immunoblotting test. Sequence analysis identified this Hs-8 protein as GAPDHS. In order to perform a control tests, a commercial mouse anti-GAPDHS MoAb was applied. Both antibodies showed similar staining patterns using immunofluorescence labelling, transmission electron microscopy and immunoblot analysis. Moreover, both Hs-8 and commercial anti-GAPDHS antibodies blocked the secondary sperm-zona pellucida binding. Generally, GAPDHS was considered mainly as sperm-specific glycolytic enzyme involved in energy production during spermatogenesis and sperm motility and its role in the sperm head was unknown. In this study, we confirmed the potential additional role of GAPDHS as a binding protein that is involved in the sperm-zona pellucida interaction

MODEL UNCERTAINTY IN ECB: AN APPLICATION OF BAYESIAN MODEL AVERAGING WITH THE EXTENDED LIST OF ECONOMIC INDICATORS

Uncertainty is one of the most important aspects of monetary policies. Recent economic and political events showed that uncertainty is omnipresent and that policymakers should take it into account in their decision-making process. Uncertainty can take many forms from data uncertainty to model uncertainty, which are very important to consider in monetary policies. Recent global financial crisis also highlighted that conventional monetary policies have their limits and that unconventional monetary policies should be considered. It also showed that the monetary policy models failed to take into account essential economic and financial variables and, therefore, don’t capture the whole economic development. The aim of this thesis is to analyze the monetary policy models with the extended list of economic and financial variables. It has the aim to extend the application of Bayesian Model Averaging in ECB monetary policies. The thesis comes to the conclusion that the inclusion of wide set of macroeconomic variables will lead to more effective monetary policy decisions.Uncertainty is one of the most important aspects of monetary policies. Recent economic and political events showed that uncertainty is omnipresent and that policymakers should take it into account in their decision-making process. Uncertainty can take many forms from data uncertainty to model uncertainty, which are very important to consider in monetary policies. Recent global financial crisis also highlighted that conventional monetary policies have their limits and that unconventional monetary policies should be considered. It also showed that the monetary policy models failed to take into account essential economic and financial variables and, therefore, don’t capture the whole economic development. The aim of this thesis is to analyze the monetary policy models with the extended list of economic and financial variables. It has the aim to extend the application of Bayesian Model Averaging in ECB monetary policies. The thesis comes to the conclusion that the inclusion of wide set of macroeconomic variables will lead to more effective monetary policy decisions

Wpływ metody rozwijania umiejętności komunikacyjnych na aktywność fizyczną dzieci z niepełnosprawnością intelektualną

Background. The purpose of the study is to evaluate the effectiveness of a selected special education method on the level of physical activity in schoolchildren with intellectual disability. Material and methods. The study was conducted on 32 children with rather similar backgrounds: all participants were Armenians with no medical history of any registered disorders and the same moderate degree of intellectual disability (IQ score = 35-49). A physical education program based on two games using the Picture Exchange Communication System (PECS) was conducted for three months by a physical educator trained in PECS. The level of habitual physical activity before and after the study period was assessed by the Physical Activity Questionnaire for children (PAIC-A) and pedometry (Omron HJ-112, Illinois, USA). Twoway ANOVA (intellectual disability × intervention) with a post hoc Bonferroni test was used to examine the differences between the pre- and post-intervention physical activity and pedometry assessment results. Results. The use of PECS resulted in an overall increase in the levels of physical activity. The results showed a significant group-by-time interaction effect for unstructured physical activity, structured exercise, organized sports, commuting to and from school, and total sedentary activities (all measured in min/week; p<0.05). Post hoc comparisons revealed a remarkable improvement in PAIC-A and pedometry scores in the intervention group (p<0.05). Conclusions. Technologies for developing communication skills are an option to increase the physical activity of children with intellectual disability. Enhanced working memory facilitates improved executive motor functions.Wprowadzenie. Celem pracy jest ocena skuteczności wybranej metody kształcenia specjalnego na poziom aktywności fizycznej u dzieci w wieku szkolnym z niepełnosprawnością intelektualną. Materiał i metody. Badaniem objęto 32 dzieci o dość podobnym pochodzeniu: wszystkie były Ormianami, bez historii medycznej jakichkolwiek zarejestrowanych zaburzeń i z takim samym umiarkowanym stopniem niepełnosprawności intelektualnej (wynik ilorazu inteligencji, IQ = 35-49). Program wychowania fizycznego oparty na dwóch grach wykorzystujących Picture Exchange Communication System (PECS) był prowadzony przez trzy miesiące przez pedagoga wychowania fizycznego przeszkolonego w zakresie PECS. Poziom nawykowej aktywności fizycznej przed i po okresie badania oceniano za pomocą kwestionariusza aktywności fizycznej dla dzieci (PAIC-A) oraz pomiaru liczby kroków (Omron HJ-112, Illinois, USA). Dwukierunkowa analiza wariancji ANOVA (niepełnosprawność intelektualna × interwencja) z testem post hoc Bonferroniego została wykorzystana do zbadania różnic między aktywnością fizyczną przed i po interwencji oraz wynikami oceny liczby kroków. Wyniki. Zastosowanie PECS spowodowało ogólny wzrost poziomu aktywności fizycznej. Wyniki wykazały istotny efekt interakcji między grupą a czasem dla nieustrukturyzowanej aktywności fizycznej, ustrukturyzowanych ćwiczeń, zorganizowanego sportu, dojazdów do i ze szkoły oraz całkowitej aktywności sedenteryjnej (wszystkie mierzone w min/tydzień; p<0,05). Porównania post hoc wykazały znaczną poprawę wyników PAIC-A i pomiaru liczby kroków w grupie interwencji (p<0,05). Wnioski. Technologie rozwijające umiejętności komunikacyjne stanowią opcję zwiększenia aktywności fizycznej dzieci z niepełnosprawnością intelektualną. Poprawiona pamięć robocza ułatwia poprawę wykonawczych funkcji motorycznych

EFFECT OF A METHOD FOR DEVELOPING COMMUNICATION SKILLS ON PHYSICAL ACTIVITY IN CHILDREN WITH INTELLECTUAL DISABILITIES

Background. The purpose of the study is to evaluate the effectiveness of a selected special education method on the level of physical activity in schoolchildren with intellectual disability. Material and methods. The study was conducted on 32 children with rather similar backgrounds: all participants were Armenians with no medical history of any registered disorders and the same moderate degree of intellectual disability (IQ score = 35-49). A physical education program based on two games using the Picture Exchange Communication System (PECS) was conducted for three months by a physical educator trained in PECS. The level of habitual physical activity before and after the study period was assessed by the Physical Activity Questionnaire for children (PAIC-A) and pedometry (Omron HJ-112, Illinois, USA). Twoway ANOVA (intellectual disability × intervention) with a post hoc Bonferroni test was used to examine the differences between the pre- and post-intervention physical activity and pedometry assessment results. Results. The use of PECS resulted in an overall increase in the levels of physical activity. The results showed a significant group-by-time interaction effect for unstructured physical activity, structured exercise, organized sports, commuting to and from school, and total sedentary activities (all measured in min/week; p<0.05). Post hoc comparisons revealed a remarkable improvement in PAIC-A and pedometry scores in the intervention group (p<0.05). Conclusions. Technologies for developing communication skills are an option to increase the physical activity of children with intellectual disability. Enhanced working memory facilitates improved executive motor functions.Wprowadzenie. Celem pracy jest ocena skuteczności wybranej metody kształcenia specjalnego na poziom aktywności fizycznej u dzieci w wieku szkolnym z niepełnosprawnością intelektualną. Materiał i metody. Badaniem objęto 32 dzieci o dość podobnym pochodzeniu: wszystkie były Ormianami, bez historii medycznej jakichkolwiek zarejestrowanych zaburzeń i z takim samym umiarkowanym stopniem niepełnosprawności intelektualnej (wynik ilorazu inteligencji, IQ = 35-49). Program wychowania fizycznego oparty na dwóch grach wykorzystujących Picture Exchange Communication System (PECS) był prowadzony przez trzy miesiące przez pedagoga wychowania fizycznego przeszkolonego w zakresie PECS. Poziom nawykowej aktywności fizycznej przed i po okresie badania oceniano za pomocą kwestionariusza aktywności fizycznej dla dzieci (PAIC-A) oraz pomiaru liczby kroków (Omron HJ-112, Illinois, USA). Dwukierunkowa analiza wariancji ANOVA (niepełnosprawność intelektualna × interwencja) z testem post hoc Bonferroniego została wykorzystana do zbadania różnic między aktywnością fizyczną przed i po interwencji oraz wynikami oceny liczby kroków. Wyniki. Zastosowanie PECS spowodowało ogólny wzrost poziomu aktywności fizycznej. Wyniki wykazały istotny efekt interakcji między grupą a czasem dla nieustrukturyzowanej aktywności fizycznej, ustrukturyzowanych ćwiczeń, zorganizowanego sportu, dojazdów do i ze szkoły oraz całkowitej aktywności sedenteryjnej (wszystkie mierzone w min/tydzień; p<0,05). Porównania post hoc wykazały znaczną poprawę wyników PAIC-A i pomiaru liczby kroków w grupie interwencji (p<0,05). Wnioski. Technologie rozwijające umiejętności komunikacyjne stanowią opcję zwiększenia aktywności fizycznej dzieci z niepełnosprawnością intelektualną. Poprawiona pamięć robocza ułatwia poprawę wykonawczych funkcji motorycznych

Molecular genetics of Mycobacterium tuberculosis resistant to aminoglycosides and cyclic peptide testing by MTBDRsl in Armenia

Objective/background: The GenoType MTBDRsl test rapidly detects resistance to ethambutol, fluoroquinolones, second-line aminoglycosides (amikacin [AMK] and kanamycin [KAN]), and cyclic peptides (capreomycin [CAP]) in Mycobacterium tuberculosis. According to data from Global Drug Resistance Surveillance Report (2007), Armenia is counted as a high-burden country for multidrug-resistant tuberculosis (MDR-TB). The estimated burden of MDR-TB in 2012 was 9.4 (7–12) and 43 (38–49) among retreatment TB cases. A total of 92 laboratory confirmed cases were reported to the World Health Organization (57 new and 35 previously treated) out of 511 cases tested for MDR-TB. Methods: A set of 77 drug-resistant TB isolates during 2011 and 2012 period, being either acid-fast bacterium positive or negative but culture-positive resistant to isoniazid, rifampin, or both according to the GenoType MTBDR plus assay, were consecutively tested using GenoType MTBDRsl. rrs gene analysis and the results from GenoType MTBDRsl were compared with phenotypic drug resistance testing. The DNA preparation method was performed as recommended by the manufacturer (Genotype MTBDR plus version 1.0 and Genotype MTBDRsl version 2.0 Hain Lifescience Nehren, Germany). Results: Aminoglycosides are key drugs for the treatment of MDR-TB. A total of 77 drug-resistant TB and four extensively drug-resistant M. tuberculosis isolates from Armenian TB patients were analyzed to characterize mutations within rrs and to compare with phenotypic drug resistance testing. Simultaneously, the following were identified: 65 (84.41%) rrs wild type (WT), 1 (1.3%) rrs WT MUT1 and MUT2 (WT; A1401G and G1484T), 1 (1.3%) rrs WT1, MUT1 (A1401G), 9 (11.7%) rrs WT1, MUT1 (A1401G), and 1 (1.3%) rrs WT1, MUT1. Mutation at position 1401 in rrs leads to resistance to KAN (7/77 = 9%), AMK (9/77 = 11.68%), and CAP (5/77 = 6.49%). Eleven (14.28%) streptomycin-resistant strains had a rrs mutation. Conclusion: Isolates with rrs structural gene mutations were cross-resistant to streptomycin, KAN, CAP, and AMK. Detection of the A1401G mutation appeared to be 100% specific for the detection of resistance to KAN and AMK. Being the first assessment, these data establish the presence of phenotypic drug-resistant and extensively drug-resistant strains using molecular profiling and are helpful in understanding aminoglycoside resistance on a molecular level