EFFICACY AND SAFETY OF SILVER NANOCOMPOSITES ON RIFAMPICIN-RESISTANT M. TUBERCULOSIS STRAINS.

BACKGROUND: Control of rifampicin-resistant tuberculosis (RR-MTB) requires novel technologies for restoring the anti-TB efficacy of priority drugs. We sought to evaluate the ability of nanotechnology application in the recovery of the anti-tuberculosis efficacy of rifampicin. METHODS: Nanocomposite- standard dose of rifampicin and 20 nm silver nanoparticles (AgNPs) suspension solution of 6 different concentrations: 0.25%; 0.5%; 1%; 2.5%; 5%; and 10%, were supplemented to 70 rifampicin-resistant mycobacterium tuberculosis (RR-MTB) isolates. The control arm consisted of 35 RR-MTB isolates and AgNPs suspension with identical concentrations. The inhibitory effect of nanocomposites was evaluated by MTB growth rate using the BACTECTM MGIT 960TM. The safety assessment of single-use AgNPs was conducted in experimental animals. RESULTS: The suppression process of AgNPs on RR-MTB isolates started with 2,5% nanocomposite solution application and full suppression was achieved in 5% and 10% nanocomposite solutions. A standard dose of rifampicin and a 2.5% solution of AgNPs increased the minimal inhibitory effect on RR-MTB by 10% (total 80%) vs the isolated use of a 2.5% solution of AgNPs (70%). An experiment on animals revealed the complete safety of a single injection of ultra-high doses of AgNPs. CONCLUSION: The study showed the potentiating effect of AgNPs in overcoming the resistance of MTB to rifampicin providing a scientific basis for further research.

Clinical outcomes among patients with tuberculous meningitis receiving intensified treatment regimens.

SETTING: National Center for Tuberculosis and Lung Diseases (NCTLD), Tbilisi, Georgia.OBJECTIVE: To determine clinical outcomes of patients with tuberculous meningitis (TBM) treated with an intensified regimen including a fluoroquinolone (FQ) and an injectable agent.DESIGN: Prospective cohort of patients aged ≥16 years initiating treatment for TBM at the NCTLD from January 2018 to December 2019. Treatment outcomes and neurologic disability at 1, 6 and 12 months after treatment initiation were assessed.RESULTS: Among 77 patients with median follow-up time of 363 days (IQR 269-374), 97% received a FQ, 62% an injectable agent, 44% linezolid and 39% a carbapenem. Fifty-seven patients (74%) successfully completed treatment, 2 (2.6%) had treatment failure, 6 (7.8%) died, and the remainder (12%) were lost to follow up. Among 11 patients treated for multidrug-resistant TBM, the median follow-up time was 467 days and one patient (8%) died. Regarding neurologic outcomes, 14/76 (18%) patients had Modified Rankin Scores of 0 at baseline, improving to 85% (56/66) and 94% (47/50) at 6 and 12 months, respectively.CONCLUSION: Intensified multidrug treatment regimens including a FQ and an injectable agent in all patients and newly implemented drugs in patients with multidrug-resistant TBM resulted in low mortality and favorable neurologic outcomes.

THE RELATIONSHIP BETWEEN TYPE-2 DIABETES AND TUBERCULOSIS.

According to the experimental and clinical investigations, innate and adaptive immune disorders play a significant role in T2-D subjects to become more susceptible to TB. It was shown that the functions of neutrophils, macrophages, NK cells and other components of innate immunity areis markedly compromised by metabolic disorders in T2-D. The number of evidences suggests that reduction in TH1:TH2 cytokines ratios may have significant influence on susceptibility of TB infection in T2-D subjects. Hormonal changes in T2-D also may increase susceptibility to TB, including 2 hormones -ghrelin and leptin that are involved to in controlling blood glucose levels related to malnutrition during TB. According Based on the experimental and clinical results resistin, being a key molecule that links obesity and TB2-D, is considered as a protein contributing contributor to the development of insulin resistance, being a key molecule that links obesity and TB2-D. Subjects with T2-D showed higher levels of resistin in serum associated with reduced possibility of human macrophages to enhance the production of reactive oxygen species (ROS) in vitro against a challenge with TB. Immunological impairment has an important role in susceptibility to TB infection for patients with T2-D. It has been revealed that IFN-y and IL-22 markedly discriminate diabetes from nondiabetic individuals. It was also established that aside from this many cytokines such as IL-17A, IFN-B, TNFα, IL-10, IL-18, IFN-Y and IL-22 are the most significant and consequently potentially related to the effects caused by diabetes caused effects in the pathogenesis of active pulmonary TB. The endothelial system plays significant role in the pathogenesis and progression of TB infection. It was demonstrated that endothelin "B" receptor antagonist leads to vasoconstriction precluding inflammatory cell infiltration in lung tissue, suggesting that ET-1 proinflammatory action involves ET "B" receptor. It was also shown that sputum endothelin-1 level is associated with active pulmonary tuberculosis and effectiveness of treatment. Reduction ins sputum ET-1 level has significant role in the assessment of anti-tuberculosis treatment efficacy. Alterations in microbiota in T2-D subjects may influence on immunity against TB infection. As it was established the amount of bacteria producing short-chain fatty acids (SCFA) markedly decrease in T2-D, and treatment with SCFA reduced induction of TFN-α, IL-10 and IL-17 cytokines in contrast to IL-6, IFN-y and IL-22, without modification of their induction after using of SCFA. Vitamin "D" deficiency in T2-D may also play a role ion the immune response against TB. A number of evidences suggest the correlation between its diminished levels and TB or TB-T2-D. In conclusion it is suggested that different risk factors, including immunological and hormonal changes as well as alterations in different cytokine production and microbiota, endothelial dysfunction and vitamin "D" deficiency are the main contributors leading to comorbidity of T2-D and in TB.

PROGNOSTIC VALUE OF PLASMA MYELOPEROXIDASE LEVEL'S AND ECHOCARDIOGRAPHIC DETERMINANTS IN CHRONIC HEART FAILURE PATIENTS.

The aim of current study was to investigate the level of Myeloperoxidase (MPO) in chronic heart failure patients with different severity and find the correlation with cardiac structural and functional markers and disease outcome. Initially eighty-six patients with different stages of CHF and ten healthy persons were included in our study. Excluded criteria were myocardial infarction or other concomitant acute diseases, renal failure (creatinine >250 µmol/L) in past 3 months. All patients underwent ECG, echocardiographic evaluation, anthropometric and other clinical examination, as well as plasma samples were collected for further analysis. MPO level (ng/ml) was measured by ELISA method, hs-CRP (mg/L) was measured by turbidimetric immunoassay and Leukocyte level (109/L) - by flow cytometry. Echocardiographic studies were performed using standard methods by parasternal and apical view. Statistical analyses were performed using IBM SPSS Statistics 16.0. After 6 month follow up we investigate our research primary outcome (mortality). The data were collected by telephone follow up from patients or members of family. Collection of information had been became possible in 79% of total cases (68 patients), from which 10 patients died: 8 cases were cardiovascular mortality, 1 case - suicide and 1 case from cancer. We found that MPO levels in the patients with CHF (mean value 9.3±7) are different from control group (mean value 4.19±2). MPO correlates with HF severity. MPO significantly were elevated in mortality group p<0,007 , as the level of hs-CRP p˂ 0.001. Changes of RV (right ventricule) sizes from normal ranges also were associated to worse outcome ( p<0.005). Depending on our data certain level of MPO (˃7.1 ng/ml) in studied patients and increased RVDD have independent importance for identification of patients with worse outcome.

INFLUENCE OF NUTRITIONAL STATE ON OUTCOME IN PATIENTS WITH CHRONIC HEART FAILURE.

The aim of present study was to investigate the nutritional status of patients with cronic heart failure and find correlation with disease outcomes. Initially 86 patients with different stages of CHF and 10 healthy persons were included in our study. Exclusion criteria were myocardial infarction or other concomitant acute diseases, renal failure (creatinine >250 µmol/L) in past 3 months. All patients underwent ECG, echocardiographic evaluation, clinical examination, as well as plasma samples were collected for further analysis. MPO level (ng/ml) was measured by ELISA method, hs-CRP (mg/L) was measured by turbidimetric immunoassay. The serum albumin level (g/L) was used as an indicator of protein reserves. The serum total cholesterol level (mg/dL) was used as a parameter of caloric depletion. For both tests colorimetric method were used. Statistical analyses were performed using IBM SPSS Statistics 16.0. After 6 month follow up we investigate CHF outcome (mortality). The data were collected by telephone follow up from patients or members of family. Collection of information had been became possible in 68 (79%) of cases. We found that MPO correlates with HF severity. 10 mortality cases were indentified, from which 8 patient died from cardiovasvular (CV) cause, all of them were males. In cardiovascular mortality group, the markers of inflammation (MPO, hs-CRP) were elevated significantly while markers of protein-energetical malnutrition were decreased (total protein, albumin, Total Colesterol, LDL-Cholesterol, HDL-Cholesterol). In our study malnutrition and inflammation are important predictors for assessment of disease prognoses in CHF patients. Additional management is necessary for these patients as well as further researches for elaboration of new treatment strategies.

[EXISTENTIAL ORGANIZATION OF BRAIN BIOELECTRICAL ACTIVITY OF THE OFFSPRING OF RATS AFTER VITAL STRESS].

An association was found between severity of behavioral disturbances in the offspring of rats that survived the stress, and changing spatial-temporal organization of the ECOG. The proportion of synchronous activity in the right frontal and left occipital regions with respect to the right nape, and between the frontal leads increases with the minimum deviations of behavior. The proportion of ahead activity increased in frontal leads; reduced the share of lagging and an increasing proportion of synchronous bioelectrical activity in relation to the right occipital region in case of serious disorders of behavior.

[Influence of maternal factor on motor behavior and lipid turnover in rats exposed to severe psychoemotional stress].

Adverse condition of rat development during the prenatal period (rearing by mother survived brain injury) or during the early post-natal period (cross fostering), causes impairment of motor behavior and a lipid turnover in adult rats. Such rats under severe stress showed decrease of adaptation (as expressed or low indexes of motor impellent behavior and lipid metabolism) in comparison with rats that were grown up by own mother. The psychoemotional stress caused the most expressed deficient behavior in males that were grown up by mothers with brain injury (decrease in serum level of HC and HDL and depressively - similar behavior).

[Features of an existential organization of a bioelectric activity of the brain at posterity from rats with a lateral injury of the brain].

Features of the existential organization of a bioelectric activity of a brain at posterity from rats with a lateral injury of the brain were investigated. The EEG analysis revealed in one month animals significantly increased the proportion of advanced and lagging decrease in the activity of the right frontal derivation towards the right occipital region. Change between structural ties in terms of the rats were correlated with the severity of behavioral disorders in the test "open field" (OF). The most significant change in the existential organization of a bioelectrical activity (the reduction of synchronous communications between the frontal derivations) recorded in rats with severe conduct disorder compared with animals of the same groups, in which the behavior was not significantly different from the norm. The violation of the behavior in the test OF was accompanied by a decrease in synchronous activity between the left frontal and right occipital region or the decline in the anticipatory activity in the right frontal region in relation to the right occipital region. By maintaining the integrity of the behavior of these parameters were increased in comparison with an intact control.

[Peculiarities of behavior of offspring of rats-ambidexters after surviving vital stress].

Psychogenic trauma inflicted to mothers (Wistar rats) during pregnancy is more tragic for establishment of psychoemotional functions in offspring than in the rat pups whose mothers survived prior to pregnancy the stress connected with threat of life. The "antenatal stress" causes in the one-month-old offspring the depression-like behavior, while the stress one month prior to conception--the increased anxiety. Disturbances of the integral behavior in the test "open field" can be caused both by the acute psychogenic trauma of mothers and by its delayed consequences. The sharp impoverishment of behavior and the more pronounced psychoemotional disturbances are realized in female individuals regardless of the terms of the action. In the male offspring the vital stress during their mothers' pregnancy produces the rougher behavioral disturbances than stress prior to conception.

Neurohumoral regulation in children with idiopathic scoliosis.

Biotesting of blood serum in children aged 8-15 with adolescent idiopathic scoliosis (0°-60° by Cobb) showed the change of neurohumoral regulation in comparison with the age norm. The degree and direction of changes in the biotesting give the indication of the progression of spinal deformity. The effectiveness of treatment can be judged by control biotesting.

[Polyunsaturated fatty acids prevent the development of locomotor disturbances caused by changes in the neurohumoral background in rat pups].

The present study shows the effectiveness of using polyunsaturated fatty acids in pregnant rats in order to prevent behavioral alterations in neonatal rats caused by the action of the pathogenic peptide factors of blood serum with perinatal damage of the central nervous system.

[The influence of the rat maternal environment on the behavior of offspring].

In Wistar's rats, the postnatal influence of maternal behaviour on the motor and emotional state and the anxiety level of the posterity born from the intact rat-mothers-ambidexter but reared by the intact mothers-ambidexter with the motor deficit, were investigated. From the first day of life, the rat-mothers with the removed part of the sense-motor cortex (right or left) nursed and reared the litter. It was discovered that the reduction of motor activity and increase of negative emotional responses occurred in experimental litter at the age of 1 month as compared with the control group (intact litter reared by the ersatz healthy rat-mothers). In the test of elevated cruciform maze, a high level of anxiety was discovered. The experimental posterity showed an emotional tension as the dominant state. Left-side lesion of the sense-motor cortex of the ersatz mother resulted in a rougher disorder in the emotional state and movement than after right-side lesion.

[Unilateral injury of sensorimotor cortex of the dominant or subdominant hemispheres of rats-mothers and emotional behavior of their offsprings in the "open field"]. / Unilateral'noe povrezhdenie sensomotornoi kory dominantnogo i subdominantnogo polusharii mozga krys-materei i povedenie potomstva v "otkrytom pole".

Neurohumoral disorders in mothers caused by brain injury, infection, hypoxia, and other pathological factors result in motor and psychoemotional disorders in children. Emotional behavior of 30-day-old offsprings of female rats with unilateral sensorimotor brain injury was studied in the "open field". Individual behavior was estimated (the probability of certain acts and significant transfers between them). Behavioral disorders in rat offsprings depended on the side (left of right) of mother's brain injury and "handedness". The right-side mother's injury turned to be more deleterious. Behavioral alterations were stronger in offsprings of ambidextral and left-handed females than in those born by right-handed males with the same sensorimotor injury.

Lateralization of brain trauma in female Wistar rats determines the immune and neurological status of offspring.

Unilateral trauma to the sensorimotor cortex in ambidextrous maternal female rats decreased the resistance of their offspring, as indicated by a reduction in the functional activity of natural killer cells. Offspring rats showed T-cell immunodeficiency regardless of the lateralization of the trauma in their mothers; this was more severe after right-sided trauma (both mature and immature T-lymphocytes were affected). The EMG pattern of evoked responses in offspring rats changed according to the lesions evoked by unilateral trauma in their mothers (in the muscles responsible for postural asymmetry). Cruder movement disturbances were also seen in offspring after right-sided trauma in mothers. Right-sided organic damage to the sensorimotor cortex in mothers evoked marked increases in negative emotionality and decreases in the motivation of orientational-investigative behavior in one-month-old offspring rats in the "open field" test.

[Lateralisation of the brain injury in female Wistar rats determines the immune and neurological status in offspring]. / Lateralizatsiia travmy mozga u krys-samok (Wistar) opredeliaet immunnyi i nevrologicheskii status potomstva.

The T-cell immune deficiency was found to be more obvious in offspring of rats with a right-side brain injury. The EMG pattern changed in accordance with the brain injury lateralisation in the muscles responsible for posture asymmetry. The offspring EMG disorders were more obvious in cases of a right-side brain injury in mothers. The right-side brain injury in mothers led to a higher negative emotionality and a diminished tentative-research behaviour in "open-field" tests in 30-day old offspring.

[Prediction of potential motor disorders in babies from neonatal serum biological test findings]. / Prognozirovanie vozmozhnykh dvigatel'nykh narushenii u detei po rezul'tatam biotesta syvorotki krovi novorozhdennykh.

To predict motor activity in the newborn, electromyographic (on rats) and ultrastructural (on chick embryos) studies were performed after administration of donor babies' sera taken in the first days of life. Morphological and electromyographic parameters were compared with late clinical symptoms. A rating schedule was developed with regards to biological test results, which allows motor disorders to be detected in the newborn.

[The embryotoxic effect in rats exposed to the blood serum of newborn infants tested for the presence of transfer factors]. / Embriotoksicheskii éffekt u krys pri vozdeistvii syvorotkoi krovi novorozhdennykh detei, testiruemykh na nalichie transfernykh faktorov.

Pregnant rats (day 14 of gestation) received intraamniotic injections of newborn human serum. Transfer factors (TF) present in injected serum produced significant embryotoxic and teratogenic effects, as well as changes in coordination of motor activity (EMG) in newborn rats. The extent of embryological disturbances and the motor activity of 33- to 34-day old recipient rats depend on TF concentration (at serum dilution 1:2) and on the state of infant's motor functions ("normal" group and "risk" group). The results obtained allow to objectively determine the pathogenic effect of serum TF on embryotoxicity and formation of motor coordination in the donor infant.