Central pontine myelinolysis in a chronic alcoholic: A clinical and brain magnetic resonance imaging follow-up

Introduction. Central pontine myelinolysis (CPM) is a noninflammatory, demyelinating lesion usually localised in the basis pontis. Chronic alcoholism is frequently associated with this condition which may have a variable clinical outcome. Until now, brain magnetic resonance imaging (MRI) follow-up in alcoholic CPM cases after alcohol withdrawal has been rarely described. Case report. We reported a 30- year-old male with a 12-year history of alcohol abuse, who presented with inability to stand and walk, nausea, vomiting and somnolence. Neurological examination revealed: impared fixation on lateral gaze, dysarthria, mild spastic quadriparesis, truncal and extremity ataxia, sock-like hypesthesia and moderate decrease in vibration sense in legs. Brain MRI showed a trident-shaped non-enhancing pontine lesion highly suggestive of CPM. After an eight-month alcoholfree follow-up period, the patient’s clinical status significantly improved, while the extent of MRI pontine lesion was merely slightly reduced. Conclusion. The presented case demonstrates that CPM in chronic alcoholics may have a benign clinical course after alcohol withdrawal, which is not necessarily associated with the reduction of lesions on brain MRI. [Projekat Ministarstva nauke Republike Srbije, br. 175031

Evaluation of an in-house developed colorimetric and other assays for PET-degrading activity

Plastic materials have become indispensable in the modern world, with their extensive use resulting in their environmental accumulation. A promising solution for overcoming this ecological threat may be found in recombinantly produced plastic-degrading enzymes. Due to the complexity of the heterogeneous catalysis occuring during enzymatic PET hydrolysis, quantifying and comparing activities of such enzymes is rendered difficult. Here, we have assessed various assays documented in existing literature, employing different preparations of the purified recombinant Ideonella sakaiensis PETase mutant W159H/S238F (expressed from commercial plasmid Addgene #112203). The investigated methods were as follows: p-nitrophenyl acetate (pNA) hydrolysis assay, bis-(2-hydrohxyethyl)-terephthalate (BHET) agar and PET agar diffusion assays, and UV absorbance monitoring after PET particle and PET bottle cut-out hydrolysis. Additionally, we introduced an indirect colorimetric assay using the indicator pyrocatechol violet (PCV). Our work reveals many advantages and problems for each of the tested methods. The pNA hydrolysis assay is the quickest, but many substances which are usually present in enzyme buffers and solutions tend to hydrolyse this compound (e.g. imidazole). It is also unspecific due to hydrolysis by other esterase enzymes. The BHET diffusion assay offers a great tool for activity comparison and estimation, with greater enzyme specificity. However, it is slow and accurate activity quantification is difficult. PET hydrolysis was conducted on in-house prepared PET particles with UV spectrophotometric measurement or by a diffusion assay. Due to the measuring wavelength (240 nm), the importance of proper blanking is critical, but accurate results can still be obtained. The sensitivity of the diffusion assay is much lower in comparison to the similar BHET assay. We also report on a modification of the phenol red indirect colorimetric assay using PCV as the indicator and PET particles as the substrate, which has not been previously described in existing literature.Dodati u SynthBio grupu

Cross-cultural adaptation and validation of the disease specific questionnaire oqlq in Serbian patients with malocclusions

Introduction. Dentofacial disorders may potentially significantly affect the quality of life. Objectives of this study were to validate translated and culturally adapted Orthognatic Quality of Life Questionnaire (OQLQ) on a cohort of Serbian patients with malocclusions. Methods. The questionnaire was validated in 111 consecutive patients with malocclusions, seen between December 2014 and February 2015 at the Clinic of Orthodontics, Faculty of Dental Medicine, University of Belgrade. Clinical validity was assessed comparing the mean scores for the four subscales of the OQLQ and mean PAR pre-treatment score. In order to assess whether the allocation of items in the subscales corresponds to their distribution in the original questionnaire, an exploratory factor analysis (principal component analysis with varimax rotation) was conducted. Results. The results of the internal consistency analysis demonstrated good relationships between the items; Cronbach's alpha coefficients for the four subscales were highly significant (p lt 0.001) (0.88-0.91). All items were significantly correlated between baseline and the retest (6 weeks after). The correlations between the PAR and all four domains of the OQLQ were all significant (p lt 0.01). The loading weights obtained in the exploratory factor analysis showed that this model revealed four factors with eigenvalue greater than 1, explaining the 64.0% of the cumulative variance. The majority of the items (86.4%) in the Serbian version of the OQLQ presented the highest loading weight in the subscales assigned by the OQLQ developer. Conclusions. The psychometric properties of the OQLQ (Serbian version) have exceptional internal consistency and reproducibility as an instrument for evaluation of dental malocclusions. Additionally, this questionnaire may be useful as a supplementary outcome measure in persons with malocclusions

Validation of the Fatigue Impact Scale in Multiple Sclerosis Patients in Serbia

Fatigue is one of the most frequent complaints of patients with multiple sclerosis (MS). The Fatigue Impact Scale (FIS), one of the 30 available fatigue questionnaires, is commonly applied because it evaluates multidimensional aspects of fatigue. The chief objectives of this study were to validate FIS and evaluate the psychometric properties of MS patients in Serbia. One hundred and twenty-one (121) MS patients and one hundred and twenty-two (122) age-, gender- and education-matched healthy control (HC) subjects completed the FIS and the Beck Depression Inventory. Internal consistency of the FIS subscales was determined using Cronbach's Alpha Coefficient. Test/retest reliability with an intra-class correlation coefficient (ICC) for each FIS subscale was performed. The total FIS score and subscale scores showed statistically significant differences between the MS patients and the HC subjects in both FIS sessions. Cronbach's Alpha was 0.966. All ICCs were statistically significant (p < 0.05). The Serbian version of this instrument may be useful as a clinical measure for fatigue and functionality in patients with MS

Autonomic symptom burden is an independent contributor to multiple sclerosis related fatigue

Objectives: To investigate a possible association between autonomic dysfunction and fatigue in people with multiple sclerosis. ----- Methods: In 70 people with multiple sclerosis early in the disease course (51 females, mean age 33.8 ± 9.1), quantitative sudomotor axon reflex tests, cardiovascular reflex tests (heart rate and blood pressure responses to the Valsalva maneuver and heart rate response to deep breathing), and the tilt table test were performed. Participants completed the Composite Autonomic Symptom Score 31, the Modified Fatigue Impact Scale, and the Epworth Sleepiness Scale, as well as the Beck Depression Inventory. Cutoff scores of ≥ 38 or ≥ 45 on the Modified Fatigue Impact Scale were used to stratify patients into a fatigued subgroup (N = 17 or N = 9, respectively). ----- Results: We found clear associations between fatigue and scores in subjective tests of the autonomic nervous system: fatigued patients scored significantly worse on Composite Autonomic Symptom Score 31, and there was a strong correlation between the Modified Fatigue Impact Scale and the Composite Autonomic Symptom Score 31 (rs = 0.607, p < 0.001). On the other hand, we found only modest associations between fatigue and scores in objective tests of the autonomic nervous system: there was a clear trend for lower sweating outputs at all measured sites, which reached statistical significance for the distal leg and foot. We found weak correlations between the Modified Fatigue Impact Scale and the Valsalva ratio (rs = - 0.306, p = 0.011), as well as between the Modified Fatigue Impact Scale and quantitative sudomotor axon reflex tests of the forearm, proximal, and distal lower leg (rs = - 0.379, p = 0.003; rs = - 0.356, p = 0.005; and rs = - 0.345, p = 0.006, respectively). A multiple regression model showed that the Composite Autonomic Symptom Score 31, Beck Depression Inventory, and Epworth Sleepiness Scale were independent predictors of fatigue (p = 0.005, p = 0.019, and p = 0.010, respectively). ----- Conclusion: These results suggest that-even early in the course of the disease-people with multiple sclerosis suffer from objective and subjective impairments of the autonomic nervous system. The results also point to an association between autonomic nervous system impairment and multiple sclerosis related fatigue

Multiple Sclerosis patients carry an increased burden of exceedingly rare genetic variants in the inflammasome regulatory genes

The role of rare genetic variation and the innate immune system in the etiology of multiple sclerosis (MS) is being increasingly recognized. Recently, we described several rare variants in the NLRP1 gene, presumably conveying an increased risk for familial MS. In the present study we aimed to assess rare genetic variation in the inflammasome regulatory network. We performed whole exome sequencing of 319 probands, comprising patients with familial MS, sporadic MS and control subjects. 62 genes involved in the NLRP1/NLRP3 inflammasome regulation were screened for potentially pathogenic rare genetic variation. Aggregate mutational burden was analyzed, considering the variants' predicted pathogenicity and frequency in the general population. We demonstrate an increased (p = 0.00004) variant burden among MS patients which was most pronounced for the exceedingly rare variants with high predicted pathogenicity. These variants were found in inflammasome genes (NLRP1/3, CASP1), genes mediating inflammasome inactivation via auto and mitophagy (RIPK2, MEFV), and genes involved in response to infection with DNA viruses (POLR3A, DHX58, IFIH1) and to type-1 interferons (TYK2, PTPRC). In conclusion, we present new evidence supporting the importance of rare genetic variation in the inflammasome signaling pathway and its regulation via autophagy and interferon-β to the etiology of MS

Validation and cross-cultural adaptation of the COMPASS-31 in Croatian and Serbian patients with multiple sclerosis

Aim To validate and cross-culturally adapt Croatian and Serbian versions of composite autonomic symptom score- 31 (COMPASS-31) for the detection of dysautonomia in patients with multiple sclerosis (MS). Methods A total of 179 patients, 67 with clinically isolated syndrome (CIS) and 112 with MS, completed the COMPASS- 31 at two MS centers in Zagreb and Belgrade between April 1 and October 31, 2016. Demographic and clinical data including age, gender, MS phenotypes, and the Expanded Disability Status Scale (EDSS) score were collected. Results The Cronbach’s alpha coefficient of COMPASS-31 total score was 0.844 for the Croatian MS sample and 0.779 for the Serbian MS sample. A joint analysis yielded Cronbach’s alpha coefficients ranging from 0.394 to 0.796, with values in four domains higher than 0.700. In Croatian and Serbian samples and the total study sample, the Cronbach’s alpha coefficient of COMPASS-31 was 0.785. Reproducibility measured by intra-class correlation coefficient (ICC) was acceptable (ICC = 0.795). With regard to the clinical validity, significant correlation was found between EDSS and the COMPASS-31 total score (P < 0.001). Furthermore, significant differences between MS phenotypes were detected for bladder and gastrointestinal domains and for the COMPASS-31 total score (P < 0.001, P = 0.005, and P = 0.027, respectively). Finally, significant differences between MS phenotypes in patients with score >0, which implies the existence of at least one of the symptoms investigated in each domain, were detected for secretomotor and bladder domains (P = 0.015 and P < 0.001, respectively). Conclusion COMPASS-31 represents a valid and acceptable self-assessment instrument for the detection of dysautonomia in MS patient

Cladribine tablets in people with relapsing multiple sclerosis : A real-world multicentric study from southeast European MS centers

Cladribine is an oral disease-modifying drug authorized by the European Medicine Agency for the treatment of highly active relapsing multiple sclerosis (MS).To provide real-world evidence of cladribine's effectiveness and safety in people with MS (pwMS).A retrospective observational multi-center, multi-national study of pwMS who were started on cladribine tablets in ten centers from five European countries.We identified 320 pwMS treated with cladribine tablets. The most common comorbidities were arterial hypertension and depression. Three patients had resolved hepatitis B infection, while eight had positive Quantiferon test prior to cladribine commencement. There were six pwMS who had malignant diseases, but all were non-active. During year 1, 91.6% pwMS did not have EDSS worsening, 86.9% were relapse-free and 72.9% did not have MRI activity. During the second year, 90.2% did not experience EDSS worsening, 86.5% were relapse-free and 75.5% did not have MRI activity. NEDA-3 was present in 58.0% pwMS in year 1 and in 54.2% in year 2. In a multivariable logistic regression model age positively predicted NEDA-3 in year 1. The most common adverse events were infections and skin-related adverse events. Lymphopenia was noted in 54.7% of pwMS at month 2 and in 35.0% at month 6. Two pwMS had a newly discovered malignant disease, one breast cancer, and one melanoma, during the first year of treatment.Our real-world data on the effectiveness and safety of cladribine tablets are comparable to the pivotal study and other real-world data with no new safety signals