Characterization of Tryptophanase from Vibrio cholerae O1

AbstractTryptophanase (Trpase) encoded by the tnaA gene catalyzes the conversion of tryptophan to indole, which is an extracellular signaling molecule detected in various bacteria including Vibrio cholerae. Indole has been demonstrated to regulate biofilm formation, drug resistance, plasmid maintenance and spore formation of bacteria. In the present study, the tnaA gene from V. cholerae O1 (VcTrpase) was cloned and expressed in E. coli BL21(DE3) tn5:tnaA (a Trpase-deficient competent). VcTrpase was purified by Ni2+-NTA chromatography. The obtained VcTrpase had a molecular mass of approximately 49 kDa, a specific activity of 3 U/mg protein, and absorption peaks at 330 and 435nm. Using a site-directed mutagenesis technique, replacement of Arg419 by Val resulted in a VcTrpase completely devoid of activity. Thus, this site can be a target for drug design for controlling V. cholerae

Validation of the Polish version of the Movement Disorder Society-Unified Parkinson’s Disease Rating Scale (MDS-UPDRS)

Background. In 2008, the Movement Disorders Society (MDS) published a new Unified Parkinson’s Disease Rating Scale (MDS-UPDRS) as the official benchmark scale for Parkinson’s Disease (PD). We have translated and validated the Polish version of the MDS-UPDRS, explored its dimensionality, and compared it to the original English one. Methods. The MDS-UPDRS was translated into Polish by a team of Polish investigators led by JS and GO. The back-translation was completed by colleagues fluent in both languages (Polish and English) who were not involved in the original translation, and was reviewed by members of the MDS Rating Scales Programme. Then the translated version of the MDS-UPDRS underwent cognitive pretesting, and the translation was modified based on the results. The final translation was approved as the Official Working Document of the MDS-UPDRS Polish version, and was tested on 355 Polish PD patients recruited at movement disorders centres all over Poland (at Katowice, Gdańsk, Łódź, Warsaw, Wrocław, and Kraków). Confirmatory and explanatory factor analyses were applied to determine whether the factor structure of the English version could be confirmed in the Polish version. Results. The Polish version of the MDS-UPDRS showed satisfactory clinimetric properties. The internal consistency of the Polish version was satisfactory. In the confirmatory factor analysis, all four parts had greater than 0.90 comparative fit index (CFI) compared to the original English MDS-UPDRS. Explanatory factor analysis suggested that the Polish version differed from the English version only within an acceptable range. Conclusions and clinical implications. The Polish version of the MDS-UPDRS meets the requirements to be designated as the Official Polish Version of the MDS-UPDRS, and is available on the MDS web page. We strongly recommend using the MDS-UPDRS instead of the UPDRS for research purposes and in everyday clinical practice.

Usefulness of Potentially Probiotic L. lactis Isolates from Polish Fermented Cow Milk for the Production of Cottage Cheese

Lactococcus lactis bacteria are used as starter cultures in cottage cheese and fermented milk production due to their acidification and contribution in the creation of the characteristic sensory features. The aim of the research was to carry out isolation, genetic identification, and verification of the probiotic properties of selected Lactococcus lactis isolates obtained from Polish fermented cow milk and apply the best strains to produce cottage cheese with good rheological and sensory characteristics. The isolates obtained were identified morphologically, biochemically, and with the use of 16S rRNA gene sequencing. After pre-screening two of the tested Lactococcus lactis strains, A13 and A14 were observed to be most tolerant to high NaCl concentrations and bile salts and to acidify milk the most. We confirmed the activity of A13 and A14 against such pathogenic strains as Escherichia coli, Salmonella enterica ssp. enterica, Salmonella enterica ssp. enterica sv. anatum, Staphylococcus aureus, and Enterococcus faecalis. As a potential industrial starter culture with probiotic potential, the selected Lactococcus lactis A13 and A14 strains produced cottage cheese quickly with good sensory (colour, smell, taste, texture) and rheological (viscosity, elasticity) properties

Usefulness of Potentially Probiotic <i>L. lactis</i> Isolates from Polish Fermented Cow Milk for the Production of Cottage Cheese

Lactococcus lactis bacteria are used as starter cultures in cottage cheese and fermented milk production due to their acidification and contribution in the creation of the characteristic sensory features. The aim of the research was to carry out isolation, genetic identification, and verification of the probiotic properties of selected Lactococcus lactis isolates obtained from Polish fermented cow milk and apply the best strains to produce cottage cheese with good rheological and sensory characteristics. The isolates obtained were identified morphologically, biochemically, and with the use of 16S rRNA gene sequencing. After pre-screening two of the tested Lactococcus lactis strains, A13 and A14 were observed to be most tolerant to high NaCl concentrations and bile salts and to acidify milk the most. We confirmed the activity of A13 and A14 against such pathogenic strains as Escherichia coli, Salmonella enterica ssp. enterica, Salmonella enterica ssp. enterica sv. anatum, Staphylococcus aureus, and Enterococcus faecalis. As a potential industrial starter culture with probiotic potential, the selected Lactococcus lactis A13 and A14 strains produced cottage cheese quickly with good sensory (colour, smell, taste, texture) and rheological (viscosity, elasticity) properties

Abies Concolor Seeds and Cones as New Source of Essential Oils—Composition and Biological Activity

The chemical composition, including the enantiomeric excess of the main terpenes, of essential oils from seeds and cones of Abies concolor was studied by chromatographic (GC) and spectroscopic methods (mass spectrometry, nuclear magnetic resonance), leading to the determination of 98 compounds. Essential oils were mainly composed of monoterpene hydrocarbons. The dominant volatiles of seed essential oil were: limonene (47 g/100 g, almost pure levorotary form) and α-pinene (40 g/100 g), while α-pinene (58 g/100 g), sabinene (11 g/100 g), and β-pinene (4.5 g/100 g) were the predominant components of the cone oil. The seed and cone essential oils exhibited mild antibacterial activity, and the MIC ranged from 26 to 30 μL/mL against all of the tested bacterial standard strains: Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, and Klebsiella pneumoniae. The cytotoxic studies have demonstrated that tested essential oils were cytotoxic to human skin fibroblasts and human microvascular endothelial cells at concentrations much lower than the MIC. The essential oils from A. concolor seeds and cones had no toxic effect on human skin fibroblasts and human microvascular endothelial cells, when added to the cells at a low concentration (0–0.075 μL/mL) and (0–1.0 μL/mL), respectively, and cultured for 24 h

Mechanical and Combustion Properties of Agglomerates of Wood of Popular Eastern European Species

The objective of the reported project was to produce wood agglomerates from popular East European species to determine their strength and combustion properties. Closed-die pellets were produced from sawdust of six types of wood common on the East European market: pine, willow, oak, poplar, birch, and beech. The properties of pellets, determined by the type of wood, were influenced by the compaction pressure and the moisture content of the sawdust. The highest average pellet density was obtained for oak sawdust, while the lowest density was obtained for poplar pellets. Expansion of pellets after removing from the die was found to be dependent on the wood species, and as expected, on compaction pressure. The pellet expansion increased after 2 h of conditioning in the laboratory and with an increase in moisture content. The highest and the smallest strength were obtained for oak pellets and for birch sawdust, respectively. The strength of the pellets increased by more than 100% with an increase in the compaction pressure from 60 MPa to 120 MPa. The average strength decreased by 65% with increasing moisture content. For all tested materials, drop resistance remained at a high level, acceptable in industrial practice. The highest calorific value of 18.97 MJ/kg was obtained for pine pellets. The highest ash value of 1.52% was obtained for willow pellets and the lowest value of 0.32% for pine pellets

Geophysical Research of Secondary Deformations in the Post Mining Area of the Glaciotectonic Muskau Arch Geopark&mdash;Preliminary Results

Underground and opencast mining adversely affects the surrounding environment. This process may continue even decades after the end of actual mineral extraction. One of the most significant effects of ceased mining are secondary deformations. Safe, new development of post-mining areas requires reliable information on potential deformation risk zones, which may be difficult to obtain due to a lack of necessary data. This study aimed to investigate and understand the secondary deformation processes in the underground mining area of the former &ldquo;Babina&rdquo; lignite mine, located in the unique glaciotectonic environment of the Muskau Arch, in western Poland. A combination of GIS-based historical mapping, geophysical 2D/3D microgravimetry, and Electrical Resistivity Tomography (ERT) measurements allowed the identification of subsidence-prone areas and the determination of potential factors of sinkhole development. The latter are associated with anthropogenic transformation of rock mass and hydrogeological conditions, by shallow underground mining. The results confirmed that multi-level mining of coal deposits in complex and complicated glaciotectonic conditions cause discontinuous deformations, and may be hazardous as long as 50 years after the end of mining operations

Montgomery‐Åsberg Depression Rating Scale factors in treatment‐resistant depression at onset of treatment: Derivation, replication, and change over time during treatment with esketamine

Abstract Objective Derive and confirm factor structure of the Montgomery‐Åsberg Depression Rating Scale (MADRS) in patients with treatment‐resistant depression (TRD) and evaluate how the factors evident at baseline change over 4 weeks of esketamine treatment. Methods Two similarly‐designed, short‐term TRANSFORM trials randomized adults to esketamine or matching placebo nasal spray, each with a newly‐initiated oral antidepressant, for 4 weeks (TRANSFORM‐1: N = 342 patients; TRANSFORM‐2: N = 223 patients). The factor structure of MADRS item scores at baseline was determined by exploratory factor analysis in TRANSFORM‐2 and corroborated by confirmatory factor analysis in TRANSFORM‐1. Change in MADRS factor scores from baseline (day 1) to the end of the 28‐day double‐blind treatment phase of TRANSFORM‐2 was analyzed using a mixed‐effects model for repeated measures (MMRM). Results Three factors were identified based on analysis of MADRS items: Factor 1 labeled affective and anhedonic symptoms (apparent sadness, reported sadness, lassitude, inability to feel), Factor 2 labeled anxiety and vegetative symptoms (inner tension, reduced sleep, reduced appetite, concentration difficulties), and Factor 3 labeled hopelessness (pessimistic thoughts, suicidal thoughts). The three‐factor structure observed in TRANSFORM‐2 was verified in TRANSFORM‐1. Treatment benefit at 24 h with esketamine versus placebo was observed on all 3 factors and continued throughout the 4‐week double‐blind treatment period. Conclusions A three‐factor structure for MADRS appears to generalize to TRD. All three factors improved over 4 weeks of treatment with esketamine nasal spray