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The synthesis of potassium 6-hydroxy-7-chloro-1,1-dimethyl-3,3-difluorobenzo-1,2,3-siloxaborolate 5b from readily available 4-bromo-2-chlorophenol was developed. This compound proved useful in various derivatizations resulting in a wide range of O-functionalized benzosiloxaboroles. Reactions of 5b with selected substituted benzoyl chlorides gave rise to a series of respective derivatives with 6-benzoate side groups attached to the benzosiloxaborole core. Furthermore, treatment of 5b with substituted benzenesufonyl chlorides afforded several benzosiloxaboroles bearing functionalized benzenesulfonate moieties at the 6 position. The synthesis of related chloropyridine-2-yloxy substituted benzosiloxaboroles was accomplished by a standard approach involving silylation/boronation of appropriate heterodiaryl ethers. Investigation of biological activity of obtained compounds revealed that some benzoate and most benzenesulfonate derivatives exhibit high activity against Gram-positive cocci such as methicillin-sensitive Staphylococcus aureus ATCC 6538P as well as methicillin-resistant S. aureus ATCC 43300 with the MIC values in the range of 0.39-3.12 mg L-1. Some benzenesulfonate derivatives showed also potent activity against Enterococcus faecalis ATCC 29212 and E. faecium ATCC 6057 with MIC = 6.25 mg L-1. Importantly, for the most promising cocci-active benzenesulfonate derivatives the obtained MIC values were far below the cytotoxicity limit determined with respect to human normal lung fibroblasts (MRC-5). For those derivatives, the obtained IC50 values were higher than 12.3 mg L-1. The results of antimicrobial activity and cytotoxicity indicate that the tested compounds can be considered as potential antibacterial agents.

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In this study, we isolated 28 yeast strains from samples of plant material and fermented food and evaluated the possibility of efficient production of 2-phenylethanol (2-PE) in the organic waste-based media supplemented with l-phenylalanine (l-Phe). We used whey, a by-product from milk processing, as a base for media, and either glucose or three by-products from sugar beet processing as a fermentable carbon source. Ten newly isolated yeast strains were capable of producing over 2 g l-1 2-PE through the l-Phe biotransformation in a batch mode in standard medium. Among them, we selected eight strains producing 2-PE in a range of 1·17-3·28 g l-1 in 72 h batch cultures in shaking flasks in whey-based media. The strains were assigned to five species of Meyerozyma caribbica, Metschnikowia chrysoperlae, Meyerozyma guilliermondii, Pichia fermentans and Saccharomyces cerevisiae. While S. cerevisiae is known to be a promising producer of 2-PE, the four latter species are poorly studied on this application. Results presented here are better than other reported values for batch cultures of unmodified yeast strains. Therefore, it seems that whey and by-products from sugar beet processing might be a good feedstock for 2-PE bioproduction. SIGNIFICANCE AND IMPACT OF THE STUDY: 2-Phenylethanol (2-PE) is an alcohol with a pleasant rosy scent, which is commonly used in the food, fragrance and cosmetic industries as an aroma compound and preservative. Promising sources of 2-PE are yeasts, but still the biotechnological route has not been economically competitive to chemical synthesis. Thus, the first challenging goal to develop biotechnological production of 2-PE is the identification of highly productive yeasts and cheap feedstock. This study demonstrates for the first time the promising production of 2-PE by selected yeasts in organic waste-based media. This could pave the way for development of a cheaper method of 2-PE bioproduction.

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The ParB protein of Pseudomonas aeruginosa is important for growth, cell division, nucleoid segregation and different types of motility. To further understand its function we have demonstrated a vital role of the hydrophobic residues in the C terminus of ParB(P.a.). By in silico modelling of the C-terminal domain (amino acids 242-290) the hydrophobic residues L282, V285 and I289 (but not L286) are engaged in leucine-zipper-like structure formation, whereas the charged residues R290 and Q266 are implicated in forming a salt bridge involved in protein stabilization. Five parB mutant alleles were constructed and their functionality was defined in vivo and in vitro. In agreement with model predictions, the substitution L286A had no effect on mutant protein activities. Two ParBs with single substitutions L282A or V285A and deletions of two or seven C-terminal amino acids were impaired in both dimerization and DNA binding and were not able to silence genes adjacent to parS, suggesting that dimerization through the C terminus is a prerequisite for spreading on DNA. The defect in dimerization also correlated with loss of ability to interact with partner protein ParA. Reverse genetics demonstrated that a parB mutant producing ParB lacking the two C-terminal amino acids as well as mutants producing ParB with single substitution L282A or V285A had defects similar to those of a parB null mutant. Thus so far all the properties of ParB seem to depend on dimerization.

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Deletions leading to complete or partial removal of ParB were introduced into the Pseudomonas aeruginosa chromosome. Fluorescence microscopy of fixed cells showed that ParB mutants lacking the C-terminal domain or HTH motif formed multiple, less intense foci scattered irregularly, in contrast to the one to four ParB foci per cell symmetrically distributed in wild-type P. aeruginosa. All parB mutations affected both bacterial growth and swarming and swimming motilities, and increased the production of anucleate cells. Similar effects were observed after inactivation of parA of P. aeruginosa. As complete loss of ParA destabilized its partner ParB it was unclear deficiency of which protein is responsible for the mutant phenotypes. Analysis of four parB mutants showed that complete loss of ParB destabilized ParA whereas three mutants that retained the N-terminal 90 aa of ParB did not. As all four parB mutants demonstrate the same defects it can be concluded that either ParB, or ParA and ParB in combination, plays an important role in nucleoid distribution, growth and motility in P. aeruginosa.

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Motor unit (MU) potentials were recorded from brachial biceps of healthy subjects aged 5.5-79 years. The subjects were subdivided into young (5.5-19 year) and adult (37.5-79 year) groups, between which single MU discharge characteristics were compared. Firing rates were in the ranges of 8.3-21.7 s(-1) (mean 12.87 s(-1)) and 5.9-18.7 s(-1) (mean 11.08 s(-1)) for young and adult groups, respectively. Standard deviations (s.d.) of interspike intervals (ISIs) were in the range 4.84-11.57 ms (mean 8.39 ms) for the young group and 4.26-12.23 ms (mean 7.76 ms) for the adult group. Both differences were statistically significant (P < 0.001). Special attention was paid to the previously developed method of ISI variability analysis, which enabled the comparison of MUs with respect to afterhyperpolarization (AHP) duration of their motoneurones (MNs). The results show that AHP duration increases gradually with increasing age, which is in line with the transformation of muscle properties towards a slower phenotype. This transformation seems to be a continuous process, covering the entire lifespan of a human being, from childhood to senescence. The results presented here are significant for their insight into the ageing process of the neuromuscular system. The age-related change in AHP duration has not been investigated previously in human studies and has been met with ambiguous results in animal studies.

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Eotaxin belongs to CC class of chemokines and is a potent eosinophil chamoattractant. Activated eosinophils are able to release many cytotoxic proteins, including eosinophil cationic protein (ECP), which has central role in allergic inflammation. The aim of this study was to assess eotaxin and ECP levels in plasma of atopic asthma patients in stable period of the disease. 19 patients with asthma and 10 healthy controls took part in this study. ELISA test was used to measure eotaxin (kits from R&D, pg/ml) and ECP (kits from Pharmacia, mg/l) levels. Plasma eotaxin level (mean +/- SD) was 176.52 +/- 50.3 (range 89-288) in asthma patients and 101.42 +/- 49.4 (range 35-206) in control group (p < 0.001). Plasma ECP concentration was 16.7 +/- 6.4 (6.3-28) and 16.8 +/- 17.1 (3.1-61.6), respectively (n.s). There was correlation between plasma eotaxin and ECP levels (Pearsons correlation co. r = +0.5, p < 0.05) and between eotaxin and FEV1 (Pearsons correlation co r = -0.4, p < 0.05) in asthma patients. We suggest that measurement of eotaxin and ECP levels as well may be useful indicator of disease.

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This study was undertaken to investigate the correlation between the serum ECP and the serum eotaxin level, and disease activity as evaluated with pulmonary function in patients with asthma or chronic obstructive pulmonary disease (COPD). 20 patients with stable asthma and 15 patients with COPD, and 15 subjects of the control group took part in this study. The analysis of ECP was performed according to the manufacturer's directions (Pharmacia Diagnostics AB, Uppsala, Sweden). The ELISA test was used to measure eotaxin levels in sserum (kits from R&D, USA). The levels of ECP were 16.9+/-6.3 microg/L in patients with asthma, 15.1+/-9.3 microg/L in patients with COPD and 11.8+/-6.2 microg/L in the control group (P<0.05). There was no significant difference in the asthma ECP level compared with the ECP level in COPD. There was a significant difference between the ECP plasma level in asthma compared with the ECP plasma level in the control group (p<0.05). The levels of eotaxin were 175.8+/-49.3 pg/mL in the control group. There was a correlation of ECP and the eotaxin level in asthma patients (r=+0.5, p<0.05). The percentage fall in FEV1 correlated with eotaxin level in asthma (r=-0.3, p<0.05) and with the eotaxin level in COPD (r=-0.5, p<0.05). Serum outcomes of eotaxin and ECP levels appear to be a useful indicator of atopic asthma, and might provide complementary data disease monitoring. Therefore, further investigations are required to clarify whether serum eotaxin measurements have a role in the clinical evaluation in COPD.

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In the early seventies, a suggestion that even in muscular dystrophy a neurogenic factor may be involved, was formulated. The argument which followed this suggestion, resulted in eventual abandoning of this concept even by its author. This discussion however has never been supported by any systematic study of motoneuron activity in muscular dystrophy. We examined an activity of motoneurons supplying brachial biceps in eight controls and 26 patients affected by Duchenne muscular dystrophy by studying single motor unit (MU) potentials picked up by fine wire bipolar electrodes. In the majority of patients, MU firing rates were higher as compared to controls and increased more rapidly with increasing force level. The relationship between standard deviation of interspike intervals and their mean value, SD(x), was shifted towards the shorter intervals and lower SDs. The numerical values describing these changes were correlated with severity of the disease. There is evidence that the break-point of the function SD(x) is correlated with motoneuron properties, in particular with after-hyperpolarization duration. In muscular dystrophy, this break-point corresponds to the shorter interspike intervals. We suggest that the motoneurons in muscular dystrophy are altered either in response to the muscle degeneration, or as a result of the disease itself.

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The activity of motoneurons supplying the brachial biceps muscle was examined in eight control subjects and 26 patients affected by Duchenne muscular dystrophy. The patients were subdivided into two groups: one whose motor units (MU) fired with normal rates (N group) and the other whose MU firing rates were higher as compared to controls (I group). Firing rates of motoneurons of patients from group I increased more rapidly with increasing force level. The relationship between the standard deviation of interspike intervals and their mean value, sigma(Tm), was shifted towards the shorter intervals and lower standard deviations in both groups of patients. The numerical values describing these changes correlated with the severity of disease. The MU recruitment was comparable for control subjects and for patients. Experimental results as well as computer simulations indicate that the break-point of the function sigma(Tm) is correlated with motoneuronal properties, and in particular with the afterhyperpolarization (AHP) duration. In muscular dystrophy this break-point corresponds to the shorter interspike intervals. Therefore, we propose that the motoneurons in muscular dystrophy are altered either in response to the muscle degeneration or as a result of the disease itself.

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