Recombinant TP-84 Bacteriophage Glycosylase-Depolymerase Confers Activity against Thermostable Geobacillus stearothermophilus via Capsule Degradation.

The TP-84 bacteriophage, which infects Geobacillus stearothermophilus strain 10 (G. stearothermophilus), has a genome size of 47.7 kilobase pairs (kbps) and contains 81 predicted protein-coding ORFs. One of these, TP84_26 encodes a putative tail fiber protein possessing capsule depolymerase activity. In this study, we cloned the TP84_26 gene into a high-expression Escherichia coli (E. coli) system, modified its N-terminus with His-tag, expressed both the wild type gene and His-tagged variant, purified the recombinant depolymerase variants, and further evaluated their properties. We developed a direct enzymatic assay for the depolymerase activity toward G. stearothermophilus capsules. The recombinant TP84_26 protein variants effectively degraded the existing bacterial capsules and inhibited the formation of new ones. Our results provide insights into the novel TP84_26 depolymerase with specific activity against thermostable G. stearothermophilus and its role in the TP-84 life cycle. The identification and characterization of novel depolymerases, such as TP84_26, hold promise for innovative strategies to combat bacterial infections and improve various industrial processes.

A Method for Isolation Bacteriophage Particles-Free Genomic DNA, Exemplified by TP-84, Infecting Thermophilic Geobacillus.

DNA purification methods are indispensable tools of molecular biology, used for many decades. Nevertheless, for certain specialized applications, the currently employed techniques are not sufficiently effective. While examining a number of the existing methods to purify the genomic DNA of the thermophilic bacteriophage TP-84, which infects Geobacillus stearothermophilus (G. stearothermophilus), we have found out that the obtained DNA is contaminated with trace amounts of infectious TP-84 particles. This was detrimental for the bacteriophage genetic manipulation purposes, as finding the recombinant TP-84 clones was essentially impossible due to the appearance of a high background of native bacteriophage plaques. Thus, we have developed a method, which enables the fast and efficient isolation of a bacteriophage genomic DNA from concentrated phage preparations, obtained using CsCl gradient ultracentrifugation, without the need to remove concentrated CsCl solutions. The method employs silica columns and mini-scale isolation of microgram amounts of high quality DNA. It is universal-the silica mini-columns from various manufacturers can be used to conduct the procedure. The purified DNA, free from infectious bacteriophage particles, is ready for further manipulations. This is particularly important for such thermophilic bacteriophages that may partially survive standard isolation procedures and contaminate the final DNA product.

Metabolic Syndrome and Cognitive Functions in Schizophrenia-Implementation of Dietary Intervention.

INTRODUCTION: The co-existence of schizophrenia and metabolic syndrome is a widely described phenomenon that contributes to the worse functioning of patients in everyday life. A relatively new area of research is the relationship between metabolic syndrome (MS) and cognitive function in patients with schizophrenia. The aim of the study was to verify the relationship between the presence of metabolic syndrome and cognitive function of patients with schizophrenia and to assess the possibility of changing cognitive function by introducing appropriate dietary intervention. MATERIALS AND METHODS: The study involved 87 individuals diagnosed with schizophrenia according to ICD-10 criteria, aged 19 to 67 years (M = 41.67; SD = 11.87). Patients were in the remission phase of schizophrenia, all using antipsychotics for pharmacological treatment. From a group of 83 patients with schizophrenia and diagnosed metabolic syndrome (according to IDF criteria) 30 patients were randomly assigned to an experimental group-with dietary intervention, 29 patients-to group without dietary intervention, 24 patients with schizophrenia without metabolic syndrome was a comparison group. All groups were evaluated for cognitive function using Stroop Test, Trail Making Test (TMT), Verbal Fluency Test, Digit Span Backwards Test. In the experimental group a dietary intervention was applied, which was to provide the examined person with a 7-day dietary plan with reduced calorie content, in compliance with the Mediterranean diet. RESULTS: After the dietary intervention there was a significant improvement in the number of errors made in the third Stroop Test (p <0.001), the time taken to complete the Point Linking Test was shortened (Test B; p = 0.005), there was an improvement in Verbal Fluency Test in "animals" category (p = 0.006) "sharp objects" category (p = 0.009), the number of repeated digits has increased in Digit Span Test in "forward" category (p = 0.001) and overall completion of the test (p = 0.021). In the group of patients with MS without dietary intervention, the results of cognitive tests remained mostly unchanged. CONCLUSIONS: Change of eating habits may be a significant element of a holistic approach to the problems of treatment of schizophrenia.

Dietary Behaviors and Metabolic Syndrome in Schizophrenia Patients.

The metabolic syndrome (MS) is highly prevalent in schizophrenia patients, resulting from both pharmacotherapy and their lifestyle. To avoid its development, the analysis of patients' eating behaviors followed by the necessary nutritional changes should become a routine element of treatment. The aim of this study is to investigate the effect of dietary habits on the course of schizophrenia and MS, cognitive performance, symptom severity, and subjective assessment of eating behaviors in schizophrenia patients. Total of 87 participants (63.2% women) aged 19 to 67 years (M = 41.67; SD = 12.87), of whom 60 met the IDF criteria for MS, completed the PANSS, the verbal fluency test, the Stroop Color-Word Test, and the digit span task, followed by a thorough nutritional interview. There were no significant differences in the dietary behaviors between investigated schizophrenia patients with and without comorbid MS. Interestingly, their eating habits compared quite favorably to those described in the literature. No associations were found between positive eating habits and other tested variables in patients with MS. They were, however, linked to lower PANSS scores in the entire sample. In addition, positive eating habits correlated with better cognitive performance and a more adequate subjective assessment of dietary habits. It would be amiss to assume that schizophrenia patients lack the ability to control their eating behaviors. Nutrition education may foster desirable dietary changes and improve the sense of agency, thus helping to reduce symptom severity and enhancing cognitive performance in this patient population.