Epitope Mapping of BmpA and BBK32 Borrelia burgdorferi Sensu Stricto Antigens for the Design of Chimeric Proteins with Potential Diagnostic Value.

Lyme disease is a tick-borne zoonosis caused by Gram-negative bacteria belonging to the Borrelia burgdorferi sensu lato (s.l.) group. In this study, IgM- and IgG-specific linear epitopes of two B. burgdorferi sensu stricto (s.s.) antigens BmpA and BBK32 were mapped using a polypeptide array. Subsequently, two chimeric proteins BmpA-BBK32-M and BmpA-BBK32-G were designed to validate the construction of chimeras using the identified epitopes for the detection of IgM and IgG, respectively, by ELISA. IgG-ELISA based on the BmpA-BBK32-G antigen showed 71% sensitivity and 95% specificity, whereas a slightly lower diagnostic utility was obtained for IgM-ELISA based on BmpA-BBK32-M, where the sensitivity was also 71% but the specificity decreased to 89%. The reactivity of chimeric proteins with nondedicated antibodies was much lower. These results suggest that the identified epitopes may be useful in the design of new forms of antigens to increase the effectiveness of Lyme disease serodiagnosis. It has also been proven that appropriate selection of epitopes enables the construction of chimeric proteins exhibiting reactivity with a specific antibody isotype.

Effect of selected lifestyle factors on the nutrition state of elderly people with metabolic syndrome.

Background: The formation and development of the metabolic syndrome (MetS) is largely caused by lifestyle factors. Many studies have shown that excessive consumption of simple carbohydrates, alcohol, salt, physical inactivity and smoking increase the risk of MetS. Objective: The aim of the study was to assess the nutritional status and its relationship with selected lifestyle factors in elderly people with MetS. Material and methods: 81 people aged 51-75 participated in the study. According to the guidelines, all tests were performed on an empty stomach. The comparison of individual quantitative variables between the groups was performed using the Student's t-test for independent measurements or using the Mann-Whitney U test. The relationship between quantitative variables was verified with the Spearman's correlation coefficient. All statistical tests were based on a significance level of p<0.05. Results: The BMI, the percentage of body fat and the waist circumference significantly exceed the norm for the population in the examined persons. The responses of respondents aged 51-65 show that 36% of people sweetened their drinks, 65.6% were salted their food, 51.6% consumed alcohol, and at the age of 66-75, respectively: 47.1%, 52.9%, 41.2%. There was no correlation between sweetening beverages and salting food and the concentration of glucose and lipids. However, a positive correlation was found between the amount of salt consumed and the heart rate (r=0.28, pp<0.05). In both age groups, statistically significant differences in the concentration of triglycerides depending on alcohol consumption or non-consumption were found. Due to the lack of precise data on the amount of alcohol consumed, the correlation between alcohol consumption and the concentration of glucose and lipids was not analyzed. Physical activity was not undertaken by 39.1% of patients aged 51-65 years and 41.2% of patients aged 66-75 years. In the group of elderly people without physical activity, a statistically significantly higher glucose concentration was found in relation to those who were physically active (130 mg/dl vs. 105 mg/dl; p=0.031). Patients aged 51-65 who engaged physical activity had statistically significantly lower body weight, BMI, waist circumference and lean body mass, which requires further studies. Conclusions: The anthropometric indices and parameters of MetS patients indicated disturbances in the nutritional status. Unhealthy lifestyle was shown mainly in the younger group of patients 51-65 years old (they sweetened drinks more often, salted dishes, consumed alcohol). Patients with metabolic syndrome did not undertake physical activity as often as recommended.

Assessment of Selected Structural Properties of High-Speed Friction Welded Joints Made of Unalloyed Structural Steel.

Commonly used S235JR structural steel, generally associated with good weldability, was joined by high-speed friction welding (HSFW). The friction welding tests were performed with a rotational speed of n = 8000 rpm and four different values of the unit pressure in the friction phase (pf) in the range of 64−255 MPa. The obtained joints were subjected to metallographic observations using an optical microscope; in selected zones of friction joints the average grain size was specified in accordance with the EN ISO 643:2012 standard; the hardness of friction joints was measured using the Vickers method. The friction-welded joint with the highest pf was EBSD-investigated. The obtained friction-welded joints resembled an hourglass, and the microstructure of individual zones of the joints differed depending on the height (axis, radius) of the observations. The generated joining conditions resulted in a significant refinement of the microstructure in the friction weld­the average grain size is about 1 µm2 (for base material it was 21 µm2). The highest increase in hardness above 340 HV0.1 was recorded in the friction weld of the welded joint with the lowest used value pressure in the friction phase. Such a sharp increase in hardness can make the resulting friction-welded joint become sensitive to dynamic or fatigue loads. The electron backscatter diffraction (EBSD) investigation confirmed the strong refinement of the microstructure in the friction-welded joint and the occurrence of the phenomenon of dynamic recrystallization (DRX). The friction weld was also characterized by a large share of high-angle boundaries (HAGBs) >80%. These results may indicate that during high-speed friction welding it is possible to create conditions like those obtained during the High-Pressure Torsion (the method used to produce UFG materials) process.

Microstructural Investigation of a Friction-Welded 316L Stainless Steel with Ultrafine-Grained Structure Obtained by Hydrostatic Extrusion.

The paper presents the microstructural investigation of a friction-welded joint made of 316L stainless steel with an ultrafine-grained structure obtained by hydrostatic extrusion (HE). Such a plastically deformed material is characterized by a metastable state of energy equilibrium, increasing, among others, its sensitivity to high temperatures. This feature makes it difficult to weld ultra-fine-grained metals without losing their high mechanical properties. The use of high-speed friction welding and a friction time of <1 s reduced the scale of the weakening of the friction joint in relation to result obtained in conventional rotary friction welding. The study of changes in the microstructure of individual zones of the friction joint was carried out on an optical microscope (OM), scanning electron microscope (SEM), transmission electron microscope (TEM) and electron backscattered diffraction (EBSD) analysis system. The correlation between the microstructure and hardness of the friction joint is also presented. The heat released during the high-speed friction welding initiated the process of dynamic recrystallization (DRX) of single grains in the heat-affected zone (HAZ). The additional occurrence of strong plastic deformations (in HAZ) during flash formation and internal friction (in the friction weld and high-temperature HAZ) contributed to the formation of a highly deformed microstructure with numerous sub-grains. The zones with a microstructure other than the base material were characterized by lower hardness. Due to the complexity of the microstructure and its multifactorial impact on the properties of the friction-welded joint, strength should be the criterion for assessing the properties of the joint.

Seroprevalence of Bartonella Species in Patients with Ocular Inflammation.

Bartonella species, vector-borne etiologic agents of many systemic or self-limited infections, are responsible for a widening spectrum of diseases in humans, including inflammatory conditions of the eye. The aim of this study was to determine whether there is any relationship between uveitis and the evidence of Bartonella spp. infection in the serum, ocular fluid, and cataract mass in patients with intraocular inflammation. Polymerase chain reaction (PCR)-based tests and DNA sequencing were performed on surgery-extracted specimens of intraocular fluid and lens mass of 33 patients. Sera from 51 patients and 101 control subjects were tested for the presence of specific antibodies against Bartonella spp. Neither IgM-class antibodies against Bartonella spp. nor Bartonella spp. DNA were detected. A specific IgG-class antibody was found in 33.3% of the patients with uveitis. The rate of positive Bartonella serology was higher among the uveitis patients than that in control subjects. This high rate may in part result from unrecognized indirect mechanisms rather than the immediate presence and multiplication of Bartonella spp. in the eyeball. Nonetheless we believe that screening for Bartonella spp. should become part of the diagnostic workup in uveitis.

Is the poly (L- lactide- co- caprolactone) nanofibrous membrane suitable for urinary bladder regeneration?

The purpose of this study was to compare: a new five-layered poly (L-lactide-co-caprolactone) (PLC) membrane and small intestinal submucosa (SIS) as a control in rat urinary bladder wall regeneration. The five-layered poly (L-lactide-co-caprolactone) membrane was prepared by an electrospinning process. Adipose tissue was harvested from five 8-week old male Wistar rats. Adipose derived stem cells (ADSCs) were seeded in a density of 3×10(6) cells/cm2 onto PLC membrane and SIS scaffolds, and cultured for 5-7 days in the stem cell culture medium. Twenty male Wistar rats were randomly divided into five equal groups. Augmentation cystoplasty was performed in a previously created dome defect. Groups: (I) PLC+ 3×10(6)ADSCs; (II) SIS+ 3×10(6)ADSCs; (III) PLC; (IV) SIS; (V) control. Cystography was performed after three months. The reconstructed urinary bladders were evaluated in H&E and Masson's trichrome staining. Regeneration of all components of the normal urinary bladder wall was observed in bladders augmented with cell-seeded SIS matrices. The urinary bladders augmented with SIS matrices without cells showed fibrosis and graft contraction. Bladder augmentation with the PLC membrane led to numerous undesirable events including: bladder wall perforation, fistula or diverticula formation, and incorporation of the reconstructed wall into the bladder lumen. The new five-layered poly (L-lactide-co-caprolactone) membrane possesses poorer potential for regenerating the urinary bladder wall compared with SIS scaffold.

Bacterial tick-borne diseases caused by Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, and Rickettsia spp. among patients with cataract surgery.

BACKGROUND: Clinical data have shown that tick-borne diseases caused by Borrelia burgdorferi sensu lato, Bartonella spp., Coxiella burnetii, and Rickettsia spp. can affect the central nervous system, including the eye. The aim of this study was to establish a relationship between the incidence of cataract and evidence of bacterial infections transmitted by ticks. MATERIAL AND METHODS: Fluid with lenticular masses from inside of the eye and blood from 109 patients were tested by PCR and sequencing. Sera from patients and the control group were subjected to serological tests to search specific antibodies to the bacteria. RESULTS: Microbiological analysis revealed the presence of Bartonella sp. DNA in intraoperative specimens from the eye in 1.8% of patients. Serological studies have shown that infections caused by B. burgdorferi sensu lato and Bartonella sp. were detected in 34.8% and 4.6% of patients with cataract surgery, respectively. CONCLUSIONS: Presence of DNA of yet uncultured and undescribed species of Bartonella in eye liquid indicates past infection with this pathogen. Specific antibodies to B. burgdorferi sensu lato and Bartonella sp. are detected more frequently in patients with cataract compared to the control group. This could indicate a possible role of these organisms in the pathological processes within the eyeball, leading to changes in the lens. Further studies are needed to identify Bartonella species, as well as to recognize the infectious mechanisms involved in cataract development.

[Concentration of sFas and sFasL in the supernatant of PBMC culture from the patients with late lyme borreliosis]. / Stfizenia bialek sFas i sFasL w hodowli komórek jednojadrzastych krwi obwodowej chorych z pózna borelioza z lyme.

OBJECTIVE: Persistent, inadequate inflammatory response present in late Lyme borreliosis may be driven by activated T lymphocytes. We estimated synthesis of extracellular proteins: soluble Fas receptor (sFas) and its ligand (sFasL), which might protect T lymphocytes from physiologic apoptosis, in the peripheral blood mononuclear cell (PBMC) culture from patients with late borreliosis. METHODS: Lyme borreliosis group (LB) consisted of 20 patients with Lyme borreliosis present for >6 months. Six patients without any active infection constituted the control group (K). PBMC were incubated for 7 days with phytohemaglutinine or suspension of Borrelia afzeli (Ba), B. garinii (Bg) and B. burgdorferi sensu stricto (Bss) spirochetes. sFas and sFasL concentrations were measured in the culture supernatant with ELISA. RESULTS: In LB mean sFasL concentration was increased significantly under stimulation with phytohemaglutinine, Ba and Bg, and, with borderline significance, with Bss, in comparison with unstimulated culture. sFas also tended to increase, which was significant with phytohemaglutinine and borderline with Bg. In K sFas and sFasL was not significantly increased under antigenic stimulation. CONCLUSIONS: Increased synthesis of antiapoptotic factors by PBMC from patients with late borreliosis incubated with B. burgdorferi antigens may suggest impaired apoptosis of T lymphocytes contributing to persistent inflammatory response in this patients.