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.

Estimated Incidence of Symptomatic Lyme Borreliosis Cases in Lublin, Poland in 2021.

Lyme borreliosis (LB), the most common tick-borne disease in Europe, is endemic to Poland. Despite public health surveillance with mandatory reporting of LB cases by physicians and laboratories, many symptomatic LB cases are not included in surveillance in Poland. We estimated the extent of the under-ascertainment of symptomatic LB cases via surveillance in the Polish province of Lublin to better understand Poland's LB burden. The number of incident symptomatic LB cases in Lublin in 2010 was estimated from two seroprevalence studies conducted among adults in Lublin, as well as estimates of the proportion of asymptomatic LB cases and the duration of LB antibody persistence. The estimated number of incident symptomatic LB cases was compared to the number of surveillance-reported cases in Lublin to derive an under-ascertainment multiplier. This multiplier was applied to the number of surveillance-reported cases in 2021 to estimate the number and population-based incidence of symptomatic LB cases in Lublin in 2021. We estimate that there are 5.9 symptomatic LB cases for every surveillance-reported LB case in Lublin. Adjusting for under-ascertainment, the estimated number of symptomatic LB cases in Lublin in 2021 was 6204 (population-based incidence: 467.6/100,000). After adjustment for under-ascertainment, the incidence of symptomatic LB in Lublin, Poland, is high.

Asymptomatic carrier of Babesia spp. among blood donors - epidemiological situation in Poland.

Introduction: Babesiosis is caused by one of several Babesia species. In Europe, B. divergens predominates in humans, while in North America it is B. microti. Babesia spp. infection in donors with a disease-free course of infection can be a major problem in blood recipients. A recipient with impaired immune system functions is at risk of full-blown development of the disease. In Poland and in most countries of the world, blood donors are not routinely tested for Babesia spp. infection. In our previous study, we detected Babesia venatorum DNA in blood donors, which was the reason for expanding the study to include more test subjects. Objective: The aim of this study was an attempt at estimating the prevalence of asymptomatic infection with Babesia spp. among blood donors from the Regional Centres for Blood Donation and Blood Treatment in Warsaw and Wroclaw. Materials and methods: The material for the study was whole blood from regular blood donors from two Regional Centre for Blood Donation and Blood Treatment in Warsaw and Wroclaw. Whole blood samples from 1,067 blood donors collected in June-July 2022 were analyzed. Blood collected directly from the donor during the blood donation procedure. All persons qualified by a doctor as a donor were selected for the study, regardless of age and sex. All subjects were informed in detail about the purpose of the study and gave their written consent. Isolation was made by using the Chelex 100 chelating resin, followed by the studying of the genetic material using the qPCR reaction. The results were analysed based on the amplification curve. Results: The protozoan Babesia spp. was not detected in the blood samples. Conclusions: The risk of blood-borne babesiosis is extremely low in Poland.

Testing of sand from recreational areas and sandpits in Warsaw, for presence of Ascaris Lumbricoides and Toxocara spp.

INTRODUCTION: Improper maintenance of the cleanliness of sand in sandboxes can be a source of human pathogenic helminths, including Toxocara spp., Enterobius vermicularis, Ascaris lumbricoides, leading to parasitic infections. AIM OF THE STUDY: The purpose of this study was to evaluate the sanitary condition of sandboxes in playgrounds and recreational areas in Warsaw for the presence of Human roundworm (Ascaris lumbricoides) and Toxocara spp. in sand. MATERIAL AND METHODS: 450 sand samples from 90 sandboxes in the city of Warsaw were analyzed. The flotation method was used for the study, and the material was evaluated using a light microscope. RESULTS. The conducted examinations did not reveal the presence of parasite eggs, indicating that hygiene rules were observed and appropriate recommendations were followed. CONCLUSIONS: The analyzed sand samples are free of the tested parasites.

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.

Phase Structure Evolution of the Fe-Al Arc-Sprayed Coating Stimulated by Annealing.

The article presents the results of research on the structural evolution of the composite Fe-Al-based coating deposited by arc spray with initial low participation of in situ intermetallic phases. The arc spraying process was carried out by simultaneously melting two different electrode wires, aluminum and low alloy steel (98.6 wt.% of Fe). The aim of the research was to reach protective coatings with a composite structure consisting of a significant participation of FexAly as intermetallic phases reinforcement. Initially, synthesis of intermetallic phases took place in situ during the spraying process. In the next step, participation of FexAly fraction was increased through the annealing process, with three temperature values, 700 °C, 800 °C, and 900 °C. Phase structure evolution of the Fe-Al arc-sprayed coating, stimulated by annealing, has been described by means of SEM images taken with a QBSD backscattered electron detector and by XRD and conversion electron Mössbauer spectroscopy (CEMS) investigations. Microhardness distribution of the investigated annealed coatings has been presented.

Microstructure of Rhenium Doped Ni-Cr Deposits Produced by Laser Cladding.

The addition of Rhenium up to 6% to Ni-Cr alloys can dramatically improve the corrosion and oxide resistance of deposited coatings at high operating temperatures. Ni-Cr+Re layers can be successfully produced using conventional powder metallurgy, high rate solidification (HRS), or magnetron sputtering methods. However, in industrial applications, high-performance deposition methods are needed, e.g., laser cladding. Laser cladding has several advantages, e.g., metallurgical bonding, narrow heat-affected zone (HAZ), low dilution, and slight thermal damage to the substrate. In this paper, a powder Ni-Cr composite with 1% (wt.) of Rhenium was produced, then deposited onto a steel substrate (16Mo3) by laser cladding to assess the micro and macrostructural properties of the obtained layers. Besides the macro and microscopic observations, scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) microanalysis of the deposit and HAZ as well as microhardness measurements have been conducted. The microstructure observations revealed four subareas of HAZ gradually changing from the fusion line towards the base material. Maximum hardness occurred in the HAZ, mainly in areas closer to the clad/substrate interface, reaching up to 350-400 HV. No sudden changes in the composition of the deposit and the area of fusion line were observed.

Anaplasmosis in Poland - analysis of 120 patients.

The aim of our study was to clarify the clinical picture of anaplasmosis through analysis of the symptoms and clinical signs presented by infected patients in a cohort of tick-bitten individuals. The study included 1375 patients with suspicion of tick-borne disease. Finally, 120 patients (8.7%) were diagnosed with anaplasmosis (HGA). Blood samples were examined by PCR for Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, Borrelia burgdorferi sensu lato, Babesia spp., and Bartonella spp.. Based on analysis of 120 patients with HGA we concluded that anaplasmosis is not as rare in Europe, as it is thought to be and often appears as a co-infection with other tick-borne pathogens. The co-infection rate of patients with A. phagocytophilum infection in tick endemic areas is high. Co-infection of A. phagocytophilum with B. burgdorferi s.l. or tick-borne encephalitis virus may influence symptom frequency. PCR together with medical history, clinical picture and basic laboratory tests is a sufficient method for the diagnosis of anaplasmosis. Doxycycline is an effective drug leading to complete recovery.

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.

Repellents as a major element in the context of prevention of tick-borne diseases

In Poland, out of the 21 species of ticks described, two species have the greatest epidemiological significance: Ixodes ricinus and Dermacentor reticulatus. Their participation in the transmission of etiologic agents of vector diseases such as Lyme disease, tick-borne encephalitis, human granulocytic anaplasmosis, babesiosis, rickettsiosis and many others is widely known. Due to the often severe and sometimes fatal course of diseases transmitted by ticks, a great deal of emphasis is placed on prophylactic activities, minimizing the possibility of biting and transfering pathogens along with the arachnid saliva. In addition to means of personal protection, including appropriate clothing, protective vaccinations, avoiding tick habitats or body checking after returning from this type of places, one of the most important elements of anti-tick prevention is the use of effective repellents. The key role of using the repellent is to discourage the arthropod from attacking and to prevent it from taking food, as a result of which there is no phenomenon of transmission of pathogens from the hematophage to the host organism. The most commonly used substances with arthropod repellent properties are: N-N-diethylm-toluamide (DEET), 3-(N-acetyl-N-butyl) aminopropionic acid ethyl ester (IR3535), icaridine, permethrin and essential oils. However, it should be remembered that no repellent ever protects 100% all the time after application - its use should be considered as one of many elements of personal protection in the prevention of tick bites.

Tick-borne pathogens Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii and Rickettsia spp. may trigger endocarditis.

BACKGROUND: Infections caused by tick-borne pathogens such as Bartonella spp., Borrelia burgdorferi s.l., Coxiella burnetii, and Rickettsia spp. are capable of causing serious lesions of the mitral and aortic valves, leading to a need for valve replacement. OBJECTIVES: The aim of the study was to determine whether such cases are sporadic or frequent. An additional goal was to establish effective diagnostic methods to detect these infections. MATERIAL AND METHODS: The study involved 148 patients undergoing valve replacement. Blood samples were drawn for serological testing. Samples of the removed mitral and aortic valves were tested with polymerase chain reaction and immunohistochemical staining. RESULTS: Specific antibodies to Bartonella spp. were detected in 47 patients (31.7%) and in 1 of the healthy controls (1%) (p < 0.05). Antibodies to B. burgdorferi spirochetes were found in 18 of the patients (12.2%) and in 6 blood donors from the control group (5.8%) (p < 0.1). Antibodies to Rickettsia spp. were detected in 12 (8.1%) and to C. burnetii phase I and II antigens in the serum of 1 patient. All the participants in the control group were seronegative to C. burnetii and Rickettsia spp. antigens. Polymerase chain reaction (PCR) tests for detection of Bartonella spp., B. burgdorferi s.l., C. burnetii and Rickettsia spp. DNA in the valve samples were all negative. Inflammation foci with mononuclear lymphoid cells in the aortic and mitral valves were seen in sections stained with hematoxiline and eozine. In sections dyed using the indirect immunofluorescence method with hyperimmune sera, Bartonella spp. and Rickettsia spp. were found. CONCLUSIONS: The results obtained indicate that laboratory diagnostics for patients with heart disorders should be expanded to include tests detecting tick-borne zoonoses such as bartonelloses, Lyme borreliosis, rickettsioses and Q fever.

Multi-Response Optimization of Electrical Discharge Machining Using the Desirability Function.

Electrical discharge machining (EDM) is a modern technology that is widely used in the production of difficult to cut conductive materials. The basic problem of EDM is the stochastic nature of electrical discharges. The optimal selection of machining parameters to achieve micron surface roughness and the recast layer with the maximal possible value of the material removal rate (MRR) is quite challenging. In this paper, we performed an analytical and experimental investigation of the influence of the EDM parameters: Surface integrity and MRR. Response surface methodology (RSM) was used to build empirical models on the influence of the discharge current I, pulse time ton, and the time interval toff, on the surface roughness (Sa), the thickness of the white layer (WL), and the MRR, during the machining of tool steel 55NiCrMoV7. The surface and subsurface integrity were evaluated using an optical microscope and a scanning profilometer. Analysis of variance (ANOVA) was used to establish the statistical significance parameters. The calculated contribution indicated that the discharge current had the most influence (over the 50%) on the Sa, WL, and MRR, followed by the discharge time. The multi-response optimization was carried out using the desirability function for the three cases of EDM: Finishing, semi-finishing, and roughing. The confirmation test showed that maximal errors between the predicted and the obtained values did not exceed 6%.

Prevalence of Borrelia Burgdorferi Sensu Lato in Ticks from the Ternopil Region in Ukraine.

INTRODUCTION: Lyme borreliosis/Lyme disease is caused by Borrelia burgdorferi and is one of the most common vector-borne diseases transmitted by ticks. MATERIAL AND METHODS: A total of 136 Ixodes ricinus ticks, collected in the Ternopil (Ukraine) region, including 126 adults (70 females and 56 males), and 10 nymphs were examined. The identification of the species and their developmental form was based on morphological characteristics. RESULTS: PCR with B5S-Bor and 23S-Bor primers resulted in Borrelia burgdorferi sensu lato DNA amplification among six ticks (4.4%). The detailed analysis based on the DNA sequencing showed the presence of DNA of Borrelia afzelii in four samples; the remaining two represented Borrelia burgdorferi sensu lato complex, although their genospecies were not determined. The research confirmed the dominance of Borrelia afzelii genospecies in the ticks from Ukraine. CONCLUSION: It seems reasonable to undertake similar research in ticks from other regions of Ukraine. Knowledge in this field can be useful for public health and planning the prevention of tick-borne diseases.

Leptospirosis in Poland in the years 2014-2017 ­ characteristics of infections and epidemiological surveillance data

OBJECTIVES: The aim of the study was the analysis of current Leptospira spp. infections in Poland on the basis of blood serum samples tests results and clinical data collected from clinicians in the Laboratory NIPH-NIH. METHODS: Clinical materials from 48 patients with clinical symptoms suggesting Leptospira spp suspected of leptospirosis from the years 2014-2017 were included to the study. Blood serum samples collected from patients were tested in Laboratory of Rickettsiae, Chlamydiae and Spirochaetes (currently Laboratory of Vector-borne Diseases) of NIPH-NIH. Levels of specific IgM and IgG antibodies to Leptospira spp. antigens were detected with the enzyme-linked immunosorbent assay (ELISA). RESULTS: Specific antibodies to Leptospira spp. were detected in 18 patients (37.5%). IgM antibodies were found in 6 patients (12.5%) and IgG antibodies were identified in 7 patients (14.6%). Both classes of antibodies of were detected in 5 patients (10.4%). The most samples for study were sent to laboratory from Masovian (13 samples) and Kuyavian-Pomeranian (11 samples) Voivodeships. Not any samples from the Lower Silesia, Lublin, Lódz, Podlaskie and Warmian-Masurian Voivodeships were received. In these patients the most common symptoms of disease were: fever, hepatitis with jaundice and renal failure. CONCLUSIONS: The number of diagnosed human leptospirosis in Poland is low in comparison to the number of cases in other countries, although the Leptospira spp. spirochetes occur in animals in the environment.

Francisella tularensis/Rickettsia spp. co-infections in patients with skin changes and lymphadenopathy.

INTRODUCTION: Tularemia and spotted fever group rickettsioses (SFG) can be transmitted by ticks and have a number of common clinical symptoms. Most characteristic are a maculopapular or vesicular rash or an eschar at the site of the tick or insect bite accompanied by painful lymph nodes. The aim of this study was to determine whether Rickettsia spp./Francisella tularensis mixed infections occurred in patients with similar symptoms who were diagnosed with either Rickettsia spp. or F. tularensis infection. MATERIAL AND METHODS: Thirty-six cases from 2011-2014, including 15 individuals with clinically and serologically recognized SFG and 21 with tularemia, were analyzed retrospectively using immunofluorescence for detection of Rickettsia spp. or ELISA for detection of F. tularensis. RESULTS: Of the 36 cases examined, specific high titers of antibodies to Rickettsia spp. were found in 1 (4.4%) patient with tularemia and specific high titers of antibodies to F. tularensis were detected in 1 (6.7%) patient with SFG. CONCLUSIONS: The results of our study show that in infections with fever, enlarged lymph nodes and skin lesions after tick and insect bites, laboratory testing of both diseases - SFG rickettsiosis and tularemia - should be implemented. Identification of F. tularensis and Rickettsia spp. mixed infections is crucial in order to administer appropriate antibiotics and to avoid treatment failure and relapse.

An outbreak of leptospirosis imported from Germany to Poland.

BACKGROUND: Leptospirosis is a zoonotic disease caused by spirochetes of the Leptospiraceae family. In both humans and animals the main route of infection is indirect contact - through water or other products contaminated with urine containing spirochetes. Infection most commonly occurs through ingestion of water or food contaminated with Leptospira spp. OBJECTIVES: The aim of the study was to characterize cases of leptospirosis imported to Poland from Germany in 2014 and to analyze methods that are helpful for making a diagnosis. MATERIAL AND METHODS: The 10 patients examined were reported as suspected leptospirosis cases on the basis of clinical symptoms and epidemiological investigations. They originated from different regions of Poland and had been working together at a strawberry plantation in the Cloppenburg district of Lower Saxony in Germany. Blood and urine samples were tested by PCR and serum samples by serology. All ELISA positive and negative cases were examined using a reference microscopic agglutination test (MAT). RESULTS: In the tested group, 6 individuals (60%) were seropositive according to the ELISA, and 2 of them were confirmed by the MAT. The PCR results for the blood and urine samples were negative. CONCLUSIONS: Using the ELISA in the diagnosis of leptospirosis allowed the disease to be identified much faster, differentiating classes of antibodies and recognizing levels of them that are too low to be detectable by the MAT.

Bactericidal Activity of Octenidine to Various Genospecies of Borrelia burgdorferi, Sensu Lato Spirochetes in Vitro and in Vivo.

The aim of our studies was to invent a reliable method for detection of the bactericidal activity of disinfectants against Borrelia burgdorferi in suspension (in vitro) and in cell line cultures (in vivo). In the suspension method, 0.01% octenidine at 20°C and 35°C was bactericidal to Borrelia afzeli; Borrelia garini, B. burgdorferi sensu stricto after 5 minutes treatment. Increase of the temperature to 35°C speed up the bactericidal effect to 1 minute. The bactericidal action of octenidine towards B. burgdorferi spirochetes growing in fibroblasts was less effective and needed a longer time to kill them than in the suspension.

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.