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The coexistence within a subcellular complex of inter-cellular proteins Ro60, responsible for preserving ncRNA quality, and Ro52, involved in intracellular proteolysis, has been a subject of ongoing debate. Employing molecular docking in tandem with experimental methods like Quartz Crystal Microbalance with Dissipation (QCM-D), Proximity Ligation Assay (PLA), and Indirect Immunofluorescence (IIF), we reveal the presence of Ro60 associating with Ro52 within the cytoplasm. This result unveils the formation of a weak transient complex with a Ka ≈ (3.7 ± 0.3) x 106 M-1, where the toroid-shaped Ro60 structure interacts with the Ro52’s Fc receptor, aligning horizontally within the PRY-SPRY domains of the Ro52’s homodimer. The stability of this complex relies on the interaction between Ro52 chain A and specific Ro60 residues, such as K133, W177, or L185, vital in the Ro60-YRNA bond. These findings bridge the role of Ro60 in YRNA management with Ro52's function in intracellular proteolysis, emphasizing the potential impact of transient complexes on cellular pathways.

The coexistence within a subcellular complex of inter-cellular proteins Ro60, responsible for preserving ncRNA quality, and Ro52, involved in intracellular proteolysis, has been a subject of ongoing debate. Employing molecular docking in tandem with experimental methods like Quartz Crystal Microbalance with Dissipation (QCM-D), Proximity Ligation Assay (PLA), and Indirect Immunofluorescence (IIF), we reveal the presence of Ro60 associating with Ro52 within the cytoplasm. This result unveils the formation of a weak transient complex with a Ka = (3.7 ± 0.3) x 106 M-1, where the toroid-shaped Ro60 structure interacts with the Ro52's Fc receptor, aligning horizontally within the PRY-SPRY domains of the Ro52's homodimer. The stability of this complex relies on the interaction between Ro52 chain A and specific Ro60 residues, such as K133, W177, or L185, vital in the Ro60-YRNA bond. These findings bridge the role of Ro60 in YRNA management with Ro52's function in intracellular proteolysis, emphasizing the potential impact of transient complexes on cellular pathways. [ABSTRACT FROM AUTHOR]

The study discussed in this article is driven by the realization that many physical processes may be understood by using applications of fractional operators and special functions. In this study, we present and examine a fractional integral operator with an I-function in its kernel. This operator is used to solve several fractional differential equations (FDEs). FDE has a set of particular cases whose solutions represent different physical phenomena. Many mathematical physics, biology, engineering, and chemistry problems are identified and solved using FDE. Specifically, a few exciting relations involving the new fractional operator with incomplete I-function (IIF) in its kernel and classical Riemann Liouville fractional integral and derivative operators, the Hilfer fractional derivative operator, and the generalized composite fractional derivate (GCFD) operator are established. The discovery and investigation of several important exceptional cases follow this. [ABSTRACT FROM AUTHOR]

The present article introduces and studies the Fredholm-type integral equation with an incomplete I-function (IIF) and an incomplete $ \bar {I} $ -function (I $ \bar {I} $ F) in its kernel. First, using fractional calculus and the Mellin transform principle, we solve an integral problem involving IIF. The idea of the Mellin transform and fractional calculus is then used to analyse an integral equation using the incomplete $ \bar {I} $ -function. This is followed by the discovery and investigation of several important exceptional cases. This article's general discoveries may yield new integral equations and solutions. The desired outcomes seem to be very helpful in resolving many real-world problems whose solutions represent different physical phenomena. And also, findings help solve introdifferential, fractional differential, and extended integral equation problems.

Background: Both enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) are available for the diagnosis of autoimmune bullous diseases (AIBD). Many studies have reported on the performance of ELISAs and concluded that ELISAs could replace IIF. This study compares the diagnostic accuracy of ELISA and IIF for the detection of autoantibodies to desmoglein 1 (DSG1), desmoglein 3 (DSG3), bullous pemphigoid antigen 2 (BP180) and bullous pemphigoid antigen 1 (BP230) to support the diagnosis of pemphigus vulgaris (PV), pemphigus foliaceus (PF) and bullous pemphigoid (BP). Methods: A literature search was performed in the PubMed database. The meta-analysis was performed using summary values and a bivariate random effect model. Results: The five included studies on PV did not demonstrate significant differences between IIF and DSG3-ELISA (sensitivity 82.3% vs. 81.6%, p = 0.9284; specificity 95.6% vs. 93.9%, p = 0.5318; diagnostic odds ratio [DOR] 101.60 vs. 67.760, p = 0.6206). The three included studies on PF did not demonstrate significant differences between IIF and DSG1-ELISA (sensitivity 80.6% vs. 83.1%, p = 0.8501; specificity 97.5% vs. 93.9%, p = 0.3614; DOR 160.72 vs. 75.615, p = 0.5381). The eight included studies on BP showed that BP230-ELISA differed significantly from both IIF on monkey esophagus (MO) and BP180-ELISA with regard to DOR (11.384 vs. 68.349, p = 0.0008; 11.384 vs. 41.699, p = 0.0125, respectively) Conclusions: Our meta-analysis shows that ELISA performs as well as IIF for diagnosing PV, PF and BP. [ABSTRACT FROM AUTHOR]

Diploid and continuous cell lines are used to propagate viral vaccines. At Human Viral Vaccine Department of Razi Vaccine and Serum Research Institute, MRC5 diploid cell is used for the development of live attenuated measles, mumps, rubella, and three types of poliovirus vaccines. Additionally, three continuous cell lines (i.e., RK13, HeLa, and Vero) are applied in quality control tests. Accordingly, cell cross-contamination can occur at cell culture labs, hence controlling the identity and specificity of cells is essential. Indirect immunofluorescence is a sensitive, specific, and simple test for cell identification. The present study was designed to develop the in-house indirect immunofluorescence test (IIF) as follows: homemade polyclonal anti-MRC5 serum was prepared in rabbits, and cross-reactive antibodies to RK13, HeLa, and Vero cells were eliminated. The diploid and continuous cell lines were fixed on Teflon slide using cold methanol and acetone. The reproducibility of the in-house IIF test was evaluated using the agreement Kappa test. The purity of the three batches of MRC5 working seed cell at Human Viral Vaccine Department of Razi institute was verified using IIF and no contamination with continuous cell lines was detected.

Feline morbillivirus (FeMV) was isolated for the first time in 2012 with an association with chronic kidney disease (CKD) suggested. This study aimed at investigating in cats from southern Italy FeMV prevalence and risk factors for exposure to FeMV, including the relationship with CKD; sequencing amplicons and analyzing phylogeny of PCR positive samples. Blood serum, K3EDTA blood and urine samples from 223 cats were investigated. Ten carcasses were also evaluated. FeMV RNA was detected in 2.4% (5/211) blood and 16.1% (36/223) urine samples. One carcass tested positive by qPCRFeMV from kidney, urinary bladder, and submandibular lymph nodes. Antibodies against FeMV were detected in 14.5% (28/193) cats. We followed up 27 cats (13 FeMV positive cats) and documented in some cases urine shedding after up to 360 days. Older and foundling cats and cats living in rescue catteries, were more frequently infected with FeMV. A significant correlation between FeMV and higher serum creatinine values or low urine specific gravity was found. FeMV positivity was significantly associated with retroviral infection, and the presence of some clinical signs apart from CKD clinicopathological markers. Our study highlights the possibility of a link between FeMV exposure and CKD and a general impairment of feline health.

Antinuclear antibody (ANA) testing is best performed using the indirect immunofluorescence (IIF) method with human epithelial type-2 (HEp-2) cells as the substrate. IIF is a subjective procedure in which HEp-2 patterns are analyzed manually from the microscope. Therefore, ANA test results greatly rely on the experience and expertise of pathologists. Hence, complete automation of the ANA test is required to avoid incorrect diagnoses. This paper represents an algorithm for the complex HEp-2 cell classification problem. The proposed algorithm used a small hybrid feature set that characterizes the texture and morphology of the HEp-2 cells along with artificial neural network (ANN). The hybrid features were extracted by breaking up the image into eight binary images. The proposed hybrid descriptors were more efficient than the popular co-occurrence matrix descriptor and local binary pattern descriptors for texture analysis. The proposed algorithm was evaluated on the ICPR 2016 IIF HEp-2 cell image dataset. The results indicated that the hybrid descriptor with an ANN approach achieved improved performance, with "96.8%" mean class accuracy. [ABSTRACT FROM AUTHOR]

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Copyright of Archives of Razi Institute is the property of Institut Razi and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: It is important to choose the correct screening titer in immunoflourescent assay for ANA. Recently, 1:160 titer is used worldwide, particularly after the suggestion of CDC (Centers for Disease Control and Prevention) and WHO (World Health Organisation). In our country, 1:100 is the mostly accepted titer for screening as a cutoff point. In this study, our aim was to check the ideal screening titer for Turkish society and investigate the convenience of suggested 1:160 screening titer for our country. Methods: Between January- February 2015, a total of 400 patients (healthy blood donors and patients with autoimmune rheumatic diseases- has been diagnosed or followed) were included in the study. Serum samples taken from donors and patients worked ANA test with two different commercial (Aesku and Euroimmun) kits by 1:100 and 1:160 screening titers. Microscobic evaluation determinated by blind and automated system (Helios). The serum samples detected positive by 1:100 and 1:160 titers worked. In the end sensivity and specificity of the ANA test was figured out. Also cut-off values of the test was figured out by ROC curve analyse. Results: In healthy group the frequency of positivity is 8%, whereas in patients group it is 92%. In healthy group distribution of ANA titer positivity is: in 1:100 titer 44%, in 1: 160 -31%, in 1: 320 -19% and in 1: 640 -6%. In healthy group in 1:160 distribution of ANA patern is: dense fine speck (dfs 70) 56%, speck 25%, nucleolar 12,5%, centromere 6.5%. In patients' group distribution of ANA titer positivity is: in 1:100-1%, in 1:160-3%, in 1:320-26%, in 1:640 25%, in 1:280-34%, in 1:2560-8% and in 1:5120-3%. In patients group in 1:160 distribution of ANA patern is: speck 55%, homogeneous 24%, nucleolar 11%, centromere 5%, nuclear dots 3% and nuclear membrane 2%. Conclusion: In our study, the ideal screening titer of 1:160 for ANA test was confirmed. The screening titer of 1:160, as proposed by CDC and WHO, is found confident to use in the laboratories in Turkey. [ABSTRACT FROM AUTHOR]

A third outbreak of Trichinella britovi with 140 people involved, occurred in Granada Spain (December 1998). The source of infection was sausage made from uninspected wild boar meat. Fifty-two patients agreed to participated in this study. An elevated eosinophil level (> 5 %) was detected in 59.6 % of patients, and persisted in most of these cases for two months. A moderate IgG response was observed. At the onset of symptoms, Western blot (WB) test detected more positive cases than Enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF). Six months from infection, ELISA revealed fewer positive cases than the other two tests. It would appear that the response to somatic antigens starts earlier than those to cuticular and excretory/secretory (ES) antigens and that the response to ES antigens is the first to decrease.

Erysipelothrix rhusiopathiae is the causative agent of erysipelas, a disease of many mammalian and avian species, mainly swine and turkeys. In cetaceans, erysipelas is considered to be the most common infection in juvenile individuals, which have not been vaccinated. Moreover, the disease manifest in both forms, the dermatologic and the acute septicemic forms, has been reported in various species of dolphins and whales. It is difficult to diagnose erysipelas by currently available approaches. Moreover, it is mainly based on culture methods and also PCR methods, which are currently being developed. At the present stage, prophylactic approaches are based on antibiotic therapy and vaccination mostly with porcine erysipelas vaccines. In the present study, an Indirect Immuno Fluorescence method for the detection of dolphin antibodies levels against E. rhusiopathiae was developed and applied in two different groups of captive bottlenose dolphins ( Tursiops truncatus) from Loro Parque (Tenerife, Canary Islands, Spain) and L'Oceanogràfic de Valencia (Valencia, Spain) in order to check the tittering levels of antibodies after application of porcine erysipelas vaccines in the studied dolphins. [ABSTRACT FROM AUTHOR]

Abstract: Human Epithelial (HEp-2) cells are commonly used in the Indirect Immunofluorescence (IIF) tests to detect autoimmune diseases. The diagnosis consists of searching and classification to specific patterns created by Anti-Nuclear Antibodies (ANAs) in the patient serum. Evaluation of the IIF test is mostly done by humans, which means that it is highly dependent on the experience and expertise of the physician. Therefore, a significant amount of research has been focused on the development of computer aided diagnostic systems which could help with the analysis of images from microscopes. This work deals with the design and development of HEp-2 cells classifier. The classifier is able to categorize pre-segmented images of HEp-2 cells into 6 classes. The core of this engine consists of the following image descriptors: Haralick features, Local Binary Patterns, SIFT, surface description and a granulometry-based descriptor. These descriptors produce vectors that form metric spaces. k-NN classification is based on aggregated distance function which combines several features together. An extensive set of evaluations was performed on the publicly available MIVIA HEp-2 images dataset which allows a direct comparison of our approach with other solutions. The results show that our approach is one of the leading classifiers when comparing with other participants in the HEp-2 Cells Classification Contest[1]. [Copyright &y& Elsevier]

Background and Objectives: Autoantibodies are immunglobulins occurred directly against autoantigens that are known as endogen antigens. Autoimmune disease is an occasion that the body begins a fight against its own cells and tissues. The antibodies that are created by the body against its own cell nuclei are called as anti-nuclear antibodies (ANA), and one of the methods used for detection and pattern of ANA is indirect immunofluorescence test (IIF). In the present study, it was aimed to determine the rate of ANA positivity and patterns of the positive specimens, and to investigate the relationship between ANA positivity and diseases in patients. Methods: ANA test results of a total of 3127 patients admitted during March 2010 to December 2012 were evaluated retrospectively. ANA test (HEp 20-10, EUROIMMUN, Germany) was used in dilution of 1:100 in IIF test. Results: A total of 494 (15.8%) resulted as ANA positive. ANA positivity rate was significantly higher in female patients than the male ones (p<0.001). The most frequent ANA patterns were coarse speckled pattern (154 patients, 31.2%), nucleolar pattern (89 patients, 18.0%), fine speckled pattern (57 patients, 11.5%), and speckled pattern (48 patients, 9.7%). ANA positivity was most commonly determined in rheumatoid arthritis (RA) (42 patients, 8.5%), systemic lupus erythematosus (SLE) (29 patients, 5.9%), and rheumatoid vasculitis (RV) (28 patients, 5.7%). The most frequent symptoms or findings were joint pain (127 patients, 26.0%) and anemia (28 patients, 5.7%). ANA positivity rates were found to be significantly higher in patients with RA (p < 0.001), with SLE (p < 0.001), and with Raynaud phenomenon (p=0.001) in comparison to the controls. Amongst the most frequent diseases evaluated, no significant differences were found between the control groups and the groups of RV (p=0.089), multiple sclerosis (p=0.374), and Sjögren syndrome (p=0.311) in terms of ANA positivity rates. Conclusions: The present study is the first study reporting the positivity rate and distribution of ANA in Bolu located in northwestern Turkey. Information about the pattern types and the distribution of the patterns according to the diseases and symptoms contribute in diagnosis of autoimmune diseases. It is observed that clinical diagnosis has been supported significantly by ANA test according to data of our study. [ABSTRACT FROM AUTHOR]

Objective: To study the therapeutic effects of low-dose amiodarone and Betaloc on hypertrophic cardiomyopathy complicated by malignant ventricular arrhythmias. Methods: Eighty-two such patients were selected and divided into a treatment group and a control group by the random number method (n=41), which were administered with low-dose amiodarone plus Betaloc and individual Betaloc respectively. Results: The treatment group had a significantly higher overall effective rate (85.4%) than the control group (65.9%) did. Based on the New York Heart Association's classification of cardiovascular disease, the treatment group mainly comprised Class III and IV patients before treatment, which were significantly relieved after treatment (P < 0.05). The heart rate was evidently decreased from (119.99±18.91) bpm to (80.98±12.34) bpm, and the incidences of premature ventricular contraction and tachycardia were significantly reduced (P < 0.05). The longest QT intervals after and before treatment were (421±32) ms and (411±35) ms respectively. The shortest QT interval after treatment [(350±36) ms] was significantly longer than that before [(307±31) ms]. The QT dispersion before treatment [(96±29) ms] was significantly higher that after [(64±17) ms] (P < 0.05). Six out of eighty two patients in the treatment group succumbed to adverse reactions (14.63%). Conclusion: Hypertrophic cardiomyopathy complicated with malignant ventricular arrhythmias can be well treated with low-dose amiodarone and Betaloc, with mitigated symptoms, improved prognosis and few adverse reactions. [ABSTRACT FROM AUTHOR]

Abstract: Predictions by various mathematical models of intracellular ice formation (proposed by Mazur, Pitt, Toner, and Karlsson, respectively) were compared to the known thermodynamic and kinetic behavior of ice formation in supercooled aqueous systems. The older models (Mazur, Pitt, and Toner) significantly underestimated the magnitude of colligative nonequilibrium freezing point depression in response to increased concentration of solutes, such as salts or cryoprotectants. Furthermore, kinetics predicted using phenomenological models (by Mazur and Pitt) exhibited implausible temperature-dependence, with the probability of intracellular ice formation being allowed to increase even at temperatures below the glass transition point. The Toner model, on the other hand, produced invalid results at temperatures below −48°C. The Karlsson model was the only model that consistently yielded realistic predictions over a wide range of temperatures and solute concentrations, especially in the presence of cryoprotectant additives. To facilitate adoption of the Karlsson model of intracellular ice nucleation, the complete set of model equations has been collected and described in detail. [Copyright &y& Elsevier]

Challenge: New Mexico Corporate Services (NMCS) recordable injuries have been averaging 2–4 per year for the last 5 years with no statistical improvement. However, we believe all NM employees must go home incident and injury free every day and just as healthy as they came to work. In addition, we have received feedback from several sources, that indicates some employees are reluctant to report injuries. These indicators showed us that continuing our current strategies, making incremental improvement and changes, would not give us the improvement desired. We needed a paradigm shift to get everyone completely engaged in the IIF (Incident & Injury Free) culture, in order to achieve true IIF results. Methods/Strategies: We formed a small (3-person) taskforce consisting of safety representatives from EHS, Site Services and CS Operations. We reviewed 5 years worth of data to determine what was injuring our people. We also decided to review all injuries, not simply those classified as recordable by OSHA standards. First we identified the types of injury information needed to get a true picture of our safety issues. We analyzed IRB (Incident Review Board) data showing us the following factors and whether any of them contributed to the injury: – Date – Incident Description – Severity – Root Cause – Type of Injury – Season – Work Group/Shift – Area – Improper evaluation of hazard – Inadequate work procedures – Incorrect Mental Model – Inadequate PPE Requirement – Failure to Follow or Unaware of PPE Requirement – Shortcut or Schedule Pressure – Last or First Day of Shift or Adjacent to Holiday – OT – Aggravate Existing Condition – Inadequate Training or Passdown – Experience in Task – Corrective Action Taken – Overall Quality of Response. Once this information was collected for all injuries in an Excel file, we graphed it several ways to help reveal trends: [-] Shift 7 had double the injuries of shift 5 [-] Night shift injuries were relatively high but lower than Shift 7 [-] Shift 5 had no severe (recordable) injuries [-] Biggest total injury type was ergonomic [-] Biggest recordable injury type (58%) was cuts/lacerations [-] Chemical exposure and inhalations resulted in no recordables [-] Biggest root cause category was behavioral [-] Biggest behavioral root cause was “Failure to Evaluate Hazards” [-] Biggest administrative controls root cause was “Inadequate Procedures” [-] Biggest engineering controls root cause was “Unrecognized Workplace Hazard” [-] Majority of injuries occurred in Spring/Summer which accounted for 86% of recordables [-] Response was inadequate or poor for 25% of injuries [-] Number of injuries by workgroup were about equal [-] Work area injuries were mostly in the CUB and Subfab but CUB injuries favored cuts/lacerations while Subfab favored ergonomic injuries These learnings and gaps were used to develop our New Mexico Site Safety Action Plan for 2005. It has also been used by individual managers and supervisors in their day-to-day business meetings, activities, and communications, for a more focused IIF message and effort. Results: ▪ Conclusions/Recommendations: Challenges are different amongst shifts. Shift culture may be stronger than workgroup culture. Work areas pose different challenges that all should be aware of and prepared for. We should prepare ourselves for challenges posed by the Spring and Summer seasons as well. We should be aware of and prepare for the different challenges and cultures characteristic of each shift. Supervisors need to spend more time in the field with their people to help them address safety issues and help make IIF a more consistent component of daily workgroup culture. Each site should analyze their data and look for their unique safety challenges... [Copyright &y& Elsevier]

Abstract: Intracellular ice formation (IIF) plays a central role in cell damage during cryopreservation. We are investigating the factors which trigger IIF in Xenopus oocytes, with and without aquaporin water channels. Here, we report differential scanning calorimeter studies of Xenopus control oocytes which do not express aquaporins. Stage I to VI oocytes (which increase progressively in size) were investigated with emphasis on stage I and II because they are translucent and can also be studied under the cryomicroscope. Measurements were made in 1, 1.5, and 2M ethylene glycol (EG) in frog Ringers plus SnoMax. A multistep freezing protocol was used in which the samples were cooled until extracellular ice formation (EIF) occurred, partially remelted, slowly recooled through the EIF temperature, and then rapidly (10°C/min) cooled. EIF in the 1, 1.5, and 2M EG occurred at −6.4, −7.8, and −8.9°C, respectively. Freezing exotherms of individual stage I–VI oocytes were readily visible. A general trend was observed in which the IIF temperature of the early stage oocytes (I–III) was well below T EIF while the later stages (IV–VI) froze at temperatures much closer to T EIF. Thus, in 1.5M EG, T IIF was −21.1, −25, and −26.6°C in stages I–III, but was −17 and −8.5°C for stage IV and V–VI. Concurrently, the percentage of oocytes in which IIF was observed fell dramatically from a high of 40 to 72% in early stages (I–III) to a low of only 7% in stage V–VI because, particularly in the later stages, IIF was hidden in the EIF exotherm. We conclude that early stage oocytes are a good model system in which to investigate modulators of IIF, but that late stage oocytes are damaged during EIF and infrequently supercool. [Copyright &y& Elsevier]

SUMMARY Antineutrophil cytoplasmic antibodies (ANCA) of IgG class have been described at high prevalence in autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC). Data on IgA class ANCA in these diseases are limited. The aim of this study was to determine the prevalence and fluorescence patterns of IgA class ANCA in AIH and PSC and to examine a relationship between the presence of IgA ANCA and clinical characteristics in these patients. Sera from 35 patients with PSC (21 with concomitant inflammatory bowel disease), 40 patients with AIH and 10 healthy controls were studied. ANCA were detected on ethanol-fixed neutrophils using an indirect immunofluorescence technique. ANCA of the IgA class were found in 20% of sera from patients with PSC and in 50% of AIH sera. The majority of AIH patients with IgA class ANCA showed a ‘classical’ perinuclear staining pattern, whereas the ‘classical’ and ‘atypical’ perinuclear fluorescence patterns were distributed equally in PSC. In sera containing IgG and IgA class ANCA simultaneously, IgG class ANCA showed an ‘atypical’ pANCA fluorescence pattern whereas IgA class ANCA produced a ‘classical’ perinuclear staining. The presence of IgA class ANCA was not associated with disease-specific clinical characteristics. IgA class ANCA are more frequently detected in sera of patients with AIH than PSC. The diversity of fluorescence patterns points to different target antigens of IgA class ANCA with distinct subcellular localizations. [ABSTRACT FROM AUTHOR]

Objective: To compare the antitumoral effects of all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9-cis RA) with those of 5-OH,11-O-hydrophenanthrene (IIF), a new derivative of retinoic acid.Materials and Methods: The effect of retinoids was tested on cell line HL-60. Cell differentiation and apoptosis were evaluated by morphological and biochemical analysis as BCL-2 protein and by DNA fragmentation assay. The ability to activate retinoic acid receptors (RAR) and/or retinoid X receptors (RXR) and to modulate gene expression was determined by transactivation assay.Results: With cell line HL-60, the antiproliferative effect of IIF was stronger than that of ATRA and 9-cis RA. Following retinoid treatment, cells appeared to differentiate and apoptotic cells were observed. The appearance of DNA laddering and a decrease in the amount of BCL-2 protein confirmed apoptosis. IIF transcriptionally activated RXR-gamma more than RAR-alpha.Conclusion: The findings indicate that IIF transcriptionally activates RXR-gamma preferentially, induces apoptosis and has a more antiproliferative activity than ATRA and 9-cis RA on cell line HL-60. [ABSTRACT FROM AUTHOR]

Anti-neutrophil cytoplasmic antibodies (ANCAs) are autoantibodies that recognize neutrophil cytoplasmic antigens. The major ANCA antigens are myeloperoxidase and proteinase 3. Necrotizing small vessel vasculitis accompanied by ANCA production is called ANCA-associated vasculitis (AAV). In addition to AAV, ANCA is sometimes produced in patients with connective tissue diseases, such as systemic lupus erythematosus, and inflammatory bowel diseases. Indirect immunofluorescence (IIF) and enzyme immunoassay (EIA) have been used to detect ANCAs. Recently, the accuracy of EIA has improved and it has become the gold standard for ANCA detection. However, IIF does not lose its role in ANCA detection because EIA cannot detect ANCAs that recognize antigens other than those coated on the plate. For IIF, neutrophil substrates prepared with two different fixations, namely, ethanol fixation and formalin fixation, are used. There is a recommended protocol for ethanol fixation but not for formalin fixation. This study prepared neutrophil substrates according to the recommended protocol for ethanol fixation and protocols in the literature and original protocols for formalin fixation and then examined ANCA specificity and how storage period would influence the number of cells, antigen distribution, and antigenicity of the substrates. As a result, the number of cells and antigen distribution did not change after storage for up to 2 months regardless of fixation protocols, whereas a time-dependent decline in ANCA antigenicity and a fixation protocol-dependent difference in ANCA specificity were observed. How neutrophils are fixed on the glass slide needs to be checked upon evaluation of ANCAs by IIF. [ABSTRACT FROM AUTHOR]

Feline morbillivirus (FeMV) was isolated for the first time in 2012 with an association with chronic kidney disease (CKD) suggested. This study aimed at investigating in cats from southern Italy FeMV prevalence and risk factors for exposure to FeMV, including the relationship with CKD; sequencing amplicons and analyzing phylogeny of PCR positive samples. Blood serum, K3EDTA blood and urine samples from 223 cats were investigated. Ten carcasses were also evaluated. FeMV RNA was detected in 2.4% (5/211) blood and 16.1% (36/223) urine samples. One carcass tested positive by qPCRFeMV from kidney, urinary bladder, and submandibular lymph nodes. Antibodies against FeMV were detected in 14.5% (28/193) cats. We followed up 27 cats (13 FeMV positive cats) and documented in some cases urine shedding after up to 360 days. Older and foundling cats and cats living in rescue catteries, were more frequently infected with FeMV. A significant correlation between FeMV and higher serum creatinine values or low urine specific gravity was found. FeMV positivity was significantly associated with retroviral infection, and the presence of some clinical signs apart from CKD clinicopathological markers. Our study highlights the possibility of a link between FeMV exposure and CKD and a general impairment of feline health. [ABSTRACT FROM AUTHOR]

Aim: Antinuclear antibodies (ANA) that bind to intracellular antigens are a distinctive feature of systemic autoimmune diseases, but also can be seen in other situations such as cancer and infections. It is claimed that the neutrophil lymphocyte ratio (NLR) is a potential marker for assessing inflammation. The relationship between patterns of antinuclear immunofluorescence antibody and NLR has not been investigated yet. In this study, we aimed to establish potential relationship among NLR and ANA immunofluorescence patterns. Material and Method: In a retrospective manner (December 2014 - February 2016), a total of 218 adult patients with rheumatologic diseases that indirect immunofluorescence ANA test were positive and have CBC test result in same day were included the study as case group randomly. According to ANA immunofluorescence patterns, patients divided to subgroups (Table 1). Inclusion criteria for control group were: no diagnosed with autoimmune diseases, ANA test were negative and have CBC test result in same day with ANA test. CBC test was performed by Mindray cell counter (BC 6800) according to manufacturer's directions and sera samples were screened for ANA by indirect immunofluorescence (IIF) in Hep-20 cells (Euroimmun). Results: The results were summarized in Table 1. There was no significant difference between case and control groups for age (year). But there was significantly difference between gender and some IIF-ANA patterns. Interestingly, the neutrophil lymphocyte ratio was significantly lower in pattern of nuclear dots when compared with control group (p=0.033). Conclusion: Although, we need well designed further studies, we think that the lower neutrophil/lymphocyte ratio may be associated with ANA patterns like 'nuclear dots'. [ABSTRACT FROM AUTHOR]

Background: Indirect immunofluorescence plays a major role in the detection of antinuclear antibodies (ANAs) and follow-up of their titers in the context of connective tissue diseases. Given the numerous unfavorable features of the conventional manual reading of HEP2 slides (need of time and expert morphologists for the reading, lack of standardization, subjectivity of the interpretation), the biomedical industry has developed automated techniques of slide preparation and microscope reading., Methods: We collected 49 sera beforehand analyzed by the conventional reading of slides. They were prepared again by QUANTA-Lyser(®) and reanalyzed in four different conditions: two dilutions of screening (1/40 and 1/80), two different systems of analysis, NOVA View(®) automated reading (INOVA Diagnostics), then confirmation by the operator, and conventional manual reading by two different qualified operators. The analysis was realized in blind of the first interpretation and clinical diagnosis. The sera were classified in four groups, on the basis of the results of the first analysis: negative sera (titer < 1/160; 11 patients), low positives (titer at 1/160; 18 patients), moderated positives (titers between 1/320 and 1/640; 10 patients), and strong positives (titers between 1/1,280 and 1/2,560; 10 patients)., Results: Among the 49 patients, 13 presented a connective tissue disease including 4 systemic scleroderma (SS), 3 rheumatoid arthritis (RA), 2 Goujerot-Sjogren (GS), 2 systemic lupus erythematosus (SLE), 1 polymyositis (PM), 1 Raynaud's syndrome (RS), and 1 CREST syndrome. One patient presented both an SLE and an SS. Regarding the screening dilution, the 1/40 dilution is less specific than the 1/80 dilution for both the systems of analysis (5.6% vs. 16.7% for the manual reading, and 27.8% vs. 50% for the automated reading). It also generates statistically more false positives (P = 0.037 for the conventional analysis and P = 0.003 for the automated system). The automated NOVA View(®) reading of slides allows a gain in specificity for both dilutions, and also statistically less false positives (P = 0.002 at the 1/40 and P = 0.0006 at the 1/80), and detriment of the sensitivity at the highest dilution (84.6% vs. 92.3% with manual reading). Thus, according to our analysis of 49 sera, the automated NOVA View(®) system of reading of slides at the dilution 1/80 seems to be a successful condition for the detection of ANAs on HEP2 cells, close to the significance (P = 0.067)., Conclusion: The automated NOVA View(®) reading of slides allows saving time, and an improvement in the standardization. Nevertheless, it requires a confirmation by a qualified operator, to interpret mixed patterns in particular., (© 2016 Wiley Periodicals, Inc.)

Aberrant mucin O-glycosylation often occurs in different cancers and is characterized by immature expression of simple mucin-type carbohydrates. At present, there are some controversial reports about the Tn antigen (GalNAcα-O-Ser/Thr) expression and there is a great lack of information about the [UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyltransferase (GalNAc-Ts)] expression in chronic lymphocytic leukemia (CLL). To gain insight in these issues we evaluated the Tn antigen expression in CLL patient samples using two Tn binding proteins with different fine specificity. We also studied the expression from 14 GalNAc-Ts genes in CLL patients by RT-PCR. Our results have provided additional information about the expression level of the Tn antigen, suggesting that a low density of Tn residues is expressed in CLL cells. We also found that GALNT11 was expressed in CLL cells and normal T cell whereas little or no expression was found in normal B cells. Based on these results, GALNT11 expression was assessed by qPCR in a cohort of 50 CLL patients. We found significant over-expression of GALNT11 in 96% of B-CLL cells when compared to normal B cells. Moreover, we confirmed the expression of this enzyme at the protein level. Finally we found that GALNT11 expression was significantly associated with the mutational status of the immunoglobulin heavy chain variable region (IGHV), [א(2)(1)=18.26; P<0.0001], lipoprotein lipase expression [א(2)(1)=13.72; P=0.0002] and disease prognosis [א(2)(1)=15.49; P<0.0001]. Our evidence suggests that CLL patient samples harbor aberrant O-glycosylation highlighted by Tn antigen expression and that the over-expression of GALNT11 constitutes a new molecular marker for CLL., (© 2013 Elsevier B.V. All rights reserved.)

Background: Direct immunofluorescence (DIF), indirect immunofluorescence (IIF), and enzyme-linked immunosorbent assay (ELISA) are used for the laboratory diagnosis of bullous pemphigoid (BP)., Objective: The diagnostic value of DIF and IIF on rabbit and monkey esophagus or human salt-split skin and commercial ELISAs was assessed., Methods: This was a single-center retrospective study where 313 patients with BP were compared with 488 control subjects., Results: DIF was the most sensitive test (90.8%) whereas sensitivities for IIF on rabbit esophagus, IIF on monkey esophagus, IIF on salt-split skin, BP180 ELISA, and BP230 ELISA were 76.0%, 73.2%, 73.3%, 72.0%, and 59.0%, respectively. The sensitivity of the serologic tests was 88.8% altogether. The specificities for DIF, IIF on rabbit esophagus, IIF on monkey esophagus, IIF on salt-split skin, BP180 ELISA, and BP230 ELISA were 98%, 96.5%, 97.1%, 100%, 94.1%, and 99.2%, respectively., Limitations: The retrospective nature of study was a limitation. Correlation of diagnostic data with clinical manifestations or disease course was not possible., Conclusions: In suspected BP, both serologic tests and DIF have to be performed because of a sensitivity issue. Although the ELISAs had a relatively low sensitivity, the serologic tests altogether almost reached the level of sensitivity of DIF. The specificities of all assays were excellent., (Copyright © 2013 American Academy of Dermatology, Inc. Published by Mosby, Inc. All rights reserved.)

Background: We determined if PR3-ANCA is a biomarker that differentiates ulcerative colitis (UC) from Crohn's disease (CrD)., Methods: A total of 946 sera were tested, including 86 granulomatosis with polyangiitis (GPA) and 491 inflammatory bowel disease (IBD) patients (283 UC and 208 CrD), 264 pathological controls (various diseases) and 105 healthy individuals. All samples were tested for PR3-ANCA by ELISA (QUANTA Flash Lite®, INOVA Diagnostics) and chemiluminescent immunoassays (CIA QUANTA Flash PR3). Conventional anti-neutrophil cytoplasmic antibody (ANCA) indirect immunofluorescence assays (IIF) was performed with NOVA Lite™ (INOVA Diagnostics)., Results: PR3-ANCA by CIA were detected in 31.1% UC vs. 1.9% CrD sera (p=2.2E-16), and by ELISA in 6% UC and 0% CrD (p=0.0003). In GPA patients, PR3-ANCA were detected in 75.6% by CIA and 61.6% by ELISA (p<0.05). PR3-ANCA by CIA were more prevalent in E3-UC compared to E1/2-UC (p<0.05), and in patients with shorter disease duration (p<0.0001). PR3-ANCA showed similar sensitivity, but significantly higher specificity (p<0.05), compared to atypical pANCA by IIF., Conclusion: The novel PR3 CIA may prove helpful in the differentiation of CrD from UC, as well as in the identification of UC patients with more extensive disease., (© 2013 Elsevier B.V. All rights reserved.)

Background & Aims: Anti-neutrophil cytoplasmic antibodies (ANCA) are a non-uniform family of antibodies recognizing diverse components of neutrophil granulocytes. ANCA formation might be induced by protracted bacterial infections or probably reflect an abnormal immune response to commensal microorganisms. Bacterial infections are common complications in cirrhosis with high incidence of episodes caused by enteric organisms, therefore, we sought to study the presence and clinical importance of ANCA in cirrhosis., Methods: Sera of 385 patients with cirrhosis of different etiologies were assayed for ANCA of IgG, IgA, IgA1, IgA2, and secretory IgA subtypes by indirect immunofluorescence and ELISAs. The control group comprised 202 patients with chronic liver diseases without cirrhosis and 100 healthy subjects. In cirrhosis, a 2-year follow-up, observational study was conducted to assess a possible association between the presence of ANCA and clinically significant bacterial infections., Results: Prevalence of ANCA IgA was significantly higher in cirrhosis (52.2%) compared to chronic liver diseases (18.6%) or healthy controls (0%, p<0.001 for both). ANCA IgA subtyping assays revealed marked increase in the proportion of IgA2 subtype (46% of total ANCA IgA) and presence of the secretory component concurrently. Presence of ANCA IgA was associated with disease-specific clinical characteristics (Child-Pugh stage and presence of ascites, p<0.001). During a 2-year follow-up period, risk of infections was higher among patients with ANCA IgA compared to those without (41.8% vs. 23.4%, p<0.001). ANCA IgA positivity was associated with a shorter time to the first infectious complication (pLogRank <0.001) in Kaplan-Meier analysis and was identified as an independent predictor in multivariate Cox-regression analysis (HR:1.74, 95% CI: 1.18-2.56, p=0.006)., Conclusions: Presence of IgA type ANCA is common in cirrhosis. Involvement of gut mucosal immune system is in center of their formation and probably reflects sustained exposure to bacterial constituents., (Copyright © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

While some of the more frequent antinuclear (auto)antibodies (ANA) patterns such as homogenous nuclear staining have been extensively studied, the prevalence and clinical significance of rare antinuclear antibody patterns are not well understood. For the purpose of this review, we defined rare patterns as patterns occurring in less than 1% of patients that test positive on indirect immunofluorescence. The prevalence of different ANA patterns was determined in 68,128 consecutive patients who attended the outpatient clinic or were hospitalized at the University Hospitals Leuven over a 14-year period (1998-2011). To avoid bias, we only included the first sample for each patient and patients who tested positive in the period 1980-1997 were excluded. There were 9268 patients who tested positive for ANA. With the exception of the clinical association of anti-multiple nuclear dots (at higher titers) and anti-nuclear envelope autoantibodies with autoimmune liver disease, there was no good clinical association of rare ANA patterns with the diagnosis of auto-immune disorders. The most important non-autoimmune cause of rare ANA patterns was carcinoma, particularly in patients with rare cell-cycle related ANAs., (Copyright © 2013. Published by Elsevier B.V.)

Glucocorticoids are given to pregnant women at risk for premature delivery to promote lung maturation. Despite reports of detrimental effects of glucocorticoids on telencephalic neural stem/progenitor cells (NSPCs), the regional and cellular expressions of the glucocorticoid receptor (GR) in various NSPC populations in the intact brain have not been thoroughly assessed. Therefore in this study we performed a detailed analysis of GR protein expression in the developing mouse ventral and dorsal telencephalon in vivo. At embryonic day 11.5 (E11.5), the majority of Pax6-positive radial glial cells (RGCs) and Tbr2-positive intermediate progenitor cells (IPCs) expressed nuclear GR, while a small number of RGCs on the apical ventricular zone (aVZ), expressed cytoplasmic GR. However, on E13.5, the latter population of RGCs increased in size, whereas abventricular NSPCs and especially neurons of the cortical plate, expressed nuclear GR. In IPCs, GR was always nuclear. A similar expression profile was observed throughout the ventral telencephalon, hippocampus and olfactory bulb, with NSPCs of the aVZ primarily expressing cytoplasmic GR, while abventricular NSPCs and mature cells primarily expressed nuclear GR. Close to birth, nuclear GR accumulated within specific cortical areas such as layer V, the subplate and CA1 area of the hippocampus. In summary, our data show that GR protein is present in early NSPCs of the dorsal and ventral telencephalon at E11.5 and primarily occupies the nucleus. Moreover, our study suggests that the subcellular localization of the receptor may be subjected to region and neurodevelopmental stage-specific regulation., (Copyright © 2013 Elsevier B.V. All rights reserved.)