Hypomethylation of CYP2E1 and DUSP22 Promoters Associated With Disease Activity and Erosive Disease Among Rheumatoid Arthritis Patients.

OBJECTIVE:Epigenetic modifications have previously been associated with rheumatoid arthritis (RA). In this study, we aimed to determine whether differential DNA methylation in peripheral blood cell subpopulations is associated with any of 4 clinical outcomes among RA patients. METHODS:Peripheral blood samples were obtained from 63 patients in the University of California, San Francisco RA cohort (all satisfied the American College of Rheumatology classification criteria; 57 were seropositive for rheumatoid factor and/or anti-cyclic citrullinated protein). Fluorescence-activated cell sorting was used to separate the cells into 4 immune cell subpopulations (CD14+ monocytes, CD19+ B cells, CD4+ naive T cells, and CD4+ memory T cells) per individual, and 229 epigenome-wide DNA methylation profiles were generated using Illumina HumanMethylation450 BeadChips. Differentially methylated positions and regions associated with the Clinical Disease Activity Index score, erosive disease, RA Articular Damage score, Sharp score, medication at time of blood draw, smoking status, and disease duration were identified using robust regression models and empirical Bayes variance estimators. RESULTS:Differential methylation of CpG sites associated with clinical outcomes was observed in all 4 cell types. Hypomethylated regions in the CYP2E1 and DUSP22 gene promoters were associated with active and erosive disease, respectively. Pathway analyses suggested that the biologic mechanisms underlying each clinical outcome are cell type-specific. Evidence of independent effects on DNA methylation from smoking, medication use, and disease duration were also identified. CONCLUSION:Methylation signatures specific to RA clinical outcomes may have utility as biomarkers or predictors of exposure, disease progression, and disease severity

PTP4A1 promotes TGFβ signaling and fibrosis in systemic sclerosis.

Systemic sclerosis (SSc) is an autoimmune disease characterized by fibrosis of skin and internal organs. Protein tyrosine phosphatases have received little attention in the study of SSc or fibrosis. Here, we show that the tyrosine phosphatase PTP4A1 is highly expressed in fibroblasts from patients with SSc. PTP4A1 and its close homolog PTP4A2 are critical promoters of TGFβ signaling in primary dermal fibroblasts and of bleomycin-induced fibrosis in vivo. PTP4A1 promotes TGFβ signaling in human fibroblasts through enhancement of ERK activity, which stimulates SMAD3 expression and nuclear translocation. Upstream from ERK, we show that PTP4A1 directly interacts with SRC and inhibits SRC basal activation independently of its phosphatase activity. Unexpectedly, PTP4A2 minimally interacts with SRC and does not promote the SRC-ERK-SMAD3 pathway. Thus, in addition to defining PTP4A1 as a molecule of interest for TGFβ-dependent fibrosis, our study provides information regarding the functional specificity of different members of the PTP4A subclass of phosphatases

Protein metabolic changes and nucleolus organizer regions activity in the lymphocytes of neonatal calves during the development of respiratory diseases

Background and Aim: Calfhood disease is an important problem in dairy farming that could cause significant effects on heifer survival and productivity and has economic and welfare effects. Total protein concentration in the blood serum could be one of the predictors of bovine respiratory disease (BRD) in newborn calves. The number of active nucleolus organizers could be used to assess the viability of the protein synthesis system in cells and tissues. We aimed for a comparative assessment of the dynamics of the main indicators of protein metabolism and nucleolus organizer regions (NORs) activity in the lymphocytes of healthy calves (Group I) and calves with BRD (Group II) during the 1st month after birth. Materials and Methods: This study included 30 calves of the red-motley Holstein breed. Venous blood samples were taken from all calves on the 1st, 7th, 14th, and 28th days after birth. Quantitative analysis of total protein (Serum total protein [STP]), immune globulin (Serum immune globulin [SIg]), urea, and creatinine in serum and transcriptionally active chromosome NORs in the interphase nuclei of lymphocytes was conducted using receiver operating characteristic analysis and factor analysis. Results: In Group I, the STP levels decreased during the 1st month of life, and in Group II, the STP levels were variable. The STP levels in both groups remained within the reference intervals. During the first 2 weeks after birth, the calves' SIg fluctuated within the statistical error limits and did not significantly differ between the groups. On the 28th day, SIg increased in both the groups (by 42.8% for Group I and 33.7% for Group II). The creatinine concentration showed a decrease but did not go beyond the range of reference values. Urea concentration in Group I markedly decreased and remained below the reference values; it did not change in Group II over the entire observation period. The number of NORs in 1-day-old calves did not significantly differ between the groups and amounted to 2.43 in Group I and 2.59 in Group II. A significant increase in the number of active NORs was found in calves in both groups at the ages of 14 and 28 days. Early BRD predictors (at 1-14 days) could not be identified among the studied indicators. The urea and creatinine concentrations and the NOR activity on day 28 after birth could be late BRD predictors. Protein metabolism in the newborn calves' organisms is regulated by three types of factors: Maintenance of a constant protein concentration in the plasma, protein decomposition, and de novo synthesis. Conclusion: There were no observed significant differences in the protein metabolism values and dynamics of indicators between healthy calves and calves with developed BRD. Alterations in the studied characteristics are the result, but not the cause of BRD. The increase in active NORs under BRD could be a favorable forecasting indicator. Protection against foreign protein and genetic material is a more important task for the organism than ensuring growth processes during the neonatal period

Retrospective diagnosis of intrauterine diselementosis in newborn calves

One of the methods in diselementosis diagnosis, i.e. in determination of the trace elements content excess, deficiency or imbalance in the organism, is screening the hair elemental composition. It is known that the tail brush hair growth in cattle begins from the 7th month of gestation. The authors suggested that the tail brush hair elemental composition in newborn calves could be used for integral evaluation of the trace elements supply to the fetus in the last months of its intrauterine development. In order to elaborate criteria for retrospective diagnosis of the intrauterine diselementosis, 125 clinically healthy 1-day-old calves (67 male and 58 female animals) were examined; they were obtained from Simmental cows with a physiological course of gestation under biogeochemical conditions of the Central Black Earth region of the Russian Federation. The concentration of 11 essential (Co, Cr, Cu, Fe, I, Li, Mn, Se, Si, V, Zn) and 8 toxic (As, B, Cd, Hg, Ni, Pb, Sn, Sr) trace elements were studied in samples of unpigmented guard hairs from the animal tail brush by inductively coupled plasma mass spectrometry (Nexion 300D, Perkin Elmer, USA). The results were expressed as mean ± standard error of mean, median, minimum, and maximum. Reference values of essential and toxic trace elements concentration in the hair of newborns were determined using the percentile scales separately for the groups of male and female animals. Group comparison was conducted using the independent samples Mann-Whitney U test. Research results demonstrated that physiologically determined the range of the trace elements concentration in the hair of newborn calves corresponded to the interval between the 25th and 75th percentiles. Values beyond the specified interval boundaries were proposed to be considered as deviations corresponding to the diselementosis

Incidence risk of bronchopneumonia in newborn calves associated with intrauterine diselementosis

Background and Aim: Macro- and micro-elements are required to ensure the normal course of biochemical processes in the development of an animal's body. Any excess, deficiency, or imbalance in chemical elements in an animal's body can cause the development of various latent or clinically expressed pathological conditions. Diselementosis in pregnant cows may lead to impaired embryo and fetal development, as well as reduced neonatal viability. The aim of this study was to analyze the content of macroelements and microelements in the blood serum of both calving cows and their calves to evaluate the relationship between indicators of mineral metabolism in the mother and newborn and to establish what role separate chemical elements play in making newborn calves more prone to bronchopneumonia. Materials and Methods: The content of potassium (K), sodium (Na), iron (Fe), Copper (Cu), Zinc (Zn), Strontium (Sr), arsenic (As), nickel (Ni), cobalt (Co), chromium (Cr), molybdenum (Mo), and selenium (Se) in the blood serum of 33 pregnant cows between 239 and 262 days of gestation and their 33 1-day old calves was determined using the Shimadzu AA6300 (Japan) atomic adsorption spectrophotometer. Calcium (Ca) and magnesium (Mg) content was determined using ion-selective electrodes from the Olympus-400 analyzer (Beckman Coulter, USA). During the 1st month of life, all calves in the sample set had some sort of respiratory diseases and seven of the calves had bronchopneumonia. Retrospectively, the samples of adult and newborn animals were divided into two groups each: Dams I – cows whose calves had uncomplicated bronchitis (n=26); Dams II – cows whose calves got bronchopneumonia (n=7); and Newborns I – calves with uncomplicated bronchitis (n=26); Newborns II – calves with bronchopneumonia (n=7). Results: The content of Ca, Mg, K, Na, Mo, and Se in dams in both groups of cows was within the reference range; the concentrations of Fe and Ni were higher than the reference range; and the concentrations of Cu, Zn, As, Co, and Cr were lower than the reference range. There were no significant differences in elemental status between the Dams I and Dams II groups. In newborn calves, the concentration of Ca and Mo corresponded to the reference range; the concentrations of Mg, Fe, Co, and Ni in both groups exceeded the reference range; and the concentrations of Cu, Zn, As, Cr, and Se were lower than the reference range. Results highlighted that there was a tendency to decrease concentration of Fe, Mo, and Se and a significant increase in the Ni concentration in calves of the Newborns II group compared with calves of the Newborns I group. It was also found that Zn, Co, Cr, and Mo actively accumulated in the body of newborn animals while the transplacental transfer of Cu, As, and Sr was limited; and transfer of Se and Ni was regulated by concentration ratios in the blood of the mother and the fetus. The excessive concentrations of Ni and Fe in the blood serum of cows and calves and the imbalance in the ratio of elements Fe–Cu–Zn, Fe–Cu–Co negatively affected erythropoiesis, formation of the immune system, and antioxidant status of the fetus and newborn. These changes were considered to be risk factors for the development of bronchopneumonia in calves. Conclusion: An excess of serum Fe and Ni and deficiency of Cu, Zn, As, Co, and Cr in cows during the gestation period can lead to similar impairments of the mineral status in newborn calves. At the systemic level, dyslementosis in combination with the influence of other adverse factors, can lead to an increased load on the respiratory and hematopoietic systems of calves during postnatal adaptation and can subsequently cause a decrease in the natural resistance of calves and development of bronchopneumonia

Extraarticular manifestations of rheumatoid arthritis in a multiethnic cohort of predominantly Hispanic and Asian patients.

We conducted a study to determine the prevalence of extraarticular manifestations (ExRA) in a cohort of predominantly Hispanic and Asian patients with rheumatoid arthritis (RA), to identify factors associated with the development of ExRA, and to compare the prevalence of ExRA between Hispanic and Asian patients. Patients with RA followed in the outpatient rheumatology clinics of a public hospital were included if they were aged ≥18 years and met the 1987 American College of Rheumatology criteria for the diagnosis of RA. We performed a cross-sectional analysis in which patients with ExRA were identified based on predefined criteria. We compared sociodemographic and clinical characteristics in patients with and without ExRA. Multivariate logistic regression was used to examine the association between sociodemographic variables, clinical characteristics, and the presence of ExRA. The prevalence of ExRA was 21.5%, and the most common manifestations were subcutaneous nodules (17.2%) and interstitial lung disease (3.6%). Hispanic patients were significantly more likely to develop ExRA than Asian patients (odds ratio, 2.53; 95% confidence interval, 1.26-5.09). The development of ExRA was also associated with disease duration, male sex, and seropositivity for serum rheumatoid factor

Validity and Responsiveness of a 10-Item Patient-Reported Measure of Physical Function in a Rheumatoid Arthritis Clinic Population.

ObjectiveWe assessed implementation of the 10-item Patient-Reported Outcomes Measurement Information System (PROMIS) physical function form (PF-10a) in routine practice in a racially and ethnically diverse population with rheumatoid arthritis (RA). Objectives were to determine feasibility of implementing PF-10a in the electronic health record (EHR) and PF-10a validity and longitudinal responsiveness.MethodsClinical and demographic data were abstracted from EHRs for all RA patients seen at a university-based rheumatology clinic between February 2013 and February 2015. We evaluated floor and ceiling (edge) effects and construct validity of PF-10a in a subgroup of patients with Health Assessment Questionnaire (HAQ) scores (n = 189). We used linear mixed-effects models to assess responsiveness of PF-10a to longitudinal changes in the Clinical Disease Activity Index (CDAI) for patients in the entire clinical cohort, with both scores recorded on at least 2 encounters (n = 326).ResultsHalf of the patients were nonwhite, and 15% were non-English speakers. Over a 2-year period, PF10a was successfully implemented; 97% of patients and 89% of encounters had at least 1 measurement performed. PF-10a had fewer ceiling (defined as best) effects than the HAQ (8% versus 22%), and convergent validity was high (r = -0.85). PF-10a was sensitive to expected differences (older versus younger patients, more versus less active disease). Longitudinal changes in PF-10a were highly associated with changes in the CDAI score (P < 0.0001).ConclusionPF-10a was feasible to implement in a diverse RA population. It strongly correlates with the HAQ but has fewer ceiling effects and is responsive to changes in RA disease activity, suggesting its validity for use in routine clinical practice

Hypomethylation of CYP2E1 and DUSP22 Promoters Associated With Disease Activity and Erosive Disease Among Rheumatoid Arthritis Patients.

OBJECTIVE:Epigenetic modifications have previously been associated with rheumatoid arthritis (RA). In this study, we aimed to determine whether differential DNA methylation in peripheral blood cell subpopulations is associated with any of 4 clinical outcomes among RA patients. METHODS:Peripheral blood samples were obtained from 63 patients in the University of California, San Francisco RA cohort (all satisfied the American College of Rheumatology classification criteria; 57 were seropositive for rheumatoid factor and/or anti-cyclic citrullinated protein). Fluorescence-activated cell sorting was used to separate the cells into 4 immune cell subpopulations (CD14+ monocytes, CD19+ B cells, CD4+ naive T cells, and CD4+ memory T cells) per individual, and 229 epigenome-wide DNA methylation profiles were generated using Illumina HumanMethylation450 BeadChips. Differentially methylated positions and regions associated with the Clinical Disease Activity Index score, erosive disease, RA Articular Damage score, Sharp score, medication at time of blood draw, smoking status, and disease duration were identified using robust regression models and empirical Bayes variance estimators. RESULTS:Differential methylation of CpG sites associated with clinical outcomes was observed in all 4 cell types. Hypomethylated regions in the CYP2E1 and DUSP22 gene promoters were associated with active and erosive disease, respectively. Pathway analyses suggested that the biologic mechanisms underlying each clinical outcome are cell type-specific. Evidence of independent effects on DNA methylation from smoking, medication use, and disease duration were also identified. CONCLUSION:Methylation signatures specific to RA clinical outcomes may have utility as biomarkers or predictors of exposure, disease progression, and disease severity

CureGN Study Rationale, Design, and Methods: Establishing a Large Prospective Observational Study of Glomerular Disease

Glomerular diseases, including minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, and immunoglobulin A (IgA) nephropathy, share clinical presentations, yet result from multiple biological mechanisms. Challenges to identifying underlying mechanisms, biomarkers, and new therapies include the rarity of each diagnosis and slow progression, often requiring decades to measure the effectiveness of interventions to prevent end-stage kidney disease (ESKD) or death. Multicenter prospective cohort study. Cure Glomerulonephropathy (CureGN) will enroll 2,400 children and adults with minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, or IgA nephropathy (including IgA vasculitis) and a first diagnostic kidney biopsy within 5 years. Patients with ESKD and those with secondary causes of glomerular disease are excluded. Clinical data, including medical history, medications, family history, and patient-reported outcomes, are obtained, along with a digital archive of kidney biopsy images and blood and urine specimens at study visits aligned with clinical care 1 to 4 times per year. Patients are followed up for changes in estimated glomerular filtration rate, disease activity, ESKD, and death and for nonrenal complications of disease and treatment, including infection, malignancy, cardiovascular, and thromboembolic events. The study design supports multiple longitudinal analyses leveraging the diverse data domains of CureGN and its ancillary program. At 2,400 patients and an average of 2 years’ initial follow-up, CureGN has 80% power to detect an HR of 1.4 to 1.9 for proteinuria remission and a mean difference of 2.1 to 3.0mL/min/1.73m2 in estimated glomerular filtration rate per year. Current follow-up can only detect large differences in ESKD and death outcomes. Study infrastructure will support a broad range of scientific approaches to identify mechanistically distinct subgroups, identify accurate biomarkers of disease activity and progression, delineate disease-specific treatment targets, and inform future therapeutic trials. CureGN is expected to be among the largest prospective studies of children and adults with glomerular disease, with a broad goal to lessen disease burden and improve outcomes