Differences In Disease Phenotype Research Articles

The burden of rheumatologic disease in Aboriginal and Torres Strait Islander Australians.

The objective of this article is to summarise the current knowledge regarding the prevalence of six rheumatological conditions in indigenous Australians - rheumatoid arthritis (RA), osteoarthritis (OA), osteoporosis (OSP), systemic lupus erythematosus (SLE), gout and musculoskeletal (MSK) pain. Online medical literature databases were searched for 'indigenous', 'Aboriginal' and 'Torres Strait Islander', as well as the names of the six conditions. Other included search terms were 'crystal', 'urate', 'arthritis' and 'arthropathy'. No limitations were placed on publication data or language. Forty-five articles examining the prevalence of the six conditions were identified. Based on the published literature, SLE appears to have a higher prevalence, while RA appears to have a lower prevalence in indigenous Australians compared to the non-indigenous community. MSK pain is prevalent, has a significant impact on indigenous people and is perceived as an important area of need. There is a paucity of data regarding these conditions in indigenous Australians. This may be impacted by the uncertainty of case ascertainment by self-report, differences in disease phenotypes and prevalence between the metropolitan compared to the rural or remote indigenous population, and difficulty with access to healthcare. Further studies in conjunction with local indigenous communities are needed to accurately determine the burden of rheumatological disease in the indigenous population. This will assist with resource and workforce planning to deliver culturally appropriate interventions. Strategies for future clinical work and research include the development and dissemination of culturally safe rheumatology resources, rheumatology training of Aboriginal Health Workers and wider integration of rheumatology clinics into community-controlled Aboriginal Health Services.

RUNX1 Germline Mutations of Autosomal-Dominant Familial Platelet Disorder Are More Common Than Expected in Adult Myeloid Leukemias Indicating Highly Variable Penetrance

Germline (GL) RUNX1 (RUNX1GL) mutations result in autosomal-dominant familial platelet disorder (FPD) and are associated with heightened risk of myeloid neoplasia (MN). Indeed about 43% of RUNX1GL carriers are expected to develop MN, by the age of 50 yrs. 1 AML progression can be accompanied by biallelic RUNX1 hits. The phenotypic heterogeneity, incomplete penetrance, and leukemogenic risk depend on mutational burden and functional impact (topology of mutations, configuration) of RUNX1 and thus evolution of MN may be delayed. 2-3 We hypothesized that a large systematic study of RUNX1GL including types and topology will elucidate the impact of various RUNX1mutations ( RUNX1MT), their configuration, and molecular associations to provide prognostic insights and mechanistic conclusions as to the pathogenesis of these mutations. We analyzed over 9000 patients from Cleveland Clinic Foundation (CCF; n=7953) and public series. 4-5 We devised a novel rational algorithm to assess of RUNX1GL variants ( Fig1A). Stringent criteria were used that prioritized exclusion of false positives: GL benign and likely benign alterations were not included, while pathogenic and likely pathogenic (P/LP) variants were further curated. Carriers of VUS RUNX1MT that displayed past medical history or family history suspicious for FPD or leukemia were classified as VUS of suspicious pathogenicity (sVUS). The remaining variants, including P/LP, were further filtered and clinically annotated ( Fig1B). GL confirmation was possible in 22 alterations, e.g., wherein our algorithm resulted in ambivalent assignment. We found 627 RUNX1 variants (GL and somatic) in 557 patients, of which 8 were sVUS and 106 (17%) were found to be GL P/LP including 3 new RUNX1GL alterations (Y281*, S141L, S410*). The mean age at presentation of RUNX1GL carriers was lower (64 yrs.) compared to patients with somatic RUNX1MT(70 yrs., P= .0053). There was no difference in disease phenotype (AML, MDS, MDS/MPN) between GL, somatic, and biallelic (somatic/somatic) RUNX1MT carriers. Of note, 1 patient with predicted RUNX1GL variants had aplastic anemia. Cytogenetic analysis revealed that 68% of RUNX1GL cases were associated with normal karyotype. Four RUNX1GL cases also displayed trisomy 21. FLT3 ITD/PTDwas more prevalent in RUNX1GL carriers (14.8%) than in those with somatic RUNX1MT(4.7%) ( P= .021). PHF6 MT were more frequent in carriers of RUNX1GL (18%) compared to somatic RUNX1 carriers (9%) ( P= .078). Patients with a second RUNX1 hit included somatic biallelic (n=52)and GL/somatic (n=6) RUNX1 MT. The most common mutations co-occuring with biallelic RUNX1 hit werethose in EZH2 (27%, P= .025), ZRSR2 (19%, P= .020), U2AF1 (21%, P= .030) and PTPN11 (16%, P= .013) compared to monoallelic RUNX1MT. We further categorized RUNX1MT according to their predicted functional effects as: i) haploinsufficient (RUNT-truncating mutations); ii) dominant negative (post-RUNT nonsense, C-terminal frameshift, and missense mutations previously described as dominant negative); and iii) RUNT missense mutations. 6-7 We analyzed 93 (25%) mutations classified as haploinsufficient, 98 (27%) as dominant negative, and 134 (36%) as RUNT missense. There was no difference in the topology between somatic and GL hits. Survival analysis indicated that there was no difference in prognosis between dominant negative, haploinsufficient, and RUNT missense mutations ( P= .59). Among RUNX1GL carriers, there was also no prognostic difference between subtypes of RUNX1GL ( P= .896). To determine whether GL mutation types convey a greater penetrance, we compared the age of patients at presentation for somatic and GL. Specifically, carries of GL dominant negative mutations were diagnosed at a younger median age (60.5 yrs.) compared to carriers of GL haploinsufficient (68 yrs.) and RUNT missense (69 yrs.). However, this trend was not statistically significant. To rule out the effect of RUNX1GL lesions on genome instability, we quantified the mutational burden per each GL variant. Compared to RUNT missense, haploinsufficient GL variants were associated with a greater number of co-occurring mutations ( P= .013) while dominant negative GL variants were not ( P= .054). In summary, our study of the topological features of RUNX1GL variants showed that dominant negative RUNX1 mutations are frequent in MN and are not associated with specific phenotypes, but rather define a variable penetrance.

Clinical Practice Patterns in IgA Nephropathy: A Global Questionnaire-Based Survey

IgA nephropathy (IgAN) displays ethnic differences in disease phenotype. We aimed to examine how this common disease is managed worldwide. An online 2-step questionnaire-based survey was conducted among nephrologists globally focusing on various management strategies used in IgAN. A total of 422 nephrologists responded to the initial survey and 339 to the follow-up survey. Of the nephrologists, 13.7% do not get MEST-C scores in biopsy reports; 97.2% of nephrologists use renin-angiotensin-aldosterone system (RAAS) blockade with angiotensin-converting-enzyme inhibitors (ACEi) / angiotensin receptor blockers (ARB) as initial treatment. Other supportive treatments commonly employed are fish oil (43.6%) and sodium-glucose co-transporter-2 (SGLT2) inhibitors (48.6%) with regional differences. Immunosuppression is generally (92.4%) initiated when proteinuria >1 g/d persists for≥3 months.Main considerations for initiating immunosuppression are level of proteinuria (87.9%), estimated glomerular filtration rate (eGFR) decline (78.7%), lack of response to RAAS blockade (57.6%) and MEST-C score (64.9%). Corticosteroids (89.1%) are universally used as first-line immunosuppression; mycophenolate mofetil is commonly used in resistant patients (49.3%). Only 30.4% nephrologist enroll patients with persistent proteinuria >1 g/d in clinical trials. Nephrologists in Europe (63.6%), North America (56.5%), and Australia (63.6%) are more likely to do so compared to South America (31.3%) and Asia (17.2%). Only 8.1% nephrologists in lower-middle income countries (LMICs) enroll patients in clinical trials, though 40% of them are aware of such trials in their nations. Although most nephrologists agree on common parameters to assess clinical severity of IgAN, use of RAAS blockade, and blood pressure control, there is heterogeneity in use of other supportive therapies and initiation of immunosuppression. There is reluctance to enroll patients in clinical trials with novel treatments, principally in LMICs.

Comparison of Disease Phenotype and Course among Elderly- and Early-Onset Inflammatory Bowel Diseases in the Middle East.

It is unknown if the clinical manifestations and phenotype of disease are comparable between early- and elderly-onset inflammatory bowel disease (IBD). We aimed to seek differences in disease phenotype, course, complications, and treatment between early- and elderly-onset IBD patients. This retrospective cohort study on registered IBD patients in the Iranian Registry of Crohn's and Colitis (IRCC) compared demographics, disease phenotype, disease activity, IBD-related surgery and medications between early- and elderly-onset IBD. A generalized linear regression model was used to investigate the relative risk of age at diagnosis adjusted for gender and disease duration for the outcomes. From 10048 IBD patients, 749 with early-onset (7.5%), and 472 (4.7%) elderly-onset IBD were enrolled: 855 (63.1%) ulcerative colitis (UC) and 366 (26.9%) Crohn's disease (CD). Left-sided colitis was more frequent among elderly-onset UC patients (P<0.001). Ileum and ileocolonic locations were the most common types in elderly-onset and early-onset CD patients, respectively. In comparison with elderly-onset UC, early-onset cases more often used prednisolone (22.1% vs. 11.4%, P=0.001), immunomodulators (44.9% vs 25.2%, P<0.001) and anti-tumor necrosis factors (TNF) (20.1% vs 11.9%, P=0.002). Elderly-onset UC patients had 0.7 times lower risk of aggressive phenotype (95%CI:0.6‒0.9, P=0.005). Early-onset CD was associated with higher use of prednisolone (27.7% vs 8.1%, P<0.001), immunomodulators (58.7% vs 41.8%, P=0.005) and anti-TNF (49.6% vs 35.4%, P=0.006). Early-onset IBD was associated with a more aggressive phenotype and higher prednisolone, immunomodulators, and anti-TNF use.

Exploring molecular markers and drug candidates for colorectal cancer through comprehensive bioinformatics analysis.

Colorectal cancer (CRC) often has a poor prognosis and identifying useful and novel agents for treating CRC is urgently required. This study aimed to examine molecular markers associated with CRC prognosis and to identify potential drug candidates. The differentially expressed genes (DEGs) of CRC in TCGA were identified. The genes associated with CRC, summarized from NCBI-gene, OMIM, and the DEGs, were used to construct a co-expression network by WGCNA. Moreover, the co-expression genes from modules of interest were used to carry out functional enrichment. A total of 2742 DEGs, including 1674 upregulated and 1068 downregulated genes, were identified. Thirteen co-expression modules were constructed with WGCNA. Brown and blue co-expression modules with significant differences in disease phenotype were found. Functional enrichment analysis showed that genes in the brown module were mainly related to cell cycle, cell proliferation, DNA replication, and RNA transport. The genes in the blue module were mainly associated with fatty acid degradation, sulfur metabolism, PPAR signaling pathway and bile secretion. In addition, both the genes in brown and blue were associated with tumor staging. Some prognostic markers and candidate small molecules drugs for CRC treatment were identified. In conclusion, we revealed molecular biomarker profiles in CRC by systematic bioinformatics analysis, constructed regulatory networks of mRNA, ncRNA and transcriptional regulators (TFs), and identified potential drugs targeting hub proteins and TFs.

Clinical and autoantibody profiles of systemic sclerosis patients: A cross-sectional study from North India.

Objectives This cross-sectional study was designed to assess the clinical profile and frequency of associated autoantibodies in all consecutive patients classified as systemic sclerosis (SSc) at Medanta-the Medicity Hospital, Gurgaon, India. Methods Between August 2017 and July 2019, we identified a total of 119 consecutive patients meeting the American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) 2013 criteria for SSc and 106 patients consented to this study. Their clinical and serological data at the time of enrolment were analysed. Results Our cohort had a mean age at symptom onset of 40 ± 13 years with a median symptom duration of 6 years. We had 76 patients (71.7%) with interstitial lung disease (ILD), which was a higher proportion compared to European cohorts. 62 patients (58.5%) had diffuse cutaneous involvement which was significantly associated with anti-Scl70 antibodies (p < 0.001), digital ulcers (p = 0.039) and the presence of ILD (p = 0.004). 65 patients (61.3%) had anti-Scl70 and 15 patients (14.2%) had anti-centromere (anti-CENP) antibodies. Scl70 positivity was associated with the presence of ILD (p < 0.001) and digital ulcers (p = 0.01). Centromere antibodies had a negative association with ILD (p < 0.001), but was a risk factor for calcinosis (p < 0.001) and pulmonary arterial hypertension (PAH) (p = 0.01). The combination of diffuse cutaneous disease and Scl70 antibodies was the strongest predictor of ILD and digital ulcers (p = 0.015). sm/RMP, RNP68 and Ku antibodies correlated with musculoskeletal involvement (p < 0.01), while all seven of the patients with Pm/Scl antibodies had ILD. Renal involvement was noted in only two patients. Limitations A single-centre study may not capture the true prevalence of disease characteristics in the population. Referral bias for patients with diffuse cutaneous disease has been noted. Data on RNA-Polymerase antibodies have not been provided. Conclusion North Indian patients have some characteristic differences in disease phenotype as compared to their Caucasian counterparts with a larger proportion of patients presenting with ILD and Scl70 antibodies. Antibodies against Ku, RNP and Pm/Scl occur in a minority of patients, but may be associated with musculoskeletal features.

Prognostic value of combined coronary CT angiography and myocardial perfusion imaging in women and men

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): State Research Funding for Turku University Hospital. Background Combined anatomical and functional imaging enables detection of non-obstructive and obstructive coronary artery disease (CAD) as well as myocardial ischemia, and also provides prognostic information. Purpose We evaluated sex differences in disease phenotype and adverse outcomes by using non-invasive combined anatomical and functional imaging in symptomatic patients with suspected CAD. Methods We retrospectively evaluated patients undergone coronary computed tomography angiography (CTA) for suspected CAD. According to local routine, patients with suspected obstructive stenosis on CTA were referred to downstream 15O-water positron emission tomography (PET) myocardial perfusion imaging to assess stress myocardial blood flow (MBF; ≤2.3 mL/g/min considered abnormal). A composite adverse endpoint was recorded, including all-cause death, myocardial infarction, and unstable angina pectoris. Results A total of 1948 patients (59% women) underwent coronary CTA of whom 657 (34%) patients underwent downstream PET perfusion imaging. During a mean follow-up of 6.8 years, 182 adverse events occurred. Women more often had normal coronary arteries (42% vs. 22%, p&amp;lt;0.001) and less often abnormal stress MBF (9% vs. 28%, p&amp;lt;0.001), as compared with men. The annual adverse event rate was lower in women versus men (1.2% vs. 1.7%, p = 0.02). Both in women and men, coronary calcification, non-obstructive CAD, and abnormal stress MBF were independent predictors of events. Abnormal stress MBF was associated with 5.0 and 5.6-fold adverse event rates in women and men, respectively. There was no statistical interaction between sex and coronary calcification, non-obstructive CAD, or abnormal stress MBF in terms of predicting adverse outcome. Conclusion Among patients evaluated for chronic chest pain, women have lower prevalence of ischemic CAD and lower rate of future adverse events. Combined coronary CTA and PET myocardial perfusion imaging predicts outcomes equally in women and men.

Antimyeloperoxidase antibodies modulate inflammatory responses and activate profibrotic pathways in human monocytes

Antimyeloperoxidase (anti-MPO) and antiproteinase 3 (anti-PR3) antibodies are found in anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV). We investigated the effect of both anti-MPO and anti-PR3 IgG on human monocytes. Peripheral blood monocytes were cultured under a range of conditions that included TLR agonists, anti-MPO IgG and anti-PR3 IgG with appropriate controls. Experiments included whole transcriptome profiling and an assessment of the role of Fc receptors.When monocytes were stimulated with LPS or R848, anti-MPO but not anti-PR3 IgG, caused a reduction in IL-10 secretion and had a profound effect on cell-surface marker expression. Anti-MPO but not anti-PR3 IgG enhanced monocyte survival in the absence of TLR stimulation. These effects depended on the Fc receptor CD32a. With TLR stimulation, the effect of anti-MPO but not anti-PR3 IgG on the transcriptional response at 6 h was variable, but we identified a core set of transcripts likely to be important. Without TLR stimulation, there was a robust effect of anti-MPO but not anti-PR3 IgG on the transcriptional response at 24 h, and there was a highly significant enrichment of genes encoding extracellular matrix and extracellular matrix-associated proteins. Analysis with nCounter confirmed many of the differentially expressed transcripts and supported a role for CD32a. These data show that anti-MPO, but not anti-PR3 IgG, from patients with AAV has wide-ranging effects on monocytes which depend on CD32a. The activation of a profibrotic transcriptional response by anti-MPO but not anti-PR3 IgG may give insights into the differences in disease phenotype.

Nlrp12 deficiency alters gut microbiota and ameliorates Faslpr -mediated systemic autoimmunity in male mice.

NLRP12 has dual roles in shaping inflammation. We hypothesized that NLRP12 would modulate myeloid cells and T cell function to control systemic autoimmunity. Contrary to our hypothesis, the deficiency of Nlrp12 in autoimmune-prone B6.Faslpr/lpr mice ameliorated autoimmunity in males but not females. Nlrp12 deficiency dampened B cell terminal differentiation, germinal center reaction, and survival of autoreactive B cells leading to decreased production of autoantibodies and reduced renal deposition of IgG and complement C3. In parallel, Nlrp12 deficiency reduced the expansion of potentially pathogenic T cells, including double-negative T cells and T follicular helper cells. Furthermore, reduced pro-inflammatory innate immunity was observed, where the gene deletion decreased in-vivo expansion of splenic macrophages and mitigated ex-vivo responses of bone marrow-derived macrophages and dendritic cells to LPS stimulation. Interestingly, Nlrp12 deficiency altered the diversity and composition of fecal microbiota in both male and female B6/lpr mice. Notably, however, Nlrp12 deficiency significantly modulated small intestinal microbiota only in male mice, suggesting that the sex differences in disease phenotype might be gut microbiota-dependent. Together, these results suggest a potential pathogenic role of NLRP12 in promoting systemic autoimmunity in males. Future studies will investigate sex-based mechanisms through which NLRP12 differentially modulates autoimmune outcomes.

Clinical and metabolic characteristics of males with early-onset androgenetic alopecia.

Background Men with early-onset androgenetic alopecia (AGA) often have an abnormal hormonal milieu. Objective To ascertain the clinico-phenotypic characteristics and the prevalence of hormonal and metabolic changes in men with early-onset AGA. Methods Consecutive male patients less than 30 years of age with a Norwood-Hamilton grade ≥3 AGA were recruited in this comparative cross-sectional study. After endocrine evaluation they were classified into two groups, that is, Group A consisting of subjects with an altered hormonal profile and Group B with normal hormonal profiles. The groups were assessed for differences in disease phenotype and severity (Norwood-Hamilton grade), insulin resistance and parameters of metabolic syndrome (ATP III guidelines). Results Altered hormonal profiles were seen in 34 of the 100 subjects with AGA, while insulin resistance and metabolic syndrome were noted in 44 and 26 respectively. Altered hormonal profiles were significantly associated with insulin resistance and severe alopecia (grade 4 and above Hamilton-Norwood Scale). Insulin resistant Group A patients had a significantly higher prevalence of severe alopecia (>grade 4) (P = 0.0036). The prevalence of metabolic syndrome was similar in both groups. Limitation The cross sectional study design was a drawback of this study. Further, a control arm without AGA was not included and the sample size of 100 was selected arbitrarily. Conclusion An altered hormonal profile and insulin resistance was noted in a third of the males with early-onset AGA. Subjects with altered hormonal profiles had a higher prevalence of insulin resistance and were likely to have severe grades of AGA.

S132 Disparity of Care Is Not Observed Across Racial and Ethnic Groups During Hospitalization for Inflammatory Bowel Disease

Background: Although Inflammatory Bowel Disease (IBD) is more common in non-Hispanic White populations, rates are increasing among non-Whites. Despite some possible differences in disease phenotypes between racial groups, disease management guidelines are identical for different groups. However, multiple studies have demonstrated that social determinants of health (SDOH) play a critical role in disease management and outcomes for patients with IBD, as patients from minority populations often have delayed diagnosis, increased surgeries, and post-surgical complications. Several factors likely contribute to these healthcare disparities, with financial compensation and access to healthcare among them. Hospital admission ideally provides a setting that removes these barriers, and therefore should not be associated with healthcare disparities, though this has not been previously studied. We sought to identify if SDOH guide inpatient management of patients with IBD. Methods: We conducted a retrospective cohort study of adult patients with IBD-related inpatient admissions to any of Northwell Health’s 21 hospitals between 2010-2018. Patients were identified by a hospital admission primary diagnosis of ulcerative colitis (UC) or Crohns disease (CD), or a secondary diagnosis of UC or CD with a primary diagnosis of diarrhea, abdominal pain, or gastrointestinal bleeding. We analyzed data on patient demographics, medical history, and inpatient management features, including the use of imaging, medications, and procedures. Biological medication was defined as the use of infliximab, adalimumab, certolizumab. Race/ethnicity was characterized as Hispanic, Asian, Black, White, other, and unknown. Insurance status was characterized as private, Medicare, or Medicaid. Self-pay patients were excluded from analysis. Variables were tested using logistic regression for binary outcomes. To control the overall Type 1 error rate for the hypothesis that there were racial disparities in inpatient management of IBD, the Holm-Bonferroni method was used to adjust the P-value thresholds for statistical significance in each of the multiple tests performed to evaluate the hypothesis. Results: In total, 1,732 patients were identified and included in the analysis. Overall, 57% (n = 985) were White, 9% Hispanic, 5% Asian, 13% Black, 6% other, 9% unknown. 55% of patients were female, 45% had Crohn’s Disease, 14% were on steroids prior to admission. 47% of patients had private insurance, with the remaining having either Medicare or Medicaid. When assessing hospital interventions and outcomes based on racial/ethnic group, there was no difference in length of stay, use of medications (corticosteroids, aminosalicylic acid formulations, opioids, biological) during hospital admission and on discharge, stool testing performed as inpatient, imaging (CT, MRI) or endoscopies performed. Conclusion(s): SDOH play a critical role in the disease course and outcomes for patients with IBD. In this large retrospective study, we found that among patients hospitalized for IBD exacerbations there was no disparity of care across racial/ethnic groups. Our findings suggest that factors such as insurance status and access to healthcare are likely the main contributors to the healthcare disparities seen in the outpatient setting. Our results reflect the experience in a large urban health care system, and further studies are needed to replicate the data in other settings.

Identification of quantitative trait loci for survival in the mutant dynactin p150Glued mouse model of motor neuron disease.

Amyotrophic lateral sclerosis (ALS) is the most common degenerative motor neuron disorder. Although most cases of ALS are sporadic, 5–10% of cases are familial, with mutations associated with over 40 genes. There is variation of ALS symptoms within families carrying the same mutation; the disease may develop in one sibling and not in another despite the presence of the mutation in both. Although the cause of this phenotypic variation is unknown, it is likely related to genetic modifiers of disease expression. The identification of ALS causing genes has led to the development of transgenic mouse models of motor neuron disease. Similar to families with familial ALS, there are background-dependent differences in disease phenotype in transgenic mouse models of ALS suggesting that, as in human ALS, differences in phenotype may be ascribed to genetic modifiers. These genetic modifiers may not cause ALS rather their expression either exacerbates or ameliorates the effect of the mutant ALS causing genes. We have reported that in both the G93A-hSOD1 and G59S-hDCTN1 mouse models, SJL mice demonstrated a more severe phenotype than C57BL6 mice. From reciprocal intercrosses between G93A-hSOD1 transgenic mice on SJL and C57BL6 strains, we identified a major quantitative trait locus (QTL) on mouse chromosome 17 that results in a significant shift in lifespan. In this study we generated reciprocal intercrosses between transgenic G59S-hDCTN1 mice on SJL and C57BL6 strains and identified survival QTLs on mouse chromosomes 17 and 18. The chromosome 17 survival QTL on G93A-hSOD1 and G59S-hDCTN1 mice partly overlap, suggesting that the genetic modifiers located in this region may be shared by these two ALS models despite the fact that motor neuron degeneration is caused by mutations in different proteins. The overlapping region contains eighty-seven genes with non-synonymous variations predicted to be deleterious and/or damaging. Two genes in this segment, NOTCH3 and Safb/SAFB1, have been associated with motor neuron disease. The identification of genetic modifiers of motor neuron disease, especially those modifiers that are shared by SOD1 and dynactin-1 transgenic mice, may result in the identification of novel targets for therapies that can alter the course of this devastating illness.

Time‐of‐Day Influence on Central Nervous System Autoimmunity

Circadian rhythms are a characteristic of most living organisms and describe biological processes experiencing an oscillation of about 24h. These rhythms regulate multiple physiological systems, including innate and adaptive immune responses. Circadian disruption is an important risk factor in neuroinflammation such as multiple sclerosis (MS), a T‐cell mediated autoimmune disease affecting around 2.8 million people worldwide. In the central nervous system (CNS), autoimmune T cells attack oligodendrocytes leading to progressive demyelination and neurological impairment. Studies performed by our lab demonstrate that the circadian clock plays an important role in the severity of experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis. However, the mechanisms responsible for this time‐of‐day difference are unknown. We investigated time‐of‐day differences in adhesion molecule expression and immune cell infiltration to the CNS that could explain differences in disease phenotype. Our results indicate a time‐of‐day‐dependent infiltration of cells into spinal cords during EAE development that correlates with expression of the adhesion molecules VCAM‐1 and ICAM‐1 in the CNS vasculature. Targeting leukocyte‐endothelial cell interactions in a time‐of‐day dependent manner with an anti‐VLA4 antibody that blocks VLA‐4/VCAM‐1 engagement reduces leukocyte infiltration and disease severity. This study shows the importance of circadian oscillations in adhesion molecule expression and cell infiltration in the CNS, and identifies an axis to target for chronotherapy purposes in MS patients.

Multi-tissue RNAseq reveals genetic and temporal differences in acute response to viral (IHNV) infection among three selected lines of rainbow trout with varying resistance

Utilizing RNA-seq, this study compared the transcriptomic responses of three improved strains (VSel, PSel, and CSel) of rainbow trout fry during acute stages of challenge with infectious hematopoietic necrosis virus (IHNV). The VSel strain has been selected for resistance against the specific strain of IHNV used in our challenge, PSel has undergone selection for utilization of plant-protein based feeds and previously has shown elevated non-specific disease resistance despite no disease related selection pressures, and the final strain, CSel, is a commercial strain that has been domesticated for several years but has not been selected for specific viral disease resistance. Following a 21-day IHNV challenge, Kaplan-Meier survival estimator curves and cumulative percent mortality (CPM) showed significant differences in IHNV resistance across strains: VSel - 19.3 ± 5.0%, PSel - 67. ± 3.03%, CSel - 94.6 ± 4.1% CPM. To evaluate acute responses to IHNV infection, whole blood, as well as samples from the kidney, liver, and intestine, were collected at 0, 4, 12, 24, and 48 h post infection (hpi). Serum lysozyme activity, a marker of non-specific innate immunity, showed strain and temporal effects during the acute infection phase with PSel showing the highest activity at 0 and 48 hpi. Differential gene expression responses were detected, with varying degrees, in all tissues, both between strains, as well as across acute timepoints within strains. The VSel strain showed upregulation for a particular subset of viral recognition genes during early infection timepoints and rather limited upregulation of immune genes later, while maintaining and reactivating metabolic pathways. The CSel strain showed a downregulation of metabolic related genes and a limited upregulation of immune genes, while the PSel strain showed similar downregulation of metabolic genes during acute infection, yet when compared to the CSel strain, showed a more robust innate immune response. Evaluation of upregulated immune response genes, as well as interferon-related genes showed the PSel strain to have the greatest number of uniquely upregulated immune genes in both the kidney and intestine, with CSel and PSel showing a similar number of such genes upregulated in liver. A moderate number of immune response genes were shared between PSel and CSel in all tissues, though both PSel and VSel showed a high number of uniquely overexpressed immune response genes in the kidney, and PSel showed the highest number of uniquely upregulated interferon related genes in the intestine. Overall, the VSel response was unique from the CSel with very little overlap in activated immune responses. Findings from this study highlight the disparity in IHNV resistance among genetic strains of rainbow trout, while identifying molecular mechanisms underlying differences in disease phenotypes. Furthermore, our results on trout strains with distinct selection backgrounds yields comparative insights into the adaptive gains brought about by selection programs for pathogen-specific disease resistance, as well as the non-specific immune enhancement associated with selection for utilization of plant-based diets.

Differences in MS clinical and epidemiological characteristics between Ashkenazi and non-Ashkenazi Jewish patients in Israel: a retrospective single center study

The prevalence and severity of Multiple Sclerosis (MS) varies across different ethnicities, with a tendency to a more severe phenotype in non-Caucasian populations. Our objective was to evaluate the differences in disease phenotype between Ashkenazi Jewish and Non-Ashkenazi Jewish patients in Israel. We conducted a single center retrospective cohort study in which subjects were assigned to Ashkenazi or Non-Ashkenazi groups according to self-reported ancestry and disease severity was assessed using the expanded disability status (EDSS), MS severity score (MSSS), progression index (PI) and MRI metrics. 330 Ashkenazi Jewish (AJ) and 207 Non-Ashkenazi Jewish patients (Non-AJ) were included. Non-AJ had a younger age of disease onset (32.7 years vs. 35.7 years, p = 0.05), with a lower proportion of females (62.3% vs. 73.3%, p = 0.01). These differences were maintained within the subgroup of Israeli native patients. Ethnicity was a significant predictor of MSSS (β = 0.601, p = 0.003), with a higher estimate than that of other epidemiological factors. To conclude, Non-AJ patients had an earlier age of onset and a more disabling disease as well as having a more balanced female to male ratio compared to AJ patients. These findings demonstrate variability of disease phenotype within Caucasian patient's dependent on their ethnicity despite equivalent access to healthcare services.

COMPARISON OF THE INFLAMMATORY BOWEL DISEASES PHENOTYPE AND COURSE IN THE ELDERLY- AND EARLY-ONSET PATIENTS

Abstract BACKGROUND AND AIMS It is unknown if clinical phenotype and course of the disease are comparable between early- and elderly-onset inflammatory bowel diseases (IBD). There is also an ongoing dispute on whether elderly-onset IBD is a separate entity. We aimed to seek differences in disease phenotype, course, complications, and treatment between early- and elderly-onset IBD patients. METHODS This retrospective cohort study on registered IBD patients in the Iranian Registry of Crohn’s and Colitis (IRCC) compared demographics, extent of ulcerative colitis (UC), Crohn’s disease (CD) location and behavior, extraintestinal manifestations, disease activity during the previous six months, and two weeks, IBD-related surgery and medications between early- and elderly-onset IBD. We defined early-onset and elderly-onset IBD as 30 years or less and 60 years and above, respectively. We used univariable analysis to investigate the association between age of diagnosis and outcomes. Variables with P-value &amp;lt;0.1 were entered into the generalized linear model to study the relative risk of age of diagnosis adjusted for gender and disease duration for the outcomes. RESULTS From 8911 IBD patients that were recruited in the IRCC from December 2017 till June 2021, 4503 were enrolled, including 4134 (46.4%) early-onset and 369 (4.1%) elderly-onset cases. Among them, 3596 (79.8%) had UC, and 907 (20.2%) had CD. Figure 1 shows relative risk of outcomes in patients with UC based on patients’ age of diagnosis adjusted for disease duration and gender. The risk of left colitis was 1.7 times higher among elderly-onset UC patients (95%CI:1.4-2.1, P&amp;lt;0.001). Early-onset IBD cases had a higher prevalence of extraintestinal manifestations (p&amp;lt;0.001). Early-onset UC cases more often used prednisolone (p=0.031), immunomodulators (p&amp;lt;0.001) and anti-tumor necrosis factors (TNF) (p=0.002). Elderly-onset UC patients had 0.7 times lower risk of active disease in the past six months in comparison with elderly-onset cases (95%CI:0.5-0.9, p=0.003). Elderly-onset CD patients had 1.7 times the risk of ilium involvement (95%CI: 1.1-2.5, p=0.010), and elderly-onset CD cases had 0.2 times the risk of stricture formation compared to early cases (95%CI:0.1-0.8, p=0.028). Early-onset CD was associated with higher use of prednisolone (p=0.024), immunomodulators (p&amp;lt;0.001) and anti-TNF (p=0.024). Disease activity during the previous six months , two weeks and IBD-related surgery were similar between the two groups. Figure 2 demonstrates relative risk of outcomes in patients with CD based on patients’ age of diagnosis adjusted for disease duration and gender. CONCLUSION Early-onset IBD was associated with a more aggressive phenotype and higher prednisolone, immunomodulators, and anti-TNF use. There is a need for a more detailed investigation of treatment complications and surgery types in future studies.

COMPARISON OF THE INFLAMMATORY BOWEL DISEASES PHENOTYPE AND COURSE IN THE ELDERLY- AND EARLY-ONSET PATIENTS

It is unknown if clinical phenotype and course of the disease are comparable between early- and elderly-onset inflammatory bowel diseases (IBD). There is also an ongoing dispute on whether elderly-onset IBD is a separate entity. We aimed to seek differences in disease phenotype, course, complications, and treatment between early- and elderly-onset IBD patients.

Studying the Function of Phytoplasma Effector Proteins Using a Chemical-Inducible Expression System in Transgenic Plants.

Phytoplasmas are bacterial pathogens that live mainly in the phloem of their plant hosts. They dramatically manipulate plant development by secreting effector proteins that target developmental proteins of their hosts. Traditionally, the effects of individual effector proteins have been studied by ectopic overexpression using strong, ubiquitously active promoters in transgenic model plants. However, the impact of phytoplasma infection on the host plants depends on the intensity and timing of infection with respect to the developmental stage of the host. To facilitate investigations addressing the timing of effector protein activity, we have established chemical-inducible expression systems for the three most well-characterized phytoplasma effector proteins, SECRETED ASTER YELLOWS WITCHES’ BROOM PROTEIN 11 (SAP11), SAP54 and TENGU in transgenic Arabidopsis thaliana. We induced gene expression either continuously, or at germination stage, seedling stage, or flowering stage. mRNA expression was determined by quantitative reverse transcription PCR, protein accumulation by confocal laser scanning microscopy of GFP fusion proteins. Our data reveal tight regulation of effector gene expression and strong upregulation after induction. Phenotypic analyses showed differences in disease phenotypes depending on the timing of induction. Comparative phenotype analysis revealed so far unreported similarities in disease phenotypes, with all three effector proteins interfering with flower development and shoot branching, indicating a surprising functional redundancy of SAP54, SAP11 and TENGU. However, subtle but mechanistically important differences were also observed, especially affecting the branching pattern of the plants.

Validity of self-reported endometriosis: a comparison across four cohorts.

How accurately do women report a diagnosis of endometriosis on self-administered questionnaires? Based on the analysis of four international cohorts, women self-report endometriosis fairly accurately with a > 70% confirmation for clinical and surgical records. The study of complex diseases requires large, diverse population-based samples, and endometriosis is no exception. Due to the difficulty of obtaining medical records for a condition that may have been diagnosed years earlier and for which there is no standardized documentation, reliance on self-report is necessary. Only a few studies have assessed the validity of self-reported endometriosis compared with medical records, with the observed confirmation ranging from 32% to 89%. We compared questionnaire-reported endometriosis with medical record notation among participants from the Black Women's Health Study (BWHS; 1995-2013), Etude Epidémiologique auprès de femmes de la Mutuelle Générale de l'Education Nationale (E3N; 1990-2006), Growing Up Today Study (GUTS; 2005-2016), and Nurses' Health Study II (NHSII; 1989-1993 first wave, 1995-2007 second wave). Participants who had reported endometriosis on self-administered questionnaires gave permission to procure and review their clinical, surgical, and pathology medical records, yielding records for 827 women: 225 (BWHS), 168 (E3N), 85 (GUTS), 132 (NHSII first wave), and 217 (NHSII second wave). We abstracted diagnosis confirmation as well as American Fertility Society (AFS) or revised American Society of Reproductive Medicine (rASRM) stage and visualized macro-presentation (e.g. superficial peritoneal, deep endometriosis, endometrioma). For each cohort, we calculated clinical reference to endometriosis, and surgical- and pathologic-confirmation proportions. Confirmation was high-84% overall when combining clinical, surgical, and pathology records (ranging from 72% for BWHS to 95% for GUTS), suggesting that women accurately report if they are told by a physician that they have endometriosis. Among women with self-reported laparoscopic confirmation of their endometriosis diagnosis, confirmation of medical records was extremely high (97% overall, ranging from 95% for NHSII second wave to 100% for NHSII first wave). Importantly, only 42% of medical records included pathology reports, among which histologic confirmation ranged from 76% (GUTS) to 100% (NHSII first wave). Documentation of visualized endometriosis presentation was often absent, and details recorded were inconsistent. AFS or rASRM stage was documented in 44% of NHSII first wave, 13% of NHSII second wave, and 24% of GUTS surgical records. The presence/absence of deep endometriosis was rarely noted in the medical records. Medical record abstraction was conducted separately by cohort-specific investigators, potentially introducing misclassification due to variation in abstraction protocols and interpretation. Additionally, information on the presence/absence of AFS/rASRM stage, deep endometriosis, and histologic findings were not available for all four cohort studies. Variation in access to care and differences in disease phenotypes and risk factor distributions among patients with endometriosis necessitates the use of large, diverse population samples to subdivide patients for risk factor, treatment response and discovery of long-term outcomes. Women self-report endometriosis with reasonable accuracy (>70%) and with exceptional accuracy when women are restricted to those who report that their endometriosis had been confirmed by laparoscopic surgery (>94%). Thus, relying on self-reported endometriosis in order to use larger sample sizes of patients with endometriosis appears to be valid, particularly when self-report of laparoscopic confirmation is used as the case definition. However, the paucity of data on histologic findings, AFS/rASRM stage, and endometriosis phenotypic characteristics suggests that a universal requirement for harmonized clinical and surgical data documentation is needed if we hope to obtain the relevant details for subgrouping patients with endometriosis. This project was supported by Eunice Kennedy Shriver National Institute of Child Health and Development grants HD48544, HD52473, HD57210, and HD94842, National Cancer Institute grants CA50385, R01CA058420, UM1CA164974, and U01CA176726, and National Heart, Lung, and Blood Institute grant U01HL154386. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. AS, SM, and KT were additionally supported by the J. Willard and Alice S. Marriott Foundation. MK was supported by a Marie Curie International Outgoing Fellowship within the 7th European Community Framework Programme (#PIOF-GA-2011-302078) and is grateful to the Philippe Foundation and the Bettencourt-Schueller Foundation for their financial support. Funders had no role in the study design, conduct of the study or data analysis, writing of the report, or decision to submit the article for publication. LA Wise has served as a fibroid consultant for AbbVie, Inc for the last three years and has received in-kind donations (e.g. home pregnancy tests) from Swiss Precision Diagnostics, Sandstone Diagnostics, Kindara.com, and FertilityFriend.com for the PRESTO cohort. SA Missmer serves as an advisory board member for AbbVie and a single working group service for Roche; neither are related to this study. No other authors have a conflict of interest to report. Funders had no role in the study design, conduct of the study or data analysis, writing of the report, or decision to submit the article for publication. N/A.

Developing an Online Portal for Determining the Genomic Signature of Archaic DNA that are Associated to Modern Human Genetic Diseases: A Meta-Analysis Study.

Mutations or introgression can cause and rise adaptive alleles of which some can be beneficial. Archaic humans lived more than 200,000 years ago in Europe and Western Asia. They were adapted to the environment and pathogens that prevailed in these locations. It can therefore be thought that modern humans obtained significant immune advantage from the archaic alleles. First, data were collected by meta-analysis from previously identified genetic diseases caused by alleles that were introgressed from archaics. Second, the in silico model portal (http://www.archaics2phenotype.xxx.edu.tr) was designed to trace the history of the Neanderthal allele. The portal also shows the current distribution of the genotypes of the selected alleles within different populations and correlates with the individuals phenotype. Our developed model provides a better understanding for the origin of genetic diseases or traits that are associated with the Neanderthal genome. The developed medicine model will help individuals and their populations to receive the best treatment. It also clarifies why there are differences in disease phenotypes in modern humans.