Race-matched Healthy Controls Research Articles

Helicobacter pylori in gastrointestinal manifestation of systemic sclerosis.

To study the prevalence of Helicobacter pylori in systemic sclerosis patients and its gastrointestinal manifestations in comparison with Helicobacter pylori-negative systemic sclerosis patients. Systemic sclerosis gastrointestinal outcome post Helicobacter pylori eradication was evaluated. In total, 70 systemic sclerosis patients and 70 age-, gender- and race-matched healthy controls had their urea breath test done. Gastrointestinal manifestations in systemic sclerosis were assessed using University of California at Los Angeles 2.0 and Leeds Dyspepsia Questionnaire questionnaires. Systemic sclerosis patients with confirmed Helicobacter pylori infection were given standard Helicobacter pylori eradication therapy. Urea breath test was repeated 6 weeks posteradication therapy with their gastrointestinal symptoms reassessed. The prevalence of Helicobacter pylori was low in both systemic sclerosis patients (10%) and healthy controls (2.9%). There was no significant difference in gastrointestinal symptoms between Helicobacter pylori-positive and Helicobacter pylori-negative systemic sclerosis patients. However, the Helicobacter pylori-positive patients reported higher median severity scores for the gastrointestinal symptoms of reflux (0.5 vs 0.35), abdominal distension (1.5 vs 0.75) and social functioning impairment score (0.5 vs 0.16) using the University of California at Los Angeles 2.0 score. The Helicobacter pylori-positive patients also indicated increased upper abdominal pain (3.0 vs 0.0), regurgitation (2.0 vs 0.0) and burping (3.0 vs 0.0), observed from the Leeds Dyspepsia Questionnaire scores. Gastrointestinal outcomes post-Helicobacter pylori eradication showed either an improvement or complete resolution of symptoms. Gastrointestinal symptoms in systemic sclerosis patients are unlikely to be caused by Helicobacter pylori in the recent years in our cohort of patients. No significant difference in gastrointestinal symptoms was seen between Helicobacter pylori-positive and Helicobacter pylori-negative systemic sclerosis patients. Helicobacter pylori can be readily tested by urea breath test to look for present infection.

P1143 Per- and poly-fluoroalkyl substances (PFAS) exposure is associated with later occurrence of inflammatory bowel disease

Abstract Background Environmental chemical exposures are increasingly acknowledged as risk factors for inflammatory bowel disease (IBD). Per- and poly-fluoroalkyl substances (PFAS), a large class of persistent fluorinated organic chemicals, widely used in the modern environment, may be implicated in IBD etiology, but data are conflicting. Methods We investigated the association of PFAS mixture concentrations in pre-diagnostic serum with adult-onset IBD in a pilot study within the pre-clinical Proteomic Evaluation and Discovery in an IBD Cohort of Tri-service Subjects (PREDICTS) study. This is a nested case-control study of military personnel with Crohn’s disease (CD), ulcerative colitis (UC), and age-, sex-, and race-matched healthy controls with serum samples obtained at four-time points 1-10 years prior to IBD diagnosis. Using untargeted liquid-chromatography high-resolution mass spectrometry analytical approach, we conducted an untargeted metabolomic analysis and semiquantitative measurement of fluorinated compounds, including legacy PFAS and emerging PFAS chemicals. We used weighted quantile sum regression models adjusted for confounders to study the association of PFAS as a mixture with the odds of CD and UC at each time point. Results Our study sample included individuals with CD, UC, and matched healthy controls (n=25, each group, 4 samples per individual). Baseline characteristics were similar across cases and controls. We identified nine well known PFAS compounds and 27 other known fluorinated compounds. The chemical mixture of fluorinated compounds, including the known PFAS, in the serum samples within one year of diagnosis was associated with higher odds of CD and UC [odds ratio (OR) 2.13, 95% CI 1.33, 3.41, and 1.76, 95% CI 1.15, 2.68, respectively] per one unit increase in decile. These associations remained consistent for the PFAS mixture measured in serum at all four time points up to 10 years prior to diagnosis for both CD and UC (Figure). Major contributors to the overall mixture indices included legacy PFAS, as well as emerging PFAS chemicals. Conclusion We observed a substantial increase in the odds of CD and UC among individuals with higher serum PFAS mixture levels up to 10 years prior to diagnosis. Studies to understand underlying mechanisms are ongoing.

Differences within the Immune Microenvironment May Account for Poorer Outcomes in African American AML Patients Compared to European American Patients with AML

Background: Acute myeloid leukemia (AML) is an aggressive hematological malignancy arising from an immature myeloid progenitor with notoriously poor outcomes. A critical factor responsible for the difficulty in treating this disease evasion of the host immune system by AML. Both the innate and adaptive immune systems have been shown to be dysfunctional in AML with poor type I interferon responses, decreased STING signaling and increased populations of granulocytic and monocytic myeloid derived suppressor cells ultimately leading to poor adaptive response. Studies in solid tumors have demonstrated that variation in immunity contributes to racial disparities in cancer and outcomes. This is felt to be due to variation in geographic ancestry leading to differences in the innate immune genome required due to the need for protection against environmental pathogens. However, no data exists comparing innate and adaptive immunity by race in patients (pts) with AML. Here we sought to characterize the immune microenvironment with treatment outcomes in racially diverse AML pts. Methods: Adults with newly diagnosed AML were identified who had corresponding cryopreserved bone marrow mononuclear cells available within Roswell Park Hematologic Procurement bank. Pts were matched as closely as possible for age, gender, cytogenetic risk per ELN 2017 and molecular profile. Transcriptional differences between African American (AA, n=26) and European American (EA, n=26) AML pts and healthy controls (n=10) were analyzed using transcriptomics data from NanoString Pan Cancer Immune Profile panels. AA and EA AML samples were compared to each other, as well as their race-matched healthy controls. The ‘Limma’ package in R was utilized for differential gene expression analysis. Research was conducted according to the Declaration of Helsinki in accordance with the Institutional Review Board regulated protocol. Results: The median age of the entire cohort (n=52) was 65 years (range: 28-92), with 28 female patients (53.8%) ( Table 1). Within the AA cohort, the majority of pts had intermediate risk cytogenetics per ELN 2017 (n=19; 73.1%, and 6 pts had FLT3 mut (26.1%). Three pts (11.5%) underwent allogeneic stem cell transplant, with a median overall survival (OS) censoring for transplant of 5 months (range; 0.5-68 months) ( Figure 1). Within the EA cohort, most pts had poor risk cytogenetics (n=15; 57.7%). Six pts had FLT3 mut (26.1%), and three pts had TP53 mut (12%). Five pts underwent allogeneic stem cell transplant (19.2%), with a median OS of 16.4 months (range; 0.8-67 months). Sixty-nine significantly (p<0.05, |logFC|>1.5) differentially expressed genes in AA vs. EA pts were identified. Gene set enrichment analysis (GSEA) revealed alterations to Interferon, Jun kinase and STAT signaling pathways in AA pts. In contrast, EA pt samples were enriched for pathways related to amyloid metabolism and other T and B cell immune pathways. Master regulator analysis using LISA highlighted transcriptional dysregulation attributed to PDGB5, CTCF, AR, and NOTCH in AA pts and to GLIS2, SMAD1 and NFE2 in EA pts. Conclusions: As previously reported, we found that a clinically matched cohort of AA AML patients experienced significantly worse OS as compared to EA counterparts. We have identified several significant alterations within the immune microenvironment between cohorts, which may potentially explain this discrepancy in survival in AA patients. These differences could potentially serve as targetable alterations for therapeutic purposes. This work represents the first of its kind evaluating the immune microenvironment by race in AML patients and are awaiting confirmation in a larger cohort.

Cerebral blood flow in patients recovered from mild COVID-19.

Cerebral hypoperfusion has been described in both severe and mild forms of symptomatic Coronavirus Disease 2019 (COVID-19) infection. The purpose of this study was to investigate global and regional cerebral blood flow (CBF) in asymptomatic COVID-19 patients. Cases with mild COVID-19 infection and age-, sex-, and race-matched healthy controls were drawn from the Aging Brain Consortium at The University of South Carolina data repository. Demographics, risk factors, and data from the Montreal Cognitive Assessment were collected. Mean CBF values for gray matter (GM), white matter (WM), and the whole brain were calculated by averaging CBF values of standard space-normalized CBF image values falling within GM and WM masks. Whole brain region of interest-based analyses were used to create standardized CBF maps and explore differences between groups. Twenty-eight cases with prior mild COVID-19 infection were compared with 28 controls. Whole-brain CBF (46.7±5.6 vs. 49.3±3.7, p=.05) and WM CBF (29.3±2.6 vs. 31.0±1.6, p = .03) were noted to be significantly lower in COVID-19 cases as compared to controls. Predictive models based on these data predicted COVID-19 group membership with a high degree of accuracy (85.2%, p<.001), suggesting CBF patterns are an imaging marker of mild COVID-19 infection. In this study, lower WM CBF, as well as widespread regional CBF changes identified using quantitative MRI, was found in mild COVID-19 patients. Further studies are needed to determine the reliability of this newly identified COVID-19 brain imaging marker and determine what drives these CBF changes.

Blunted sympathetic transduction to blood pressure in females with relapsing-remitting multiple sclerosis

Approximately 1 million people in the United States are living with multiple sclerosis (MS) with nearly 75% of those being female. MS is a progressive demyelinating neurological disease of the central nervous system associated with a high prevalence of orthostatic hypotension. Our laboratory has previously shown that individuals with MS have a reduced ability to increase mean arterial pressure during simulated hypotension which was attributed to a blunted vascular response. The purpose of this project was to investigate how spontaneous bursts of muscle sympathetic nerve activity (MSNA) induce end-organ response(s) and ultimately evoke a change in blood pressure (BP) on a heartbeat-to-heartbeat basis in females with MS, a process known as sympathetic transduction to BP. We tested the hypothesis that females with MS have reduced resting sympathetic transduction to BP compared to sex-, height-, weight-, age- and race-matched healthy controls. Beat-to-beat BP (Finometer) and MSNA (peroneal microneurography) were continuously recorded during 10 minutes of quiet supine rest in 9 females with relapsing-remitting MS (age: 39.6 ± 3.3 years; Expanded Disability Status Scale &lt; 6.0) and 10 healthy female controls (age: 35.4 ± 2.1 years). Total vascular conductance (TVC) was estimated using modelflow. Sympathetic transduction was determined by identifying the peak change in diastolic BP and the nadir reduction of TVC over 10 cardiac cycles following spontaneous MSNA bursts using signal averaging. No differences were detected in resting mean arterial pressure (MS: 88 ± 3 mmHg; controls: 85 ± 2; P = 0.42) or MSNA burst frequency (MS: 26 ± 3 bursts/min; controls: 19 ± 3 bursts/min; P = 0.11) between groups. Peak increases in diastolic BP following MSNA bursts were lower in females with MS (1.2 ± 0.2 mmHg) compared to controls (2.5 ± 0.6 mmHg; P = 0.039). Whereas no differences were observed in nadir reductions in TVC (MS: -1.7 ± 0.5 ml/min/mmHg; controls: -3.0 ± 0.8 ml/min/mmHg; P = 0.11). These preliminary data suggest that sympathetic transduction to BP is blunted in females with MS. Reduced sympathetic transduction to BP may contribute to the high occurrence of orthostatic hypotension in this clinical population. Further investigation is warranted to help better understand the potential mechanism(s) underlying lower sympathetic transduction in females with MS. This work was supported by American Heart Association Predoctoral Fellowship (Award #915655, Trotter) and American College of Sports Medicine Doctoral Research Grant (Award # 19-01141, Trotter) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Sickle Cell Disease Patient Plasma Sensitizes iPSC-Derived Sensory Neurons from Sickle Cell Disease Patients

Individuals living with sickle cell disease (SCD) experience severe recurrent acute and chronic pain. The underlying neurobiology of the complex pain of SCD is not well understood. Limited and often ineffective treatment options and multifactorial healthcare disparities create an immediate need to develop non-opioid based SCD pain therapies. However, in order to develop novel therapies, we must better understand the neurobiological mechanisms underlying SCD pain. Although humanized mouse models of SCD do exist, growing evidence indicates there are striking differences in gene expression and functional responsiveness between human and mouse dorsal root ganglia (DRG) sensory neurons. Human DRG tissues are extremely limited in availability, postmortem tissues only provide static, endpoint molecular signatures, and obtaining DRGs from patients with SCD is untenable. These are all barriers to gaining mechanistic insight into pathogenic pain processes. Human induced pluripotent stem cell (iPSC)-derived sensory neurons (iSNs) have recently been used to model the altered function and expression of molecules in native DRG neurons in chronic pain disorders. These iPSC-derived SNs express canonical markers of human nociceptors and exhibit functional responses to noxious stimuli similar to those properties of native mammalian nociceptors. The objective of this study was to utilize iSNs differentiated from SCD patient iPSCs as well as age- and race-matched healthy controls (HCs) to characterize baseline receptor expression and function of SCD iSNs compared to HC iSNs. Our goal is to assess the intrinsic properties of SCD nociceptors that may contribute to the persistent sensitization of nociceptors in SCD patients with pain. To this end, both HC and SCD iSNs expressed transcripts for canonical markers of mature sensory neurons (NF200, CGRP), immune receptors (CCR1/2/3/5, CD4, CRLF2, CSF2Ra/b), and functional ion channels (HCN1, Nav1.7, P2X3/4, CACNA2D1, CHRNA9) (Table 1). Further, they expressed transcripts for ion channels relevant to SCD pain (Piezo1, Piezo2, TRPA1, TRPC5, TRPM3, TRPM8, TRPV1, TRPV4). We confirmed the expression of Peripherin, CGRP, TRPV1 at the protein level using immunocytochemistry. There were no significant differences in expression for any of these targets in iSNs derived from SCD patients compared to those from HCs. However, we did identify trends for higher endothelin 1 receptor type A (ETRa) and CC motif chemokine ligand 2 (CCL2) transcript production by SCD iSNs compared to HC iSNs (Table 1). Additionally, we observed robust responses with no significant differences in baseline function between HC and SCD iSNs when stimulated with known agonists of human SNs (KCl, Glutamate, ab-meATP, capsaicin) in traditional or high-throughput calcium imaging studies. To begin defining extrinsic mechanisms that may sensitize human SCD SNs, we tested the effects of SCD patient plasma (n=4 from SCD baseline [SS BL] and acute pain [SS IP]- paired samples, n=4 Black controls [HC]) on iSN hypersensitivity by exposing iSNs in vitro to human plasma diluted 1:10 in maintenance media for 30 minutes before analyzing calcium flux. Plasma samples from SCD patients experiencing acute pain (SS IP) were found to significantly increase SCD iSN response to capsaicin (p<0.05) compared to HC iSNs treated with SS IP plasma, while race-matched HC plasma or SCD patient plasma at baseline pain states (SS BL) did not induce this effect (Figure 1). Excitingly, these findings reproduce results identified in native DRG neurons from SCD mice and suggest that iSNs derived from patient iPSCs can be used in parallel with native SCD neurons from mouse models to investigate mechanisms underlying the chronic sensory neuron sensitization in SCD. They also identify potential intrinsic mechanisms of SCD pain which may extend beyond a blood-based disease pathology. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Visual Pathway Involvement in NMDA Receptor Encephalitis: A Clinical, Optical Coherence Tomography, and 18-Fluorodeoxyglucose PET/CT Approach.

Anti-NMDA receptor (NMDAR) encephalitis patients have been reported to exhibit visual dysfunction without retinal thinning. The objective of our study was to examine the involvement of the visual pathway structure and function in anti-NMDAR encephalitis by assessing postrecovery visual function and retinal structure, and acute-phase occipital cortex function. In this cross-sectional study, patients diagnosed with anti-NMDAR encephalitis per consensus criteria underwent postrecovery visual acuity (VA) testing and optical coherence tomography (OCT) with automated retinal layer segmentation. Clinical data and acute-phase brain 18F-fluorodeoxyglucose (FDG) PET/CT (performed within 90 days of symptom onset, assessed qualitatively and semi-quantitatively) were retrospectively analyzed. VA and OCT measures were compared between anti-NMDAR and age, sex, and race-matched healthy controls (HC). When available, FDG-PET/CT metabolism patterns were analyzed for correlations with VA, and OCT measures. A total of 16 anti-NMDAR (32 eyes) and 32 HC (64 eyes) were included in the study. Anti-NMDAR exhibited lower low-contrast VA (2.5% contrast: -4.4 letters [95% CI; -8.5 to -0.3]; P = 0.04, 1.25% contrast: -6.8 letters [95%CI; -12 to -1.7]; P = 0.01) compared with HC, but no differences were found on OCT-derived retinal layer thicknesses. Acute-phase FDG-PET/CT medial occipital cortex metabolism did not correlate with follow-up low-contrast VA or ganglion cell/inner plexiform layer thickness (GCIPL) (n = 7, 2.5% contrast: r = -0.31; P = 0.50, 1.25% contrast: r = -0.34; P = 0.45, GCIPL: r = -0.04; P = 0.94). Although the visual system seems to be involved in anti-NMDAR encephalitis, no retinal structural or occipital cortex functional abnormalities seem to be responsible for the visual dysfunction. When detected acutely, occipital lobe hypometabolism in anti-NMDAR encephalitis does not seem to associate with subsequent retrograde trans-synaptic degenerative phenomena, potentially reflecting reversible neuronal/synaptic dysfunction in the acute phase of the illness rather than neuronal degeneration.

Hypomethylation of miR-17-92 cluster in lupus T cells and no significant role for genetic factors in the lupus-associated DNA methylation signature

ObjectivesLupus T cells demonstrate aberrant DNA methylation patterns dominated by hypomethylation of interferon-regulated genes. The objective of this study was to identify additional lupus-associated DNA methylation changes and determine the...

The Role and Therapeutic Target of Base Excision Repair Genes in Multiple Myeloma (MM)

Background: Dysregulated DNA repair genes play a pathological role in diverse forms of malignancies, including multiple myeloma (MM). High expression of some DNA repair enzymes has been reported in various tumors and associated with their progression and drug resistance. Here, we investigated the association between MM and base excision repair (BER) system, which is known as a DNA repair pathway for the processing of small base lesions derived from oxidation and alkylation damages. To elucidate the roles of BER in the pathogenesis of MM, we investigated the distribution of functional polymorphisms of BER, the expression level of BER genes, and the effectiveness of BER inhibitors and knockdown BER genes for MM cell lines.Material and methods: Genotyping of OGG1 Ser326Cys, MUTYH Gln324His, APE1 Asp148Glu, XRCC1 Arg399Gln, and PARP1 Val762Ala in 146 MM patients and 202 race-matched healthy controls was determined by using the PCR-restriction fragment length polymorphism (PCR-RFLP) method. Mononuclear cells were separated from bone marrow aspirates and CD138+ cells were collected using EasySep. The mRNA expression of BER genes in MM cell lines (KMM1, KMS12PE, KMSE12BM, KMS11, RPMI8226, and OPM2), primary MM cells, and normal plasma cells was assessed by real-time PCR method. The protein level of BER genes was assessed by western blot analysis. BER inhibitors, including APE1 inhibitor and PARP inhibitor were used. Inducible knockdown system was used for suppressed OGG1 expression. Genotype and allele frequencies were compared between the study groups using Χ2-test. Probability values <0.05 were considered statistically significant. This study was approved by the Institutional Review Board of Gunma University Hospital.Results: The PARP1 Val762Ala single nucleotide polymorphism, which is reported to increase cancer risk, was associated with susceptibility to MM. In the patient group, the Val/Val genotype frequency was lower (32.9% compared with 43.6%, P = 0.044) and the Val/Ala genotype frequency was higher (56.8% compared with 43.1%, P = 0.011) than those in the control groups. In contrast to PARP1 polymorphism, the distributions of other BER polymorphisms, including OGG1 Ser326Cys, MUTYH Gln324His, APE1 Asp148Glu, and XRCC1 Arg399Gln, showed no significant difference between MM patients and control groups. Next, we examined the association between the polymorphisms and the clinical characteristics of MM patients. No significant difference was observed in the allele frequency as well as in sex, age, other clinical settings, and overall survival. The BER genes, such as OGG1, APE1, and PARP1, were detected in all MM cell lines. Protein expression of OGG1, APE1, and PARP1 correlated with mRNA expression in MM cell lines. The mRNA levels of OGG1, APE1, and PARP1 of malignant plasma cells of MM patients were significantly higher than those of normal plasma cells (p < 0.05). The mRNA levels of OGG1, APE1, and PARP1 genesof plasma cells of newly diagnosed MM patients were significantly higher than those of refractory or relapsed MM patients. We exposed MM cell lines to increasing concentrations of APE-1 inhibitor, methoxamine, and PARP1-selective inhibitor, BYK204165, which demonstrated potent and selective growth inhibition in vitro. To determine the role of OGG1 gene in MM growth and survival, we suppressed its expression using an inducible knockdown system and observed significant inhibition of MM cell lines (KMS11 and KMM1) proliferation.Conclusion: The PARP1 Val762Ala polymorphism may be associated with the susceptibility to MM. High expression of BER genes such as OGG1, APE1, and PARP1 was detected in malignant plasma cells, and the inhibition of these BER genes reduced MM cell growth. According to these results, inhibition of BER genes may be a potential therapeutic target of MM. DisclosuresMurakami:Sanofi: Honoraria; Fujimoto: Honoraria; BMS: Honoraria; Ono: Honoraria; Takeda: Honoraria; Celgene: Honoraria.

Racial differences in the limb skeletal muscle transcriptional programs of patients with critical limb ischemia.

Critical limb ischemia (CLI) is the most severe manifestation of peripheral artery disease (PAD) and is characterized by high rates of morbidity and mortality. As with most severe cardiovascular disease manifestations, Black individuals disproportionately present with CLI. Accordingly, there remains a clear need to better understand the reasons for this discrepancy and to facilitate personalized therapeutic options specific for this population. Gastrocnemius muscle was obtained from White and Black healthy adult volunteers and patients with CLI for whole transcriptome shotgun sequencing (WTSS) and enrichment analysis was performed to identify alterations in specific Reactome pathways. When compared to their race-matched healthy controls, both White and Black patients with CLI demonstrated similar reductions in nuclear and mitochondrial encoded genes and mitochondrial oxygen consumption across multiple substrates, indicating a common bioenergetic paradigm associated with amputation outcomes regardless of race. Direct comparisons between tissues of White and Black patients with CLI revealed hemostasis, extracellular matrix organization, platelet regulation, and vascular wall interactions to be uniquely altered in limb muscles of Black individuals. Among traditional vascular growth factor signaling targets, WTSS revealed only Tie1 to be significantly altered from White levels in Black limb muscle tissues. Quantitative reverse transcription polymerase chain reaction validation of select identified targets verified WTSS directional changes and supports reductions in MMP9 and increases in NUDT4P1 and GRIK2 as unique to limb muscles of Black patients with CLI. This represents a critical first step in better understanding the transcriptional program similarities and differences between Black and White patients in the setting of amputations related to CLI and provides a promising start for therapeutic development in this population.

IL-11 induces surface marker expression and migration of T-lymphocytes

Abstract Objective To study the phenotypic and migratory changes of PBMCs following IL-11 stimulation Background Multiple sclerosis (MS) is presumably an autoimmune disease in which immune cells mediate neuroinflammation and neurodegeneration in central nervous system. Cytokines play a significant role in orchestrating the inflammatory response during the disease. Our lab has previously shown that in untreated MS patients, IL-11 is upregulated in the CSF and serum in comparison to matched healthy controls (HCs). Previous study has shown that CD4+IL11+ cells are accumulated in the CSF and the brain MS lesions, suggesting that they play a role in the lesion formation. Methods 14 untreated RRMS patients and age-, sex- and race-matched HCs were enrolled in the study upon signing an Institutional Review Board consent forms. Results By using flow cytometry we characterized IL-11-secreting CD4+ lymphocytes and show that they have increased expression of CCR6, CCR4, ICAM-1, IL-11RA, GM-CSF, and IFNγ in comparison to CD4+IL-11− cells (p values&amp;lt;0.03). In vitro studies further showed that stimulation of PBMCs with recombinant human IL-11, upregulated the expression of CCR4 and CCR6 (p values&amp;lt;0.03, 1.2 and 1.4 fold changes) in CD4+ gated cells. In a trans-well system we observed that IL-11 can act as a chemoattractant for anti-CD3/CD28 mAb stimulated PBMCs and increase their migration (p value=0.04, migration index=1.2) in both patients and HCs. Conclusion We observed that IL-11 induces migratory phenotype in CD4+ lymphocytes in patients with RRMS, which can be explored as a therapeutic target in early RRMS.

PARP1 V762A polymorphism affects the prognosis of myelodysplastic syndromes.

Myelodysplastic syndromes (MDS), caused by various genetic mutations in hematopoietic stem cells, are associated with highly variable outcomes. Poly (ADP-ribose) polymerase-1 (PARP1) plays an important role in DNA damage repair and contributes to the progression of several types of cancer. Here, we investigated the impact of PARP1 V762A polymorphism on the susceptibility to and prognosis of MDS. Samples collected from 105 MDS patients and 202 race-matched healthy controls were subjected to polymerase chain reaction-restriction fragment length polymorphism for genotyping. The allele and genotype frequencies of PARP1 V762A did not differ between MDS patients and the control group. However, MDS patients with the PARP1 V762A non-AA genotype, which is associated with high gene activity, had shorter overall survival rates (P=.01) than those with the AA genotype. Multivariate analysis of overall survival also revealed PARP1 V762A non-AA genotype as a poor prognostic factor (P=.02). When patients were analyzed according to treatment history, the PARP1 V762A non-AA genotype was only associated with poor survival in patients who had received treatment (P=.02). PARP1 V762A polymorphism may be an independent prognostic factor for MDS, and a predictive biomarker for MDS treatment.

Characterizing the Skin and Gut Microbiome of Alopecia Areata Patients

Background: Alopecia areata (AA) is caused by autoimmune attack of the hair follicle. The exact pathogenesis is unknown, but hypotheses include innate immunity imbalance, environmental exposures, genetic predisposition, and possibly the microbiome. The objective of this study was to characterize the skin and gut microbiome of AA patients, and compare microbial composition to healthy individuals. Methods: This was a pilot, case-control study. Scalp and fecal microbiome samples were collected from 25 AA patients, and 25 age, gender, and race-matched healthy controls in Southern California with no significant difference in demographic characteristics. After library preparation and identification of bacterial and fungal taxonomy, multivariant analysis was performed to compare AA and healthy microbiomes. Results: The AA scalp microbiome was significant for decreased Clostridia and Malasseziomycetes, and the gut microbiome was significant for decreased Bacteroidia and increased Bacilli (p&lt;0.05) compared to healthy controls. Conclusions: The composition of the AA bacterial and fungal, scalp and gut microbiome is significantly different than healthy individuals. Future directions include using this data to characterize microbial changes associated with AA patient diet, relating to disease severity, and predicting disease progression, prognosis and/or therapeutic response.

Increased Adiposity and Reduced Lean Body Mass in Patients with Short Bowel Syndrome.

Few studies have examined the metabolic consequences of short bowel syndrome (SBS) and its effects on body composition in adults. We hypothesized that body composition of SBS patients is altered compared to a normal age-, race-, and sex-matched population, regardless of parenteral nutrition (PN) dependence. To compare the body composition of adult patients with SBS to age-, sex-, and race-matched healthy controls. Twenty patients with SBS underwent body composition analysis using the GE Lunar iDXA scanner. Patients were age-, sex-, and race-matched to controls from the National Health and Nutrition Examination Survey (1999-2004). Mean differences in body mass index, fat-free mass, fat mass, percent body fat, visceral adipose tissue mass and volume, and bone mineral density were measured. Statistical analysis was performed using SAS 9.4 software. Fifty-five percent of subjects had a history of PN use, and 30% were current PN users. Mean percent body fat for SBS patients was 35.1% compared to 30.9% for healthy controls (p = 0.043). Fat-free mass was reduced in SBS (p = 0.007). Patients with reduced bone mass had a trend toward significantly more years of PN exposure compared to those with normal bone mass (p = 0.094), and a trend toward older age (p = 0.075). SBS is associated with increased percent body fat and reduced fat-free mass, suggesting that improved dietary and therapeutic interventions are needed to restore normal metabolic indices and avoid risk of metabolic syndrome in SBS patients.

Foveal avascular zone morphology and parafoveal capillary perfusion in sickle cell retinopathy

Background/aimsTo assess foveal avascular zone (FAZ) morphology and parafoveal capillary perfusion in patients with various stages of sickle cell retinopathy (SCR) using optical coherence tomography angiography (OCT-A).MethodsThis is a multi-institutional...

(184) - The effects of pain-related anxiety on hypnosis treatment responses in adults with and without sickle cell disease

Pain is the most common symptom in sickle cell disease (SCD) and individuals with SCD exhibit pain hypersensitivity. Hypnosis can have beneficial effects on pain modulation, but it is unknown whether pain-related anxiety—a cognitive-affective factor also associated with pain modulation—attenuates the effects of hypnosis in individuals with SCD. This study examined if pain-related anxiety alters the effects of hypnosis on laboratory pain responsivity in adults with and without SCD. The sample included 14 adults with SCD and 14 race-matched healthy controls. Participants completed the Pain Anxiety Symptoms Scale (PASS-20) at baseline. Pain tolerance (°C), threshold (°C), and intensity (0–100) were collected during a standardized thermal pain protocol before and during a hypnosis session. Change in pain outcomes indicated participants' responses to hypnosis. Nonparametric tests were used and a median split determined within-group levels of pain-related anxiety. Pain-related anxiety did not significantly differ across SCD and non-SCD groups, and was not associated with pre-hypnosis pain tolerance or threshold in either group. In adults with SCD who had lower levels of pain-related anxiety, pain tolerance increased 5.68°C during hypnosis compared to 1.97°C in those with high anxiety (Z = -1.98, p < .05). The effect on pain threshold approached significance (1.97°C vs. -.38°C, Z = -1.85, p = .06). There was no effect of pain-related anxiety on change in pain responsivity in healthy controls. Results partially align with other pain anxiety research and suggest that higher pain-related anxiety may be particularly important to consider in individuals with SCD as it may attenuate pain modulatory effects of hypnosis. Further, in individuals with higher pain-related anxiety, a single hypnosis session may be insufficient to reduce modulatory effects of anxiety on pain responsivity. Thus, examination of long-term treatment effects in a larger sample is needed to confirm underlying mechanisms affecting hypnosis treatment responses in individuals SCD. Funding: NHLBI-1U54HL117718.

Increased circulating leukocyte-derived microparticles in ischemic cerebrovascular disease

ObjectivesCirculating leukocyte-derived microparticles act as proinflammatory mediators that reflect vascular inflammation. In this study, we examined the hypothesis that the quantity of leukocyte-derived microparticles is increased in patients with ischemic cerebrovascular diseases, and investigated utility of various phenotypes of leukocyte-derived microparticles as specific biomarkers of vascular inflammation injury. Additionally we focused on identifying leukocyte-derived microparticles that may be correlated with stroke severity in acute ischemic stroke patients. MethodsThe plasma concentration of leukocyte-derived microparticles obtained by a series of centrifugations of 76 consecutive patients with ischemic cerebrovascular diseases and 70 age-, sex-, and race-matched healthy controls were determined by flow cytometry. ResultsSignificantly elevated numbers of leukocyte (CD45+), monocyte (CD14+), lymphocyte (CD4+), granulocyte (CD15+) derived microparticles were found in the plasma samples of patients ischemic cerebrovascular diseases, compared to healthy controls (p<0.05). Furthermore, the plasma levels of CD14+ microparticles were significantly correlated with stroke severity (r=0.355, p=0.019), cerebral vascular stenosis severity (r=0.255, p=0.025) and stroke subtype (r=0.242, p=0.036). No association with stroke was observed for other leukocyte-derived phenotypes. ConclusionsThese results demonstrate that circulating leukocyte-derived microparticles amounts are increased in patients with ischemic cerebrovascular diseases, compared with healthy controls. As proinflammatory mediators, leukocyte-derived microparticles may contribute to vascular inflammatory and the inflammatory process in acute ischemic stroke. Levels of CD14+ microparticles may be a promising biomarker of ischemic severity and outcome of stroke in the clinic.

Thromboelastographic characterization of the activated clotting system in children with sickle cell trait or sickle cell disease

BackgroundRecent epidemiological evidence suggests sickle cell disease (SCD) and sickle cell trait (SCT) is a risk factor for venous thromboembolism. The increased in-vivo markers of thrombin generation support the notion that such patients are in a chronic hypercoagulable state. In an attempt to better understand the underlying mechanism, global hemostatic assays including thrombin generation assay (TGA) and thromboelastography (TEG) have been utilized by several groups, but thus far, have shown inconsistent results either due to small sample size or technical differences. ObjectivesGlobal hemostatic characterization of children with SCD or SCT by using TGA and modified TEG methods. Materials and MethodsIn this pilot study, we obtained TGA, TEG and other hemostatic data on specimens from 13 patients with SCD, 14 with SCT and 12 race-matched healthy controls (NC). ResultsR time and K time with modified TEG methods were significantly shorter in SCD when compared to SCT and NC. Alpha and MA did not show any significant differences between the groups. There was no difference seen between SCT and NC. TGA profiles did not show any difference between the three groups. As expected the in-vivo markers of thrombin generation and activation of fibrinolysis including D dimer and thrombin-antithrombin complexes were significantly higher in SCD subjects as compared to SCT and NC. ConclusionThe modified TEG methods are able to detect the activated coagulation system for the SCD population but a larger and more homogenous SCT cohort needs to be studied for more conclusive results.

Tumor necrosis factor-alpha gene promoter methylation in Japanese adults with chronic periodontitis and rheumatoid arthritis.

Over-expression of tumor necrosis factor-alpha (TNF-α) plays a pathological role in chronic periodontitis (CP) and rheumatoid arthritis (RA), which might be regulated by the epigenetic mechanism. The aim of the present study was to evaluate whether there is a unique methylation profile of the TNF-α gene promoter in blood cells of individuals with CP and RA. The study participants consisted of 30 Japanese adults with RA (RA group), 30 race-matched adults with CP only (CP group) and 30 race-matched healthy controls (H group). Genomic DNA isolated from peripheral blood was modified by sodium bisulfite and analyzed, by direct sequencing, to investigate DNA methylation of the TNF-α gene promoter region. The level of TNF-α produced in mononuclear cells stimulated with Porphyromonas gingivalis lipopolysaccharide was determined using ELISA. Twelve cytosine-guanine dinucleotide (CpG) motifs were identified in the TNF-α promoter fragment from -343 to +57bp. The CP group showed a significantly higher methylation rate and frequency at -72bp than the H group (p<0.01). The RA group exhibited significantly higher methylation rates at seven CpG motifs (-302, -163, -119, -72, -49, -38 and +10bp), and significantly higher methylation frequencies at six CpG motifs (-163, -119, -72, -49, -38 and +10bp), than the H group (p<0.01 for all comparisons). The levels of TNF-α produced were significantly different between individuals with and without methylation at -163bp (p=0.03). These results suggest that the hypermethylated status of CpG motifs in the TNF-α gene promoter in blood cells may be unique to Japanese adults with CP and RA.

Do clinical features of Lesch-Nyhan disease correlate more closely with hypoxanthine or guanine recycling?

Lesch-Nyhan disease (LND) is a rare, X-linked recessive neurodevelopmental disorder caused by deficiency of hypoxanthine-guanine phosphoribosyltransferase (HGprt), an enzyme in the purine salvage pathway. HGprt has two functions; it recycles hypoxanthine and guanine. Which of these two functions is more relevant for pathogenesis is unclear because some evidence points to hypoxanthine recycling, but other evidence points to guanine recycling. In this study, we selectively assayed hypoxanthine (Hprt) and guanine (Gprt) recycling in skin fibroblasts from 17 persons with LND, 11 with an attenuated variant of the disease (LNV), and 19 age-, sex-, and race-matched healthy controls (HC). Activity levels of both enzymes differed across groups (p < 0.0001), but only Gprt distinguished patients with LND from those with LNV (p < 0.05). Gprt also showed slightly stronger correlations than Hprt with 13 of 14 measures of the clinical phenotype, including the severity of dystonia, cognitive impairment, and behavioral abnormalities. These findings suggest that loss of guanine recycling might be more closely linked to the LND/LNV phenotype than loss of hypoxanthine recycling.