NIH Score Research Articles

Tertiary Lymphoid Organs in Takayasu Arteritis.

The role of B cells in the pathogenesis of Takayasu arteritis (TA) is controversial. We aimed to study the presence of tertiary lymphoid organs (TLOs) in the aortic wall of TA patients. Hematoxylin and eosin-stained sections from aorta specimens from patients with TA were screened for TLOs. The presence of B cell aggregates (CD20), follicular dendritic cells (FDCs, CD21), and high endothelial venules (HEVs, PNAd) was investigated by immunohistochemistry. Immune cells from the adventitial layer of one patient were characterized by flow cytometry. Demographic, medical history, laboratory, imaging, treatment, and follow-up data were extracted from medical records. Aorta specimens from Bentall procedures were available from seven patients (5 females, aged 22-57 years) with TA. Surgical treatment was performed at TA diagnosis (n = 4) or at a median of 108 months (84-156) after TA diagnosis. Disease was active at surgery in four patients according to NIH score. B cell aggregates-TLOs containing HEVs were observed in the adventitia of all but one patient. Of note, ectopic follicles containing CD21(+) FDCs were found in all patients (4/4) with increased aortic (18)F-fluoro-deoxyglucose (FDG) uptake before surgery but were absent in all but one patients (2/3) with no FDG uptake. In addition, flow cytometry analysis confirmed the accumulation of memory/germinal center-like B cells in the adventitial layer and showed the presence of antigen-experienced T follicular helper cells. Ectopic lymphoid neogenesis displaying functional features can be found in the aortic wall of a subset of patients with active TA. The function of these local B cell clusters on the pathogenesis of TA remains to be elucidated.

Abstract WP439: Who Benefits From CT-Angiography Previous Intravenous Thrombolysis?

Background: Endovascular treatment (EVT) is an effective treatment in strokes with persistent large artery occlusion despite previous intravenous thrombolisis (IVT) as rescue treatment. Performing computer tomography angiography (CTA) before IVT could allow early activation of neurointerventional teams; however routine CTA could delay unnecessary door-to-needle time of IVT and may be infeasible. Methods: We reviewed stroke code activations between May 2011 and June 2015 in our comprehensive stroke center and divided into groups based on NIHSS and patency of arterial occlusion according to non-enhanced CT on admission (dense artery sign or dot sign) and baseline CTA. We assessed patients treated with IVT and selected to EVT according to results in CTA post-IVT. We analyze percentage of recanalization or migration of thrombus after IVT alone and variables associated to successful treatment. Results: Of 2856 stroke codes registered during the study period 1810 were diagnosis of ischemic strokes. We treated 520 patients with IVT, 202 had a radiological evidence of large artery occlusion (55%M1, 32% M2, 5%TICA, 5%ICA, 3% basilar). Thirty-two percent of patients showed changes in CTA carried out after IVT(17% successfully recanalized, 15% distal migration of thrombus) so they were not selected to endovascular treatment. There were significant difference between M1 and M2 occlusion regarding changes in CTA after IVT (23% vs 70%; p<0.001). In multivariate logistic regression a baseline score NIH<10 was associated with higher percentage of recanalization with rtPA despite signs of large vessel occlusion (78% vs 32%; p:0.001). In receiver operating characteristic analysis higher baseline NIH was associated with persistent occlusion after IVT (area under curve=0.79;95% CI, 0.6-0.9; P:0.001) with optima threshold of 10 ( Sensivity 84%, Specificity 74%). Conclusions: We consider defer CTA angiography until after IVT in stroke code patients with moderate clinical impairment (NIH<10) or M2-segment occlusion, because they achieve a high percentage of arterial recanalization. CTA previous IVT could be unnecessary, provide unreliable information and delay IVT in that clinical group but could be useful to plan EVT in patients with higher NIH scores.

AB0568 Biologics in Takayasu Arteritis: Preliminary Data from the French Registry

ObjectivesThe aim of this registry is to determine: (1) the real-life use of various biologics in Takayasu arteritis in France; (2) to compare the efficacy of different biologics among them;...

Cytokine gene polymorphisms and severity of CF lung disease

BackgroundThe search for modifier genes to explain inconsistencies in cystic fibrosis (CF) genotype–phenotype relationships has yielded mixed results. In a previous cross-sectional study from our centre the clinical effect (as described by FEV1, BMI z-score, admitted days and NIH score) of single nucleotide polymorphisms (SNPs) of four cytokine genes (IL-8, TNF-α, IL-1β and IL-10) was examined in 158 children with CF. No association between cytokine genotype and any biological outcome measure was found. In this present study a cross-sectional and longitudinal examination of this relationship was undertaken to test the hypothesis that pro-inflammatory SNPs would affect longitudinal changes in CF lung disease. MethodsUsing the cohort examined in our earlier study we performed both longitudinal and cross-sectional data analyses examining the relationship between SNPs (TNF-α, IL-8, IL-10 and IL-1β) and clinical outcome measurements. In the first part of this current study, lung function data (annual decline of FEV1 percent predicted) was compared with the cytokine genotype over a 13year period. In the second part of this current study multiple regression was used to assess associations between clinical outcomes (best FEV1 percent predicted and BMI at the age of 10years) and alleles of cytokine genes, adjusting for gender, CF genotype and lung infection status. ResultsA total of 152 patients with CF were analysed in the longitudinal study and data from 130 patients at the age of 10years were analysed in the cross-sectional study. There was evidence for an association between pro-inflammatory SNPs of the IL-8, IL-10 and IL-1β genes and more severe lung disease. Multiple regression of the longitudinal data with a total of 10,956 lung function measurements showed an additional annual decline of the percentage predicted FEV1 of −1.15 (IL-8, p<0.001), −0.24 (IL-10, p=0.049) and −0.41 (IL-1β, p<0.001) for patients with any of the pro-inflammatory alleles. None of the cross-sectional data showed a significant association between the cytokine genotypes and the clinical outcomes. ConclusionPro-inflammatory alleles of three cytokine genotypes, IL-8, IL-10 and IL-1β, appear to be associated with slightly more severe lung disease in patients with CF over a 13year period. Further studies are required to exclude influence of confounders on the severity of lung disease.

Edge assay: kinetic analysis of reagents affecting cell clumping

Cells in clumps are often more dangerous than single cells in a cancer setting and in infectivity. We present a fine‐tuned method to test reagents that may affect cell clumping. In each experiment two 0.2ml droplets of distilled water were placed on glass microscope slides to which 1% formaldehyde fixed yeast (Saccharomyces cerevisiae) in distilled water (pH adjusted to 4.0) were added at dilutions that resulted in 8–25 single yeast and clumped yeast cells that were observed at the edge of the droplets, providing a standardized reference point for counting single vs clumped (two or more) yeast. Using birchwood toothpicks, various concentrations of solid reagents (salts, sugars, complex carbohydrates, amino acids) (1–10 mg) were added to the experimental drops and nothing was added to the control drops. Numbers of single and clumped yeast were counted before addition of reagents, immediately after addition and at 20, 40, 60 min after reagent additions in 1179 trials (21 C). The numbers of single yeast and yeast clumps were observed with a light microscope (100–200 X) and recorded at each step. The percentages of single yeast in experimentals and controls were plotted showing SEM error bars and t‐tests were performed at 20,40,60 min comparing the percentage of single yeast in experimentals vs controls. P values of less that 0.05 suggested that the percentages of single cells in the experimentals were significantly different than in the controls. The results show that the precise analysis that this assay affords is able to detect very small yet statistically significant differences in reagent effects and could lead to identification of reagents that may be further developed into anti‐clumping drugs (Supported by NSF Presidential Award 0731633, NIH SCORE, RISE, MARC, the Joseph Drown Foundation, the Sidney Stern Memorial Trust).

Abstract TP383: Use of Oral Anticoagulants in FAST-MAG Patients

Background: Many patients being evaluated acutely in the field for acute ischemic stroke will be on anticoagulation, usually for atrial fibrillation. Ischemic stroke related to atrial fibrillation is more severe than that of other etiologies and there is the added risk of intracranial hemorrhage in these patients. We describe rates anticoagulant and clinical characteristics use among stroke patients being evaluated by paramedics &lt;2 hours from symptom onset. Methods: The Field Administration Stroke Therapy-Magnesium (FAST-MAG) is a phase 3 NIH-funded, randomized, placebo-control study of prehospital initiated magnesium sulfate within 2 hours of stroke onset. Data on demographics and examination was obtained from paramedic run sheets and medical records. Results: Of the 1116 consecutive, mean age was 70 (SD 13) years, 40% were women, 23% Hispanic Ethnicity, 77% white race, and 23% had ICH on initial imaging. A total of 114 (10%) were on anticoagulation at EMS evaluation, most often for known atrial fibrillation (n= 83) and/or valvular heart disease (n=44). Those on anticoagulation were less likely to present with ICH (16% vs 25%). Those on anticoagulation were often found to have an INR less than 2 (n=77 66%), 26 were therapeutic with an INR between 2 and 3, and 11 had an INR &gt; 3. NIH scores were higher at the time of ED arrival (11.16 vs 9.87) and at the 24hour mark (8.70 vs 7.97). Additionally, patients with a sub- therapeutic INR (0-2) had the highest observed NIHSS (mean 12.03). No association between high INR (over 3.0) and intracerebral hemorrhage was seen (p=0.752). Conclusion: Patients on anticoagulation therapy most often present with ischemic stroke. A sub therapeutic INR led to greater severity of stroke.

A case of carotid dissection having significant benefit from thrombolytic treatment

Ischemic stroke due to carotid artery dissection is more common among young people and it accounts about 2-5% of all strokes. A 56 year old left landed male patient was evaluated for left-sided hemiparesis and aphasia in the emergency department. His cranial CT was assessed normal and NIH score was assessed 24 then he undergone thrombolytic therapy. His NIH score was decreased to 14 after 24 hours. At extracranial dupplex USG, cranial MRI and MR-Angiography, done the same day there were findings for carotid dissection. At MR-anjio repeated 5 days later, partial recanalization were seen. At control cranial CT only striocapsuler infarct was seen. There was no any evidence of bleeding as complication. Here we present a rare case of extracranial carotid artery dissection case undergone IV thrombolytic therapy and want to take attention that in carotid dissection IV thrombolytic therapy is quiet safe and there is no difference of complication such as hemorrhage and the prognosis is no different then those with no carotid dissection.

Recommended Tools for Joint Chronic Graft-Versus-Host Disease: Results From the Chronic Gvhd Consortium

AbstractAbstract 464 Background:Assessing joint chronic GVHD needs to be accomplished reliably, simply and in a clinically meaningful way. To determine the optimal tool for assessing joint GVHD, we evaluated 2 NIH recommended joint tools (Table), a photographic range of motion (P-ROM) scale, and 7 other NIH recommended tools (Lee symptom scale, 10-point overall symptoms, FACT-G, SF36, Human Activities Profile, walk test and grip test). Methods:Patients ≥ age 2 with systemically treated chronic GVHD ≤ 3 years after hematopoietic cell transplantation were eligible for a prospective multicenter observational study. Incident and prevalent cases were included. At follow-up visits every 3–6 months, the clinician (MD) and patient (PT) rated separately their perception of change in joint GVHD on an 8-pt scale, which was collapsed into improved, stable or worse categories. Linear mixed models were used to correlate change in each tool with MD or PT-perceived change (improved vs. stable or worse vs. stable) in joint GVHD status. Results:Nine sites in the Consortium enrolled 567 participants through December 2011. Joint involvement, as defined by NIH joint/fascia score ≥1, was present at enrollment in 164 (29%) patients and included wrists (64%), ankles (47%), shoulders (35%) and elbows (30%). Joint involvement at enrollment was associated with longer duration of chronic GVHD, high-dose total body irradiation, higher symptom burden, lower quality of life (QOL), similar activity profile and similar physical function, compared to those without joint involvement. Change in joint GVHD status was examined for 652 paired visits when joint involvement was documented in the previous or current visit. In the later visits, both MDs and PTs more often reported improvement (44% and 45%) than worsening (5% and 11%). Tools that correlated with both MD and PT-perceived joint improvement were NIH joint/fascia score, Hopkins fascia score and SF36-PCS. Tools that correlated with both MD and PT-perceived joint worsening were P-ROM total score, NIH joint/fascia score, Hopkins fascia score, Lee muscle/joint subscale, Lee symptom overall score, 10-point overall symptoms and FACT-G. Among the 3 joint/fascia tools (Figure), for MD-perceived improvement, estimated change in the NIH score and P-ROM score was slightly larger than in the Hopkins score. For PT-perceived improvement, estimated change was similar for NIH and Hopkins scores. In contrast, for both MD and PT-perceived worsening, estimated change for the P-ROM score was significantly larger than in the other tools. Conclusion:Joint involvement with chronic GVHD is frequent and associated with increased symptom burden and decreased QOL. Our results support the combined use of NIH joint/fascia score and P-ROM scale to assess joint GVHD. The NIH score better reflects joint improvement and the P-ROM scale better reflects joint worsening. The more objective P-ROM scale is insensitive to PT-perceived joint improvement possibly because unlike the other two joint assessment tools, it does not incorporate tightness with or without activities of daily living.TableJoint/fascia toolsToolExplanationClinically meaningful change*NIH joint/fascia scoreA 4-pt (0-3) composite measure of ROM, tightness and activities of daily living.1 pointHopkins fascia scoreA 4-pt (0-3) measure of tightness.1 pointP-ROM total scoreThe sum of the 7-pt (1-7) wrist, shoulder, elbow scales plus the 4-pt (1-4) ankle scale (Carpenter, Blood 2011). Maximum 25 points.1 point*Derived from original design. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Impact of prior acute GVHD on chronic GVHD outcomes: a chronic graft versus host disease consortium study

Chronic graft-versus-host disease (CGVHD) is one of the most debilitating complications post hematopoietic cell transplant (HCT). Several studies have evaluated factors affecting survival in CGVHD,1–8 most utilizing the older definition of CGVHD where patients with any symptoms of GVHD (acute or chronic) may be classified as CGVHD if symptomatic beyond day 100 post HCT. However, the new NIH consensus guidelines have re-defined criteria for CGVHD diagnosis and staging9 based on actual symptoms at presentation and excludes patients with persistent, recurrent or late AGVHD who were previously considered to have CGVHD. The CGVHD consortium follows a large multi-center prospectively enrolled cohort of CGVHD patients utilizing NIH consensus guidelines for assessments.10 Several reports from this consortium have identified factors affecting survival including global severity, thrombocytopenia, KPS and the overlap syndrome.11,12 Prior AGVHD has been associated with a higher mortality in patients who develop CGVHD,1,3 indicating that these patients may be more debilitated and hence suffer worse outcome. However, the clinical and prognostic importance of prior AGVHD on subsequent CGVHD in the current era using NIH diagnostic and staging criteria is unknown. We evaluated the significance of prior AGVHD on CGVHD presentation, overall response and survival in a contemporary cohort of CGVHD patients prospectively enrolled by the CGVHD consortium. Cases enrolled in the consortium could be incident (enrollment < 3 months after diagnosis) or prevalent (enrollment ≥3 months after diagnosis).10 Detailed CGVHD assessments were performed reflecting the recommendations of the NIH Consensus Conference,9,13 and are published in the cohort study rationale and design summary.10 Prior AGVHD was considered present if maximum grade I-IV was reported. Comparisons were performed between two groups: Prior AGVHD and no prior AGVHD. Statistical comparisons between groups were made with the two-sample t-test for continuous variables, and chi-squared test for categorical variables. Spearman correlation coefficients (ρ, rho) and simple kappa statistics were used to describe rank correlation and agreement between acute and CGVHD overall severity scores and skin, GI, and liver severity. Survival was estimated by the Kaplan-Meier method.14 Overall survival was calculated from enrollment, and non-relapse mortality (NRM) was defined as death without prior relapse. Failure-free survival (FFS) was defined as survival without prior relapse or adding new systemic medications to treat CGVHD. Cox regression15 was used for multivariate analysis of survival relative to presence or absence of prior acute GVHD and other risk factors. The total cohort included 496 patients: 329 with prior AGVHD and 167 with no prior AGVHD (de-novo CGVHD) (table 1). The prior AGVHD and de-novo CGVHD groups did not differ with respect to time between HCT and CGVHD diagnosis, or time between HCT and enrollment. The proportion of patients with classic and overlap CGVHD and the distribution of CGVHD global severity (mild, moderate and severe CGVHD), using clinician or patient reported and NIH scores, was similar. These observations support the hypothesis that CGVHD in patients with prior AGVHD does not differ fundamentally from de-novo CGVHD. In particular, prior AGVHD was not associated with AGVHD manifestations at the time of CGVHD diagnosis (overlap syndrome). There were some differences between prior AGVHD and de-novo CGVHD groups: Myeloablative conditioning, a mismatched or unrelated donor, and thrombocytopenia were more frequent in those with prior AGVHD. No further differences were observed between the two groups, including frequency of organ involvement, functional assessments or patient reported outcomes. Table 1 Patient and CGVHD characteristics

The myth of the ‘unaffected’ side after unilateral stroke: Is reorganisation of the non‐infarcted corticospinal system to re-establish balance the price for recovery?

BackgroundBilateral changes in the hemispheric reorganisation have been observed chronically after unilateral stroke. Our hypotheses were that activity dependent competition between the lesioned and non-lesioned corticospinal systems would result in persisting asymmetry and be associated with poor recovery. MethodsEleven subjects (medium 6.5years after stroke) were compared to 9 age-matched controls. The power spectral density (PSD) of the sensorimotor electroencephalogram (SM1-EEG) and electromyogram (EMG) and corticomuscular coherence (CMC) were studied during rest and isometric contraction of right or left opponens pollicis (OP). Global recovery was assessed using NIH score. FindingsThere was bilateral loss of beta frequency activity in the SM1-EEGs and OP-EMGs in strokes compared to controls. There was no difference between strokes and controls in symmetry indices estimated between the two corticospinal systems for SM1-EEG, OP-EMG and CMC. Performance correlated with preservation of beta frequency power in OP-EMG in both hands. Symmetry indices for the SM1-EEG, OP-EMG and CMC correlated with recovery. InterpretationSignificant changes occurred at both cortical and spinomuscular levels after stroke but to the same degree and in the same direction in both the lesioned and non-lesioned corticospinal systems. Global recovery correlated with the degree of symmetry between corticospinal systems at all three levels — cortical and spinomuscular levels and their connectivity (CMC), but not with the absolute degree of abnormality. Re‐establishing balance between the corticospinal systems may be important for overall motor function, even if it is achieved at the expense of the non-lesioned system.

Evaluation of prognostic factors among patients with chronic graft-versus-host disease

Chronic graft-versus-host disease (cGVHD) is a major complication after allogeneic stem cell transplantation with an adverse effect on both mortality and morbidity. In 2005, the National Institute of Health proposed new criteria for diagnosis and classification of chronic graft-versus-host disease for clinical trials. New sub-categories were recognized such as late onset acute graft-versus-host disease and overlap syndrome. We evaluated the prognostic impact of the new sub-categories as well as the clinical scoring system proposed by the National Institute of Health in a retrospective, multicenter study of 820 patients undergoing allogeneic stem cell transplantation between 2000 and 2006 at 3 different institutions. Patients were retrospectively categorized according to the National Institute of Health criteria from patients' medical histories. As far as the new sub-categories are concerned, in univariate analysis diagnosis of overlap syndrome adversely affected the outcome. Also, the number of organs involved for a cut-off value of 4 significantly influenced both cGVHD related mortality and survival. In multivariate analysis, in addition to NIH score, platelet count and performance score at the time of cGVHD diagnosis, plus gut involvement, significantly influenced outcome. These 3 variables allowed us to develop a simple score system which identifies 4 subgroups of patients with 84%, 64%, 43% and 0% overall survival at five years after cGVHD diagnosis (score 0: HR=15.96 (95% CI: 6.85-37.17), P<0.001; score 1: HR=5.47 (95% CI: 2.6-11.5), P<0.001; score 2: HR=2.8 (95% CI: 1.32-5.93), P=0.007). In summary, we have identified a powerful and simple tool to discriminate different subgroups of patients in terms of chronic graft-versus-host disease related mortality and survival.

Abstract 2284: Reliability and Validity of the MGHS Dysphagia Screening Tool for Stroke Patients

Background Dysphagia occurs in approximately 55% of acute stroke patients and is associated with aspiration pneumonia. Clinical practice guidelines recommend that all patients who are admitted with the diagnosis of stroke or suspected stroke should be screened for dysphagia utilizing a validated screening tool prior to receiving any food, fluid, or medication by mouth. Purpose The purpose of the study is to determine the reliability and validity of the Marquette General Health Systems Dysphagia Screen for identifying patients at risk for aspiration. Method This non-randomized study utilized a collaborative tracking tool that compared results of the Nursing Dysphagia Screen (NDS) to the Speech Language Pathologist Swallow Evaluation (SLPSE). Two Registered Nurses rated each patient utilizing the NDS and then the patient was rated on the SLPSE. The NDS includes10 items: and the Speech Language Pathologist portion of the tool consisted of 8 items. Results One hundred subjects ranged in age from 20-94 (M=70.31, SD=14.6). There were more males (n=58) than females (n=42). The majority were admitted to the medical unit (64%), followed by cardiac (14%), ICU (12%) and CCU (10%). Length of stay ranged from 1-15 days (M=3.73, SD=2.8). NIH scores ranged from 0-20 (M=6.8, SD=7.4). Kappa was calculated to determine inter-rater reliability. For 63 pairs of ratings between two staff nurses, Kappa=.81 indicates strong reliability. Scores on the Nursing Dysphagia Screen were compared to 86 Speech Language Pathologist Swallow Evaluations. The Phi Coefficient was not significant indicating that the nurses’ and speech therapists ratings’ were similar. In addition, when there were differences it was false positives by nurses. Conclusion The MGHS NDS is a quick screening tool used by nursing in an effort to prevent aspiration. Prior to implementing the use of the tool, it was important to establish the reliability and validity of the tool. Results indicate that there is strong inter-rater reliability. Validity was established given that there were no significant difference in ratings between the nurses and speech therapists. It appears that the MGHS NDS is an effective early screening tool for dysphagia.

Fluorescence Studies of Local Environmental Heterogeneity Around Guanine Residues of the Telomeric DNA Quadruplex Conformation

Formation and stabilization of G-quadruplexed DNA formed by the guanine-rich human telomeric sequence d(TTAGGG)4 (HT4), has been shown to inhibit the abnormal activity of telomerase in tumor cells, making stabilization of this DNA secondary structure a target for potential cancer therapeutics. To investigate the effect of local environment on the guanine residues of the G-quadruplex, we previously designed several fluorescence analogs of the HT4 sequence with incorporation of 6-methyl-8-(2-deoxy-D-ribofuranosyl) isoxanthopterin (6MI) at varying guanine-tetrad positions: G1, G4, and G11, which show fluorescence sensitivity to quadruplex folding. In the current studies, we have collected decay-associated spectra (DAS) for the fluorescence lifetime components (τlong, τmedium, and τshort) resolved for each of the mono-labeled sequences. Multi-exponential decay profiles suggest local environmental heterogeneity surrounding the guanine residues in both the folded and unfolded states. Interestingly, the DAS for τlong observed for guanine positions G1 and G11, which are located at the 5’- and 3’-ends, respectively, of the quadruplex-forming sequence, showed significant (10nm) red-wavelength shifts on quadruplex folding. In contrast no DAS shifts are observed for the G4 position, located in the loop region of the folded quadruplex. Thus, in addition to base stacking of the guanine residues with neighboring bases, guanine positions located at G1 and G11, on average, may also be exposed to a more polar environment with quadruplex folding, possibly due to conformational heterogeneity as they are located near the termini of the telomeric sequence. The solvent accessibility of these positions may make them key docking sites for designing ligands that can stabilize the folded conformation of the G-quadruplex.This work was supported by NIH SCORE Grant S06-GM076168-01 (LD).

Disturbance of B-Cell Homeostasis In Chronic Graft-Versus-Host Disease of the Lung

AbstractAbstract 900 Introduction:Bronchiolitis obliterans syndrome (BOS) characterized by the new development of fixed airflow obstruction after allogeneic hematopoietic cell transplantation (HCT) is an increasingly recognized manifestation of chronic graft-versus-host disease (cGVHD) affecting up to 30% of patients. BOS is associated with increased non-relapse mortality and poor patient outcome after HCT. Its cause is currently unknown but allorecognition of lung antigens is the suspected etiology since BOS has been observed in states of alloimmunity such as cGVHD and after lung transplantation. Currently, the diagnosis of BOS is based on radiologic findings in high resolution expiratory chest CTs (HR-CT) and pulmonary function tests (PFTs). For early diagnosis of BOS biomarkers would be highly desirable allowing efficient treatment of early disease before progression to irreversibility is observed. B-cells are of importance in cGVHD and excess of B-cell activation factor (BAFF), increased CD19+CD21- immature transitional B-cells and dysgammaglobulinemia characterize cGVHD. Objective:We investigated circulating B-cell subpopulations in cGVHD patients to define novel cellular biomarkers for diagnosis of BOS and scoring its severity. Patients and methods:One hundred and thirty-five patients (74 males, 61 females) with a median age of 46 (range, 20–65) years, given myeloablative (n=76) or reduced-intensity (n=59) conditioning for related (n=108) or unrelated donor (n=27) HCT were prospectively evaluated for presence of cGVHD from day 100 after HCT and thereafter serially every 3 months for 2 years. At the same time points PFTs, multicolor flow cytometric analyses of peripheral blood (PB) cells stained for CD19, CD21, CD27, CD10, CD38, IgM, IgD, CD3, CD4, CD8, and CD56, measurements of serum immunoglobulin levels and BAFF were performed. BOS and its severity were defined according to the NIH consensus criteria. All patients with BOS except 2 had HR-CTs of the lung, bronchoalveolar lavage and transbronchial biopsies performed. Results:After a mean follow-up of 250 (range, 100–640) days after HCT 35 of 135 (26%) patients developed BOS. Other manifestations of cGVHD included skin in 72%, eyes in 58%, oral mucosa in 50%, and liver in 65% of patients. Severity of BOS consisted of NIH score I (FEV1=60-79%) in 14 (40%), NIH score II (FEV1=40-59%) in 16 (46%), and NIH score III (FEV<39%) in 5 (14%) patients, respectively. On day 100 after HCT serum IgG levels (860 vs 540 mg/dl, p=0.001) and CD4+ T-cells (17 vs 27%, p<0.0001) were significantly different between patients not experiencing cGVHD (n=63) and patients developing BOS (n=35) during follow-up. Furthermore, a significant difference between serum IgG levels (540 vs 780 mg/dl, p=0.009), CD4+ T-cells (27% vs 22%, p=0.04) and CD8+ T-cells (34% vs 42%, p=0.04) was observed between patients developing BOS and others experiencing cGVHD without lung manifestations (n=37). In patients with newly diagnosed BOS significantly higher BAFF levels (8.3 vs 4.0 ng/ml, p=0.0003), CD19+CD21- immature transitional B-cells (19.5 vs 9.8%, p=0.01), BAFF/CD19+ ratios (0.35 vs 0.05, p=0.008), BAFF/CD21- ratios (1.33 vs 0.43, p=0.01), CD19+CD21-/CD19+CD27+ ratios (4.7 vs 1.9, p=0.04) and CD4+ T-cells (24.6 vs 19.6%, p=0.03) compared to patients without evidence of cGVHD were observed. Furthermore, patients with newly diagnosed BOS had a significant deficiency of CD19+ B-cells (160 vs 254 × 106/L, p=0.027) and CD19+CD27+ memory B-cells (72 vs 154 × 106/L, p=0.01), and significant lower serum IgG levels (570 vs 1260 mg/dl, p<0.001) compared to patients without cGVHD assessed at the same time point after HCT. A comparison of BOS patients with others presenting cGVHD without lung involvement at the same time point after HCT (day 250) revealed significant differences in BAFF levels (8.3 vs 4.4 ng/ml, p<0.0001), BAFF/CD19+ ratios (0.35 vs 0.06, p=0.005) and BAFF/CD21- ratios (1.33 vs 0.517, p=0.012), respectively. Numbers of B-cell subsets and BAFF levels did not differ between lung severity scores, but mild BOS patients had significantly higher BAFF levels (6.4 vs 2.8 ng/ml, p=0.015) compared to patients with mild cGVHD without lung involvement. Conclusion:Disturbance of B-cell homeostasis is more pronounced in cGVHD of the lung compared to other main organ manifestations. B-cell subpopulation numbers and excess of BAFF can serve as novel cellular biomarkers for BOS. Disclosures:No relevant conflicts of interest to declare.

Diagnostic approach to the hyper-IgE syndromes: Immunologic and clinical key findings to differentiate hyper-IgE syndromes from atopic dermatitis

Hyper-IgE syndromes (HIES) are primary immunodeficiency disorders characterized by Staphylococcus aureus abscesses, recurrent pneumonia, increased serum IgE levels, and eczema. The association of heterozygous signal transducer and activator of transcription 3 (STAT3) mutations with autosomal dominant (AD)-HIES allows the differentiation of AD-HIES from disorders associated with eczema and increased serum IgE levels, such as other primary immunodeficiencies and atopic dermatitis. To facilitate early diagnosis of AD-HIES to initiate appropriate therapy. The clinical phenotype (suggested by a National Institutes of Health [NIH] score of >or=40 points), STAT3 genotype, and T(H)17 cell counts were compared in a cohort of 78 patients suspected of having HIES. Heterozygous STAT3 missense mutations and in-frame deletions were identified in 48 patients, all but 2 with an NIH score >or=40 points. Patients with STAT3 mutations with HIES showed significantly lower T(H)17 cell counts compared with patients with wild-type STAT3 and control subjects. Only 1 patient with wild-type STAT3 had both an NIH score >or=40 points and abnormal T(H)17 cell counts (<or=0.2% of CD4(+) cells), with this exception being identified with a homozygous dedicator of cytogenesis 8 protein (DOCK8) mutation. Pathologic shedding of primary teeth was present in 3 patients with wild-type STAT3 and 33 patients with STAT3 mutations. Internal abscesses and severe infections were exclusively seen in patients with STAT3 mutations, who also had increased pneumatocele formation and skeletal or connective tissue manifestations compared with patients with wild-type STAT3. We expanded the number of STAT3 mutations and validated that the NIH score sensitively identifies patients with HIES. Based on our patient cohort, we propose key findings that, when combined with T(H)17 cell numbers, predict patients with AD-HIES with STAT3 mutations, supporting early diagnosis of AD-HIES.

795 THE ROLE OF MEPARTRICIN IN CATEGORY III CHRONIC NONBACTERIAL PROSTATITIS/ CHRONIC PELVIC PAIN SYNDROME (CPPS): A RANDOMIZED PROSPECTIVE PLACEBO-CONTROLLED TRIAL

You have accessJournal of UrologyInfections/Inflammation of the Genitourinary Tract: Prostate & Genitalia1 Apr 2010795 THE ROLE OF MEPARTRICIN IN CATEGORY III CHRONIC NONBACTERIAL PROSTATITIS/ CHRONIC PELVIC PAIN SYNDROME (CPPS): A RANDOMIZED PROSPECTIVE PLACEBO-CONTROLLED TRIAL Aldo Franco De Rose, Paolo Traverso, Tommaso Montanaro, and Giorgio Carmignani Aldo Franco De RoseAldo Franco De Rose More articles by this author , Paolo TraversoPaolo Traverso More articles by this author , Tommaso MontanaroTommaso Montanaro More articles by this author , and Giorgio CarmignaniGiorgio Carmignani More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1471AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Primary outcome is verify the efficacy of mepartricin versus placebo with regard to the improvement of symptoms in patients with CPPS. Secondary outcome is record hormonal assessment of these patients. METHODS 30 patients with CPPS have been included in our study and randomized in two groups of 15 subjects each. Group A patients were treated with mepartricin (80mg daily). Group B patients with placebo. All patients underwent the treatment for 60 days. At the beginning and at the end of therapy, all patients were evaluated using a standardized history, physical examination, LH, FSH, testosterone, beta estradiol dosages and a NIH-CPSI questionnaire. RESULTS We observed a decrease in total NIH score from 24.0 to 6.0 in group A, and from 25.0 to 19.0 in group B, showing about a 70% and 18% improvement in Group A and Group B, respectively. A statistically significant decrease was observed with regard to pain (from 12.0 to 3.0 and from 10.0 to 7.0, respectively) and to quality of life (from 10.0 to 4.0 and from 10.0 to 9.0). No significant difference was observed in urinary dysfunctions. LH, FSH, and testosterone values were similar in both groups before and after treatment, whereas 17 beta estradiol levels were significantly lower in Group A with respect to Group B, at the end of the study. CONCLUSIONS Mepartricin, as showed in the past, provides a significant symptomatic improvement in men with CPPS, versus the treatment with placebo. In this study we tested 80 mg a day, with an higher response than a previous trial performed with 40 mg. From this data we can confirm the role of mepartricin to decrease estrogen plasmatic levels and their concentration in the prostate, perhaps related to the recorded clinical improvement. Genoa, Italy© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e311 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Aldo Franco De Rose More articles by this author Paolo Traverso More articles by this author Tommaso Montanaro More articles by this author Giorgio Carmignani More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Conformational effects induced by YC‐1 and carbon monoxide binding to soluble guanylate cyclase

Soluble guanylate cyclase binds the gaseous ligands NO and CO. Active sGC starts a signaling cascade by converting GTP to cGMP, resulting in vasodilation and neurotransmission. NO activates sGC at least 200‐fold, while CO‐sGC only marginally increases activity. However, in the presence of the allosteric effector YC‐1, the CO complex is activated close to that of the NO complex. Our objective is to understand the molecular mechanism of how YC‐1 binding induces sGC activation by CO. We used MCD to probe the electronic environment of the heme sensing site, and FRET to probe conformational changes as a function of YC‐1 binding. Our findings indicate that the heme is not affected by YC‐1 binding, and that the conformational effects of CO/YC‐1 resemble those caused by NO. These results suggest that a region outside of the heme environment can bind small molecules and induce conformational changes required for sensitization to CO. This work was supported by the following grants: NIH SCORE GM52588(RE) and NIH MCMHD MD000544 (RE), and fellowships NIH MBRS MARC 5T34 GM08574 (IL) and NIH RISE R25‐GM59298 (JK)

Very Early Rehabilitation or Intensive Telemetry after Stroke: A Pilot Randomised Trial

Background: Stroke patients are more likely to make a good recovery if they receive care in a well-organised stroke unit. However, there are uncertainties about how best to provide such care. We studied 2 key aspects of early stroke unit care: early active mobilisation (EM) and automated monitoring (AM) for physiological complications such as hypoxia. Methods: This was an observer-blinded, factorial (2 × 2) pilot randomised controlled trial recruiting stroke patients within 36 h of symptom onset. The patients were randomised to 1 of 4 nurse-led treatment protocols: (a) standard stroke unit care, (b) EM, (c) AM or (d) combined EM and AM. The primary outcome was the Rankin score at 3 months. We also report the data on feasibility and safety. Results: We randomised 32 patients (mean age = 65 years; mean baseline modified NIH score = 6). On unadjusted comparisons, the EM patients were significantly (p < 0.05) more likely to mobilise very early (within 1 h of randomisation) and to achieve walking by day 5 and were less likely to develop complications of immobility. The AM group was significantly (p < 0.05) more likely to have pre-defined physiological complication events detected. All these associations remained, but were less statistically significant, after correcting for age, baseline NIH score and co-interventions. There were no significant safety concerns. Discussion: We have demonstrated the feasibility of implementing EM and AM for physiological complications in a randomised controlled trial. Larger trials are warranted to determine whether these interventions have clinical benefits.

G-Quadruplex Folding Observed by two Photon Fluorescence Correlation Spectroscopy and Dual Time Scale

G-rich DNA sequences are known to fold upon addition of salt into a stacked well defined configuration called a quadruplex. A fluorescently labeled 5′-FAM −24-mer G-quadruplex sequence was used to explore the variation of diffusion coefficients at extremely low, low and high KCl concentrations. We found a shift in the diffusion coefficient of about 10μm2/sec toward faster diffusion from extremely low to high KCl concentrations. This shift can be related to the compact structure formed by the G-quadruplex. We have also used a fluorescent guanosine analog, 6MI, to label a 24mer that has shown folding behavior at high KCl concentrations. To explore this further, we have added in excess a sequence that complements the G-rich region to deter the formation of the G-quadruplex. The diffusion coefficient also increased from the unfolded, low KCl concentration to the high salt, G-quadruplex structure. We have constructed a dual-timescale (ps TCSPC and uS-mS FCS) photon correlation system and we are using it to explore linked changes in the fluorophores' lifetimes and the translational diffusion coefficients as they move between low and high salt environments. Part of this work was supported by NIH SCORE Grant S06 GM 060654.

Local Dynamic Studies of Guanine Residues within the Human Telomeric DNA G-Quadruplexed Conformation

Formation and stabilization of guanine-rich G-quadruplexed DNA conformations can inhibit the abnormal activity of the enzyme telomerase in tumor cells, making it a target for potential cancer therapeutics. To study the effect individual guanine residues have on the folding process and stabilization of the G-quadruplex conformations, the fluorescence of HT4 oligonucleotides incorporating the fluorescent guanine analog 6-methyl-8-(2-deoxy-D-ribofuranosyl) isoxanthopterin (6MI) into different tetrads of the quadruplex (G1,G4,G5,G9 and G11) were investigated. This guanine probe exhibits changes in fluorescence intensity sensitive to base-stacking and hydrogen-bonding. Fluorescence intensities quench for G4, G5, G9 and G11, and de-quench for G1 when each 6MI-labelled oligonucleotide folds to the G-quadruplex conformation with addition of K+-ion. This suggests stronger base-stacking interactions with neighboring bases for G4, G5, G9 and G11, compared with G1 located on the 5’-end. Fluorescence intensity peaks observed for G1 and G11 also show significant red wavelength shifts with folding. This suggests these guanine positions may be more exposed to a polar environment within the folded state. Fluorescence lifetime studies of the labeled quadruplex sequences reveal that the observed intensity quenching arises predominantly from fast (sub-nanosecond) quasi-static self-quenching. This self-static quenching apparently arises from the proximity of 6MI to neighboring bases. The “dark” component (Adark) dominates the decay behavior for both the folded and unfolded conformations of each 6MI-labeled sequence. With folding, the contribution of Adark increases for all labeled oligonucleotide sequences as the conformation is now more compact. This effect is greatest for those 6MI replacements located in the loop regions. Overall these studies suggest individual guanines play different roles in the stabilization of G-quadruplex structures. This work was supported by NIH SCORE Grant S06 GM 060654.