Recurrent Angiolymphoid Hyperplasia with Eosinophilia of the Parotid.

Angiolymphoid hyperplasia with eosinophilia (ALHE) is a benign vasculoproliferative disorder. It usually affects young adults presenting as papules or nodules involving the skin of head and neck region and rarely involves extracutaneous sites. ALHE involving parotid is rare and can be a diagnostic dilemma as it mimics a parotid neoplasm. This is a case of a 23 year old male presenting with a recurrent swelling over the left parotid region post surgery. Ultrasonography revealed a vascular soft tissue lesion in the preauricular region suggestive of a benign lesion. Contrast enhanced magnetic resonance imaging showed a hyperintense lesion involving the superficial lobe of the left parotid gland. Patient underwent superficial parotidectomy and histopathologically was diagnosed to have ALHE. Very few cases have been reported and this case is highlighted as the facial nerve was enmeshed by the intraparotid lesion which was a surgical challenge.

Comparison of Two High Dose Rate Brachytherapy Regimes.

Background: Two HDR brachytherapy regimes were compared, 9.5 Gray per fraction for two fractions and 7.5 Gray per fraction for three fractions. Materials and Methods: A total of 80 patients with histologically evident squamous cell carcinoma cervix were taken in the current research after randomization. Radiotherapy dose delivered was 50 Gray/25#/5 weeks with concomitant chemotherapy with weekly cisplatin 35 mg/m2. Following external chemoradiation, patients were randomized into two arms. In Arm A, 40 patients were given high dose rate (HDR) brachytherapy of weekly 7.5 Gray in three fractions over 3 weeks. In Arm B, 40 patients were given high dose rate (HDR) brachytherapy of weekly 9.5 Gray in two fractions over 2 weeks. BED and LQED had been calculated, and the evaluation of response and consequences was examined. Results: In Arm A, BED to point A was 99.38 ± 0.00 and EQD2 to point A was 82.81 ± 0.00. In Arm B, BED to point A was 97.05 ± 0.00 and EQD2 to point A was 80.89 ± 0.00. With respect to rectum in Arm A, BED rectum was 108.66 ± 11.43 and EQD2 rectum was 65.07 ± 6.84. In Arm B, BED rectum was 107 ± 10.83 and EQD2 rectum was 64.21 ± 6.49. Similarly in Arm A, BED bladder was 107.86 ± 10.23 and EQD2 bladder was 64.59 ± 6.13. In arm B, BED bladder was 104.14 ± 10.79 and EQD2 bladder was 62.36 ± 6.46. Conclusion: In a follow-up of 6 months, no statistical significance in terms of local control as well as complications rates in both the arms. Our research demonstrates that two fractions of HDR are comparable with three fractions of HDR.

CD26lowPD-1+ CD8 T cells are terminally exhausted and associated with leukemia progression in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a devastating blood cancer with poor prognosis. Novel effective treatment is an urgent unmet need. Immunotherapy targeting T cell exhaustion by blocking inhibitory pathways, such as PD-1, is promising in cancer treatment. However, results from clinical studies applying PD-1 blockade to AML patients are largely disappointing. AML is highly heterogeneous. Identification of additional immune regulatory pathways and defining predictive biomarkers for treatment response are crucial to optimize the strategy. CD26 is a marker of T cell activation and involved in multiple immune processes. Here, we performed comprehensive phenotypic and functional analyses on the blood samples collected from AML patients and discovered that CD26lowPD-1+ CD8 T cells were associated with AML progression. Specifically, the percentage of this cell fraction was significantly higher in patients with newly diagnosed AML compared to that in patients achieved completed remission or healthy controls. Our subsequent studies on CD26lowPD-1+ CD8 T cells from AML patients at initial diagnosis demonstrated that this cell population highly expressed inhibitory receptors and displayed impaired cytokine production, indicating an exhaustion status. Importantly, CD26lowPD-1+ CD8 T cells carried features of terminal exhaustion, manifested by higher frequency of TEMRA differentiation, increased expression of transcription factors that are observed in terminally exhausted T cells, and high level of intracellular expression of granzyme B and perforin. Our findings suggest a prognostic and predictive value of CD26 in AML, providing pivotal information to optimize the immunotherapy for this devastating cancer.

Vagus nerve stimulation primes platelets and reduces bleeding in hemophilia A male mice.

Deficiency of coagulation factor VIII in hemophilia A disrupts clotting and prolongs bleeding. While the current mainstay of therapy is infusion of factor VIII concentrates, inhibitor antibodies often render these ineffective. Because preclinical evidence shows electrical vagus nerve stimulation accelerates clotting to reduce hemorrhage without precipitating systemic thrombosis, we reasoned it might reduce bleeding in hemophilia A. Using two different male murine hemorrhage and thrombosis models, we show vagus nerve stimulation bypasses the factor VIII deficiency of hemophilia A to decrease bleeding and accelerate clotting. Vagus nerve stimulation targets acetylcholine-producing T lymphocytes in spleen and α7 nicotinic acetylcholine receptors (α7nAChR) on platelets to increase calcium uptake and enhance alpha granule release. Splenectomy or genetic deletion of T cells or α7nAChR abolishes vagal control of platelet activation, thrombus formation, and bleeding in male mice. Vagus nerve stimulation warrants clinical study as a therapy for coagulation disorders and surgical or traumatic bleeding.

Real-world evidence of the safety and survival with CD19 CAR-T cell therapy for relapsed/refractory solid organ transplant-related PTLD.

The use of CD19 chimeric antigen receptor T-cell (CAR-T) therapy for relapsed/refractory solid organ transplantation (SOT)-related post-transplant lymphoproliferative disorder (PTLD) is not well studied. We conducted a multicentre, retrospective analysis of adults with relapsed/refractory SOT-associated PTLD. Among 22 relapsed/refractory SOT-PTLD patients, the pathology was monomorphic B cell. Prior SOTs included 14 kidney (64%), three liver (14%), two heart (9%), one intestinal (5%), one lung (5%), and one pancreas after kidney transplant (5%). The median time from SOT to PTLD diagnosis was 107 months. Pre-CAR-T bridging therapy was used in 55% of patients, and immunosuppression was stopped completely before CAR-T infusion in 64%. Eighteen (82%) patients experienced cytokine release syndrome: one (5%) each grade (G) 3 and G4. The immune effector cell-associated neurotoxicity syndrome was observed in 16 (73%) patients: six (27%) G3 and two (9%) G4. The overall response rate was 64% (55% complete response). Three patients (14%) experienced allograft rejection after CAR-T. The two-year progression-free survival and overall survival rates were 35% and 58%, respectively. Additionally, the achievement of CR post-CAR-T was strongly associated with survival. Collectively, the safety and efficacy of CD19 CAR-T therapy in relapsed/refractory SOT-related PTLD appeared similar to pivotal CAR-T data, including approximately one-third of patients achieving sustained remission.

High-grade B-cell lymphoma, not otherwise specified: a multi-institutional retrospective study.

In this multi-institutional retrospective study, we examined the characteristics and outcomes of 160 patients with high-grade B-cell lymphoma, not otherwise specified (HGBL-NOS)-a rare category defined by high-grade morphologic features and lack of MYC rearrangements with BCL2 and/or BCL6 rearrangements ("double hit"). Our results show that HGBL-NOS tumors are heterogeneous: 83% of patients had a germinal center B-cell immunophenotype, 37% a dual-expressor immunophenotype (MYC and BCL2 expression), 28% MYC rearrangement, 13% BCL2 rearrangement, and 11% BCL6 rearrangement. Most patients presented with stage IV disease, a high serum lactate dehydrogenase, and other high-risk clinical factors. Most frequent first-line regimens included dose-adjusted cyclophosphamide, doxorubicin, vincristine, and etoposide, with rituximab and prednisone (DA-EPOCH-R; 43%); rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP; 33%); or other intensive chemotherapy programs. We found no significant differences in the rates of complete response (CR), progression-free survival (PFS), or overall survival (OS) between these chemotherapy regimens. CR was attained by 69% of patients. PFS at 2 years was 55.2% and OS was 68.1%. In a multivariable model, the main prognostic factors for PFS and OS were poor performance status, lactate dehydrogenase >3 × upper limit of normal, and a dual-expressor immunophenotype. Age >60 years or presence of MYC rearrangement were not prognostic, but patients with TP53 alterations had a dismal PFS. Presence of MYC rearrangement was not predictive of better PFS in patients treated with DA-EPOCH-R vs R-CHOP. Improvements in the diagnostic criteria and therapeutic approaches beyond dose-intense chemotherapy are needed to overcome the unfavorable prognosis of patients with HGBL-NOS.

Post-transplant cyclophosphamide alters immune signatures and leads to impaired T cell reconstitution in allogeneic hematopoietic stem cell transplant.

Despite the increased usage of post-transplant cyclophosphamide (PTCy) in allogeneic hematopoietic stem cell transplantation (allo-HSCT), our knowledge of immune reconstitution post-allo-HSCT in the setting of PTCy is limited. Adequate immune reconstitution is the key to a successful transplant. In this study, we aim to investigate the effect of PTCy on the reconstitution of each immune component; more focus was placed on the immunophenotype and functions of T cells. Using blood samples from patients who underwent allo-HSCT under regimens containing PTCy (n = 23) versus those who received no PTCy (n = 14), we examined the impact of PTCy on the post-transplant immune response. We demonstrated a distinct T cell immune signature between PTCy versus non-PTCy group. PTCy significantly delayed T cell reconstitution and affected the T cell subsets by increasing regulatory T cells (Treg) while reducing naïve T cells. In addition, we observed remarkable enhancement of multiple inhibitory receptors (TIGIT, PD-1, TIM-3, CD38, CD39) on both CD4+ and CD8+ T cells on day 30 post-transplantation in patients who received PTCy. Importantly, upregulation of PD-1 on CD8 T cells was persistent through day 180 and these T cells were less functional, manifested by reduced cytokine production upon anti-CD3/CD28 stimulation. Furthermore, we found a significant correlation of T cell immune phenotypes to clinical outcome (disease relapse and GVHD) in patients who received PTCy. Our novel findings provide critical information to understand the mechanism of how PTCy impacts immune reconstitution in allo-HSCT and may subsequently lead to optimization of our clinical practice using this treatment.

Re-examining attention capture at irrelevant (ignored?) locations.

In 2018, Ruthruff and Gaspelin used a modified spatial cuing paradigm in which targets were presented at two locations while abrupt-onset cues could be presented at four locations. They found that performance following cues presented at irrelevant locations was no worse than following no cue or following a centrally presented cue. They concluded, as conveyed by the title of their article (Immunity to Attentional Capture at Ignored Locations) that a spatial attentional control setting had eliminated capture of attention. This conclusion was reached by comparing response time to targets on cue-absent versus irrelevant cues condition. We administered the exact same task in Experiment 1 and observed that responses on irrelevant trials were faster compared with cue absent trials providing support for the "immunity to attention capture claim" made by Ruthruff and Gaspelin (2018). However, cue absent trials may not be the most appropriate baseline condition as they lack the alerting benefit provided by cue-present trials. Thus, equivalent response times (RTs) on trials with absent cues and irrelevant cues observed in Ruthruff and Gaspelin (2018) could have been due to the lack of this alerting benefit. We tested this in Experiment 2 by additionally including a warning beep on every trial as an alerting signal. With this methodological change, we observed that responses were slower on irrelevant trials compared with the cue absent trials suggesting interference from cues at irrelevant locations. This study underscores the importance of using the appropriate baseline while testing attention capture. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Aeromonas hydrophila RIT668 and Citrobacter portucalensis RIT669-Potential Zoonotic Pathogens Isolated from Spotted Turtles.

Antimicrobial resistance (AMR) is one of the biggest challenges of the 21st century, and biofilm formation enables bacteria to resist antibiotic at much higher concentrations than planktonic cells. Earlier, we showed that the Gram-negative Aeromonas hydrophila RIT668 and Citrobacter portucalensis RIT669 (closely related to C. freundii NBRC 12681) from infected spotted turtles (Clemmys guttata), formed biofilms and upregulated toxin expression on plastic surfaces, and were predicted to possess multiple antibiotic resistance genes. Here, we show that they each resist several antibiotics in the planktonic phase, but were susceptible to neomycin, and high concentrations of tetracycline and cotrimoxazole. The susceptibility of their biofilms to neomycin and cotrimoxazole was tested using the Calgary device. For A. hydrophila, the minimum inhibitory concentration (MIC) = 500-1000, and the minimum biofilm eradication concentration (MBEC) > 1000 µg/mL, using cotrimoxazole, and MIC = 32.3-62.5, and MBEC > 1000 µg/mL, using neomycin. For C. freundii MIC = 7.8-15.6, and, MBEC > 1000 µg/mL, using cotrimoxazole, and MIC = 7.8, and MBEC > 1000 µg/mL, using neomycin. Both A. hydrophila and C. portucalensis activated an acyl homoserine lactone (AHL) dependent biosensor, suggesting that quorum sensing could mediate biofilm formation. Their multidrug resistance in the planktonic form, and weak biofilm eradication even with neomycin and cotrimoxazole, indicate that A. hydrophila and C. portucalensis are potential zoonotic pathogens, with risks for patients living with implants.

Expression of a Shiga-Like Toxin during Plastic Colonization by Two Multidrug-Resistant Bacteria, Aeromonas hydrophila RIT668 and Citrobacter freundii RIT669, Isolated from Endangered Turtles (Clemmys guttata).

Aeromonas hydrophila RIT668 and Citrobacter freundii RIT669 were isolated from endangered spotted turtles (Clemmys guttata). Whole-genome sequencing, annotation and phylogenetic analyses of the genomes revealed that the closest relative of RIT668 is A. hydrophila ATCC 7966 and Citrobacter portucalensis A60 for RIT669. Resistome analysis showed that A. hydrophila and C. freundii harbor six and 19 different antibiotic resistance genes, respectively. Both bacteria colonize polyethylene and polypropylene, which are common plastics, found in the environment and are used to fabricate medical devices. The expression of six biofilm-related genes-biofilm peroxide resistance protein (bsmA), biofilm formation regulatory protein subunit R (bssR), biofilm formation regulatory protein subunit S (bssS), biofilm formation regulator (hmsP), toxin-antitoxin biofilm protein (tabA) and transcriptional activator of curli operon (csgD)-and two virulence factors-Vi antigen-related gene (viaB) and Shiga-like toxin (slt-II)-was investigated by RT-PCR. A. hydrophila displayed a >2-fold increase in slt-II expression in cells adhering to both polymers, C. freundii adhering on polyethylene displayed a >2-fold, and on polypropylene a >6-fold upregulation of slt-II. Thus, the two new isolates are potential pathogens owing to their drug resistance, surface colonization and upregulation of a slt-II-type diarrheal toxin on polymer surfaces.

Estimating policy-relevant health effects of ambient heat exposures using spatially contiguous reanalysis data.

BACKGROUND: Regional National Weather Service (NWS) heat advisory criteria in New York State (NYS) were based on frequency of heat events estimated by sparse monitoring data. These may not accurately reflect temperatures at which specific health risks occur in large geographic regions. The objectives of the study were to use spatially resolved temperature data to characterize health risks related to summertime heat exposure and estimate the temperatures at which excessive risk of heat-related adverse health occurs in NYS. We also evaluated the need to adjust current heat advisory threshold and messaging based on threshold temperatures of multiple health outcomes. METHODS: We assessed the effect of multi-day lag exposure for maximum near-surface air temperature (Tmax) and maximum Heat Index derived from the gridded National Land Data Assimilation System (NLDAS) reanalysis dataset on emergency department (ED) visits/ hospitalizations for heat stress, dehydration, acute kidney failure (AKF) and cardiovascular diseases (CVD) using a case-crossover analysis during summers of 2008-2012. We assessed effect modification using interaction terms and stratified analysis. Thresholds were estimated using piecewise spline regression. RESULTS: We observed an increased risk of heat stress (Risk ratio (RR) = 1.366, 95% confidence interval (CI): 1.347, 1.386) and dehydration (RR = 1.024, 95% CI: 1.021, 1.028) for every 1 °C increase in Tmax on the day of exposure. The highest risk for AKF (RR = 1.017, 95% CI: 1.014, 1.021) and CVD (RR = 1.001, 95% CI: 1.000, 1.002) were at lag 1 and 4 respectively. The increased risk of heat-health effects persists up to 6 days. Rural areas of NYS are at as high a risk of heat-health effects as urban areas. Heat-health risks start increasing at temperatures much lower than the current NWS criteria. CONCLUSION: Reanalysis data provide refined exposure-response functions for health research, in areas with sparse monitor observations. Based on this research, rural areas in NYS had similar risk for health effects of heat. Heat advisories in New York City (NYC) had been reviewed and lowered previously. As such, the current NWS heat advisory threshold was lowered for the upstate region of New York and surrounding areas. Enhanced outreach materials were also developed and disseminated to local health departments and the public.

Accessibility of cooling centers to heat-vulnerable populations in New York State.

Introduction: Spending a few hours to cool down in a cooling center reduces the impact of heat on health. But limited or lack of accessibility of these facilities is often a barrier to their utilization. The objective of this study was to assess accessibility of the cooling centers to heat-vulnerable populations in New York State (NYS) by various modes of transportation. Methods: We estimate the proximity of 377 cooling centers to general and heat-vulnerable populations in NYS (excluding New York City (NYC)) and determine their accessibility via walking, public transportation and driving. Distances between tract populations and nearest cooling center, and between cooling centers and public transportation stops were estimated. Accessibility in four metropolitan regions was determined via public transportation while accessibility in heat-vulnerable rural areas was estimated via driving. Results: Distances to nearest cooling center ranged from 0 to 53.2 miles with only a third of NYS population within walking distance (0.5 miles) of a cooling center. About 51% of heat-vulnerable tracts were within 0.5 miles, with an average distance of 2.4 miles to the nearest cooling center. Within the four metro politan regions 80% of cooling centers within 0.5 miles of a public transportation stop. All cooling centers in heat-vulnerable tracts were accessible via public transportation. In rural heat-vulnerable tracts, driving distances averaged at about 18 miles. Conclusions: In urban areas many residents were not within walking distance of a cooling center, but most, and nearly all in the most heat-vulnerable areas, were within walking distance of public transportation to a cooling center. In rural locations distances were longer, and accessibility is a greater issue. Cooling centers can be a valuable resource for general and heat-vulnerable populations during an extreme heat event. When planning and implementing cooling centers, it is therefore important to improve accessibility and address other barriers that can hamper their utilization.

Surveying Local Health Departments and County Emergency Management Offices on Cooling Centers as a Heat Adaptation Resource in New York State.

Local agencies in New York State (NYS) set up cooling centers to provide relief from summer-time heat especially for people with limited access to air-conditioning. We aimed to determine cooling center locations in NYS, and explore county agencies' involvement in organizing and promoting utilization of cooling centers. We conducted a survey among county health and emergency preparedness offices in NYS (excluding NYC) and explored official county websites. We identified 377 cooling centers, mostly in metropolitan areas of NYS. Although 47 % of counties listed locations online, only 29 % reported locations via survey. Radio (90 %) and internet (84 %) were popular for information dissemination. Air-conditioning was available at all indoor cooling center facilities. Cooling centers in 13 % of the counties were accessible by either public transportation or shuttles arranged by the facility. About 38 % counties do not consider cooling centers important in their region or promote informal cooling centers. More than a third of New York counties had neither cooling centers nor plans to establish a cooling center as extreme heat was not perceived as a threat in their region.

Iconic Native Culture Cues Inhibit Second Language Production in a Non-immigrant Population: Evidence from Bengali-English Bilinguals.

We examined if iconic pictures belonging to one's native culture interfere with second language production in bilinguals in an object naming task. Bengali-English bilinguals named pictures in both L1 and L2 against iconic cultural images representing Bengali culture or neutral images. Participants named in both "Blocked" and "Mixed" language conditions. In both conditions, participants were significantly slower in naming in English when the background was an iconic Bengali culture picture than a neutral image. These data suggest that native language culture cues lead to activation of the L1 lexicon that competed against L2 words creating an interference. These results provide further support to earlier observations where such culture related interference has been observed in bilingual language production. We discuss the results in the context of cultural influence on the psycholinguistic processes in bilingual object naming.

Associations between summertime ambient pollutants and respiratory morbidity in New York City: comparison of results using ambient concentrations versus predicted exposures.

Epidemiological analyses of air quality often estimate human exposure from ambient monitoring data, potentially leading to exposure misclassification and subsequent bias in estimated health risks. To investigate this, we conducted a case-crossover study of summertime ambient ozone and fine particulate matter (PM(2.5)) levels and daily respiratory hospitalizations in New York City during 2001-2005. Comparisons were made between associations estimated using two pollutant exposure metrics: observed concentrations and predicted exposures from the EPA's Stochastic Human Exposure and Dose Simulation (SHEDS) model. Small, positive associations between interquartile range mean ozone concentrations and hospitalizations were observed and were strongest for 0-day lags (hazard ratio (HR)=1.013, 95% confidence interval (CI): 0.998, 1.029) and 3-day lags (HR=1.006, 95% CI: 0.991, 1.021); applying mean predicted ozone exposures yielded similar results. PM(2.5) was also associated with admissions, strongest at 2- and 4-day lags, with few differences between exposure metrics. Subgroup analyses support recognized sociodemographic differences in concentration-related hospitalization risk, whereas few inter-stratum variations were observed in relation to SHEDS exposures. Predicted exposures for these spatially homogenous pollutants were similar across sociodemographic strata, therefore SHEDS predictions coupled with the case-crossover design may have masked observable heterogeneity in risks. However, significant effect modification was found for subjects in the top exposure-to-concentration ratio tertiles, suggesting risks may increase as a consequence of infiltration or greater exposure to outdoor air.

Returning to health visiting practice: completing the circle.

One strategic health authority, NHS London, initiated a pilot return to health visiting/nursing practice scheme in London in 2010. This paper reports on the experiences of the first three cohorts of returnees on the City University London programme, one of the London programmes, and the adaptations that have been made to the programme to help provide returnees with the theory base and practice experience to equip them to work in today's health visiting. Written evaluation forms were completed by the returnees and information gathered from their application forms. This information was supplemented for Cohort 1 with some interviews with practice teachers and lecturers and a mid-stage questionnaire to the returnees. Of the 54 students in the three cohorts over half were still on one or both Nursing and Midwifery Council registers, which had not been anticipated at the start of the programme and led to modifications to the programme after Cohort 1 with an increase in the health visiting specific content. The returnees had a wide range of experience to bring back to health visiting reflecting the fact that a large number had been out of health visiting for more than 11 years. The evaluation shows that providing support by the university to the practice placement areas; ensuring that the taught element is current and useful to health visiting practice and having a relevant but not too onerous assessment process are critical.

Stressful events, smoking exposure and other maternal risk factors associated with gestational diabetes mellitus.

The incidence of gestational diabetes mellitus (GDM) has increased significantly in the last few decades in the US. Understanding its risk factors is imperative for the prevention of GDM and its sequelae, but the roles of behavioural risk factors such as stressful events and smoking on GDM are generally not well understood. Using data obtained from the New York State (NYS) Pregnancy Risk Assessment Monitoring System survey for 2004-06 and the NYS birth certificates, we examined relationships between GDM, stressful events and smoking among 2690 women who had live singleton births and did not have pre-pregnancy diabetes. After adjustment for risk factors such as maternal age, race/ethnicity, pre-pregnancy body mass index, hypertension, as well as smoking exposure, education, parity, and gestation at first visit for prenatal care, we found that having five or more stressful events 12 months before the baby was born was significantly associated with GDM (OR = 2.49, [95% CI 1.49, 4.16]). In another model, having any stressful event(s) other than 'moved to a new address' 12 months before the baby was born was also moderately associated with GDM (OR = 1.38, [95% CI 1.04, 1.85]). Smoking exposure, assessed by combining maternal smoking and second-hand smoke exposure into six levels, had no significant association with GDM, and did not show a dose-response pattern. The present study suggests that stressful events during pregnancy may be an independent risk factor for GDM. Future studies of GDM should include this common, but potentially modifiable risk factor in analyses.

Oligomerization of the E. coli core RNA polymerase: formation of (α2ßß'ω)2-DNA complexes and regulation of the oligomerization by auxiliary subunits.

In this work, using multiple, dissimilar physico-chemical techniques, we demonstrate that the Escherichia coli RNA polymerase core enzyme obtained through a classic purification procedure forms stable (α(2)ßß'ω)(2) complexes in the presence or absence of short DNA probes. Multiple control experiments indicate that this self-association is unlikely to be mediated by RNA polymerase-associated non-protein molecules. We show that the formation of (α(2)ßß'ω)(2) complexes is subject to regulation by known RNA polymerase interactors, such as the auxiliary SWI/SNF subunit of RNA polymerase RapA, as well as NusA and σ(70). We also demonstrate that the separation of the core RNA polymerase and RNA polymerase holoenzyme species during Mono Q chromatography is likely due to oligomerization of the core enzyme. We have analyzed the oligomeric state of the polymerase in the presence or absence of DNA, an aspect that was missing from previous studies. Importantly, our work demonstrates that RNA polymerase oligomerization is compatible with DNA binding. Through in vitro transcription and in vivo experiments (utilizing a RapA(R599/Q602) mutant lacking transcription-stimulatory function), we demonstrate that the formation of tandem (α(2)ßß'ω)(2)-DNA complexes is likely functionally significant and beneficial for the transcriptional activity of the polymerase. Taken together, our findings suggest a novel structural aspect of the E. coli elongation complex. We hypothesize that transcription by tandem RNA polymerase complexes initiated at hypothetical bidirectional "origins of transcription" may explain recurring switches of the direction of transcription in bacterial genomes.

RapA, Escherichia coli RNA polymerase SWI/SNF subunit-dependent polyadenylation of RNA.

In this work, we describe RapA-dependent polyadenylation of model RNA substrates and endogenous, RNA polymerase-associated nucleic acid fragments. We demonstrate that the Escherichia coli RNA polymerase obtained through the classic purification procedure carries endogenous RNA oligonucleotides, which, in the presence of ATP, are polyriboadenylated in a RapA-dependent manner by an accessory poly(rA) polymerase. RNA polymerase isolated from poly(A) polymerase- (PAP-) and polynucleotide phosphorylase- (PNP-) deficient E. coli strain lacks accessory (rA)(n)-synthetic activity. Experiments with reconstituted RNA polymerase-PAP and RNA polymerase-PNP mixtures suggest that RapA enables the polyadenylation by PAP of RNA polymerase-associated RNA. Mutations disrupting RapA's ATP-hydrolytic function disrupt RapA-dependent polyadenylation, and the rapA(-)E. coli strain displays a measurable reduction in RNA polyadenylation. RapA-dependent polyadenylation can also be modulated by mutations in the section of RapA's SWI/SNF domain linked to interaction with single-stranded nucleic acid. We have developed enzymatic assays in which model, synthetic RNAs are polyriboadenylated in a RapA-dependent manner. Taken together, our results are consistent with RapA acting as an RNA polymerase-associated, ATP-dependent RNA translocase. Our work further links RapA to RNA remodeling and provides new mechanistic insights into the functional interaction between RNA polymerase and RapA.

Agreement between self-report and birth certificate for gestational diabetes mellitus: New York State PRAMS.

This study examined agreement (concordance or convergent validity) between self-report and birth certificate for gestational diabetes. Study population was 2,854 women who had live births 2-6 months earlier and responded to a questionnaire from the New York State Pregnancy Risk Assessment Monitoring System (PRAMS) survey, 2004-2006. Agreement between self-report and birth certificate was assessed for the study population overall, and for subgroups defined by race, age, education, marital status, number of previous live births, time of first prenatal care, and birth weight of the newborn. A total of 258 women self-reported gestational diabetes, while birth certificates indicated that 138 women had gestational diabetes. For the study population overall, percent agreement was 93.8% and Kappa was 0.53. Due to the moderate bias index (68.2% overall, ranged from 33.3 to 100% in subgroups) and the high skewed prevalence index (91.8% overall, ranged from 70.7 to 97.5% in subgroups), we determined Prevalence-Adjusted and Bias-Adjusted Kappa (PABAK) was a better measure of agreement. PABAK was 0.88 overall, indicating very good agreement. PABAK was uniformly high in all subgroups. The highest PABAK was found among women aged 25 years and younger (0.93), and the lowest PABAK was among Asian women (0.79). Although the absence of a gold standard for gestational diabetes hinders assessment of criterion validity, high PABAK measures suggest that self-reporting by PRAMS respondents is feasible for identifying cases of gestational diabetes for surveillance and population-based epidemiologic research.