Regulation of gene expression by chromatin boundary elements

Boundary elements consisting of barriers and insulators are genomic sequence elements that along with their associated DNA-binding proteins block the spread of heterochromatin into euchromatic regions or prevent the targeted activation of promoters from distal/proximal enhancers, respectively. In Saccharomyces cerevisiae, the deletion of RPD3, a histone deacetylase, results in an extended SIR protein-mediated silencing effect bypassing a tRNAthr barrier element adjacent to the cryptic mating locus, HMRa. We mutagenized rpd3Δ strains and identified suppressor mutants through a genetic screen that no longer displayed this enhanced silencing effect. Our results identified BRE1 and BRE2, which are either directly or indirectly responsible for the tri-methylation of histone H3K4 and H3K79, as effectors of the rpd3Δ extended silencing effect at HMRa. We hypothesize that the increased silencing effect in rpd3Δ mutants is the result of a redistribution of SIR proteins which become concentrated at the HMRa region in response to a global change in the acetylation and/or methylation state of histones contingent on RPD3, BRE1, and BRE2. ETC, or Extra-TFIIIC, sites are genomic elements which bind the RNA Polymerase III transcription factor, TFIIIC. ETC sites contain B-box promoter sequences normally associated with RNA Polymerase III promoters, and their locations are over-represented between divergently transcribed RNA Polymerase II genes. Our results show that the transcription of TFC6, which codes for a DNA-binding component of TFIIIC, is auto-regulated by TFIIIC which binds to the ETC6 site in the TFC6 promoter region. Inhibition of TFIIIC binding to the ETC6 site results in increased TFC6 expression from its own promoter, and transcription of TFC6 is inversely correlated with TFIIIC binding to the ETC6 site. The TFC6 promoter is also down-regulated when its own gene product is over-expressed. We present here a novel function of gene regulation where a Pol III transcription factor directly (auto) regulates a Pol II gene. Our results also point to how this regulation might be mediated by an insulator-like function of TFIIIC which can implicate the functionality of Extra-TFIIIC sites in other eukaryotes

The Neural Structures Expressing Perceptual Hysteresis in Visual Letter Recognition

AbstractPerception can change nonlinearly with stimulus contrast, and perceptual threshold may depend on the direction of contrast change. Such hysteresis effects in neurometric functions provide a signature of perceptual awareness. We recorded brain activity with functional neuroimaging in observers exposed to gradual contrast changes of initially hidden visual stimuli. Lateral occipital, frontal, and parietal regions all displayed both transient activations and hysteresis that correlated with change and maintenance of a percept, respectively. Medial temporal activity did not follow perception but increased during hysteresis and showed transient deactivations during perceptual transitions. These findings identify a set of brain regions sensitive to visual awareness and suggest that medial temporal structures may provide backward signals that account for neural and, thereby, perceptual hysteresis

Evaluation of a national universal coverage campaign of long-lasting insecticidal nets in a rural district in north-west Tanzania.

\ud \ud Insecticide-treated nets (ITN) are one of the most effective measures for preventing malaria. Mass distribution campaigns are being used to rapidly increase net coverage in at-risk populations. This study had two purposes: to evaluate the impact of a universal coverage campaign (UCC) of long-lasting insecticidal nets (LLINs) on LLIN ownership and usage, and to identify factors that may be associated with inadequate coverage. In 2011 two cross-sectional household surveys were conducted in 50 clusters in Muleba district, north-west Tanzania. Prior to the UCC 3,246 households were surveyed and 2,499 afterwards. Data on bed net ownership and usage, demographics of household members and household characteristics including factors related to socio-economic status were gathered, using an adapted version of the standard Malaria Indicator Survey. Specific questions relating to the UCC process were asked. The proportion of households with at least one ITN increased from 62.6% (95% Confidence Interval (CI) = 60.9-64.2) before the UCC to 90.8% (95% CI = 89.0-92.3) afterwards. ITN usage in all residents rose from 40.8% to 55.7%. After the UCC 58.4% (95% CI = 54.7-62.1) of households had sufficient ITNs to cover all their sleeping places. Households with children under five years (OR = 2.4, 95% CI = 1.9-2.9) and small households (OR = 1.9, 95% CI = 1.5-2.4) were most likely to reach universal coverage. Poverty was not associated with net coverage. Eighty percent of households surveyed received LLINs from the campaign. The UCC in Muleba district of Tanzania was equitable, greatly improving LLIN ownership and, more moderately, usage. However, the goal of universal coverage in terms of the adequate provision of nets was not achieved. Multiple, continuous delivery systems and education activities are required to maintain and improve bed net ownership and usage.\ud \u

Unexpected expression of carbonic anhydrase I and selenium‐binding protein as the only major non‐heme proteins in erythrocytes of the subterranean mole rat (Spalax ehrenbergi)

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/117190/1/feb2s0014579398006905.pd

Comparing the Effectiveness of Malaria Vector-Control Interventions Through a Mathematical Model

Although some malaria-control programs are beginning to combine insecticide-treated nets (ITNs) and indoor residual spraying (IRS), little is known about the effectiveness of such combinations. We use a mathematical model to compare the effectiveness of ITNs and IRS with dichlorodiphenyltrichloroethane (DDT) or bendiocarb, applied singly and in combination, in an epidemiological setting based in Namawala, Tanzania, with Anopheles gambiae as the primary vector. Our model indicates that although both IRS (with DDT) and ITNs provide personal protection, humans with only ITNs are better protected than those with only IRS, and suggests that high coverage of IRS with bendiocarb may interrupt transmission, as can simultaneous high coverage of ITNs and IRS with DDT. When adding a second vector-control intervention, it is more effective to cover the unprotected population first. Although our model includes some assumptions and approximations that remain to be addressed, these findings should be useful for prioritizing and designing future field research

The Efficacy of Crotalidae Polyvalent Immune Fab (Ovine) Antivenom Versus Placebo Plus Optional Rescue Therapy on Recovery From Copperhead Snake Envenomation: A Randomized, Double-Blind, Placebo-Controlled, Clinical Trial

Study objective: Copperhead snake (Agkistrodon contortrix) envenomation causes limb injury resulting in pain and disability. It is not known whether antivenom administration improves limb function. We determine whether administration of antivenom improves recovery from limb injury in patients envenomated by copperhead snakes. Methods: From August 2013 through November 2015, we performed a multicenter, randomized, double-blind, placebo- controlled, clinical trial to evaluate the effect of ovine Crotalidae polyvalent immune Fab (ovine) (CroFab; FabAV) antivenom therapy on recovery of limb function in patients with copperhead snake envenomation at 14 days postenvenomation. The study setting was 18 emergency departments in regions of the United States where copperhead snakes are endemic. Consecutive patients aged 12 years or older with mild- to moderate-severity envenomation received either FabAV or placebo. The primary outcome was limb function 14 days after envenomation, measured by the Patient-Specific Functional Scale. Additional outcomes included the Patient-Specific Functional Scale at other points; the Disorders of the Arm, Shoulder, and Hand, Lower Extremity Functional Scale, and Patient’s Global Impression of Change instruments; grip strength; walking speed; quality of life (Patient-Reported Outcomes Measurement Information System Physical Fucntion-10); pain; and analgesic use. Results: Seventy-four patients received study drug (45 FabAV, 29 placebo). Mean age was 43 years (range 12 to 86 years). Fifty-three percent were men, 62% had lower extremity envenomation, and 88% had mild initial severity. The primary outcome, the least square mean Patient-Specific Functional Scale score at 14 days postenvenomation, was 8.6 for FabAV-treated subjects and 7.4 for placebo recipients (difference 1.2; 95% confidence interval 0.1 to 2.3; P1�4.04). Additional outcome assessments generally favored FabAV. More FabAV-treated subjects experienced treatment- emergent adverse events (56% versus 28%), but few were serious (1 in each group). Conclusion: Treatment with FabAV reduces limb disability measured by the Patient-Specific Functional Scale 14 days after copperhead envenomation. [Ann Emerg Med. 2017;70:233-244.

Antivenom Treatment Is Associated with Fewer Patients using Opioids after Copperhead Envenomation

Introduction Copperhead envenomation causes local tissue destruction, leading people to seek treatment for the pain and swelling. First-line treatment for the pain is opioid medications. There is rising concern that an initial opioid prescription from the emergency department (ED) can lead to long-term addiction. This analysis sought to determine whether use of Fab antivenom (FabAV) for copperhead envenomation affected opioid use. Methods We performed a secondary analysis using data from a randomized clinical trial designed to determine the effect of FabAV on limb injury recovery following mild to moderate copperhead envenomation. Opioid use was a defined secondary outcome in the parent trial. Patients were contacted after discharge, and data were obtained regarding medications used for pain and the patients’ functional status. This analysis describes the proportion of patients in each treatment group reporting opioid use at each time point. It also assesses the interaction between functional status and use of opioids. Results We enrolled 74 patients in the parent trial (45 received FabAV, 29 placebo), of whom 72 were included in this secondary analysis. Thirty-five reported use of any opioids after hospital discharge. A smaller proportion of patients treated with FabAV reported opioid use: 40.9% vs 60.7% of those in the placebo group. The proportion of patients using opioids remained smaller in the FabAV group at each follow-up time point. Controlling for confounders and interactions between variables, the model estimated that the odds ratio of using opioids after hospital discharge among those who received placebo was 5.67 times that of those who received FabAV. Patients who reported higher baseline pain, those with moderate as opposed to mild envenomation, and females were more likely to report opioid use at follow-up. Patients with ongoing limitations to functional status had an increased probability of opioid use, with a stronger association over time. Opioid use corresponded with the trial’s predefined criteria for full recovery, with only two patients reporting opioid use in the 24 hours prior to achieving full limb recovery and no patients in either group reporting opioid use after full limb recovery. Conclusion In this study population, the proportion of patients using opioids for pain related to envenomation was smaller in the FabAV treatment group at all follow-up time points

The validity, reliability and minimal clinically important difference of the patient specific functional scale in snake envenomation

Objective: Valid, reliable, and clinically relevant outcome measures are necessary in clinical studies of snake envenomation. The aim of this study was to evaluate the psychometric (validity and reliability) and clinimetric (minimal clinically important difference [MCID]) properties of the Patient-Specific Functional Scale (PSFS) in snakebite envenomation. Methods: We performed a secondary analysis of two existing snakebite trials that measured clinical outcomes using the PSFS as well as other quality of life and functional assessments. Data were collected at 3, 7, 10, and 17 days. Reliability was determined using Cronbach’s alpha for internal consistency and the intraclass correlation coefficient (ICC) for temporal stability at 10 and 17 days. Validity was assessed using concurrent validity correlating with the other assessments. The MCID was evaluated using the following criteria: (1) the distribution of stable patients according to both standard error of measurement (SEM) and responsiveness techniques, and (2) anchor-based methods to compare between individuals and to detect discriminant ability of a positive change with a receiver operator characteristic (ROC) curve and optimal cutoff point. Results: A total of 86 patients were evaluated in this study. The average PSFS scores were 5.37 (SD 3.23), 7.95 (SD 2.22), and 9.12 (SD 1.37) at 3, 7, and 10 days, respectively. Negligible floor effect was observed (maximum of 8% at 3 days); however, a ceiling effect was observed at 17 days (25%). The PSFS showed good reliability with an internal consistency of 0.91 (Cronbach’s alpha) (95% CI 0.88, 0.95) and a temporal stability of 0.83 (ICC) (95% CI 0.72, 0.89). The PSFS showed a strong positive correlation with quality of life and functional assessments. The MCID was approximately 1.0 for all methods. Conclusions: With an MCID of approximately 1 point, the PSFS is a valid and reliable tool to assess quality of life and functionality in patients with snake envenomation