A content analysis of hand hygiene materials targeting elementary-age children.

Millions of dollars have been spent on the design and dissemination of educational materials to improve handwashing to prevent infectious diseases. School-age children have been the focus of many of these efforts; yet little is known about the content of these materials. This study uses content analysis to examine the theoretical and motivational trends as well as the communication approach used in a sample of hand hygiene intervention materials targeting elementary-age children. Two trained coders analyzed 144 communication materials. Study results indicate that educational materials infrequently exhibit information consistent with theories of communication for behavior change, commonly use fear-based messaging, and rarely recommend using technology in the design of the interventions. Implications for future research and the design of more strategic, child-focused hand hygiene interventions are discussed.

Rapid Decrease in KRT14 and TP53 mRNA Expression in the Buccal Mucosa of Patients Receiving Total-Body Irradiation for Allogeneic Stem Cell Transplantation.

The only curative treatment option for relapsed patients with acute myeloid leukemia (AML) is allogeneic stem cell transplantation. Depletion of hematopoietic stem cells and leukemic blast cells is achieved through the systemic administration of DNA damaging agents, including total-body irradiation (TBI) prior to transplantation. Since other tissues are radiosensitive, the identification of biomarkers could facilitate the management of additional toxicities. Buccal keratinocytes are readily accessible and could provide a source of cells for RNA analysis. In this study, we obtained miRNAs and mRNAs from daily buccal swabs collected from patients undergoing allogeneic stem cell transplantation. Unexpectedly, there was no prominent p53-induced mRNA or miRNA response in these samples, despite the fact that the p53 pathway is a well-characterized radiation-inducible response. Instead, the expression of mRNAs encoding p53 and cytokeratin 14 (TP53 and KRT14, respectively) decreased precipitously within hours of the first radiation treatment. These patients went on to develop oral mucositis, however, it is unclear whether TP53 and/or KRT14 expression are predictive of this adverse event. Larger scale analysis of buccal epithelial samples from patients undergoing allogeneic stem cell transplantation appears to be warranted.

Paving the future: finding suitable ISMB venues.

The International Society for Computational Biology, ISCB, organizes the largest event in the field of computational biology and bioinformatics, namely the annual international conference on Intelligent Systems for Molecular Biology, the ISMB. This year at ISMB 2012 in Long Beach, ISCB celebrated the 20th anniversary of its flagship meeting. ISCB is a young, lean and efficient society that aspires to make a significant impact with only limited resources. Many constraints make the choice of venues for ISMB a tough challenge. Here, we describe those challenges and invite the contribution of ideas for solutions.

The anti-apoptotic role for p53 following exposure to ultraviolet light does not involve DDB2.

The p53 tumour suppressor is a transcription factor that can either activate or repress the expression of specific genes in response to cellular stresses such as exposure to ultraviolet light. The p53 protein can exert both pro- and anti-apoptotic effects depending on cellular context. In primary human fibroblasts, p53 protects cells from UV-induced apoptosis at moderate doses but this is greatly affected by the nucleotide excision repair (NER) capacity of the cells. The damage-specific DNA binding protein 2 (DDB2) is involved in NER and is associated with xeroderma pigmentosum subgroup E (XP-E). Importantly, DDB2 is also positively regulated by the p53 protein. To study the potential interplay between DDB2 and p53 in determining the apoptotic response of primary fibroblasts exposed to UV light, the expression of these proteins was manipulated in primary normal and XP-E fibroblast strains using human papillomavirus E6 protein (HPV-E6), RNA interference and recombinant adenoviruses expressing either p53 or DDB2. Normal and XP-E fibroblast strains were equally sensitive to UV-induced apoptosis over a broad range of doses and disruption of p53 in these strains using HPV-E6 or RNA interference led to a similar increase in apoptosis following exposure to UV light. In contrast, forced expression of p53 or DDB2 did not affect UV-induced apoptosis greatly in these normal or XP-E fibroblast strains. Collectively, these results indicate that p53 is primarily protective against UV-induced apoptosis in primary human fibroblasts and this activity of p53 does not require DDB2.

Enhanced accumulation of A2E in individuals homozygous or heterozygous for mutations in BEST1 (VMD2).

Best vitelliform macular dystrophy (BMD) is an autosomal dominant inherited macular degenerative disease caused by mutations in the gene BEST1 (formerly VMD2). Prior reports indicate that BMD is characterized histopathologically by accumulation of lipofuscin in the retinal pigment epithelium (RPE). However, this accumulation has not been quantified and the chemical composition of lipofuscin in BMD has not been examined. In this study we characterize the histopathology of a donor eye from a rare individual homozygous for a mutation (W93C) in BEST1. We find that this individual's disease was not any more severe than has been described for heterozygotes. We then used this tissue to quantify lipofuscin accumulation by enriching intracellular granules from RPE cells on sucrose gradients and counting the granules in each density fraction. Granules from the homozygous donor eye as well as a donor eye from an individual heterozygous for the mutation T6R were compared with age-matched control eyes. Interestingly, the least dense fraction, representing classical lipofuscin granules was either not present or significantly diminished in the BMD donor eyes and the autoflourescence associated with lipofuscin had shifted to denser fractions. However, a substantial enrichment for granules in fractions of higher density was also noted in the BMD samples. Inspection of granules from the homozygous donor eye by electron microscopy revealed a complex abnormal multilobular structure. Analysis of granules by HPLC indicated a approximately 1.6- and approximately fourfold overall increase in A2E in the BMD eyes versus age-matched control eyes, with a shift of A2E to more dense granules in the BMD donor eyes. Despite the increase in A2E and total intracellular granules, the RPE in the homozygous donor eyes was relatively well preserved. Based on these data we conclude that the clinical and histopathologic consequences to the homozygous donor were not any more severe than has been reported previously for individuals who are established or presumptive heterozygotes. We find that A2E is a component of the lipofuscin accumulated in BMD and that it is more abundant than in control eyes suggesting that the etiology of BMD is similar to Stargardt's disease and Stargardt-like macular dystrophy. Finally, the changes we observe in the granules suggest that the histopathology and eventual vision loss associated with BMD may be due to defects in the ability of the RPE to fully degrade phagocytosed photoreceptor outer segments.

Myocilin-associated exosomes in human ocular samples.

Mutations in myocilin result in ocular hypertension, likely due to decreased drainage of aqueous humor through the trabecular meshwork. Since less myocilin is found in the aqueous humor of those with disease-causing mutations, understanding myocilin's role in the aqueous humor is of clinical importance. Recently, myocilin was shown to exit cultured trabecular meshwork cells in association with shed vesicles called exosomes. To examine relevance of this finding in a physiological setting, the present study examined three different types of ocular samples for the presence of myocilin-associated exosomes. Using differential centrifugation steps, we found myocilin associated with exosomes isolated from effluent collected from human anterior segments in organ culture and aqueous humor obtained from human cadaveric eyes or from patients undergoing excisional surgery. Similar to results with cultured cells, myocilin associated predominately with exosomes in fresh samples, appeared mostly soluble at later times, and had biochemical properties (density of 1.13-1.19 g/ml in linear sucrose gradient) similar to those characteristics of exosomes. These data indicate that exosomes are present and may facilitate the transport of myocilin into the extracellular space of human ocular cells.

Coiled-coil targeting of myocilin to intracellular membranes.

Mutations in myocilin (MYOC) associate with glaucoma and ocular hypertension. Unfortunately, the specific role of MYOC, a widely expressed protein of unknown function, in ocular hypertension is unknown. Since MYOC localizes both to intracellular membranes and to the cytosol, we tested the hypothesis that MYOC is a cytosolic protein that associates with cellular membranes via its coiled-coil domain. Using green fluorescent protein (GFP) chimeras in expression and metabolic labeling studies, we observed that MYOC's putative signal peptide failed to traffic GFP into the secretory machinery and out of transfected cells. Next, we tested which of MYOC's three folding domains were responsible for targeting. In cell fractionation and immunofluorescence microscopy studies, the coiled-coil, but not the helix-turn-helix or olfactomedin domains, was necessary and sufficient to target GFP chimeras to cell membranes. Interestingly, a vesicular phenotype required sequential addition of the helix-turn-helix and olfactomedin domains to the coiled-coil. Taken together, these data indicate that the coiled-coil domain, not the putative signal sequence, is responsible for the targeting of MYOC to the secretory machinery.

Temperature dependence of T-type calcium channel gating.

T-type calcium channel isoforms are expressed in a multitude of tissues and have a key role in a variety of physiological processes. To fully appreciate the physiological role of distinct channel isoforms it is essential to determine their kinetic properties under physiologically relevant conditions. We therefore characterized the gating behavior of expressed rat voltage-dependent calcium channels (Ca(v)) 3.1, Ca(v)3.2, and Ca(v)3.3, as well as human Ca(v)3.3 at 21 degrees C and 37 degrees C in saline that approximates physiological conditions. Exposure to 37 degrees C caused significant increases in the rates of activation, inactivation, and recovery from inactivation, increased the current amplitudes, and induced a hyperpolarizing shift of half-activation for Ca(v)3.1 and Ca(v)3.2. At 37 degrees C the half-inactivation showed a hyperpolarizing shift for Ca(v)3.1 and Ca(v)3.2 and human Ca(v)3.3, but not rat Ca(v)3.3. The observed changes in the kinetics were significant but not identical for the three isoforms, showing that the ability of T-type channels to conduct calcium varies with both channel isoform and temperature.

Weak, physiologically patterned magnetic fields do not affect maze performance in normal rats, but disrupt seized rats normalized with ketamine: possible support for a neuromatrix concept?

The concept of a neuromatrix as a determinant of behavior proposes that complex neuroelectromagnetic patterns supported by specific spatial configurations of neurons underlie the generation of behaviors. When the pattern of neuronal connectivity is changed, as occurs during limbic epilepsy, neuroelectromagnetic patterns change in parallel to sustain behavioral output. Thus, a testable prediction of the neuromatrix concept is that the "normal" behaviors of animals with markedly reorganized neuroelectromagnetic patterns are vulnerable to specific stimuli that are ineffective when applied to a normal population. Because rats treated with ketamine after being induced to seize with pilocarpine exhibit behaviors indistinguishable from those of control populations despite marked changes in brain structure, they represent an ideal population in which to examine this hypothesis. Ketamine-treated pilocarpine-seized rats and normal rats were exposed continuously either to a complex sequence magnetic field or to control conditions during the acquisition of a radial arm maze task for 8 consecutive days. After 14 days of subsequent exposure to a frequency-modulated field (7-500 nT), during which time there was no training, the rats that had been induced to seize and had been exposed continuously to this magnetic configuration exhibited conspicuously slower response durations per arm than rats that had been induced to seize and exposed to control conditions or normal rats that had been exposed to either magnetic fields or control conditions. Thus, the behaviors of rats who have sustained multiple, discrete injuries throughout the brain may be seriously disrupted by the appropriate pattern of exogenous weak magnetic fields. Our results represent the first empirical support for the concept of the neuromatrix.

Complex magnetic fields enable static magnetic field cue use for rats in radial maze tasks.

Male Wistar rats were trained in an eight-arm radial maze task (two sessions per day, delayed-non-matching-to-sample) that included an intramaze static magnetic field "cue" (185 microT) specific to the entrance point of one of the arms. Rats were exposed daily for 60 min to a complex magnetic field waveform (theta-burst pattern, 200-500 nT), presented with several different interstimulus intervals (ISIs), either immediately following training sessions or immediately preceding testing sessions. Application of the theta-burst stimulus with a 4000 ms ISI significantly improved the rats' memory for the arm of the radial maze whose position was indicated by the presence of a static magnetic field cue. Reference memory errors were homogeneously distributed among all eight arms of the maze for sham-exposed rats, and among the other seven arms of the maze for complex magnetic field-treated rats. These results suggest that static magnetic field cues may be salient orienting cues even in a microenvironment such as a radial maze, but their use as a cue during maze learning in rats is dependent on whole-body application of a specific time-varying complex magnetic field.

Sudden death in epileptic rats exposed to nocturnal magnetic fields that simulate the shape and the intensity of sudden changes in geomagnetic activity: an experiment in response to Schnabel, Beblo and May.

To test the hypothesis that sudden unexplained death (SUD) in some epileptic patients is related to geomagnetic activity we exposed rats in which limbic epilepsy had been induced to experimentally produced magnetic fields designed to simulate sudden storm commencements (SSCs). Prior studies with rats had shown that sudden death in groups of rats in which epilepsy had been induced months earlier was associated with the occurrence of SSCs and increased geomagnetic activity during the previous night. Schnabel et al. [(2000) Neurology 54:903-908] found no relationship between SUD in human patients and geomagnetic activity. A total of 96 rats were exposed to either 500, 50, 10-40 nT or sham (less than 10 nT) magnetic fields for 6 min every hour between midnight and 0800 hours (local time) for three successive nights. The shape of the complex, amplitude-modulated magnetic fields simulated the shape and structure of an average SSC. The rats were then seized with lithium and pilocarpine and the mortality was monitored. Whereas 10% of the rats that had been exposed to the sham field died within 24 h, 60% of the rats that had been exposed to the experimental magnetic fields simulating natural geomagnetic activity died (P<.001) during this period. These results suggest that correlational analyses between SUD in epileptic patients and increased geomagnetic activity can be simulated experimentally in epileptic rats and that potential mechanisms might be testable directly.

Leptin-induced satiation mediated by abdominal vagal afferents.

Leptin is a hormone secreted into the systemic blood primarily by white adipose tissue. However, leptin also is synthesized and stored by cells in the gastric mucosa. Because gastric mucosal leptin is secreted in response to ingestion of a meal, we hypothesized that it might contribute to satiation (meal termination) by acting on gastrointestinal vagal afferent neurons. To test whether leptin is capable of acutely reducing short-term food intake, we measured consumption of a liquid meal (15% sucrose) following low-dose leptin administration via the celiac artery, which perfuses the upper gastrointestinal tract. Leptin (1, 3, 10 mug) was infused via a chronically implanted, nonocclusive celiac arterial catheter or via a jugular vein catheter with its tip in the right cardiac atrium. Fifteen percent sucrose intake was then measured for 30 min. We found that leptin dose dependently inhibited sucrose intake when infused through the celiac catheter but not when infused into the general circulation via a jugular catheter. Plasma leptin concentrations in the general circulation following celiac arterial or jugular leptin infusions were not significantly different. Celiac arterial leptin infusion did not reduce meal size in vagotomized or capsaicin-treated rats. Finally, we also found that reduction of meal size by celiac leptin infusion was markedly enhanced when coinfused with cholecystokinin, a gastrointestinal satiety peptide whose action depends on vagal afferent neurons. Our results support the hypothesis that leptin contributes to satiation by a mechanism dependent on gastrointestinal vagal afferent innervation of the upper gastrointestinal tract.

Lithium ion "cyclotron resonance" magnetic fields decrease seizure onset times in lithium-pilocarpine seized rats.

The cyclotron resonance equation predicts that the frequency of an applied magnetic field that might optimally interact with a single ion species may be computed as a function of the charge-to-mass ratio of the ion and the strength of the background static magnetic field. The present study was undertaken to discern the applicability of this equation for optimizing lithium ion utilization in the rat, as inferred by the predicted magnetic "ion resonance "field-induced shift of lithium's dose-dependent curve for seizure onset times (SOTs) when combined with the cholinergic agent pilocarpine. Groups of rats were administered 1.5 thru 3 mEq/kg lithium chloride (in 0.5 mEq/kg increments) and exposed to reference conditions or to one of three intensities (70 nanoTesla, 0.8 microTesla, or 25 microTesla) of a 85 Hz magnetic field calculated to resonate with lithium ions given the background static geomagnetic field of approximately 38,000 nanoTesla (0.38 Gauss). A statistically significant quadratic relationship for SOT as a function of magnetic field intensity (irrespective of lithium dose) was noted: this U-shaped function was characterized by equal SOTs for the reference and 25 microTesla groups, with a trend toward shorter SOTs for the 70 nanoTesla and 0.8 microTesla groups. Although not predicted by the equations, this report extends other findings suggestive of discrete intensity windows for which magnetic field frequencies derived from the cyclotron ion resonance equation may affect ion activity.