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Context: Peer assessment/feedback (PAF) is a pedagogic technique that falls under the peer assisted learning umbrella. Peer assessment is defined as students judging the level or quality of a fellow student's understanding or work and peer feedback involves providing information on what was performed correct and incorrect in order to improve understanding. Documented benefits of PAF include enhanced learning, improved critical thinking, self-assessment, increased reflective thinking and collaboration. Objective: Examine the effects of mandatory PAF after training on psychomotor skill performance. Design: Prospective cohort study. Setting: Undergraduate athletic training program in a mid-sized public university in the southeast. Subjects/Participants: Fifty-seven sophomore athletic training students enrolled in a lower extremity orthopedic evaluation course over two years participated in the intervention and completed all required parts of the study. A control group consisted of 53 students that took the course during the previous two years; the course was constructed the same, with the exception of no PAF training and no mandatory PAF. Intervention: Participants in the intervention group completed a 2 hour workshop that covered proper mechanisms to assess, provide feedback, and receive feedback. Training also included analysis of videotaped scenarios and practice scenarios. Participants were also required to complete PAF of the skills prior to the practical exams. Main Outcome Measures: The critical dependent variable was participation in the PAF training and mandatory PAF prior to practical exams. Scores of 4 practical exams were compared through 4 separate independent samples t-test using SPSS. Results: The control group performed better on all four 60 point practical exams. The differences were statistically significant for exams 1 and 3. Exam 1 covered neurological and ankle [t(108)=4.95, p=0.00, means: experimental 52.80±5.24, control 47.85±5.25]. Exam 3 covered the hip [t(108)=2.27, p=0.03, means: experimental 51.40±5.00, control 49.02±6.75]. There were no statistical significant differences for exams 2 and 4. Exam 2 covered knee and patellofemoral joint [t(108)=1.87, p=0.07, means: experimental 52.35±5.00, control 50.66±4.46]. Exam 4 was a cumulative practical exam [t(108)=4.95, p=0.08, means: experimental 51.77±6.25, control 51.36±5.58]. Conclusions: PAF training and mandatory PAF improved practical exam grades for 2 of the 4 exams; PAF did not hinder student learning. PAF can be a helpful tool to increase the amount that a student practices skill, and increase the amount of feedback students receive. This can be especially with helpful with large classes. [ABSTRACT FROM AUTHOR]

A controlled chamber method using continuous gold trap atomic fluorescence spectroscopy (AFS) (Tekran 2537X) for the analysis of Hg(0) emissions from moderate mass rock samples was developed and tested. A series of black shale and other bedrock samples from Nova Scotia, Canada, were used to test the method and its reproducibility. Hg(0) emissions at 170°C were measured to quantify both free surficial Hg(0) and Hg(0) that had penetrated the rock structure. High volumes of chamber air (45 L) were sampled using 30 min collection times to achieve detectable elemental mercury (Hg(0)) emissions. We found higher percentage masses of Hg(0) were released (1.1%-4.1% of total Hg mass present) in black shale samples as compared to granite and basalt samples from the same region (0.0%-0.3% released) over 350 h of continuous analysis time. The pseudo first order emission rate constants ranged from 0.015-0.245 h -1 (mean 0.063 h -1 , standard deviation (SD) 0.102) for the black shale samples analyzed and was 0.004 h -1 for the granite sample. The 24-h zero-order emission rate constants ranged between 0.41 and 3.54 ng h -1 (mean 1.4 ng h -1 , SD 1.3) for the black shale samples analyzed and were ~ 0.01 ng h -1 for the granite and basalt samples. This technique has useful implications for examining rock properties and Hg(0) emission rates., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: Near-road ambient air pollution concentrations that are affected by vehicle emissions are typically characterized by substantial spatial variability with respect to distance from the roadway and temporal variability based on the time of day, day of week, and season. The goal of this work is to identify variables that explain either temporal or spatial variability based on case studies for a freeway site and an urban intersection site. The key hypothesis is that dispersion modeling of near-road pollutant concentrations could be improved by adding estimates or indices for site-specific explanatory variables, particularly related to traffic. Based on case studies for a freeway site and an urban intersection site, the specific aims of this project are to (1) develop and test regression models that explain variability in traffic-related air pollutant (TRAP) ambient concentration at two near-roadway locations; (2) develop and test refined proxies for land use, traffic, emissions and dispersion; and (3) prioritize inputs according to their ability to explain variability in ambient concentrations to help focus efforts for future data collection and model development., The key pollutants that are the key focus of this work include nitrogen oxides (NO x ), carbon monoxide (CO), black carbon (BC), fine particulate matter (PM 2.5 ; PM ≤ 2.5 μm in aerodynamic diameter), ultrafine particles (UFPs; PM ≤ 0.1 μm in aerodynamic diameter), and ozone (O 3 ). NO x , CO, and BC are tracers of vehicle emissions and dispersion. PM 2.5 is influenced by vehicle table emissions and regional sources. UFPs are sensitive to primary vehicle emissions. Secondary particles can form near roadways and on regional scales, influencing both PM 2.5 and UFP concentrations. O 3 concentrations are influenced by interaction with NO x near the roadway. Nitrogen dioxide (NO 2 ), CO, PM 2.5 , and O 3 are regulated under the National Ambient Air Quality Standards (NAAQS) because of demonstrated health effects. BC and UFPs are of concern for their potential health effects. Therefore, these pollutants are the focus of this work., Methods: The methodological approach includes case studies for which variables are identified and assesses their ability to explain either temporal or spatial variability in pollutant ambient concentrations. The case studies include one freeway location and one urban intersection. The case studies address (1) temporal variability at a fixed monitor 10 meters from a freeway; (2) downwind concentrations perpendicular to the same location; (3) variability in 24-hour average pollutant concentrations at five sites near an urban intersection; and (4) spatiotemporal variability along a walking path near that same intersection., The study boundary encompasses key factors in the continuum from vehicle emissions to near-road exposure concentrations. These factors include land use, transportation infrastructure and traffic control, vehicle mix, vehicle (traffic) flow, on-road emissions, meteorology, transport and evolution (transformation) of primary emissions, and production of secondary pollutants, and their resulting impact on measured concentrations in the near-road environment. We conducted field measurements of land use, traffic, vehicle emissions, and near-road ambient concentrations in the vicinity of two newly installed fixed-site monitors. One is a monitoring station jointly operated by the U.S. Environmental Protection Agency (U.S. EPA) and the North Carolina Department of Environmental Quality (NC DEQ) on I-40 between Airport Boulevard and I-540 in Wake County, North Carolina. The other is a fixed-site monitor for measuring PM 2.5 at the North Carolina Central University (NCCU) campus on E. Lawson Street in Durham, North Carolina. We refer to these two locations as the freeway site and the urban site, respectively. We developed statistical models for the freeway and urban sites., Results: We quantified land use metrics at each site, such as distances to the nearest bus stop. For the freeway site, we quantified lane-by-lane total vehicle count, heavy vehicle (HV) count, and several vehicle-activity indices that account for distance from each lane to the roadside monitor. For the urban site, we quantified vehicle counts for all 12 turning movements through the intersection. At each site, we measured microscale vehicle tailpipe emissions using a portable emission measurement system., At the freeway site, we measured the spatial gradient of NO x , BC, UFPs, and PM, quantified particle size distributions at selected distances from the roadway and assessed partitioning of particles as a function of evolving volatility. We also quantified fleet-average emission factors for several pollutants., At the urban site, we measured daily average concentrations of nitric oxide (NO), NO x , O 3 , and PM 2.5 at five sites surrounding the intersection of interest; we also measured high resolution (1-second to 10-second averages) concentrations of O 3 , PM 2.5 , and UFPs along a pedestrian transect. At both sites, the Research LINE-source (R-LINE) dispersion model was applied to predict concentration gradients based on the physical dispersion of pollution., Statistical models were developed for each site for selected pollutants. With variables for local wind direction, heavy-vehicle index, temperature, and day type, the multiple coefficient of determination (R 2 ) was 0.61 for hourly NO x concentrations at the freeway site. An interaction effect of the dispersion model and a real-time traffic index contributed only 24% of the response variance for NO x at the freeway site. Local wind direction, measured near the road, was typically more important than wind direction measured some distance away, and vehicle-activity metrics directly related to actual real-time traffic were important. At the urban site, variability in pollutant concentrations measured for a pedestrian walk-along route was explained primarily by real-time traffic metrics, meteorology, time of day, season, and real-world vehicle tailpipe emissions, depending on the pollutant. The regression models explained most of the variance in measured concentrations for BC, PM, UFPs, NO, and NO x at the freeway site and for UFPs and O 3 at the urban site pedestrian transect., Conclusions: Among the set of candidate explanatory variables, typically only a few were needed to explain most of the variability in observed ambient concentrations. At the freeway site, the concentration gradients perpendicular to the road were influenced by dilution, season, time of day, and whether the pollutant underwent chemical or physical transformations. The explanatory variables that were useful in explaining temporal variability in measured ambient concentrations, as well as spatial variability at the urban site, were typically localized real-time traffic-volume indices and local wind direction. However, the specific set of useful explanatory variables was site, context (e.g., next to road, quadrants around an intersection, pedestrian transects), and pollutant specific. Among the most novel of the indicators, variability in real-time measured tailpipe exhaust emissions was found to help explain variability in pedestrian transect UFP concentrations. UFP particle counts were very sensitive to real-time traffic indicators at both the freeway and urban sites. Localized site-specific data on traffic and meteorology contributed to explaining variability in ambient concentrations. HV traffic influenced near-road air quality at the freeway site more so than at the urban site. The statistical models typically explained most of the observed variability but were relatively simple. The results here are site-specific and not generalizable, but they are illustrative that near-road air quality can be highly sensitive to localized real-time indicators of traffic and meteorology., (© 2022 Health Effects Institute. All rights reserved.)

Malaria is caused by the protozoan parasite Plasmodium , which undergoes a complex life cycle in a human host and a mosquito vector. The parasite's cyclic GMP (cGMP)-dependent protein kinase (PKG) is essential at multiple steps of the life cycle. Phosphoproteomic studies in Plasmodium falciparum erythrocytic stages and Plasmodium berghei ookinetes have identified proteolysis as a major biological pathway dependent on PKG activity. To further understand PKG's mechanism of action, we screened a yeast two-hybrid library for P. falciparum proteins that interact with P. falciparum PKG (PfPKG) and tested peptide libraries to identify its phosphorylation site preferences. Our data suggest that PfPKG has a distinct phosphorylation site and that PfPKG directly phosphorylates parasite RPT1, one of six AAA + ATPases present in the 19S regulatory particle of the proteasome. PfPKG and RPT1 interact in vitro , and the interacting fragment of RPT1 carries a PfPKG consensus phosphorylation site; a peptide carrying this consensus site competes with the RPT1 fragment for binding to PfPKG and is efficiently phosphorylated by PfPKG. These data suggest that PfPKG's phosphorylation of RPT1 could contribute to its regulation of parasite proteolysis. We demonstrate that proteolysis plays an important role in a biological process known to require Plasmodium PKG: invasion by sporozoites of hepatocytes. A small-molecule inhibitor of proteasomal activity blocks sporozoite invasion in an additive manner when combined with a Plasmodium PKG-specific inhibitor. Mining the previously described parasite PKG-dependent phosphoproteomes using the consensus phosphorylation motif identified additional proteins that are likely to be direct substrates of the enzyme., (Copyright © 2018 American Society for Microbiology.)

Background: Transanal hemorrhoidal dearterialization (THD) is a recently developed procedure to minimize postoperative pain from hemorrhoidectomy. This technique utilizes Doppler signals to aid ligation of hemorrhoidal arteries followed by mucopexy of redundant mucosa if needed. The aim of the present study was to assess patient satisfaction after THD., Methods: This is a retrospective cohort study of patients who underwent THD at three different sites from April 2007 through October 2010. All procedures were performed in ambulatory settings according to protocol. Telephone surveys were conducted after a minimum of 1-month follow-up to assess patients' satisfaction on a scale of 1-10. Patients were asked whether the procedure had alleviated their symptoms. Patients were asked to recall duration of pain and time from surgery to return to work., Results: Between April 2007 and October 2010, 216 patients with grade III-IV hemorrhoids underwent THD. There were 165 males and 61 females. Average age was 52.2 ± 14.2 years. All patients were discharged the same day after meeting ambulatory surgery center discharge criteria. Postoperative difficulty urinating occurred in 37 (17 %) patients, and six of them required temporary urinary catheterization. Transitory postoperative bleeding was reported by 38 (18 %) patients. Transitory incontinence to stool and flatus occurred in 18 (9 %) and 16 patients (8 %), respectively. Pelvic muscle spasms occurred in 21 (10 %) patients. Median follow-up was 23 months (range 1-42 months) with 143 (66 %) having at least 9 months between procedure and interview. Mean patient satisfaction was 8.5 ± 0.7 (on a scale of 1-10 with 10 being the best), and 91.5 % of patients felt the procedure had "helped" them. Average number of days with discomfort was 6.7 ± 2.1. Patients returned to work after an average of 10.3 ± 3.2 days. Our study is limited by lack of long-term follow-up and by retrospective complication assessment., Conclusions: Patient satisfaction with THD performed in ambulatory settings is high. Our data support performance of this procedure in an ambulatory setting.

Presacral bleeding is a dreaded complication of pelvic surgery. Rapid and effective control of such bleeding is important to avoid potentially life-threatening outcomes. Various methods for controlling presacral bleeding, all with only limited success, have been described in the literature. We report the alternative technique of using the argon beam coagulator (ABC) to control presacral bleeding. We demonstrate its efficacious use in both open surgery and a laparoscopic case. Our approach involved applying an argon beam at bone setting directly to the bleeders and using a "point and shoot" technique. We found that ABC is a simpler, equally effective and expeditious way of addressing presacral bleeding. To the best of our knowledge, there has been only one previously reported case in the literature of the use of ABC to control presacral bleeding.

Purpose: Until recently the treatment verification for patients undergoing fast neutron therapy at our facility was accomplished through a combination of neutron beam portal films aligned with a graticule mounted on an orthronormal x-ray tube. To eliminate uncertainty with respect to the relative positions of the x-ray graticule and the therapy beam, we have developed a graticule which is placed in the neutron beam itself., Methods: For a graticule to be visible on the portal film, the attenuation of the neutron beam by the graticule landmarks must be significantly greater than that of the material in which the landmarks are mounted. Various materials, thicknesses, and mounting points were tried to gain the largest contrast between the graticule landmarks and the mounting material., Results: The final design involved 2 inch steel pins of 0.125 inch diameter captured between two parallel plates of 0.25 inch thick clear acrylic plastic. The distance between the two acrylic plates was 1.625 inches, held together at the perimeter with acrylic sidewall spacers. This allowed the majority of length of the steel pins to be surrounded by air. The pins were set 1 cm apart and mounted at angles parallel to the divergence of the beam dependent on their position within the array. The entire steel pin and acrylic plate assembly was mounted on an acrylic accessory tray to allow for graticule alignment., Conclusion: Despite the inherent difficulties in attenuating fast neutrons, our simple graticule design produces the required difference of attenuation between the arrays of landmarks and the mounting material. The graticule successfully provides an in-beam frame of reference for patient portal verification., (© 2012 American Association of Physicists in Medicine.)

Purpose: Intensity Modulated Neutron Radiotherapy (IMNRT) has been commissioned for clinical use. The number of allowable segments in IMNRT plans is limited by MLC speed. Quality and deliverability of static IMNRT treatment plans using the TG-119 test suite were evaluated to establish guidelines for the number of segments per plan., Methods: Treatment plans were created and optimized to specified constraints for all cases in the TG-119 test suite using the Varian Eclipse TPS. A 4MV photon beam with similar penetration characteristics as the fast neutron beam was used as a surrogate for this optimization. Final dose calculations were performed using an in-house TPS commissioned for neutron dose calculations. Following optimization, MLC segments were created for three ranges of total plan complexity - very limited (15-23 segments), limited (24- 31 segments), and unlimited. Calculated DVHs were then compared for compliance with TG-119 dose constraints. The estimated time of delivery for plans in each range was calculated based on known delivery parameters., Results: The prostate case passes all constraints for each complexity level. All other plans fail to meet at least one constraint for one or more of the complexity levels. For all cases combined, the very limited, limited, and unlimited complexity levels meet 16, 17, and 19 of 23 total dose constraints, respectively. The mean estimated delivery time for the very limited, limited, and unlimited plans is 34 minutes (range: 27-39), 40 minutes (range: 34-45), and 68 minutes (range: 53-81) respectively, neglecting any delay due to therapists entering the treatment room., Conclusion: IMNRT plan quality is limited by current MLC capabilities. IMNRT plans should be limited to 25 segments to ensure a reasonable treatment time of 45 minutes. Even with this small number of segments, we were able to meet most dose constraints set forth in TG-119., (© 2012 American Association of Physicists in Medicine.)

Elevated immunoglobulin free light chain (FLC) level and abnormal FLC ratio are commonly seen in multiple myeloma (MM) and have prognostic implications. We hypothesized that presence of immunoglobin heavy chain (IgH) translocations leads to unbalanced production of light chains and more extreme abnormalities of FLC, and may explain the prognostic value of FLC. We studied 314 patients with newly diagnosed MM enrolled in a phase III trial, in whom results of fluorescence in situ hybridization testing and data on serum FLC levels were available. Cytogenetic analyses and FLC estimates were performed on stored samples and results were correlated with clinical data. The median ratio (FLC ratio) and the absolute difference (FLC diff) between the involved and uninvolved FLC were higher among those with IgH translocations, especially t(14;16). In multivariate analysis, the prognostic value of FLC estimates on progression-free and overall survival were independent of high-risk IgH translocations t(4;14) and t(14;16). A combination of the risk factors; either abnormal FLC estimate and/or the presence of high-risk IgH translocation, achieved better prognostic stratification. We conclude that patients with IgH translocations have higher FLC levels and abnormal ratios, but the prognostic effect of FLC is only partially explained by translocation status. A system including both these risk factors allows better prediction of outcome.

It is well established that B-Raf signaling through the MAP kinase (ERK) pathways plays a prominent role in regulating cell proliferation but how it does this is not completely understood. Here, we show that B-Raf serves a physiological role during mitosis in human somatic cells. Knockdown of B-Raf using short interfering RNA (siRNA) resulted in pleiotropic spindle abnormalities and misaligned chromosomes in over 80% of the mitotic cells analyzed. A second B-Raf siRNA gave similar results suggesting these effects are specific to downregulating B-Raf protein. In agreement with these findings, a portion of B-Raf was detected at the spindle structures including the spindle poles and kinetochores. Knockdown of C-Raf (Raf-1) had no detectable effects on spindle formation or chromosome alignment. Activation of the spindle assembly checkpoint was found to be dependent on B-Raf as evident by the inability of checkpoint proteins Bub1 and Mad2 to localize to unattached kinetochores in HeLa cells treated with B-Raf siRNA. Consistent with this, live-cell imaging microscopy showed that B-Raf-depleted cells exited mitosis earlier than control non-depleted cells. Finally, we provide evidence that B-Raf signaling promotes phosphorylation and kinetochore localization of the mitotic checkpoint kinase Mps1. Blocking B-Raf expression, ERK activity, or phosphorylation at Ser-821 residue perturbed Mps1 localization at unattached kinetochores. Thus, our data implicates a mitotic role for B-Raf in regulating spindle formation and the spindle checkpoint in human somatic cells.

We have used genomic tiling arrays to identify transcribed regions throughout the human genome. Analysis of the mapping results of RNA isolated from five cell/tissue types, NB4 cells, NB4 cells treated with retinoic acid (RA), NB4 cells treated with 12-O-tetradecanoylphorbol-13 acetate (TPA), neutrophils, and placenta, throughout the ENCODE region reveals a large number of novel transcribed regions. Interestingly, neutrophils exhibit a great deal of novel expression in several intronic regions. Comparison of the hybridization results of NB4 cells treated with different stimuli relative to untreated cells reveals that many new regions are expressed upon cell differentiation. One such region is the Hox locus, which contains a large number of novel regions expressed in a number of cell types. Analysis of the trinucleotide composition of the novel transcribed regions reveals that it is similar to that of known exons. These results suggest that many of the novel transcribed regions may have a functional role.

A new super-oxidized water (SOW) product, Microcyn, was tested for in vitro antimicrobial and antiviral activities. The effectiveness of this neutral-pH SOW at killing Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Candida albicans in pure culture was evaluated. One millilitre (approximately 10(8)colony-forming units/mL) of each micro-organism was subjected to 9 mL Microcyn or sterile water at room temperature for 30s. Under these conditions, a log(10) reduction factor of 8 in the level of all pathogens occurred in the treatment samples. In addition, results of tests with three batches of Microcyn exposed to Bacillus atrophaeus spores for 5 min demonstrated complete inactivation of the spores within 2-3 min (log(10) reduction factor >4). The effectiveness of Microcyn in reducing human immunodeficiency virus-1 (HIV-1) on hard surfaces (glass) was also evaluated in compliance with Environmental Protection Agency requirements for virucidal claims. After exposure of the tested surfaces to Microcyn for 5 min without agitation, there was a log(10) reduction factor >3 in the viral load as measured by both cytopathic effect and antigen p24 of HIV-1 production in MT-2 cultures. Microcyn activity against adenoviral vector type 5 was also analysed under simulated laboratory in-use conditions with viral suspensions. In order to increase the sensitivity of the test, the fluorescent light emitted by AdGFP-infected cells was measured with the use of a flow cytometer. A log(10) reduction factor >3 in the viral load was achieved after a 5-min exposure to Microcyn under these strict conditions. These results show that Microcyn exerts a wide antimicrobial spectrum with major advantages over acidic SOWs, including neutral pH, lower free active chlorine (51-85 ppm) and long shelf life (1 year).

Introduction: Kaiser Permanente of the Mid-Atlantic States (KPMAS) is collaborating with the Electronic Surveillance System for Early Notification of Community-Based Epidemics II (ESSENCE II) program to understand how managed-care data can be effectively used for syndromic surveillance., Objectives: This study examined whether KPMAS nurse advice hotline data would be able to predict the syndrome diagnoses made during subsequent KPMAS office visits., Methods: All nurse advice hotline calls during 2002 that were linked to an outpatient office visit were identified. By using International Classification of Diseases, Ninth Revision (ICD-9) codes, outpatient visits were categorized into seven ESSENCE II syndrome groups (coma, gastrointestinal, respiratory, neurologic, hemorrhagic, infectious dermatologic, and fever). Nurse advice hotline calls were categorized into ESSENCE II syndrome groups on the basis of the advice guidelines assigned. For each syndrome group, the sensitivity, specificity, and positive predictive value of hotline calls were calculated by using office visits as a diagnostic standard. For matching syndrome call-visit pairs, the lag (i.e., the number of hours that elapsed between the date and time the patient spoke to an advice nurse and the date and time the patient made an office visit) was calculated., Results: Of all syndrome groups, the sensitivity of hotline calls for respiratory syndrome was highest (74.7%), followed by hotline calls for gastrointestinal syndrome (72.0%). The specificity of all nurse advice syndrome groups ranged from 88.9% to 99.9%. The mean lag between hotline calls and office visits ranged from 8.3 to 50 hours, depending on the syndrome group., Conclusions: The timeliness of hotline data capture compared with office visit data capture, as well as the sensitivity and specificity of hotline calls for detecting respiratory and gastrointestinal syndromes, indicate that KPMAS nurse advice hotline data can be used to predict KPMAS syndromic outpatient office visits.

gamma-Tubulin is essential for microtubule nucleation in yeast and other organisms; whether this protein is regulated in vivo has not been explored. We show that the budding yeast gamma-tubulin (Tub4p) is phosphorylated in vivo. Hyperphosphorylated Tub4p isoforms are restricted to G1. A conserved tyrosine near the carboxy terminus (Tyr445) is required for phosphorylation in vivo. A point mutation, Tyr445 to Asp, causes cells to arrest prior to anaphase. The frequency of new microtubules appearing in the SPB region and the number of microtubules are increased in tub4-Y445D cells, suggesting this mutation promotes microtubule assembly. These data suggest that modification of gamma-tubulin is important for controlling microtubule number, thereby influencing microtubule organization and function during the yeast cell cycle.

Haploid cells of the budding yeast Saccharomyces cerevisiae respond to mating pheromones by arresting their cell-division cycle in G1 and differentiating into a cell type capable of locating and fusing with mating partners. Yeast cells undergo chemotactic cell surface growth when pheromones are present above a threshold level for morphogenesis; however, the morphogenetic responses of cells to levels of pheromone below this threshold have not been systematically explored. Here we show that MATa haploid cells exposed to low levels of the alpha-factor mating pheromone undergo a novel cellular response: cells modulate their division patterns and cell shape, forming colonies composed of filamentous chains of cells. Time-lapse analysis of filament formation shows that its dynamics are distinct from that of pseudohyphal growth; during pheromone-induced filament formation, daughter cells are delayed relative to mother cells with respect to the timing of bud emergence. Filament formation requires the RSR1(BUD1), BUD8, SLK1/BCK1, and SPA2 genes and many elements of the STE11/STE7 MAP kinase pathway; this response is also independent of FAR1, a gene involved in orienting cell polarization during the mating response. We suggest that mating yeast cells undergo a complex response to low levels of pheromone that may enhance the ability of cells to search for mating partners through the modification of cell shape and alteration of cell-division patterns.

To facilitate studies of the yeast proteome, we cloned 5800 open reading frames and overexpressed and purified their corresponding proteins. The proteins were printed onto slides at high spatial density to form a yeast proteome microarray and screened for their ability to interact with proteins and phospholipids. We identified many new calmodulin- and phospholipid-interacting proteins; a common potential binding motif was identified for many of the calmodulin-binding proteins. Thus, microarrays of an entire eukaryotic proteome can be prepared and screened for diverse biochemical activities. The microarrays can also be used to screen protein-drug interactions and to detect posttranslational modifications.

Research has documented the positive effects that t'ai chi has on the well-being of elders. However, the reasons that elders practice t'ai chi have not been explored. The purpose of this study was to describe the facilitators and barriers to t'ai chi practice in elderly populations. Taiwanese community-dwelling elders (40 of whom practiced t'ai chi and 40 who did not) aged 65 and older, matched on age and gender, were interviewed. Results showed that encouragement from others was the most important factor for elders to practice t'ai chi, whereas positive health outcomes were the reason they continued to practice it. Most of the non-t'ai chi group participants had never thought about practicing t'ai chi because they felt they were too weak to practice. Because t'ai chi can be helpful in promoting the well-being of elders, it is essential for health care professionals to overcome the barriers to its uses in elderly populations.

Tai chi, a type of low-intensity exercise, has received growing attention in both eastern and western cultures, especially its use with the most rapidly increasing segment of the population-elders. Previous research findings further supported the idea that tai chi is appropriate for elderly populations and helps promote their well-being. In this article, the beneficial effects of tai chi for elders are summarized, resources to increase awareness about the exercise are provided, and ways to promote tai chi in elderly populations are suggested.

A genome-wide screen of 4168 homozygous diploid yeast deletion strains has been performed to identify nonessential genes that participate in the bipolar budding pattern. By examining bud scar patterns representing the sites of previous cell divisions, 127 mutants representing three different phenotypes were found: unipolar, axial-like, and random. From this screen, 11 functional classes of known genes were identified, including those involved in actin-cytoskeleton organization, general bud site selection, cell polarity, vesicular transport, cell wall synthesis, protein modification, transcription, nuclear function, translation, and other functions. Four characterized genes that were not known previously to participate in bud site selection were also found to be important for the haploid axial budding pattern. In addition to known genes, we found 22 novel genes (20 are designated BUD13-BUD32) important for bud site selection. Deletion of one resulted in unipolar budding exclusively from the proximal pole, suggesting that this gene plays an important role in diploid distal budding. Mutations in 20 other novel BUD genes produced a random budding phenotype and one produced an axial-like budding defect. Several of the novel Bud proteins were fused to green fluorescence protein; two proteins were found to localize to sites of polarized cell growth (i.e., the bud tip in small budded cells and the neck in cells undergoing cytokinesis), similar to that postulated for the bipolar signals and proteins that target cell division site tags to their proper location in the cell. Four others localized to the nucleus, suggesting that they play a role in gene expression. The bipolar distal marker Bud8 was localized in a number of mutants; many showed an altered Bud8-green fluorescence protein localization pattern. Through the genome-wide identification and analysis of different mutants involved in bipolar bud site selection, an integrated pathway for this process is presented in which proximal and distal bud site selection tags are synthesized and localized at their appropriate poles, thereby directing growth at those sites. Genome-wide screens of defined collections of mutants hold significant promise for dissecting many biological processes in yeast.

This study examined the stressors and stress levels, loneliness, and depression experienced by Taiwanese rural elders and determine if differences in the level of stress were associated with specific person-environmental characteristics. The relationship between stress and depression was also explored. A total of 201 older rural community residents participated in the study. Findings showed that rural elders experienced a number of stressors related to health and family issues. Differences in the degree of stressfulness of individuals differed by gender, educational level, living arrangement, and socioeconomic status (SES). Findings in this study also showed that there was a high association among stress level, depressive symptoms, and mood status. The findings of this study will assist community health policy-makers in determining the need for health-related services for rural elders experiencing these problems.

Adherence with prescribed therapies is a key factor in maintaining health in persons with chronic illnesses. The purpose of this descriptive study (which is one segment of a larger study being conducted to develop decision rules for early detection of infection or rejection in posttransplant patients) is to explore facilitators and barriers to adherence with home monitoring. Use of latent analysis revealed that the facilitators to adherence were health promotion, commitment to research, and following advice. Barriers to adherence were poor health status, laziness, and time conflict. Health professionals can use this information to plan strategies to promote adherence to home monitoring.

This unit provides comprehensive protocols for the use of insertional libraries generated by shuttle mutagenesis. From the basic protocol, a small aliquot of insertional library DNA may be used to mutagenize yeast, producing strains containing a single transposon insertion within a transcribed and translated region of the genome. This transposon-mutagenized bank of yeast strains may be screened for any desired mutant phenotype. Alternatively, since the transposon contains a reporter gene lacking its start codon and promoter, transposon-tagged strains may also be screened for specific patterns of gene expression. Strains of interest may be characterized by vectorette PCR (protocol provided) in order to locate the precise genomic site of transposon insertion within each mutant. A method by which Cre/lox recombination may be used to reduce the transposon in yeast to a small insertion element encoding an epitope tag is described. This tag serves as a tool by which transposon-mutagenized gene products may be analyzed further (e.g., localized to a discrete subcellular site).

During the early stages of budding, cell wall remodeling and polarized secretion are concentrated at the bud tip (apical growth). The CBK1 gene, encoding a putative serine/threonine protein kinase, was identified in a screen designed to isolate mutations that affect apical growth. Analysis of cbk1Delta cells reveals that Cbk1p is required for efficient apical growth, proper mating projection morphology, bipolar bud site selection in diploid cells, and cell separation. Epitope-tagged Cbk1p localizes to both sides of the bud neck in late anaphase, just prior to cell separation. CBK1 and another gene, HYM1, were previously identified in a screen for genes involved in transcriptional repression and proposed to function in the same pathway. Deletion of HYM1 causes phenotypes similar to those observed in cbk1Delta cells and disrupts the bud neck localization of Cbk1p. Whole-genome transcriptional analysis of cbk1Delta suggests that the kinase regulates the expression of a number of genes with cell wall-related functions, including two genes required for efficient cell separation: the chitinase-encoding gene CTS1 and the glucanase-encoding gene SCW11. The Ace2p transcription factor is required for expression of CTS1 and has been shown to physically interact with Cbk1p. Analysis of ace2Delta cells reveals that Ace2p is required for cell separation but not for polarized growth. Our results suggest that Cbk1p and Hym1p function to regulate two distinct cell morphogenesis pathways: an ACE2-independent pathway that is required for efficient apical growth and mating projection formation and an ACE2-dependent pathway that is required for efficient cell separation following cytokinesis. Cbk1p is most closely related to the Neurospora crassa Cot-1; Schizosaccharomyces pombe Orb6; Caenorhabditis elegans, Drosophila, and human Ndr; and Drosophila and mammalian WARTS/LATS kinases. Many Cbk1-related kinases have been shown to regulate cellular morphology.

Study Objectives: Incomplete follow-up can bias interpretation of data that are collected in longitudinal studies. We noted that many patients failed to return for follow-up in a study of effect of lung volume reduction surgery (LVRS) on quality of life (QOL). Accordingly, we designed this investigation to determine the reasons patients dropped out, and to assess differences between those who continued in the study (attendees) and those who did not (nonattendees)., Design: Telephone survey., Subjects: Patients with advanced emphysema who had undergone LVRS and had previously agreed to participate in a longitudinal QOL study., Results: No differences were found with regard to age, gender, preoperative pulmonary function, or oxygen use between attendees and nonattendees. Long-term mortality in nonattendees (27%) was considerably greater than that seen in attendees (3%, p < 0.05). Distance from the hospital, financial burden, and living out of the region were the most common reasons cited by surviving nonattendees for their failure to return for follow-up., Conclusions: Studies reporting the long-term mortality after LVRS can be biased in the direction of underestimating the true value if they are compromised by incomplete follow-up.

The genomic revolution is undeniable: in the past year alone, the term 'genomics' was found in nearly 500 research articles, and at least 6 journals are devoted solely to genomic biology. More than just a buzzword, molecular biology has genuinely embraced genomics (the systematic, large-scale study of genomes and their functions). With its facile genetics, the budding yeast Saccharomyces cerevisiae has emerged as an important model organism in the development of many current genomic methodologies. These techniques have greatly influenced the manner in which biology is studied in yeast and in other organisms. In this review, we summarize the most promising technologies in yeast genomics.

The soothing, repetitive movement of rocking and swinging is well known to anyone who has enjoyed a porch swing or rocking chair. Positive effects of rocking have been reported. However, investigations on its effect in people with dementia are limited. The purposes of this quasi-experimental, repeated-measures design study were to measure the effects of a glider swing on emotions, relaxation, and aggressive behaviors in a group of nursing home residents with dementia (n = 30). Data were obtained during a 5-day baseline phase, a 10-day intervention phase, and a 5-day posttreatment phase. Subjects were placed on the glider for 20 minutes each day during the intervention phase. The results of the study indicate that the glider intervention significantly improved emotions and relaxation. The most noted changes were found after 10 minutes of swinging. However, no differences were found in aggressive behaviors.

The completion of the DNA sequence of the budding yeast Saccharomyces cerevisiae resulted in the identification of a large number of genes. However, the function of most of these genes is not known. One of the best ways to determine gene function is to carry out mutational and phenotypic analysis. In recent years, several approaches have been developed for the mutational analysis of yeast genes on a large scale. These include transposon-based insertional mutagenesis, and systematic deletions using PCR-based approaches. These projects have produced collections of yeast strains and plasmid alleles that can be screened using novel approaches. Analysis of these collections by the scientific community promises to reveal a great deal of biological information about this organism.

In the past, studies of protein activities have focused on studying a single protein at a time, which is often time-consuming and expensive. Recently, with the sequencing of entire genomes, large-scale proteome analysis has begun. Arrays of proteins have been used for the determination of subcellular localization, analysis of protein-protein interactions and biochemical analysis of protein function. New protein-microarray technologies have been introduced that enable the high-throughput analysis of protein activities. These have the potential to revolutionize the analysis of entire proteomes.

Proteins interact with genomic DNA to bring the genome to life; and these interactions also define many functional features of the genome. SBF and MBF are sequence-specific transcription factors that activate gene expression during the G1/S transition of the cell cycle in yeast. SBF is a heterodimer of Swi4 and Swi6, and MBF is a heterodimer of Mbpl and Swi6 (refs 1, 3). The related Swi4 and Mbp1 proteins are the DNA-binding components of the respective factors, and Swi6 mayhave a regulatory function. A small number of SBF and MBF target genes have been identified. Here we define the genomic binding sites of the SBF and MBF transcription factors in vivo, by using DNA microarrays. In addition to the previously characterized targets, we have identified about 200 new putative targets. Our results support the hypothesis that SBF activated genes are predominantly involved in budding, and in membrane and cell-wall biosynthesis, whereas DNA replication and repair are the dominant functions among MBF activated genes. The functional specialization of these factors may provide a mechanism for independent regulation of distinct molecular processes that normally occur in synchrony during the mitotic cell cycle.

The rapid advances in the Human Genome Project and genomic technologies have produced massive amounts of data populated in a large number of network-accessible databases. These technological advances and the associated data can have a great impact on biomedicine and healthcare. To answer many of the biologically or medically important questions, researchers often need to integrate data from a number of independent but related genome databases. One common practice is to download data sets (text files) from various genome Web sites and process them by some local programs. One main problem with this approach is that these programs are written on a case-by-case basis because the data sets involved are heterogeneous in structure. To address this problem, we define metadata that maps these heterogeneously structured files into a common eXtensible Markup Language (XML) structure to facilitate data interoperation. We illustrate this approach by interoperating two sets of essential yeast genes that are stored in two yeast genome databases (MIPS and YPD).

Intervention studies for persons with dementia present many challenges. This article describes challenges encountered in conducting a glider swing intervention study for persons with dementia and the strategies used to manage the problems encountered. The overall purpose of a quasi-experimental study was to explore the effects of a glider swing intervention on emotions, relaxation, and aggressive behaviors in nursing home residents with dementia. Suggestions for investigators conducting intervention studies for persons with dementia are provided.

Consumer interest in and use of complementary/alternative therapies has increased exponentially in the past decades. Although many of the over 1800 therapies have not been used in the delivery of nursing care, a number of these therapies have a long tradition of use in nursing. Additionally, nurses have conducted research on selected complementary therapies. Nursing is in an excellent position to be a leader in integrating these therapies into the Western biomedical health model and in continuing the research that will provide a better scientific base for the use of complementary therapies.

The purpose of this study was to determine the effect on clinical outcomes for newly admitted nursing home residents when advanced practice gerontological nurses (APNs) worked with staff to implement scientifically based protocols for incontinence, pressure ulcers, depression, and aggressive behavior. Use of APNs in this manner differs from the usual way APNs have been used in nursing homes, in which their primary focus has been to augment the physician's role. The APN treatment was randomly assigned to two nursing homes and usual care was assigned to a third. Trajectories from admission to 6 months revealed that residents with APN input into their care (n = 86) experienced significantly greater improvement or less decline in incontinence, pressure ulcers, and aggressive behavior, and they had higher mean composite trajectory scores compared with residents receiving usual care (n = 111). Significantly less deterioration in affect was noted in cognitively impaired residents in the treatment group. Findings suggest that APNs can be effective links between current scientific knowledge about clinical problems and nursing home staff.

We have developed a novel protein chip technology that allows the high-throughput analysis of biochemical activities, and used this approach to analyse nearly all of the protein kinases from Saccharomyces cerevisiae. Protein chips are disposable arrays of microwells in silicone elastomer sheets placed on top of microscope slides. The high density and small size of the wells allows for high-throughput batch processing and simultaneous analysis of many individual samples. Only small amounts of protein are required. Of 122 known and predicted yeast protein kinases, 119 were overexpressed and analysed using 17 different substrates and protein chips. We found many novel activities and that a large number of protein kinases are capable of phosphorylating tyrosine. The tyrosine phosphorylating enzymes often share common amino acid residues that lie near the catalytic region. Thus, our study identified a number of novel features of protein kinases and demonstrates that protein chip technology is useful for high-throughput screening of protein biochemical activity.

The role of gamma-tubulin in microtubule nucleation is well established, however, its function in other aspects of microtubule organization is unknown. The carboxy termini of alpha/beta-tubulins influence the assembly and stability of microtubules. We investigated the role of the carboxy terminus of yeast gamma-tubulin (Tub4p) in microtubule organization. This region consists of a conserved domain (DSYLD), and acidic tail. Cells expressing truncations lacking the DSYLD domain, tail or both regions are temperature sensitive for growth. Growth defects of tub4 mutants lacking either or both carboxy-terminal domains are suppressed by the microtubule destabilizing drug benomyl. tub4 carboxy-terminal mutants arrest as large budded cells with short bipolar spindles positioned at the bud neck. Electron microscopic analysis of wild-type and CTR mutant cells reveals that SPBs are tightly associated with the bud neck/cortex by cytoplasmic microtubules in mutants lacking the tail region (tub4-delta 444, tub4-delta 448). Mutants lacking the DSYLD residues (tub4-delta 444, tub4-delta DSYLD) form many cytoplasmic microtubules. We propose that the carboxy terminus of Tub4p is required for re-organization of the microtubules upon completion of nuclear migration, and facilitates spindle elongation into the bud.

Dehydrated hereditary stomatocytosis (DHS) is a rare genetic disorder of red cell permeability to cations, leading to a well-compensated hemolytic anemia. DHS was shown previously to be associated in some families with a particular form of perinatal edema, which resolves in the weeks following birth and, in addition, with pseudohyperkalemia in one kindred. The latter condition was hitherto regarded as the separate entity, "familial pseudohyperkalemia." DHS and familial pseudohyperkalemia are thought to stem from the same gene, mapping to 16q23-q24. This study screened 8 French and 2 American families with DHS. DHS appeared to be part of a pleiotropic syndrome in some families: DHS + perinatal edema, DHS + pseudohyperkalemia, or DHS + perinatal edema + pseudohyperkalemia. If adequately attended to, the perinatal edema resolved spontaneously after birth. Logistic regression showed that increased mean corpuscular volume and mean corpuscular hemoglobin concentration were the parameters best related to DHS. In patients in whom cation fluxes were investigated, the temperature dependence of the monovalent cation leak exhibited comparable curves. Specific recombination events consistently suggested that the responsible gene lies between markers D16S402 and D16S3037 (16q23-q24). The 95% confidence limits (Z(max) >/= 3.02) spanned almost the complete 9-cM interval between these 2 markers.