Clusters of drug involvement in Panama: results from Panama's 1996 National Youth Survey.
This report is based on the first epidemiological investigation of clustering of tobacco, alcohol, inhalant, and other drug involvement within individual schools using data from Panama's 1996 National Youth Survey on Alcohol and Drug Use. Clustering was estimated with the Alternating Logistic Regression method. Adjusted estimates of pair-wise cross-product ratios (PWCPR), a measure of clustering, show modest clustering (i.e. PWCPR>1.0) at the school level for tobacco smoking (PWCPR=1.41; 95% confidence interval, CI=1.22-1.64), alcohol consumption (PWCPR=1.33; 95% CI=1.22-1.45), use of inhalants, (PWCPR=1.35; 95% CI=1.07-1.69), and other drug use (PWCPR=1.38; 95% CI=1.14-1.68). These findings provide preliminary evidence that the odds of drug use among school-attending youths increase when another youth in the same school uses drugs, and suggest a new line of research on within-school diffusion that should include the identification of school-level factors that contribute to student drug use.


Foodborne Illness Outbreaks at Retail Establishments - National Environmental Assessment Reporting System, 16 State and Local Health Departments, 2014-2016.
State and local public health departments report hundreds of foodborne illness outbreaks each year to CDC and are primarily responsible for investigations of these outbreaks. Typically, investigations involve epidemiology, laboratory, and environmental health components. Health departments voluntarily report epidemiologic and laboratory data from their foodborne illness outbreak investigations to CDC through the Foodborne Disease Outbreak Surveillance System (FDOSS); however, minimal environmental health data from outbreak investigations are reported to FDOSS. 2014-2016. In 2014, CDC launched the National Environmental Assessment Reporting System (NEARS) to complement FDOSS surveillance and to use these data to enhance prevention efforts. State and local health departments voluntarily report data from their foodborne illness outbreak investigations of retail food establishments. These data include characteristics of foodborne illness outbreaks (e.g., agent), characteristics of establishments with outbreaks (e.g., number of meals served daily), food safety policies and practices of these establishments (e.g., glove use policies), and characteristics of outbreak investigations (e.g., timeliness of investigation activities). NEARS is the only available data source that includes characteristics of retail establishments with foodborne illness outbreaks. During 2014-2016, a total of 16 state and local public health departments reported data to NEARS on 404 foodborne illness outbreaks at retail establishments. The majority of outbreaks with a suspected or confirmed agent were caused by norovirus (61.1%). The majority of outbreaks with identified contributing factors had at least one factor associated with food contamination by a worker who was ill or infectious (58.6%). Almost half (47.4%) of establishments with outbreaks had a written policy excluding ill workers from handling food or working. Approximately one third (27.7%) had a written disposable glove use policy. Paid sick leave was available for at least one worker in 38.3% of establishments. For most establishments with outbreaks (68.7%), environmental health investigators initiated their component of the investigation soon after learning about the outbreak (i.e., the same day) and completed their component in one or two visits to the establishment (75.0%). However, in certain instances, contacting the establishment and completing the environmental health component of the investigation occurred much later (>8 days). Most outbreaks reported to NEARS were caused by norovirus, and contamination of food by workers who were ill or infectious contributed to more than half of outbreaks with contributing factors; these findings are consistent with findings from other national outbreak data sets and highlight the role of workers in foodborne illness outbreaks. The relative lack of written policies for ill workers and glove use and paid sick leave for workers in establishments with outbreaks indicates gaps in food safety practices that might have a role in outbreak prevention. The environmental health component of the investigation for most outbreaks was initiated quickly, yet the longer initiation timeframe for certain outbreaks suggests the need for improvement. Retail establishments can reduce viral foodborne illness outbreaks by protecting food from contamination through proper hand hygiene and excluding workers who are ill or infectious from working. NEARS data can help prioritize training and interventions for state and local food safety programs and the retail food establishment industry by identifying gaps in food safety policies and practices and types of establishments vulnerable to outbreaks. Improvement of certain outbreak investigation practices (e.g., delayed initiation of environmental health investigations) can accelerate identification of the agent and implementation of interventions. Future analysis comparing establishments with and without outbreaks will contribute knowledge about how establishments' characteristics and food safety policies and practices relate to foodborne illness outbreaks and provide information to develop effective prevention approaches.


Reverse polarity optical-OFDM (RPO-OFDM): dimming compatible OFDM for gigabit VLC links.
Visible light communications (VLC) technology permits the exploitation of light-emitting diode (LED) luminaries for simultaneous illumination and broadband wireless communication. Optical orthogonal frequency-division multiplexing (O-OFDM) is a promising modulation technique for VLC systems, in which the real-valued O-OFDM baseband signal is used to modulate the instantaneous power of the optical carrier to achieve gigabit data rates. However, a major design challenge that limits the commercialization of VLC is how to incorporate the industry-preferred pulse-width modulation (PWM) light dimming technique while maintaining a broadband and reliable communication link. In this work, a novel signal format, reverse polarity O-OFDM (RPO-OFDM), is proposed to combine the fast O-OFDM communication signal with the relatively slow PWM dimming signal, where both signals contribute to the effective LED brightness. The advantages of using RPO-OFDM include, (1) the data rate is not limited by the frequency of the PWM signal, (2) the LED dynamic range is fully utilized to minimize the nonlinear distortion of the O-OFDM communication signal, and (3) the bit-error performance is sustained over a large fraction of the luminaire dimming range. In addition, RPO-OFDM offers a practical approach to utilize off-the-shelf LED drivers. We show results of numerical simulations to study the trade-offs between the PWM duty cycle, average electrical O-OFDM signal power, radiated optical flux as well as human perceived light.


Diffusion of medication drop-boxes in North Carolina from 2007 to 2016.
A permanent drug donation box ("drop-box") is one strategy implemented in communities across the United States to reduce the availability of excess controlled medications, including prescription opioids, for diversion. The objective of this study was to examine correlates of the diffusion and implementation of drop-boxes in North Carolina. We assessed the number and location of drop-boxes implemented in North Carolina. Cox proportional hazards models were used to examine covariates associated with drop-box implementation in NC counties (n = 100) between 2007 and 2016. There were 311 drop-boxes implemented in 91 (out of 100) counties. Most drop-boxes were in law enforcement agencies (78.8%) and a growing number were in pharmacies (14.5%). Counties with a higher percentage of whites, more educated residents, a substance abuse prevention coalition, higher rates of controlled medications dispensed and prescription opioid overdose, and that were Appalachian were more likely to be early adopters. Rural counties were less likely to have a drop-box. In the multivariate model, only higher rate of controlled medicines dispensed was significant. A growing number of drop-boxes are being implemented in law enforcement offices and pharmacies. Given that communities with higher rates of controlled medication dispensing likely have the highest need for disposal opportunities, it is promising that they are early adopters of drop-boxes. Future research should assess the effectiveness of drop-boxes as they become more widespread in a variety of locations.


Electrogastrography in experimental pigs: the influence of gastrointestinal injury induced by dextran sodium sulphate on porcine gastric erythromycin-stimulated myoelectric activity.
Electrogastrography (EGG) is a non-invasive investigation of gastric myoelectrical activity. The aim of study was to evaluate the impact of erythromycin on EGG in gastrointestinal toxic injury induced by dextran sodium sulphate (DSS) in experimental pigs. The experiments were carried out on 12 adult pigs (weighing 30-35 kg). EGG was recorded using Digitrapper equipment (Synectics Medical AB, Stockholm). Running spectrum activity was used for EGG evaluation. There were two groups of animals: Group I: 6 controls with erythromycin administration (1,600 mg intragastrically); Group II: 6 animals treated with DSS (for 5 days, 0.25 g/kg per day in a dietary bolus) followed by erythromycin administration. Baseline and subsequent six separate 30-minute EGG-recordings (from time 0 to 360 min) were accomplished in each animal. A total of 84 records were analysed. Baseline dominant frequency of slow waves was fully comparable in both groups. In Group I, there was a significant increase in dominant frequency after erythromycin administration (maximum between 240-360 min). There was a flat non-significant and delayed increase in dominant frequency after erythromycin administration in Group II. The difference between Group I and II at particular time intervals was not significant but a diverse trend was evident. EGG recording enables us to register a gastric myoelectrical effect of prokinetic drugs. Erythromycin induced a significant increase in the dominant frequency of slow waves. DSS caused toxic injury to the porcine gastrointestinal tract responsible for the delayed and weaker myoelectrical effect of erythromycin in experimental animals.


Seasonal influenza vaccine compliance among hospital-based and nonhospital-based healthcare workers.
Influenza vaccination among nonhospital healthcare workers (HCWs) is imperative, but only limited data are available for factors affecting their compliance. To examine the factors influencing influenza vaccine compliance among hospital and nonhospital HCWs. Design and Setting. A vaccine compliance questionnaire was administered to HCWs working in myriad healthcare settings in March-June 2011. Online and paper surveys were used to assess compliance with the 2010/2011, 2009/2010, and H1N1 influenza vaccines and to examine factors that predicted the uptake of the 2010/2011 seasonal influenza vaccine. In all, 3,188 HCWs completed the survey; half of these ([Formula: see text]) reported no hospital work time. Compliance rates for all 3 vaccines were significantly higher ([Formula: see text]) among hospital versus nonhospital HCWs. In logistic regression stratified by hospital versus nonhospital setting, and when controlling for demographics and past behavior, the determinants of vaccination against the 2010/2011 seasonal influenza among nonhospital-based HCWs included having a mandatory vaccination policy, perceived importance, no fear of vaccine adverse effects, free and on-site access, and perceived susceptibility to influenza. Determinants of hospital-based HCW vaccine compliance included having a mandatory vaccination policy, belief that HCWs should be vaccinated every year, occupational health encouragement, perceived importance of vaccination, on-site access, and no fear of vaccine adverse effects. The strongest predictor of compliance for both worker groups was existence of a mandatory vaccination policy. The reasons for vaccine uptake among nonhospital-based versus hospital-based HCWs differed. Targeted interventions should be aimed at workers in these settings to increase their vaccine compliance, including implementing a mandatory vaccination policy.


Metastasis-associated protein 1 (MTA1) is transferred by exosomes and contributes to the regulation of hypoxia and estrogen signaling in breast cancer cells.
Exosomes are small membrane-bound vesicles that contribute to tumor progression and metastasis by mediating cell-to-cell communication and modifying the tumor microenvironment at both local and distant sites. However, little is known about the predominant factors in exosomes that contribute to breast cancer (BC) progression. MTA1 is a transcriptional co-regulator that can act as both a co-activator and co-repressor to regulate pathways that contribute to cancer development. MTA1 is also one of the most up-regulated proteins in cancer, whose expression correlates with cancer progression, poor prognosis and increased metastatic potential. We identified MTA1 in BC exosomes by antibody array and confirmed expression of exosome-MTA1 across five breast cancer cells lines. Ectopic expression of tdTomato-tagged MTA1 and exosome transfer were examined by fluorescent microscopy. CRISPR/Cas9 genetic engineering was implemented to knockout MTA1 in MCF7 and MDA-MB-231 breast cancer cells. Reporter assays were used to monitor hypoxia and estrogen receptor signaling regulation by exosome-MTA1 transfer. Ectopic overexpression of tdTomato-MTA1 in BC cell lines demonstrated exosome transfer of MTA1 to BC and vascular endothelial cells. MTA1 knockout in BC cells reduced cell proliferation and attenuated the hypoxic response in these cells, presumably through its co-repressor function, which could be rescued by the addition of exosomes containing MTA1. On the other hand, consistent with its co-activator function, estrogen receptor signaling was enhanced in MTA1 knockout cells and could be reversed by addition of MTA1-exosomes. Importantly, MTA1 knockout sensitized hormone receptor negative cells to 4-hydroxy tamoxifen treatment, which could be reversed by the addition of MTA1-exosomes. This is the first report showing that BC exosomes contain MTA1 and can transfer it to other cells resulting in changes to hypoxia and estrogen receptor signaling in the tumor microenvironment. These results, collectively, provide evidence suggesting that exosome-mediated transfer of MTA1 contributes to BC progression by modifying cellular responses to important signaling pathways and that exosome-MTA1 may be developed as a biomarker and therapeutic target for BC.


The proteasome inhibitor PS-341 inhibits growth and induces apoptosis in Bcr/Abl-positive cell lines sensitive and resistant to imatinib mesylate.
Imatinib mesylate (IM) is the choice treatment for Bcr/Abl-positive malignancies. Emergence of resistance to IM warrants the exploration of novel well-tolerated anticancer agents. We intended to evaluate the effect of PS-341 on proliferation, survival, and cellular events in Bcr/Abl-positive cells sensitive and resistant to IM, and to investigate the effect of PS-341 and IM in conjunction. Bcr/Abl-positive cell lines sensitive (p210Bcr/Abl KBM5, p210Bcr/Abl KBM7, and p190Bcr/Abl Z-119) or resistant (KBM5-R) to IM were treated with PS-341 alone or in combination with IM. The effect on cell growth was determined using the MTT assay. Cell-cycle analysis was performed by propidium iodide staining. Apoptosis was evaluated by measurement of sub-G1 DNA content, annexin V binding, and caspase 3 activity assays. Levels of apoptotic proteins, P-IkBalpha, Bcr/Abl, and phosphorylated Bcr/Abl were determined by western blotting. NF-kappaB activity was evaluated by electromobility gel shift assays. PS-341 exerted growth inhibition effects in IM-sensitive and -resistant cells. This phenomenon correlated with accumulation of cells in the G2/M phase of cell cycle; transient downregulation of NFkappaB DNA binding activity; downregulation of Bcl-xL; activation of caspase 3, induction of apoptosis; inhibition of expression and phosphorylation of Bcr/Abl. Sequential combination of PS-341 followed by IM demonstrated a synergistic pro-apoptotic effect in IM-sensitive cells; concomitant exposure was antagonistic. PS-341 suppresses growth and induces apoptosis in Bcr/Abl-positive cells sensitive and resistant to IM. The use of PS-341 should be explored in patients with chronic myelogenous leukemia resistant to IM. Trials of combinations of PS-341 and IM require cautious design.


Resistance of Eisenia fetida (Oligochaeta) to cadmium after long-term exposure.
The reliance on earthworms as test organisms in risk assessment studies of polluted environments raises the question whether they can evolve resistance, e.g., by adaptation to specific toxicants. Protection criteria may be biased if sensitivity data from adapted populations are used. Increased resistance to the heavy metal cadmium has not yet been determined for terrestrial Oligochaeta. Eisenia fetida was exposed to a sublethal concentration of cadmium sulfate for more than 10 generations. Clitellate worms from this culture were used in experiments to determine the extent of possible tolerance for the heavy metal. Preexposed animals as well as worms with no previous history of exposure to cadmium were exposed to a control substrate without cadmium and also to two substrates with 600 and 1200 microg g-1 cadmium. Changes in biomass, cocoon production, and hatching success were monitored. The results obtained indicated that in both substrates in which cadmium was present the preexposed worms performed better than the unexposed worms with respect to growth rate but not reproductively. In the substrate without cadmium the preexposed worms exhibited signs of poisoning after a few weeks. Preexposed and unexposed worms were also exposed to concentrations of 1500 to 4000 microg g-1 cadmium sulfate in an artisol medium for a period of 2 weeks. The preexposed worms survived higher concentrations of cadmium than the unexposed group and some specimens from the unexposed group had a gross increase in body fluids. It is concluded that worms with a long-term history of exposure to the metal developed resistance to cadmium.


Random phenotypic variation of yeast (Saccharomyces cerevisiae) single-gene knockouts fits a double pareto-lognormal distribution.
Distributed robustness is thought to influence the buffering of random phenotypic variation through the scale-free topology of gene regulatory, metabolic, and protein-protein interaction networks. If this hypothesis is true, then the phenotypic response to the perturbation of particular nodes in such a network should be proportional to the number of links those nodes make with neighboring nodes. This suggests a probability distribution approximating an inverse power-law of random phenotypic variation. Zero phenotypic variation, however, is impossible, because random molecular and cellular processes are essential to normal development. Consequently, a more realistic distribution should have a y-intercept close to zero in the lower tail, a mode greater than zero, and a long (fat) upper tail. The double Pareto-lognormal (DPLN) distribution is an ideal candidate distribution. It consists of a mixture of a lognormal body and upper and lower power-law tails. If our assumptions are true, the DPLN distribution should provide a better fit to random phenotypic variation in a large series of single-gene knockout lines than other skewed or symmetrical distributions. We fit a large published data set of single-gene knockout lines in Saccharomyces cerevisiae to seven different probability distributions: DPLN, right Pareto-lognormal (RPLN), left Pareto-lognormal (LPLN), normal, lognormal, exponential, and Pareto. The best model was judged by the Akaike Information Criterion (AIC). Phenotypic variation among gene knockouts in S. cerevisiae fits a double Pareto-lognormal (DPLN) distribution better than any of the alternative distributions, including the right Pareto-lognormal and lognormal distributions. A DPLN distribution is consistent with the hypothesis that developmental stability is mediated, in part, by distributed robustness, the resilience of gene regulatory, metabolic, and protein-protein interaction networks. Alternatively, multiplicative cell growth, and the mixing of lognormal distributions having different variances, may generate a DPLN distribution.


When and how to use Q methodology to understand perspectives in conservation research.
Understanding human perspectives is critical in a range of conservation contexts, for example, in overcoming conflicts or developing projects that are acceptable to relevant stakeholders. The Q methodology is a unique semiquantitative technique used to explore human perspectives. It has been applied for decades in other disciplines and recently gained traction in conservation. This paper helps researchers assess when Q is useful for a given conservation question and what its use involves. To do so, we explained the steps necessary to conduct a Q study, from the research design to the interpretation of results. We provided recommendations to minimize biases in conducting a Q study, which can affect mostly when designing the study and collecting the data. We conducted a structured literature review of 52 studies to examine in what empirical conservation contexts Q has been used. Most studies were subnational or national cases, but some also address multinational or global questions. We found that Q has been applied to 4 broad types of conservation goals: addressing conflict, devising management alternatives, understanding policy acceptability, and critically reflecting on the values that implicitly influence research and practice. Through these applications, researchers found hidden views, understood opinions in depth and discovered points of consensus that facilitated unlocking difficult disagreements. The Q methodology has a clear procedure but is also flexible, allowing researchers explore long-term views, or views about items other than statements, such as landscape images. We also found some inconsistencies in applying and, mainly, in reporting Q studies, whereby it was not possible to fully understand how the research was conducted or why some atypical research decisions had been taken in some studies. Accordingly, we suggest a reporting checklist.


Do mandibular cross-sectional properties and dental microwear give similar dietary signals?
Previous animal experimental work evaluating the effects of dietary consistency on mastication was generally limited to studies of either mandibular structure or rates and types of tooth wear. Control groups fed hard diets (HD) consistently exhibited increased cortical remodeling and/or bone strength when compared to groups fed soft diets (SD). Results of tooth-wear studies showed faster rates of tooth wear in HD animals. This study evaluates the effects of dietary differences on both mandibular structural morphology and postcanine dental microwear in the same animals. We examined mandibles and dentitions from eight miniature swine, raised from 4 weeks to 9 months of age on HD and SD (n = 4, each group). Mandibular structural properties were calculated from peripheral quantitative computed tomography slices at the dp3-dp4 and dp4-M1 junctions. Dental microwear analysis was performed on mandibular lingual crushing facets of dp4 and M1, using photomicrographs of high-resolution casts taken at 500x magnification in a scanning electron microscope. Our results suggest that between the dp3-dp4 contact, HD animals have mandibles that are stronger and more rigid mediolaterally than SD animals. At the dp4-M1 contact, HD animals have mandibles that are stronger and more rigid mediolaterally, dorsoventrally, and in torsion than SD animals. Dental microwear results indicate that SD pigs have higher incidences of pitting and more overall microwear features on their premolars than do HD pigs, yet there are no significant differences in molar microwear morphology between the dietary groups. Near-significant correlations exist between pit size and dorsoventral bending strength, but only for HD pigs. These results suggest that dietary consistency significantly affects both mandibular structure and dental microwear, yet direct correlations between the two are complicated by a number of factors.


A Bayesian model for cluster detection.
The detection of areas in which the risk of a particular disease is significantly elevated, leading to an excess of cases, is an important enterprise in spatial epidemiology. Various frequentist approaches have been suggested for the detection of "clusters" within a hypothesis testing framework. Unfortunately, these suffer from a number of drawbacks including the difficulty in specifying a p-value threshold at which to call significance, the inherent multiplicity problem, and the possibility of multiple clusters. In this paper, we suggest a Bayesian approach to detecting "areas of clustering" in which the study region is partitioned into, possibly multiple, "zones" within which the risk is either at a null, or non-null, level. Computation is carried out using Markov chain Monte Carlo, tuned to the model that we develop. The method is applied to leukemia data in upstate New York.


Inhibition of human converting enzyme in vitro by a novel tripeptide analog.
We have studied inhibition of homogeneous human converting enzyme by a new inhibitor, a ketomethylene derivative of the blocked tripeptide substrate, Bz-Phe-Gly-Pro (ketoACE). KetoACE inhibited the hydrolysis of Hip-His-Leu and Hip-Phe-Arg at different concentrations (I50 values were 4 X 10(-8) M and 2 X 10(-7) M, respectively). Kinetic studies indicated that ketoACE inhibits the hydrolysis of both substrates by a similar, non-competitive mechanism. At the lowest enzyme concentration tested, using 3H-Hip-Gly-Gly as substrate, the I50 of ketoACE was 6 X 10(-9) M. KetoACE protected a functional tyrosine residue in the active site of human converting enzyme from modification with N-acetylimidazole. It is proposed that there are alternate (hydrophobic) binding sites for both inhibitors and substrates in the active site of human converting enzyme. It should be possible to develop other high-affinity inhibitors of this class that bind to hydrophobic sites and do not require metal binding via a sulfhydryl group.


Changes in the Healthy Beverage Index in Response to an Intervention Targeting a Reduction in Sugar-Sweetened Beverage Consumption as Compared to an Intervention Targeting Improvements in Physical Activity: Results from the Talking Health Trial.
The recently developed Healthy Beverage Index (HBI) was designed to evaluate overall beverage intake quality (including total fluid consumption and beverage calories), yet no known intervention studies have assessed longitudinal changes to the HBI. The objective of this investigation was to assess changes in HBI scores in response to a sugar-sweetened beverage (SSB) reduction trial as compared to a physical activity comparison group. Participants were enrolled into a six-month, community-based, controlled behavioral trial and randomized into either a SSB reduction group (SIPsmartER) or a physical activity group (MoveMore). Correlations and multilevel mixed-effects linear regression with intention-to-treat analyses are presented. Total HBI score significantly increased for SIPsmartER (n = 149) (mean increase = 7.5 points (5.4, 9.7), p ≤ 0.001) and MoveMore (n = 143) (mean increase = 3.4 points (1.6, 5.2), p ≤ 0.001) participants, with a significant between group effect (p ≤ 0.05), over the six-month intervention. Other significant changes in HBI components for SIPsmartER included increased SSB and total beverage calorie scores, and decreased low-fat milk and diet soda scores. Changes in total HBI scores were significantly correlated with changes in total Healthy Eating Index-2010 scores (r = 0.15, p ≤ 0.01). Our findings suggest that individual HBI component scores, beyond the SSB component, are influenced by intervention strategies that primarily focus on SSB reduction.


Sex differences in morbidity and mortality.
Women have worse self-rated health and more hospitalization episodes than men from early adolescence to late middle age, but are less likely to die at each age. We use 14 years of data from the U.S. National Health Interview Survey to examine this paradox. Our results indicate that the difference in self-assessed health between women and men can be entirely explained by differences in the distribution of the chronic conditions they face. This is not true, however, for hospital episodes and mortality. Men with several smoking-related conditions--including cardiovascular disease and certain lung disorders--are more likely to experience hospital episodes and to die than women who suffer from the same chronic conditions, implying that men may experience more-severe forms of these conditions. While some of the difference in mortality can be explained by differences in the distribution of chronic conditions, an equally large share can be attributed to the larger adverse effects of these conditions on male mortality. The greater effects of smoking-related conditions on men's health may be due to their higher rates of smoking throughout their lives.


Station-length requirements for reliable performance-based examination scores.
To directly compare the generalizability of medical students' performance scores under systematically varied station times in two surgery end-of-clerkship performance-based examinations. The participants were 36 third-year students randomly assigned to the first two rotations of the core surgery clerkship during 1991-92 at Southern Illinois University School of Medicine. The students rotated through a 12-station examination that employed standardized patients (SPs). In the first rotation, the student took six five-minute stations and six ten-minute stations. In the second rotation, the time lengths were reversed for the same stations. The students' total scores were based on (1) subscores on checklists that were completed by the SPs and (2) subscores on the students' written responses to short questions about each station (these responses were provided at station couplets that were five minutes long, regardless of station length). Generalizability coefficients were computed from the pooled rotation results to provide reliabilities for scores from the two station lengths. Generalizability decreased in the ten-minute stations, mostly attributable to less variability among students' performances. The checklist subscores accounted for most of this variability, while couplet subscores remained stable between station lengths. The longer station length actually decreased the generalizability of the scores by decreasing the variability among students' performances; thus, allocating different times to stations can affect the score reliability, as well as impact on the overall testing time, of performance-based examinations.


Pharmacologic aspects of pentamidine.
Pentamidine is an aromatic diamidino compound synthesized originally for the therapy of trypanosomiasis. The pharmacologic effects of pentamidine vary, depending on its route of administration. In animals, the dominant effects have been a precipitous, transitory drop in blood pressure after injection and renal toxicity following repeated administration. To avoid the possibility of immediate toxic reactions associated with iv administration, we now usually give the drug im to humans. Further interest in pentamidine has been stimulated by its usefulness in the treatment of interstitial pneumonia caused by Pneumocystis carinii. In some patients receiving antineoplastic or immunosuppressive therapy who have superimposed P. carinii pneumonia, pentamidine may cause serious renal toxicity. Distribution and excretion studies in animals indicate pentamidine is deposited in tissues, with the greatest concentration in the kidneys, and gradually eliminated over a prolonged period. The mechanism of action of pentamidine against P. carinii or the means whereby fixation in tissues and subsequent toxicity occur have not been elucidated. Recent investigations to help clarify these points indicate that pentamidine inhibits dihydrofolate reductase in all tissues studied both in vitro and in vivo. In addition, pentamidine interacts and forms water-insoluble products with specific nucleotides and nucleic acids.


Risk factors of chronic hepatitis in antiretroviral-treated HIV infection, without hepatitis B or C viral infection.
Increasing rates of non-AIDS defining illnesses, and in particular liver diseases, have been found after the initiation of highly active antiretroviral therapy. However, there is little evidence concerning the risk factors for and clinical characteristics of liver disease in antiretroviral (ARV)-treated HIV infection, in the absence of hepatitis B or C viral co-infection. A nested case-control study of HIV infected volunteers, matched by starting date of anti-retroviral treatment, was conducted in a Thai cohort studied from Nov 2002 - July 2012. Cases were defined as those subjects with an elevated alanine aminotransferase (ALT ≥ 40 IU/L) at two consecutive visits six months apart, while controls were defined as individuals who never demonstrated two consecutive elevated ALT results and had a normal ALT result (< 40 IU/L) at their last visit. Both groups had normal ALT levels prior to ARV initiation. Clinical demographics and risk factors for chronic hepatitis including HIV-related illness, ARV treatment and metabolic diseases were collected and analyzed. Conditional logistic regression was used to determine risk factors for chronic hepatitis in HIV infection. A total of 124 matched pairs with HIV infection were followed over 3,195 person-years. The mean age (±SD) was 33.0 ± 7.3 years, with 41.1% of subjects being male. The incidence of chronic hepatitis was 5.4 per 100 person-years. The median time from initiation of ARV to chronic hepatitis was 1.3 years (IQR, 0.5-3.5). From univariate analysis; male sex, plasma HIV-1 RNA level > 5 log 10 copies/ml, metabolic syndrome at baseline visit, high BMI > 23 kg/m(2), abnormal HDL cholesterol at time of ALT elevation and treatment experience with NNRTI plus boosted PI were selected (p value < 0.2) to the final model of multivariate analysis. Male sex had 3.1 times greater risk of chronic hepatitis than the females by multivariate analysis (adjusted OR, 95% CI: 3.1, 1.5-6.3, p =0.002). High BMI ≥ 23 kg/m(2) was also associated with 2.4 times greater risk of chronic hepatitis (adjusted OR, 95% CI: 2.4, 1.2-4.8, p = 0.01). Chronic hepatitis in ARV-treated HIV-infected patients is common and may lead to a major health care problem. Male sex and high BMI ≥ 23 kg/m(2) carry higher risks for developing chronic hepatitis in this study. Therefore, these patients should be closely monitored for long-term hepatotoxicity.


[Use of Anterolateral Thigh Free Flap in the Treatment of a Sternal Keloid].
Hypertrophic and keloid scars represent hyperproliferative disorders that can have a significant impact on patients' lives. The authors present the case of a 53 years-old male with a sternal keloid after excision of a sebaceous cyst and multiple sessions of steroid infiltration, with worsening of the lesion. The patient underwent complete excision of the scar and reconstruction with an anterolateral thigh flap - ALT. The postoperative period was uneventful, with no signs of relapse. Keloid scar treatment in sternal area implies a reconstruction with no tension, in order to avoid relapse. Treatment of this type of scars is complex and a challenge to the plastic surgeon. In this case, a microvascular tissue transfer was used to reconstruct the large defect that ultimately resulted after scar excision, in a tension-free manner.


Molecular species identification of bushmeat recovered from the Serengeti ecosystem in Tanzania.
Bushmeat harvesting and consumption represents a potential risk for the spillover of endemic zoonotic pathogens, yet remains a common practice in many parts of the world. Given that the harvesting and selling of bushmeat is illegal in Tanzania and other parts of Africa, the supply chain is informal and may include hunters, whole-sellers, retailers, and individual resellers who typically sell bushmeat in small pieces. These pieces are often further processed, obscuring species-identifying morphological characteristics, contributing to incomplete or mistaken knowledge of species of origin and potentially confounding assessments of pathogen spillover risk and bushmeat offtake. The current investigation sought to identify the species of origin and assess the concordance between seller-reported and laboratory-confirmed species of origin of bushmeat harvested from in and around the Serengeti National Park in Tanzania. After obtaining necessary permits, the species of origin of a total of 151 bushmeat samples purchased from known intermediaries from 2016 to 2018 were characterized by PCR and sequence analysis of the cytochrome B (CytB) gene. Based on these sequence analyses, 30%, 95% Confidence Interval (CI: 24.4-38.6) of bushmeat samples were misidentified by sellers. Misreporting amongst the top five source species (wildebeest, buffalo, impala, zebra, and giraffe) ranged from 20% (CI: 11.4-33.2) for samples reported as wildebeest to 47% (CI: 22.2-72.7) for samples reported as zebra although there was no systematic bias in reporting. Our findings suggest that while misreporting errors are unlikely to confound wildlife offtake estimates for bushmeat consumption within the Serengeti ecosystem, the role of misreporting bias on the risk of spillover events of endemic zoonotic infections from bushmeat requires further investigation.


pmar1/phb homeobox genes and the evolution of the double-negative gate for endomesoderm specification in echinoderms.
In several model animals, the earliest phases of embryogenesis are regulated by lineage-specific genes, such as Drosophila bicoid Sea urchin (echinoid) embryogenesis is initiated by zygotic expression of pmar1, a paired-class homeobox gene that has been considered to be present only in the lineage of modern urchins (euechinoids). In euechinoids, Pmar1 promotes endomesoderm specification by repressing the hairy and enhancer of split C (hesC) gene. Here, we have identified the basal echinoid (cidaroid) pmar1 gene, which also promotes endomesoderm specification but not by repressing hesC A further search for related genes demonstrated that other echinoderms have pmar1-related genes named phb Functional analyses of starfish Phb proteins indicated that, similar to cidaroid Pmar1, they promote activation of endomesoderm regulatory gene orthologs via an unknown repressor that is not HesC. Based on these results, we propose that Pmar1 may have recapitulated the regulatory function of Phb during the early diversification of echinoids and that the additional repressor HesC was placed under the control of Pmar1 in the euechinoid lineage. This case provides an exceptional model for understanding how early developmental processes diverge.


Clinical Telemedicine Utilization in Ontario over the Ontario Telemedicine Network.
Northern Ontario is a region in Canada with approximately 775,000 people in communities scattered across 803,000 km(2). The Ontario Telemedicine Network (OTN) facilitates access to medical care in areas that are often underserved. We assessed how OTN utilization differed throughout the province. We used OTN medical service utilization data collected through the Ontario Health Insurance Plan and provided by the Ministry of Health and Long Term Care. Using census subdivisions grouped by Northern and Southern Ontario as well as urban and rural areas, we calculated utilization rates per fiscal year and total from 2008/2009 to 2013/2014. We also used billing codes to calculate utilization by therapeutic area of care. There were 652,337 OTN patient visits in Ontario from 2008/2009 to 2013/2014. Median annual utilization rates per 1,000 people were higher in northern areas (rural, 52.0; urban, 32.1) than in southern areas (rural, 6.1; urban, 3.1). The majority of usage in Ontario was in mental health and addictions (61.8%). Utilization in other areas of care such as surgery, oncology, and internal medicine was highest in the rural north, whereas primary care use was highest in the urban south. Utilization was higher and therapeutic areas of care were more diverse in rural Northern Ontario than in other parts of the province. Utilization was also higher in urban Northern Ontario than in Southern Ontario. This suggests that telemedicine is being used to improve access to medical care services, especially in sparsely populated regions of the province.


Triangulating laser profilometer as a navigational aid for the blind: optical aspects.
We propose a navigational aid approach for the blind that relies on active optical profilometry with real-time electrotactile interfacing on the skin. Here we are concerned with the optical parts of this system. We point out the particular requirements the profilometer must meet to meet the needs of blind people. We show experimentally that an adequate compromise is possible that consists of a compact class I IR laser-diode triangulation profilometer with the following characteristics: 30-cm to 30-m range, 1-deg angular resolution, 20-ms acquisition time per measure of distance, 60° angular scanning field.


Diagnosis of patients with syncope in emergency medicine.
Syncope is a common medical problem, which accounts for 3.5% of emergency room visits. It includes diverse clinical etiologies, from cardiac origin with poor prognosis to reflex-mediated hypotension with benign clinical course. Trauma is not rare in patients with syncope, e.g. bruises or lacerations on the head and face were recognized in 17%. Accordingly, patients presenting with syncope may have a risk of sudden cardiac death, or risk of trauma if the episodes are repetitive. It is a physicians's task to diagnose syncope and identify its cause. The history is an essential part of the initial evaluation. Electrocardiography (ECG) is necessary in every case. However, organic disease as a cause is identified in 20% of syncope in routine work-up, and the remaining cases are reflex induced hypotension (neurocardiogenic syncope), misdiagnosed arrhythmia, coronary spasm or unknown. Recent investigations suggest the usefulness of provocation to diagnose neurocardiogenic syncope. Two important non-invasive provocations to reproduce syncope are carotid sinus compression and tilt table test. In patients with syncope who visited emergency room, tilt test was positive in 25%, whereas the response to carotid sinus compression was positive in 21%. One or both of the tests was positive in 39%, suggesting that neurally-mediated syncope accounts for a significant number of patients.


Intestinal parasitic infections among long-term-residents and settled immigrants in Qatar in the period 2005 to 2011.
The expanding economy of Qatar in the last two decades has attracted immigrants, often from countries with poor socio-economic levels. Many arrive with patent intestinal parasitic infections, and recent analyses have indicated consistently rising trends in the prevalence of some infections. Here, we examined 18,563 hospital records of subjects in Qatar seeking medical assistance for a variety of ailments, combining data from 2009 to 2011 with the earlier dataset from 2005 to 2008 to enable trends to be identified across a 7-year period. We found that 8.6% were infected with one or more species of parasites, however in contrast to the earlier period (2005-2008), in the latter 3 years there were falling trends of prevalence providing some optimism that parasitic infections among the resident immigrants have begun to decline. We identified also geographic regions from which resident workers still maintain a relatively high prevalence of helminth infections despite their long-term residence in Qatar.


Defining the role of the nurse in population-based cancer screening programs.
Nurses are pivotal in cancer prevention and early detection, but the nurse's role in cancer screening programs has been described only in very general terms without specification of activities needed to develop the role. To identify the set of activities that compose the role of the cancer screening nurse, the authors of the current article performed a critical descriptive literature review to document nursing involvement in cancer screening, covering articles published from 2000-2012. A total of 726 potentially relevant studies were identified, and 22 of those were included in the review. Nurses carry out follow-up, coordinate treatment, ensure continuity throughout the process, provide up-to-date and pertinent information to facilitate patient knowledge and choice, work to ensure coordination among the various levels of care, provide ongoing training, lead research and publications concerning daily practice, and collaborate in investigation oriented toward early detection. The literature revealed that the nurse's role in cancer screening involves case management as the main activity as well as, exceptionally, carrying out diagnostic tests.


The Heme-Lys Cross-Link in Cytochrome P460 Promotes Catalysis by Enforcing Secondary Coordination Sphere Architecture.
Cytochrome (cyt) P460 is a c-type monoheme enzyme found in ammonia-oxidizing bacteria (AOB) and methanotrophs; additionally, genes encoding it have been found in some pathogenic bacteria. Cyt P460 is defined by a unique post-translational modification to the heme macrocycle, where a lysine (Lys) residue covalently attaches to the 13' meso carbon of the porphyrin, modifying this heme macrocycle into the enzyme's eponymous P460 cofactor, similar to the cofactor found in the enzyme hydroxylamine oxidoreductase. This cross-link imbues the protein with unique spectroscopic properties, the most obvious of which is the enzyme's green color in solution. Cyt P460 from the AOB Nitrosomonas europaea is a homodimeric redox enzyme that produces nitrous oxide (N2O) from 2 equiv of hydroxylamine. Mutation of the Lys cross-link results in spectroscopic features that are more similar to those of standard cyt c' proteins and renders the enzyme catalytically incompetent for NH2OH oxidation. Recently, the necessity of a second-sphere glutamate (Glu) residue for redox catalysis was established; it plausibly serves as proton relay during the first oxidative half of the catalytic cycle. Herein, we report the first crystal structure of a cross-link deficient cyt P460. This structure shows that the positioning of the catalytically essential Glu changes by approximately 0.8 Å when compared to a cross-linked, catalytically competent cyt P460. It appears that the heme-Lys cross-link affects the relative position of the P460 cofactor with respect to the second-sphere Glu residue, therefore dictating the catalytic competency of the enzyme.


Pyrogenic molecular markers: linking PAH with BPCA analysis.
Molecular characterization of pyrogenic organic matter (PyOM) is of great interest to understand the formation and behavior of these increasingly abundant materials in the environment. Two molecular marker methods have often been used to characterize and trace PyOM: polycyclic aromatic hydrocarbon (PAH) and benzenepolycarboxylic acid (BPCA) analysis. Since both methods target pyrogenic polycyclic compounds, we investigated the linkages between the two approaches using chars that were produced under controlled conditions. Rye and maize straws and their analogues charred at 300, 400 and 500 °C, respectively, were thus analyzed with both methods. Moreover, we also measured BPCAs directly on the lipid extracts, on which PAHs were analyzed, and on the respective extraction residues, too. Both methods revealed important features of the chars, in particular the increasing degree of aromatic condensation with increasing highest heating temperature (HTT). The overlap between the two methods was identified in the lipid fraction, where the proportion of benzenetricarboxylic acids (B3CAs) correlated with PAH abundance. The results confirmed the validity and complementarity of the two molecular marker methods, which will likely continue to play a crucial role in PyOM research due to the recent developments of compound-specific PAH and BPCA stable carbon (δ(13)C) and radiocarbon ((14)C) isotope methods.


Predictors of indoor smoking at young children's homes--a cross-sectional study.
The aim of the study was to determine factors associated with indoor smoking in homes (ISIH) using a sample of households with at least one child aged 3 or younger and at least one smoking adult. In a defined German region, all households (n = 3,570) with a child aged 3 or younger were invited to participate in a study that tested the efficacy of an intervention for reducing exposure to environmental tobacco smoke. In 1,282 households, at least one parent reported daily smoking. Among these, 917 (71.5 %) participated in the study. ISIH was defined as smoking 'in specific rooms only' or 'everywhere'. Cross-sectional data were analysed using regression analysis. Among the households, 37.5 % reported ISIH. ISIH was more likely if the youngest child had not visited a nursery (OR, 1.81; CI, 1.21-2.70) and if no private outdoor area was present (OR = 4.38, CI, 2.64-7.25). Lower household education level and partly unemployment in dual-parent households were associated with ISIH. Fostering nursery attendance and availability of a private outdoor area may protect young children living in household with smoking parents from environmental tobacco smoke.


Comparison of three magnetic nanoparticle tracers for sentinel lymph node biopsy in an in vivo porcine model.
Breast cancer staging with sentinel lymph node biopsy relies on the use of radioisotopes, which limits the availability of the procedure worldwide. The use of a magnetic nanoparticle tracer and a handheld magnetometer provides a radiation-free alternative, which was recently evaluated in two clinical trials. The hydrodynamic particle size of the used magnetic tracer differs substantially from the radioisotope tracer and could therefore benefit from optimization. The aim of this study was to assess the performance of three different-sized magnetic nanoparticle tracers for sentinel lymph node biopsy within an in vivo porcine model. Sentinel lymph node biopsy was performed within a validated porcine model using three magnetic nanoparticle tracers, approved for use in humans (ferumoxytol, with hydrodynamic diameter d H =32 nm; Sienna+(®), d H =59 nm; and ferumoxide, d H =111 nm), and a handheld magnetometer. Magnetometer counts (transcutaneous and ex vivo), iron quantification (vibrating sample magnetometry), and histopathological assessments were performed on all ex vivo nodes. Transcutaneous "hotspots" were present in 12/12 cases within 30 minutes of injection for the 59 nm tracer, compared to 7/12 for the 32 nm tracer and 8/12 for the 111 nm tracer, at the same time point. Ex vivo magnetometer counts were significantly greater for the 59 nm tracer than for the other tracers. Significantly more nodes per basin were excised for the 32 nm tracer compared to other tracers, indicating poor retention of the 32 nm tracer. Using the 59 nm tracer resulted in a significantly higher iron accumulation compared to the 32 nm tracer. The 59 nm tracer demonstrated rapid lymphatic uptake, retention in the first nodes reached, and accumulation in high concentration, making it the most suitable tracer for intraoperative sentinel lymph node localization.


A model for Escherichia coli DNA polymerase III holoenzyme assembly at primer/template ends. DNA triggers a change in binding specificity of the gamma complex clamp loader.
The gamma complex of the Escherichia coli DNA polymerase III holoenzyme assembles the beta sliding clamp onto DNA in an ATP hydrolysis-driven reaction. Interactions between gamma complex and primer/template DNA are investigated using fluorescence depolarization to measure binding of gamma complex to different DNA substrates under steady-state and presteady-state conditions. Surprisingly, gamma complex has a much higher affinity for single-stranded DNA (K(d) in the nM range) than for a primed template (K(d) in the microM range) under steady-state conditions. However, when examined on a millisecond time scale, we find that gamma complex initially binds very rapidly and with high affinity to primer/template DNA but is converted subsequently to a much lower affinity DNA binding state. Presteady-state data reveals an effective dissociation constant of 1.5 nM for the initial binding of gamma complex to DNA and a dissociation constant of 5.7 microM for the low affinity DNA binding state. Experiments using nonhydrolyzable ATPgammaS show that ATP binding converts gamma complex from a low affinity "inactive" to high affinity "active" DNA binding state while ATP hydrolysis has the reverse effect, thus allowing cycling between active and inactive DNA binding forms at steady-state. We propose that a DNA-triggered switch between active and inactive states of gamma complex provides a two-tiered mechanism enabling gamma complex to recognize primed template sites and load beta, while preventing gamma complex from competing with DNA polymerase III core for binding a newly loaded beta.DNA complex.


[Cutaneous leishmaniasis cases in Nizip, Turkey after the Syrian civil war].
Cutaneous leishmaniasis (CL), seen endemically in many countries, is a widespread protozoon disease all around the world. The neighboring countries of Turkey namely Iran, Iraq and Syria are highly endemic regions for CL, and more than 98% of the cases in Turkey are reported from South and Southeastern Anatolian regions. The aim of this study was to detect the prevalence of CL in Nizip, a district of Gaziantep province of southeastern Turkey, for three and half year period and to call attention to the dramatic increase of CL cases observed after the Syrian civil war. A total of 416 samples obtained from clinically suspected CL patients (of them 341 were Syrian refugees) who were admitted to Nizip State Hospital between January 1st 2010 and March 19th 2013 were included in the study. Lesion samples were collected according to the notice issued by Turkish Ministry of Health and Giemsa-stained smears were examined under the microscope (x1000). Samples from 77 patients (18.5%) yielded positive results with the observation of Leishmania amastigote forms. Fourty-seven (61%) of patients were female and 30 (39%) were male. Of the positive patients 52 (67.5%) belonged to 0-19 age group, 13 (16.9%) 20-39 and 12 (15.6%) 40-60 age groups. In the evaluation of the lesion characteristics, 33 (43%) patients had single and 44 (57%) had multiple lesions with a distribution mainly on face, arm and lower extremities, in a decreasing order. The period of time for the development of the lesions varied from 1.5 month to one year with the mean value of 3.4 months. There was no statistically significant relationship between the age and gender of patients, and the characteristics (quantity, distribution and time of occurence) of lesions (p> 0.05). The number of domestic and Syrian CL cases detected in Nizip in the years of 2010, 2011, 2012 and 2013 (the first three months) were as follows; 1 and 0, 2 and 0, 7 and 0, 5 and 62, respectively. So a total of 62 (80.5%) and 15 (19.5%) of CL patients were found to be Syrian refugees and Turkish citizens, respectively. Since the number of the cases admitted to the hospital was significantly low in comparison to the total population of refugees living in the camps, it was assumed that the real incidence of CL was much higher than determined. The data obtained in this study revealed that Nizip and the surroundings which have already had favourable climate and vector potential for CL, exhibited a higher threat for the spread of the disease following the hosting of the refugees. Thus implementation of effective prevention and control measures should be taken into consideration implemented in that specific area.


Intraperitoneal gemcitabine pharmacokinetics: a pilot and pharmacokinetic study in patients with advanced adenocarcinoma of the pancreas.
The pyrimidine analogue gemcitabine (2', 2'-difluorodeoxycitidine, dFdC) is active against pancreatic cancer, and its high clearance (CL(tb)) and low incidence of local toxicity make it an excellent candidate for evaluation as intraperitoneal (IP) therapy. We designed a dosing schema that used multiple sequential exchanges of a peritoneal dialysate containing dFdC in an effort to produce prolonged IP dFdC exposure. As part of a study involving multi-modality therapy for advanced pancreatic adenocarcinoma, patients were treated with four 6-h IP dwells of dFdC (50 mg/m(2) in 2 l) over a 24-h period. A second 24-h cycle of IP dFdC therapy was repeated 1 week later. Each exchange of dialysate contained 50 mg/m(2) dFdC in 2 l of commercial 1.5% dextrose dialysis solution. Plasma and peritoneal fluid were analyzed by HPLC to determine concentrations of dFdC and its inactive metabolite 2', 2' difluorodeoxyuridine (dFdU). Clinical data were recorded to note drug toxicity and response. Nine patients underwent IP dFdC therapy, and eight were able to receive two cycles. There were no recorded significant toxicities. Low plasma dFdC concentrations (<1 microg/ml) were present transiently in seven of nine patients, and dFdC was not detectable in the plasma of the other two. Plasma dFdU concentrations were low but increased gradually until 12 h and then declined little if any. IP dFdC concentrations declined rapidly, and dFdC was seldom measurable prior to administration of the next scheduled 6-h dwell. dFdU concentrations in peritoneal fluid were very low (<0.5 microg/ml) throughout treatment. The mean area under the concentration versus time curve (AUC) for dFdC in peritoneal fluid was 182 microg/ml x h, which was approximately 70x the AUC of dFdC reported in the ascites of a patient undergoing systemic dFdC therapy. IP dFdC was well tolerated, and no significant toxicities were noted. The rapid decrease in peritoneal dFdC concentrations and low concentrations of IP dFdU imply almost total absorption of IP-administered dFdC. Little, if any, dFdC could be detected in plasma, but the steady-state plasma dFdU concentrations also imply absorption and inactivation of virtually all IP-administered dFdC. These findings are consistent with the known high CL(tb) and low incidence of local toxicity of dFdC and argue for its further evaluation as a drug for IP therapy.


Transcriptional profiling of aluminum toxicity and tolerance responses in maize roots.
Aluminum (Al) toxicity is a major factor limiting crop yields on acid soils. There is considerable genotypic variation for Al tolerance in most common plant species. In maize (Zea mays), Al tolerance is a complex phenomenon involving multiple genes and physiological mechanisms yet uncharacterized. To begin elucidating the molecular basis of maize Al toxicity and tolerance, a detailed temporal analysis of root gene expression under Al stress was performed using microarrays with Al-tolerant and Al-sensitive genotypes. Al altered the expression of significantly more genes in the Al-sensitive genotype, presumably as a result of more severe Al toxicity. Nevertheless, several Al-regulated genes exhibited higher expression in the Al-tolerant genotype. Cell wall-related genes, as well as low phosphate-responsive genes, were found to be regulated by Al. In addition, the expression patterns of genes related to Al-activated citrate release indicated that in maize this mechanism is probably regulated by the expression level and/or function of the citrate transporter. This study is the first comprehensive survey of global transcriptional regulation under Al stress. The results described here will help to further our understanding of how mechanisms of Al toxicity and tolerance in maize are regulated at the transcriptional level.


Partners in School: An Implementation Strategy to Promote Alignment of Evidence-Based Practices Across Home and School for Children with Autism Spectrum Disorder.
When parents and teachers align their practices across home and school, it may optimize services for children with autism spectrum disorder (ASD). Partners in School is a multi-faceted implementation strategy designed to improve ASD services in schools. The goal is to increase parents' and teachers' use of evidence-based practices (EBPs) and to align those EBPs across settings. We piloted Partners in School with 49 parent-teacher dyads to assess administration and the factors associated with reported fidelity to the model. Specifically, we measured the number of intervention steps both parents and teachers completed (reported alignment) and the characteristics associated with intervention alignment. Partners in School involves parent-teacher participation in a pre-consultation interview, an in-person consultation meeting, active implementation of the same EBPs in their respective settings, and a post-consultation interview. Parents and teachers also completed surveys pre- and post-consultation. On average, parents and teachers completed approximately five EBP steps on their own in their respective settings (i.e., at home or at school). Of these five steps, parents and teachers both completed three of the same EBPs steps, on average. Different factors were related to reported alignment for parents versus teachers; however, a similarity noted for both parents and teachers was that communication variables were associated with reported alignment. Our findings indicate the important role of communication in aligning stakeholders for ASD service delivery models.


Prescribing of psychostimulant medications for attention deficit hyperactivity disorder in children: differences between clinical specialties.
To examine differences in psychostimulant prescribing between paediatricians and child/adolescent psychiatrists for treating children with attention deficit hyperactivity disorder (ADHD) in Western Australia. Using whole-population prescribing data, logistic and linear regressions were used to model the number of children (aged 2-17 years) treated with psychostimulants between August 2003 and December 2004 for ADHD and medication dose prescribed by clinical specialty, controlling for age, sex, body weight, and other medication use. Mean number of patients treated by specialty; associations between prescriber specialty and patient characteristics; associations between stimulant dose and patient characteristics and prescriber specialty. 54 paediatricians and 23 child/adolescent psychiatrists prescribed stimulant medications for children with ADHD. The mean number of patients treated (per prescriber) was 159.8 (range, 1-1977) for paediatricians and 34.3 (range, 1-166) for psychiatrists. Boys were 32% more likely to be treated with stimulants by paediatricians (P = 0.002). Psychiatrists were 2.9 times (95% CI, 2.4-3.3; P < 0.001) more likely than paediatricians to treat patients with multiple psychotropic medications. When controlled for all other factors, psychiatrists prescribed higher stimulant doses (4.5 mg/day greater; 95% CI, 2.0-7.0 mg/day; P < 0.001) than paediatricians. Treatment of children with stimulant medicines for ADHD differed between clinical specialties. Paediatricians treated more patients per prescriber, a greater proportion of boys, and a younger age demographic, but relied less on combined psychotropic pharmacotherapy and prescribed lower stimulant doses than psychiatrists.


Sorted consecutive local binary pattern for texture classification.
In this paper, we propose a sorted consecutive local binary pattern (scLBP) for texture classification. Conventional methods encode only patterns whose spatial transitions are not more than two, whereas scLBP encodes patterns regardless of their spatial transition. Conventional methods do not encode patterns on account of rotation-invariant encoding; on the other hand, patterns with more than two spatial transitions have discriminative power. The proposed scLBP encodes all patterns with any number of spatial transitions while maintaining their rotation-invariant nature by sorting the consecutive patterns. In addition, we introduce dictionary learning of scLBP based on kd-tree which separates data with a space partitioning strategy. Since the elements of sorted consecutive patterns lie in different space, it can be generated to a discriminative code with kd-tree. Finally, we present a framework in which scLBPs and the kd-tree can be combined and utilized. The results of experimental evaluation on five texture data sets--Outex, CUReT, UIUC, UMD, and KTH-TIPS2-a--indicate that our proposed framework achieves the best classification rate on the CUReT, UMD, and KTH-TIPS2-a data sets compared with conventional methods. The results additionally indicate that only a marginal difference exists between the best classification rate of conventional methods and that of the proposed framework on the UIUC and Outex data sets.


Comprehensive miRNA expression analysis in peripheral blood can diagnose liver disease.
miRNAs circulating in the blood in a cell-free form have been acknowledged for their potential as readily accessible disease markers. Presently, histological examination is the golden standard for diagnosing and grading liver disease, therefore non-invasive options are desirable. Here, we investigated if miRNA expression profile in exosome rich fractionated serum could be useful for determining the disease parameters in patients with chronic hepatitis C (CHC). Exosome rich fractionated RNA was extracted from the serum of 64 CHC and 24 controls with normal liver (NL). Extracted RNA was subjected to miRNA profiling by microarray and real-time qPCR analysis. The miRNA expression profiles from 4 chronic hepatitis B (CHB) and 12 non alcoholic steatohepatitis (NASH) patients were also established. The resulting miRNA expression was compared to the stage or grade of CHC determined by blood examination and histological inspection. miRNAs implicated in chronic liver disease and inflammation showed expression profiles that differed from those in NL and varied among the types and grades of liver diseases. Using the expression patterns of nine miRNAs, we classified CHC and NL with 96.59% accuracy. Additionally, we could link miRNA expression pattern with liver fibrosis stage and grade of liver inflammation in CHC. In particular, the miRNA expression pattern for early fibrotic stage differed greatly from that observed in high inflammation grades. We demonstrated that miRNA expression pattern in exosome rich fractionated serum shows a high potential as a biomarker for diagnosing the grade and stage of liver diseases.


Comparison of biofilm formation between major clonal lineages of methicillin resistant Staphylococcus aureus.
Epidemic methicillin-resistant S. aureus (MRSA) clones cause infections in both hospital and community settings. As a biofilm phenotype further facilitates evasion of the host immune system and antibiotics, we compared the biofilm-forming capacities of various MRSA clones. Seventy-six MRSA classified into 13 clones (USA300, EMRSA-15, Hungarian/Brazilian etc.), and isolated from infections or from carriers were studied for biofilm formation under static and dynamic conditions. Static biofilms in microtitre plates were quantified colorimetrically. Dynamic biofilms (Bioflux 200, Fluxion, USA) were studied by confocal laser-scanning and time-lapse microscopy, and the total volume occupied by live/dead bacteria quantified by Volocity 5.4.1 (Improvision, UK). MRSA harbouring SCCmec IV produced significantly more biomass under static conditions than SCCmec I-III (P = 0.003), and those harbouring SCCmec II significantly less than those harbouring SCCmec I or III (P<0.001). In the dynamic model, SCCmec I-III harbouring MRSA were significantly better biofilm formers than SCCmec IV (P = 0.036). Only 16 strains successfully formed biofilms under both conditions, of which 13 harboured SCCmec IV and included all tested USA300 strains (n = 3). However, USA300 demonstrated remarkably lower percentages of cell-occupied space (6.6%) compared to the other clones (EMRSA-15 = 19.0%) under dynamic conditions. Time-lapse microscopy of dynamic biofilms demonstrated that USA300 formed long viscoelastic tethers that stretched far from the point of attachment, while EMRSA-15 consisted of micro-colonies attached densely to the surface. MRSA harbouring SCCmec types IV and I-III demonstrate distinct biofilm forming capacities, possibly owing to their adaptation to the community and hospital settings, respectively. USA300 demonstrated abundant biofilm formation under both conditions, which probably confers a competitive advantage, contributing to its remarkable success as a pathogen.


Antibody response to porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural proteins and implications for diagnostic detection and differentiation of PRRSV types I and II.
To further characterize the humoral immune response of pigs to porcine reproductive and respiratory syndrome virus (PRRSV), direct enzyme-linked immunosorbent assays (ELISA) were used to study the kinetics of antibody responses directed against PRRSV nonstructural proteins in pigs experimentally exposed to the virus. The highest immunoreactivities were against nsp1, nsp2, and nsp7. Using the recombinant nsp7 as an antigen, we validated a dual ELISA for the simultaneous detection and differentiation of serum antibodies against type I and type II PRRSV. Receiver operating characteristic analysis based on 1,334 known-positive and 1,357 known-negative samples showed good specificity (98.3% to type I and 99.3% to type II) and sensitivity (97.4% for type I and 99.8% for type II). To differentiate type I and type II PRRSV, 470 sera originating from experimentally inoculated pigs were tested, and positive sera were correctly differentiated in 469 of 470 samples. The capability of the nsp7 dual ELISA to detect serum antibody responses from pigs infected with various genetically different field strains was determined. The nsp7 dual ELISA possessed 97.6% agreement with the Idexx HerdChek PRRS 2XR ELISA. In further testing of Idexx ELISA suspected false-positive samples, the nsp7 dual ELISA resolved 98% of the samples as negative. Taken together, these results indicate that the nsp7 dual ELISA can be used as a differential test for PRRSV serology with high levels of sensitivity and specificity. This ELISA offers an additional tool for routine or follow-up diagnostics, as well as having substantial value in epidemiological surveys and outbreak investigations.


Choroid plexus tumors: an institutional series of 25 patients.
Choroid plexus tumors (CPT) are rare neoplasms that pose considerable treatment challenges. This study reviews a single institute's experience with 25 patients of CPT and attempts to contribute to the general body of knowledge on CPT. A retrospective analysis of the case records of 25 patients operated for CPT since January 1998 and having a minimum of 1 year follow-up. The study group included 12 (48%) cases of choroid plexus papilloma (CPP), 09 (36%) cases of choroid plexus carcinoma (CPC) and 4 cases of atypical CPP. The mean age at presentation was 18.6 years (range, 6 months to 54 years; SD, 18.7) and a male preponderance was noted (17:8). Raised intracranial pressure was the commonest presenting symptom (72%). The tumors were distributed as follows: lateral ventricle (16; 64%), fourth ventricle (5; 20%), fourth ventricle with cerebellopontine angle extension (3; 12%), and third ventricle (1; 4%). A complete surgical excision was achieved in 11 cases of CPP and 8 cases of CPC. Operative complications include pneumocephalus (40%), focal deficits (36%), subdural effusion (32%), and persistent hydrocephalus requiring shunt (24%). All patients with CPP had a good outcome at the end of a mean follow-up of 5.4 years, whereas the median survival for patients with CPCs who underwent a subtotal resection with adjuvant therapy was 36 months. CPTs include a spectra ranging from CPP to CPC. Radiologic and histologic characterization of these tumors is difficult and newer immunohistochemical and genetic studies should be done to differentiate them from each other. Total excision offers a good prognosis and should be attempted for all forms of CPTs. CPPs carry a good prognosis, and adjuvant therapy is not indicated even after partial excision. CPCs and atypical CPCs carry a poor prognosis, and adjuvant therapy improves survival marginally after total excision. Spinal drop metastases are common for CPC and screening of the spine for possible metastasis should be part of the routine preoperative and postoperative investigation protocol.


Clinical outcomes of a furosemide infusion protocol in edematous patients in the intensive care unit.
Many critically ill patients have severe volume overload due to vigorous fluid resuscitation. Optimal fluid management strategies to clear tissue edema are unclear. To assess safety and effectiveness of a clinical application of a furosemide infusion protocol in edematous critically ill patients. A prospective, cohort study of consecutive adult critically ill patients who received furosemide infusion by protocol from June 2003 to July 2004. The mean total dose of furosemide was 2240 mg. The mean cumulative fluid balance therapy was -3376 mL. Electrolyte values in the critical laboratory range were 3.3% for potassium, 0.2% for sodium, and no critical values for magnesium. The mean change in creatinine level was +0.2 mg/dL during furosemide infusion therapy, but the mean creatinine level returned to baseline by 3 days after the furosemide infusion. A minimum mean arterial pressure less than 55 mm Hg occurred 12% of the time during the furosemide infusion. Furosemide infusion therapy was associated with moderately negative cumulative fluid balances, electrolyte shifts, and mild transient worsening of renal function.


Evidence for the recent origin of a bacterial protein-coding, overlapping orphan gene by evolutionary overprinting.
Gene duplication is believed to be the classical way to form novel genes, but overprinting may be an important alternative. Overprinting allows entirely novel proteins to evolve de novo, i.e., formerly non-coding open reading frames within functional genes become expressed. Only three cases have been described for Escherichia coli. Here, a fourth example is presented. RNA sequencing revealed an open reading frame weakly transcribed in cow dung, coding for 101 residues and embedded completely in the -2 reading frame of citC in enterohemorrhagic E. coli. This gene is designated novel overlapping gene, nog1. The promoter region fused to gfp exhibits specific activities and 5' rapid amplification of cDNA ends indicated the transcriptional start 40-bp upstream of the start codon. nog1 was strand-specifically arrested in translation by a nonsense mutation silent in citC. This Nog1-mutant showed a phenotype in competitive growth against wild type in the presence of MgCl2. Small differences in metabolite concentrations were also found. Bioinformatic analyses propose Nog1 to be inner membrane-bound and to possess at least one membrane-spanning domain. A phylogenetic analysis suggests that the orphan gene nog1 arose by overprinting after Escherichia/Shigella separated from the other γ-proteobacteria. Since nog1 is of recent origin, non-essential, short, weakly expressed and only marginally involved in E. coli's central metabolism, we propose that this gene is in an initial stage of evolution. While we present specific experimental evidence for the existence of a fourth overlapping gene in enterohemorrhagic E. coli, we believe that this may be an initial finding only and overlapping genes in bacteria may be more common than is currently assumed by microbiologists.


Research on the nutritional characteristics of medium-chain fatty acids.
Medium-chain fatty acids (MCFA) have attracted attention as part of a healthy diet, because they are absorbed and transported directly into the liver via the portal vein, metabolized rapidly by beta-oxidation, and increase diet-induced thermogenesis. Because medium-chain triacylglycerols (MCT) containing only MCFA has a few weak points as frying oils, we have developed medium- and long-chain triacylglycerols (MLCT). MLCT is produced by lipase-catalyzed enzymatic transesterification. Recently, long-term clinical trials have demonstrated that MLCT and MCT result in less body fat-deposition. MLCT oil (Trade name: Healthy Resseta) is safe and can be substitute for common edible vegetable oils. Healthy Resseta has been approved as FOSHU (Food for Specified Health Use), by the Ministry of Health, Labour and Welfare of Japan in December 2002 for use as a cooking oil with a suppressing effect on body fat accumulation. Healthy Resseta is widely sold in Japan.


Nondiagnostic fine-needle aspirations of the thyroid: is the risk of malignancy higher?
Nondiagnostic fine-needle aspirations (FNAs) pose a dilemma in the management of patients with thyroid nodules. In most cases, these patients undergo either repeat FNA or surgical resection. However, a significant number of patients will only be observed, assuming that the risk of malignancy is low. Therefore, the purpose of this study was to determine whether the risk of malignancy is higher in patients with thyroid nodules and nondiagnostic FNAs. We reviewed reports from 4286 consecutive FNA biopsies performed on patients with thyroid nodules at our institution between 2002 and 2010. We divided FNAs into two categories: diagnostic and nondiagnostic. We collected demographic, follow-up, and pathology data from both groups and then analyzed them with analysis of variance and chi-square tests. Of the 4286 FNAs, 259 were classified as nondiagnostic (6%). We saw no significant differences in age or gender between patients with diagnostic versus nondiagnostic FNAs. Of the patients with nondiagnostic FNAs, 62 underwent diagnostic thyroidectomy (24%), 74 had a repeat FNA (29%), and 123 had observation only (47%); thus, 136 patients had a cytologic or pathologic diagnosis. Patients with nondiagnostic FNAs had a significantly higher rate of all types of thyroid cancer, compared with those with diagnostic FNAs (12% versus 5%, respectively; P < 0.001). Impressively, the chance of papillary thyroid cancer was twofold higher in patients with nondiagnostic FNAs. The percentage of nondiagnostic FNA at our institution during this period (6%) was relatively low. However, the incidence of malignancy in these patients was significantly higher. Therefore, we recommend that patients with thyroid nodules and nondiagnostic FNAs undergo either repeat biopsy or diagnostic thyroidectomy.


Lineage Switch Between B-Lymphoblastic Leukemia and Acute Myeloid Leukemia Intermediated by "Occult" Myelodysplastic Neoplasm: Two Cases of Adult Patients With Evidence of Genomic Instability and Clonal Selection by Chemotherapy.
Lineage switch occurs in rare leukemias, and the mechanism is unclear. We report two cases of B-lymphoblastic leukemia (B-ALL) relapsed as acute myeloid leukemia (AML). Retrospective review of clinical and laboratory data. Complex cytogenetic abnormalities were detected in B-ALL for both cases with subclone heterogeneity. Postchemotherapy marrow biopsies showed trilineage hematopoiesis without detectable B-ALL. Cytogenetics in both showed stemline abnormalities. The cases were considered "occult" myelodysplastic syndrome (MDS) preceding B-ALL. The patients relapsed 6.5 and 9 months following induction, respectively. Case 1 relapsed as AML-M5 initially, was treated as such, and then relapsed again as B-ALL. Case 2 relapsed as AML-M6. Cytogenetics demonstrated persistent abnormalities. Both patients died soon after relapse. Lineage switch between B-ALL and AML could be intermediated by occult MDS. A pluripotent progenitor likely undergoes neoplastic transformation, resulting in a genomically unstable clone. This leads to a repertoire of heterogeneous subclones that may be selected by chemotherapy. Lineage switch heralds a dismal clinical outcome.


Assessing violence risk in stalking cases: a regression tree approach.
Advances in the field of risk assessment have highlighted the importance of developing and validating models for problematic or unique subgroups of individuals. Stalking offenders represent one such subgroup, where fears of and potential for violence are well-known and have important implications for safety management. The present study applies a Classification and Regression Tree (CART) approach to a sample of stalking offenders in order to help further the process of identifying and understanding risk assessment strategies. Data from 204 stalking offenders referred for psychiatric evaluation to a publicly-funded clinic were used to develop and assess putative risk factors. A series of nested models were used to generate tree algorithms predicting violence in this sample of offenders. Both simplified and more extensive models generated high levels of predictive accuracy that were roughly comparable to logistic regression models but much more straightforward to apply in clinical practice. Jack-knifed cross-validation analyses demonstrated considerable shrinkage in the CART, although the models were still comparable to many other actuarial risk assessment instruments. Logistic regression models were much more resilient to cross-validation, with relatively modest loss in predictive power.


Anemia and iron homeostasis in a cohort of HIV-infected patients in Indonesia.
Anemia is a common clinical finding in HIV-infected patients and iron deficiency or redistribution may contribute to the development of low hemoglobin levels. Iron overload is associated with a poor prognosis in HIV and Hepatitis C virus infections. Iron redistribution may be caused by inflammation but possibly also by hepatitis C co-infection. We examined the prevalence of anemia and its relation to mortality in a cohort of HIV patients in a setting where injecting drug use (IDU) is a main mode of HIV transmission, and measured serum ferritin and sTfR, in relation to anemia, inflammation, stage of HIV disease, ART and HCV infection. Patient characteristics, ART history and iron parameters were recorded from adult HIV patients presenting between September 2007 and August 2009 in the referral hospital for West Java, Indonesia. Kaplan-Meier estimates and Cox's regression were used to assess factors affecting survival. Logistic regression was used to identity parameters associated with high ferritin concentrations. Anemia was found in 49.6% of 611 ART-naïve patients, with mild (Hb 10.5 -12.99 g/dL for men; and 10.5-11.99 g/dL for women) anemia in 62.0%, and moderate to severe anemia (Hb < 10.5 g/dL) in 38.0%. Anemia remained an independent factor associated with death, also after adjustment for CD4 count and ART (p = 0.008). Seroprevalence of HCV did not differ in patients with (56.9%) or without anemia (59.6%). Serum ferritin concentrations were elevated, especially in patients with anemia (p = 0.07) and/or low CD4 counts (p < 0.001), and were not related to hsCRP or HCV infection. Soluble TfR concentrations were low and not related to Hb, CD4, hsCRP or ART. HIV-associated anemia is common among HIV-infected patients in Indonesia and strongly related to mortality. High ferritin with low sTfR levels suggest that iron redistribution and low erythropoietic activity, rather than iron deficiency, contribute to anemia. Serum ferritin and sTfR should be used cautiously to assess iron status in patients with advanced HIV infection.


The substitutability of physical and social warmth in daily life.
Classic and contemporary research on person perception has demonstrated the paramount importance of interpersonal warmth. Recent research on embodied cognition has shown that feelings of social warmth or coldness can be induced by experiences of physical warmth or coldness, and vice versa. Here we show that people tend to self-regulate their feelings of social warmth through applications of physical warmth, apparently without explicit awareness of doing so. In Study 1, higher scores on a measure of chronic loneliness (social coldness) were associated with an increased tendency to take warm baths or showers. In Study 2, a physical coldness manipulation significantly increased feelings of loneliness. In Study 3, needs for social affiliation and for emotion regulation, triggered by recall of a past rejection experience, were subsequently eliminated by an interpolated physical warmth experience. Study 4 provided evidence that people are not explicitly aware of the relationship between physical and social warmth (coldness), as they do not consider a target person who often bathes to be any lonelier than one who does not, with all else being equal. Together, these findings suggest that physical and social warmth are to some extent substitutable in daily life and that this substitution reflects an unconscious self-regulatory mechanism.


Use and Acceptability of HIV Self-Testing Among First-Time Testers at Risk for HIV in Senegal.
HIV Self-Testing (HIVST) aims to increase HIV testing coverage and can facilitate reaching the UNAIDS 90-90-90 targets. In Senegal, key populations bear a disproportionate burden of HIV and report limited uptake of HIV testing given pervasive stigma and criminalization. In these contexts, HIVST may represent a complementary approach to reach populations reporting barriers to engagement with existing and routine HIV testing services. In this study, 1839 HIVST kits were distributed in Senegal, with 1149 individuals participating in a pre-test questionnaire and 817 participating in a post-test questionnaire. Overall, 46.9% (536/1144) were first-time testers and 26.2% (300/1144) had tested within the last year; 94.3% (768/814) reported using the HIVST, and 2.9% (19/651) reported a reactive result which was associated with first-time testers (p = 0.024). HIVST represents an approach that reached first-time testers and those who had not tested recently. Implementation indicators suggest the importance of leveraging existing community structures and programs for distribution.


Cellular mechanisms of alpha herpesvirus egress: live cell fluorescence microscopy of pseudorabies virus exocytosis.
Egress of newly assembled herpesvirus particles from infected cells is a highly dynamic process involving the host secretory pathway working in concert with viral components. To elucidate the location, dynamics, and molecular mechanisms of alpha herpesvirus egress, we developed a live-cell fluorescence microscopy method to visualize the final transport and exocytosis of pseudorabies virus (PRV) particles in non-polarized epithelial cells. This method is based on total internal reflection fluorescence (TIRF) microscopy to selectively image fluorescent virus particles near the plasma membrane, and takes advantage of a virus-encoded pH-sensitive probe to visualize the precise moment and location of particle exocytosis. We performed single-particle tracking and mean squared displacement analysis to characterize particle motion, and imaged a panel of cellular proteins to identify those spatially and dynamically associated with viral exocytosis. Based on our data, individual virus particles travel to the plasma membrane inside small, acidified secretory vesicles. Rab GTPases, Rab6a, Rab8a, and Rab11a, key regulators of the plasma membrane-directed secretory pathway, are present on the virus secretory vesicle. These vesicles undergo fast, directional transport directly to the site of exocytosis, which is most frequently near patches of LL5β, part of a complex that anchors microtubules to the plasma membrane. Vesicles are tightly docked at the site of exocytosis for several seconds, and membrane fusion occurs, displacing the virion a small distance across the plasma membrane. After exocytosis, particles remain tightly confined on the outer cell surface. Based on recent reports in the cell biological and alpha herpesvirus literature, combined with our spatial and dynamic data on viral egress, we propose an integrated model that links together the intracellular transport pathways and exocytosis mechanisms that mediate alpha herpesvirus egress.


THE CULTIVATION OF AMOEBAE IN PURE CULTURE UPON AUTOLYZED TISSUES.
Since the purpose of this paper is to record the cultivation of amoebae upon autolyzed tissue without bacterial association, the morphological characteristics, life cycle, means of differentiating species, and pathogenicity of the protozoa have been omitted. These subjects will be considered in later publications. The result of this study proves that some species of amoebae from liver abscesses and the human intestine can be cultivated upon various autolyzed tissues of man and some of the lower animals without a symbiotic microorganism. Their cultivation from liver abscesses upon such bacteria-free autolyzed tissue indicates that their multiplication in these lesions depends upon some product or products in the process of dissociation of the liver cells. That such a process exists in amoebic liver abscesses cannot be questioned when histological and biochemical studies are made of such lesions, and this explains not only why the multiplication of the parasites in the organ occurs, but suggests the probable origin of the lesion. It has long been known that tissue kept for several days in a perfectly aseptic condition and at body heat, or preferably at slightly higher temperature undergoes softening and final disintegration of its cells. Wells and others who have made a thorough study of this phenomenon find that different enzymic actions take place in this process: thus in the liver they find that soluble nitrogen compounds are greatly increased, the nucleoproteids are altered by nuclease, the purin bases are freed and in their turn acted upon by the guanase and adenase, the fats are split and fatty acids set free, the glycogen gives rise to glucose and undergoes further splitting. lecithin is cleaved, and allied bodies appear, and there is a marked appearance of cholin and cholesterin. Similar changes varying only in degree occur in the process of autolysis of other tissues. Furthermore, Duval in his experiments upon the cultivation of Bacillus leprae found that the initial multiplication was accomplished when the specific organism was in symbiosis with an associated bacterium capable of hydrolyzing the leprous tissue. In later experiments he noted that the products of split proteins supply what is actually required for the growth of this particular obligate cell parasite, and that while this end is reached with bacteria through their proteolytic action equally good results can be obtained with tissue free from contaminating microörganisms provided that it is allowed to autolyze. The separation of amoebae cultivated from the human intestine from their bacterial symbiont, and their development upon various autolyzed tissues indicate that it is not the bacterium that is essential for the life of these protozoa, but the action of the living bacteria upon the protein contained in the media. This would explain the failure of many investigators to cultivate amoebae with dead bacteria or bacterial filtrates. Mention has been made that the autolyzed tissue used in the cultivation experiments gave a distinct acid reaction. The multiplication of amoebae upon a medium with such a reaction appears contradictory to the findings of Musgrave and Clegg, Walker, and others, who have emphasized the necessity of an alkaline medium for the successful cultivation of amoebae with a bacterial symbiont, though in accord with what is known to be the reaction of the contents from amoebic liver abscesses and of the bloody stools in intestinal amoebiosis. The fact should not be lost sight of that, in the cultivation of amoebae, these authors lay stress upon the selectiveness of amoebae for a special microörganism. A comparison of their work with our own results indicates that bacteria known to possess strong hydrolyzing properties, e. g., Vibrio cholerae, Bacillus coli, Bacillus subtilis, Vibrio proteus (Finkler-Prior), etc., furnish the best symbionts to the amoebae. It is well known that these bacteria growing upon gelatin or blood serum liquify the medium and alter its reaction to a marked degree of acidity. This acidity, for the most part, results from dissociation of the protein molecule contained in the media, for Wiener has shown that autolysis does not begin until the normal alkalinity of the tissues has been neutralized by the production of organic acids. Since experiments show that the development of the amoebae is scant or completely arrested if the autolyzed tissue is smeared upon an acid agar base or upon agar with an alkalinity higher than 1 per cent., the limit within which multiplication can occur must be small. It is possible that this explains why amoebae are so few in the center of liver abscesses where the acidity is very high and so plentiful in or near its walls where the normal tissue juices furnish a controlling influence over the acidity of the autolyzing tissue. It explains also why a neutral or a 1 to 1.5 per cent. alkaline medium is essential for the cultivation of amoebae with a bacterial symbiont. Here the necessity for a medium with an alkaline reaction seems necessary, as bacteria, especially those having high hydrolyzing properties, develop a marked acidity in the medium. If the medium possesses an initial acidity the limit is either quickly reached or already present and no multiplication occurs. On the other hand, a medium with a reaction too alkaline either inhibits the growth of symbiotic microörganisms, or neutralizes the acid products as rapidly as they are formed by the associated bacterium. Whether the amoebae cultivated from liver abscesses and from the intestinal canal upon diverse autolyzed tissues are able to produce lesions similar to those from which they were isolated, or whether they are non-pathogenic species which are accidental contaminators to those responsible for frank lesions, remains still to be determined. Experiments bearing upon these points, as well as on many others made possible by this work, are now under way.


Combined use of CSF NfL and CSF TDP-43 improves diagnostic performance in ALS.
To determine the diagnostic and prognostic significance of neurofilament light chain (NfL), TAR DNA-binding protein 43 (TDP-43), and total tau (t-tau) in cerebrospinal fluid (CSF) and plasma of patients with amyotrophic lateral sclerosis (ALS) and to investigate whether the combined use of those biomarker candidates can improve their diagnostic performance. This was a single-center, prospective, longitudinal study. CSF and plasma samples were collected at the time of enrollment from a discovery cohort of 29 patients with ALS and 29 age-matched controls without neurodegenerative disease. In a validation cohort, there were 46 patients with ALS, and 46 control (not age-matched) patients with motor weakness resulting from neuromuscular diseases. NfL, TDP-43, and t-tau levels in CSF and plasma were measured using ultrasensitive single molecule assay (Simoa) technology. The following findings were reproducibly observed among the discovery and validation cohorts: increased levels of CSF NfL, plasma NfL, and CSF TDP-43 in ALS compared with control groups; shorter survival associated with higher levels of CSF and plasma NfL. When the CSF NfL and CSF TDP-43 levels were combined, the areas under the ROC curves (AUC) were slightly improved relative to AUCs for each biomarker alone. CSF and plasma NfL may not only serve as diagnostic biomarkers but also provide a measure of disease progression. CSF TDP-43 is also useful as a diagnostic biomarker of ALS, but has no prognostic value. The combined use of CSF NfL and CSF TDP-43 may be a useful biomarker for the diagnosis of ALS.


Global forecasts of urban expansion to 2030 and direct impacts on biodiversity and carbon pools.
Urban land-cover change threatens biodiversity and affects ecosystem productivity through loss of habitat, biomass, and carbon storage. However, despite projections that world urban populations will increase to nearly 5 billion by 2030, little is known about future locations, magnitudes, and rates of urban expansion. Here we develop spatially explicit probabilistic forecasts of global urban land-cover change and explore the direct impacts on biodiversity hotspots and tropical carbon biomass. If current trends in population density continue and all areas with high probabilities of urban expansion undergo change, then by 2030, urban land cover will increase by 1.2 million km(2), nearly tripling the global urban land area circa 2000. This increase would result in considerable loss of habitats in key biodiversity hotspots, with the highest rates of forecasted urban growth to take place in regions that were relatively undisturbed by urban development in 2000: the Eastern Afromontane, the Guinean Forests of West Africa, and the Western Ghats and Sri Lanka hotspots. Within the pan-tropics, loss in vegetation biomass from areas with high probability of urban expansion is estimated to be 1.38 PgC (0.05 PgC yr(-1)), equal to ∼5% of emissions from tropical deforestation and land-use change. Although urbanization is often considered a local issue, the aggregate global impacts of projected urban expansion will require significant policy changes to affect future growth trajectories to minimize global biodiversity and vegetation carbon losses.