Covalent binding of antitumor benzoacronycines to double-stranded DNA induces helix opening and the formation of single-stranded DNA: unique consequences of a novel DNA-bonding mechanism.
The majority of DNA-binding small molecules known thus far stabilize duplex DNA against heat denaturation. A high, drug-induced increase in the melting temperature (Tm) of DNA is generally viewed as a good criterion to select DNA ligands and is a common feature of several anticancer drugs such as intercalators (e.g., anthracyclines) and alkylators (e.g., ecteinascidin 743). The reverse situation (destabilization of DNA to facilitate its denaturation) may be an attractive option for the identification of therapeutic agents acting on the DNA structure. We have identified the tumor-active benzoacronycine derivative S23906-1 [(+/-)-cis-1,2-diacetoxy-6-methoxy-3,3,14-trimethyl-1,2,3,14-tetrahydro-7H-benzo[b]pyrano[3,2]acridin-7-one] as a potent DNA alkylating agent endowed with a helicase-like activity. Using complementary molecular approaches, we show that covalent binding to DNA of the diacetate compound S23906-1 and its monoacetate analogue S28687-1 induces a marked destabilization of the double helix with the formation of alkylated ssDNA. The DNA-bonding properties and effects on DNA structure of a series of benzoacronycine derivatives, including the dicarbamate analogue S29385-1, were studied using complementary biochemical (electromobility shift assay, nuclease S1 mapping) and spectroscopic (fluorescence and Tm measurements) approaches. Alkylation of guanines in DNA by S28687-1 leads to a local denaturation of DNA, which becomes susceptible to cleavage by nuclease S1 and significantly decreases the Tm of DNA. The drug also directly alkylates single-strand DNA, but mass spectrometry experiments indicate that guanines in duplexes are largely preferred over single-stranded structures. This molecular study expands the repertoire of DNA-binding mechanisms and provides a new dimension for DNA recognition by small molecules.


Genetic Analysis of Root-to-Shoot Signaling and Rootstock-Mediated Tolerance to Water Deficit in Tomato.
Developing drought-tolerant crops is an important strategy to mitigate climate change impacts. Modulating root system function provides opportunities to improve crop yield under biotic and abiotic stresses. With this aim, a commercial hybrid tomato variety was grafted on a genotyped population of 123 recombinant inbred lines (RILs) derived from Solanumpimpinellifolium, and compared with self- and non-grafted controls, under contrasting watering treatments (100% vs. 70% of crop evapotranspiration). Drought tolerance was genetically analyzed for vegetative and flowering traits, and root xylem sap phytohormone and nutrient composition. Under water deficit, around 25% of RILs conferred larger total shoot dry weight than controls. Reproductive and vegetative traits under water deficit were highly and positively correlated to the shoot water content. This association was genetically supported by linkage of quantitative trait loci (QTL) controlling these traits within four genomic regions. From a total of 83 significant QTLs, most were irrigation-regime specific. The gene contents of 8 out of 12 genomic regions containing 46 QTLs were found significantly enriched at certain GO terms and some candidate genes from diverse gene families were identified. Thus, grafting commercial varieties onto selected rootstocks derived from S.pimpinellifolium provides a viable strategy to enhance drought tolerance in tomato.


An evaluation of intra-operative and post-operative blood loss in total knee arthroplasty at the National Orthopaedic Hospital, Lagos.
Total knee replacement is a rewarding and reliable procedure, producing a lasting relief to severe knee pains. However, significant blood loss usually in the post-operative period may be a challenge, necessitating prompt restoration of circulating blood volume to minimize morbidity and mortality. The aim of this study was to evaluate blood loss after total knee replacement. A prospective study of blood loss after Total Knee Arthroplasty in 53 patients in the National Orthopaedic Hospital, Lagos. Consecutive patients with established indications, presenting for total knee arthroplasty were recruited into the study after obtaining their consent. The mean intra-operative blood loss was 342.4 mls, with a range of 50 - 1500 mls. The mean post-operative blood loss and total blood loss were 603.6 mls and 940.3 mls respectively, showing a strong positive correlation (r = 0.884, p < 0.01). The average pre-operative and post-operative haemoglobin concentration were 12.5 ± 1.2 g/dl and 9.8 ± 0.9 g/dl, respectively. The mean haemoglobin loss was 2.6 ± 1.2 g/dl (r = 0.46, p < 0.001). Post-operative blood loss as measured by suction drainage, is a good predictor of total blood loss, showing a strong and positive correlation.


Paradoxical Roles of Oxidative Stress Response in the Digestive System before and after Carcinogenesis.
Oxidative stress is recognized as a cancer-initiating stress response in the digestive system. It is produced through mitochondrial respiration and induces DNA damage, resulting in cancer cell transformation. However, recent findings indicate that oxidative stress is also a necessary anticancer response for destroying cancer cells. The oxidative stress response has also been reported to be an important step in increasing the anticancer response of newly developed molecular targeted agents. Oxidative stress might therefore be a cancer-initiating response that should be downregulated in the precancerous stage in patients at risk of cancer but an anticancer cell response that should not be downregulated in the postcancerous stage when cancer cells are still present. Many commercial antioxidant agents are marketed as "cancer-eliminating agents" or as products to improve one's health, so cancer patients often take these antioxidant agents. However, care should be taken to avoid harming the anticancerous oxidative stress response. In this review, we will highlight the paradoxical effects of oxidative stress and antioxidant agents in the digestive system before and after carcinogenesis.


Gender differences in quality of life among cardiac patients.
Prior studies of quality of life among cardiac patients have examined mostly men. This study evaluated gender differences in quality of life and examined the degree to which social support was associated with quality of life. A sample of 536 patients (35% women) was recruited during a 14-month period from the inpatient cardiology service of a University-based hospital. Participants completed assessments at baseline and at 3-month intervals over the subsequent 12 months, for a total of 5 assessments. Measures at each assessment included quality of life [Mental Component Score (MCS) and Physical Component Score (PCS) from the Medical Outcomes Study--Short Form 36] and social support [Interpersonal Support Evaluation List--Short Form]. A total of 410 patients completed the baseline assessment and at least one follow-up, and were included in the data analyses. Linear mixed effects modeling of the MCS score revealed a significant effect of gender (p =.028) and time (p <.001), as well as a significant interaction of gender by social support (p =.009). Modeling of the PCS revealed a significant effect of gender (p =.010) and time (p <.001). Women with cardiac disease indicated significantly lower quality of life than men with cardiac disease over the course of a 12-month longitudinal follow-up. Social support, especially a sense of belonging or companionship, was significantly associated with emotional quality of life (MCS) among women. Strategies to increase social support may be important for health and well-being of women with cardiac disease.


Summary health statistics for u.s. Adults: national health interview survey, 2011.
Objectives-This report presents health statistics from the 2011 National Health Interview Survey (NHIS) for the civilian noninstitutionalized adult population, classified by sex, age, race and Hispanic origin, education, family income, poverty status, health insurance coverage, marital status, and place and region of residence. Estimates (frequencies and percentages) are presented for selected chronic conditions and mental health characteristics, functional limitations, health status, health behaviors, health care access and utilization, and human immunodeficiency virus testing. Percentages and percent distributions are presented in both age-adjusted and unadjusted versions. Data Source-NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics. In 2011, data were collected on 33,014 adults in the Sample Adult questionnaire. The conditional response rate was 81.6%, and the final response rate was 66.3%. The health information for adults in this report was obtained from one randomly selected adult per family. In very rare instances where the sample adult was not able to respond for himself or herself, a proxy was used. Highlights-In 2011, 61% of adults aged 18 and over had excellent or very good health. Eleven percent of adults had been told by a doctor or other health professional that they had heart disease, 24% had been told on two or more visits that they had hypertension, 9% had been told that they had diabetes, and 22% had been told that they had some form of arthritis, rheumatoid arthritis, gout, lupus, or fibromyalgia. Nineteen percent of adults were current smokers, and 21% were former smokers. Based on estimates of body mass index, 34% of adults were overweight and 28% were obese.


Impact of gonadectomy on blood pressure regulation in ageing male and female rats.
Sexual dimorphism in blood pressure has been associated with differential expression of the angiotensin II (AII) receptors and with activity of the nervous system. It is generally accepted that ageing affects kidney function as well as autonomic nervous system and hormonal balance. Given that hypertension is more prevalent in men than women until women reach their seventh decade, we hypothesised that females would be relatively protected from adverse effects of ageing compared to males and that this would be mediated by the protective effect of ovarian steroids. Intact and gonadectomised male and female normotensive Wistar rats aged 6, 12 and 18 months were used to study renal function, blood pressure, heart rate, and blood pressure variability. We observed that intact females had lower levels of proteinuria and higher (12.5%) creatinine clearance compared to intact males and that this difference was abolished by castration but not by ovariectomy. Ovariectomy resulted in a change by 9% in heart rate, resulting in similar cardiovascular parameters to those observed in males or gonadectomised males. Spectral analysis of systolic blood pressure revealed that high-frequency power spectra were significantly elevated in the females vs. males and were reduced by ovariectomy. Taken altogether, the results show that females are protected from age-related declining renal function and to a lesser extent from rising blood pressure in comparison to males. Whilst ovariectomy had some deleterious effects in females, the strongest effects were associated with gonadectomy in males, suggesting a damaging effect of male hormones.


Competitive disruption of the tumor-promoting function of membrane type 1 matrix metalloproteinase/matrix metalloproteinase-14 in vivo.
Membrane type 1 matrix metalloproteinase (MT1-MMP) is a potent modulator of the pericellular environment and promotes tumor cell invasion and proliferation in many types of tumor. The activation of proMMP-2 and processing of collagen I by MT1-MMP have been thought to be important for its tumor-promoting function. These activities can be inhibited by mutant forms of MT1-MMP lacking the catalytic domain. However, the effect of such dominant-negative mutants has never been evaluated in vivo. Various mutants lacking the catalytic domain (dCAT) were prepared and confirmed to inhibit MT1-MMP activity in human fibrosarcoma HT1080 cells, and tumor cells expressing these mutants were implanted s.c. into nude mice to monitor tumor formation. Only the membrane-anchored form of a dCAT construct through the transmembrane domain [dCAT(1)] showed potent antitumor activity not only in HT1080 cells but also in gastric carcinoma MKN28 and MKN45 cells expressing MT1-MMP. A soluble form of dCAT lacking the transmembrane domain did not show such activity. The expression of dCAT(1) in MKN28 or MKN45 further prevented the metastatic spread of tumor cells into the peritoneal cavity; however, dCAT(1) showed no effect against TMK-1, another gastric carcinoma cell line expressing no MT1-MMP. It is of note that the tumorigenicity of TMK-1 cells enhanced by MT1-MMP overexpression was, in turn, canceled by the additional expression of dCAT(1). Thus, MT1-MMP expressed in tumor cells seems to play a pivotal role in tumor growth in mice. The results also suggest new possibilities to abrogate the tumor-promoting function of MT1-MMP other than the conventional protease inhibitor-based approach.


Food safety practices among Norwegian consumers.
An informed consumer can compensate for several potential food safety violations or contaminations that may occur earlier in the food production chain. However, a consumer can also destroy the work of others in the chain by poor food handling practices, e.g., by storing chilled ready-to-eat foods at abusive temperatures. To target risk-reducing strategies, consumer groups with high-risk behavior should be identified. The aim of this study was to identify demographic characteristics associated with high-risk food handling practices among Norwegian consumers. More than 2,000 randomly selected Norwegian consumers were surveyed, and the results were analyzed with a risk-based grading system, awarding demerit points for self-reported food safety violations. The violations were categorized into groups, and an ordinary multiple linear regression analysis was run on the summarized demerit score for each group and for the entire survey group as a whole. Young and elderly men were identified as the least informed consumer groups with the most unsafe practices regarding food safety. Single persons reported poorer practices than those in a relationship. People with higher education reported poorer practices than those with lower or no education, and those living in the capital of Norway (Oslo) reported following more unsafe food practices than people living elsewhere in Norway. Men reported poorer food safety practices than women in all categories with two exceptions: parboiling raw vegetables before consumption and knowledge of refrigerator temperature. These findings suggest that risk-reducing measures should target men, and a strategy is needed to change their behavior and attitudes.


Panax ginseng: a systematic review of adverse effects and drug interactions.
Panax ginseng C. A. Meyer is a perennial herb native to Korea and China and has been used as an herbal remedy in eastern Asia for thousands of years. Modern therapeutic claims refer to vitality, immune function, cancer, cardiovascular diseases, improvement of cognitive and physical performance and sexual function. A recent systematic review of randomised controlled trials found that the efficacy of ginseng root extract could not be established beyond doubt for any of these indications. In order to obtain a balanced assessment of the therapeutic value of P. ginseng it is also necessary to consider the safety profile. In view of the extremely widespread use of P. ginseng it seems important to ask whether this herbal medicine involves health risks for the consumer. This review was conducted as a systematic attempt to document and evaluate all the available safety data on P. ginseng root extracts. Systematic searches were performed in five electronic databases and the reference lists of all papers located were checked for further relevant publications. All articles containing original data on adverse events and drug interactions with P. ginseng were included. Information was also requested from 12 manufacturers of ginseng preparations, the spontaneous reporting schemes of the WHO and national drug safety bodies. No language restrictions were imposed. Data from clinical trials suggest that the incidence of adverse events with ginseng monopreparations is similar to that with placebo. The most commonly experienced adverse events are headache, sleep and gastrointestinal disorders. The possibility of more serious adverse events is indicated in isolated case reports and data from spontaneous reporting schemes; however, causality is often difficult to determine from the evidence provided. Combination products containing ginseng as one of several constituents have been associated with serious adverse events and even fatalities. Interpretation of these cases is difficult as ingredients other than P. ginseng may have caused the problems. Possible drug interactions have been reported between P. ginseng and warfarin, phenelzine and alcohol. Collectively, these data suggest that P. ginseng monopreparations are rarely associated with adverse events or drug interactions. The ones that are documented are usually mild and transient. Combined preparations are more often associated with such events but causal attribution is usually not possible.


Epigenetic Modification of Mitochondrial DNA in the Development of Diabetic Retinopathy.
Retinal mitochondria are dysfunctional in diabetes, and mitochondrial DNA (mtDNA) is damaged and its transcription is compromised. Our aim was to investigate the role of mtDNA methylation in the development of diabetic retinopathy. Effect of high glucose (20 mM) on mtDNA methylation was analyzed in retinal endothelial cells by methylation-specific PCR and by quantifying 5-methylcytosine (5mC). Dnmt1 binding at the D-loop and Cytb regions of mtDNA was analyzed by chromatin immunoprecipitation. The role of mtDNA methylation in transcription and cell death was confirmed by quantifying transcripts of mtDNA-encoded genes (Cytb, ND6, and CoxII) and apoptosis, using cells transfected with Dnmt1-small interfering RNA (siRNA), or incubated with a Dnmt inhibitor. The key parameters were validated in the retinal microvasculature from human donors with diabetic retinopathy. High glucose increased mtDNA methylation, and methylation was significantly higher at the D-loop than at the Cytb and CoxII regions. Mitochondrial accumulation of Dnmt1 and its binding at the D-loop were also significantly increased. Inhibition of Dnmt by its siRNA or pharmacologic inhibitor ameliorated glucose-induced increase in 5mC levels and cell apoptosis. Retinal microvasculature from human donors with diabetic retinopathy presented similar increase in D-loop methylation and decrease in mtDNA transcription. Hypermethylation of mtDNA in diabetes impairs its transcription, resulting in dysfunctional mitochondria and accelerated capillary cell apoptosis. Regulation of mtDNA methylation has potential to restore mitochondrial homeostasis and inhibit/retard the development of diabetic retinopathy.


Tourniquet Conversion: A Recommended Approach in the Prolonged Field Care Setting.
Life-saving interventions take precedence over diagnostic maneuvers in the Care Under Fire stage of Tactical Combat Casualty Care. The immediate threat to life with an actively hemorrhaging extremity injury is addressed with the liberal and proper use of tourniquets. The emphasis on hemorrhage control has and will continue to result in the application of tourniquets that may not be needed past the Care Under Fire stage. As soon as tactically allowable, all tourniquets must be reassessed for conversion. Reassessment of all tourniquets should occur as soon as the tactical situation permits, but no more than 2 hours after initial placement. This article describes a procedure for qualified and trained medical personnel to safely convert extremity tourniquets to local wound dressings, using a systematic process in the field setting.


Lenalidomide plus dexamethasone for relapsed multiple myeloma in North America.
Lenalidomide, an oral immunomodulatory drug that is similar to thalidomide but has a different safety profile, has clinical activity in relapsed or refractory multiple myeloma. Patients in the United States and Canada who had received at least one previous therapy for multiple myeloma but who required additional treatment were randomly assigned to receive either 25 mg of lenalidomide or placebo on days 1 to 21 of a 28-day cycle. Both groups also received 40 mg of oral dexamethasone on days 1 to 4, 9 to 12, and 17 to 20 for the first four cycles. After the fourth cycle, 40 mg of dexamethasone was administered only on days 1 to 4. Safety, clinical response, time to progression, and overall survival were assessed. We assigned 177 patients to the lenalidomide group and 176 to the placebo group. Complete, near-complete, or partial responses occurred in 108 patients (61.0%) in the lenalidomide group and in 35 patients (19.9%) in the placebo group (P<0.001); complete responses occurred in 14.1% and 0.6%, respectively (P<0.001). The median time to progression was 11.1 months in the lenalidomide group and 4.7 months in the placebo group (P<0.001). Median overall survival times in the two groups were 29.6 months and 20.2 months, respectively (P<0.001). Grade 3 or 4 adverse events were reported in 85.3% of the lenalidomide group and in 73.1% of the placebo group; these events resulted in study discontinuation in 19.8% and 10.2%, respectively. Grade 3 or 4 neutropenia and venous thromboembolism were more common in the lenalidomide group than in the placebo group (41.2% vs. 4.6% and 14.7% vs. 3.4%, respectively; P<0.001 for both comparisons). Lenalidomide plus dexamethasone is superior to placebo plus dexamethasone in patients with relapsed or refractory multiple myeloma. (ClinicalTrials.gov number, NCT00056160 [ClinicalTrials.gov].).


Regional characterization of longitudinal DT-MRI to study white matter maturation of the early developing brain.
The human brain undergoes rapid and dynamic development early in life. Assessment of brain growth patterns relevant to neurological disorders and disease requires a normative population model of growth and variability in order to evaluate deviation from typical development. In this paper, we focus on maturation of brain white matter as shown in diffusion tensor MRI (DT-MRI), measured by fractional anisotropy (FA), mean diffusivity (MD), as well as axial and radial diffusivities (AD, RD). We present a novel methodology to model temporal changes of white matter diffusion from longitudinal DT-MRI data taken at discrete time points. Our proposed framework combines nonlinear modeling of trajectories of individual subjects, population analysis, and testing for regional differences in growth pattern. We first perform deformable mapping of longitudinal DT-MRI of healthy infants imaged at birth, 1 year, and 2 years of age, into a common unbiased atlas. An existing template of labeled white matter regions is registered to this atlas to define anatomical regions of interest. Diffusivity properties of these regions, presented over time, serve as input to the longitudinal characterization of changes. We use non-linear mixed effect (NLME) modeling where temporal change is described by the Gompertz function. The Gompertz growth function uses intuitive parameters related to delay, rate of change, and expected asymptotic value; all descriptive measures which can answer clinical questions related to quantitative analysis of growth patterns. Results suggest that our proposed framework provides descriptive and quantitative information on growth trajectories that can be interpreted by clinicians using natural language terms that describe growth. Statistical analysis of regional differences between anatomical regions which are known to mature differently demonstrates the potential of the proposed method for quantitative assessment of brain growth and differences thereof. This will eventually lead to a prediction of white matter diffusion properties and associated cognitive development at later stages given imaging data at early stages.


Beyond Average Information: How Q-Methodology Enhances Course Evaluations in Anatomy.
Course evaluations can be used for curriculum improvement and have the potential to better the student learning experience. However, because most are based on Likert scales and open-ended feedback, understanding diversity in student opinion and uncovering optimal options for course change and improvement are often difficult. Alternatively, Q-methodology can be used to investigate patterns of thought within a group and may offer greater potential for course reform. This manuscript offers a tutorial-based explanation of the three components of Q-methodology studies (1) survey instrument development, (2) data collection, and (3) analysis and interpretation, then demonstrates, via case study, the use of Q-methodology to evaluate a fourth-year undergraduate pathoanatomy course. The goal of this article is to enable the reader to broadly apply Q-methodology in other courses to gain insight and feedback beyond that offered by traditional Likert scale methods. As demonstrated through the pathoanatomy case study, Q-methodology highlights groups (denoted by factors) of like-minded students that share opinions, preferences, and values. Overall, Q-methodology analyses support course instructors in identifying areas of course strength and improvement in an evidence-based way. This alternative to traditional Likert scales represents a promising solution to ongoing course evaluation limitations.


[Personality disorders in smokers: a review].
The association between psychopathology and nicotine dependence in smokers has been a relevant topic in recent years. Nevertheless, little is known about personality disorders in smokers. The aim of this article is to review research published nationally and internationally which analyzes the relation between tobacco use and personality disorders. Our review permits us to affirm that research on the presence of Axis II disorders in smokers is rather scarce. Of 12 studies analyzed, it can be concluded that the prevalence of personality disorders in smokers is highly variable (between 9% and 45%), and that, in the majority of the studies, smoking is associated first with the presence of cluster C disorders (dependent, avoidant and obsessive-compulsive), and second, with cluster B disorders (histrionic, narcissistic, borderline and antisocial). Finally, we note the various limitations of previous studies and stress the need to better understand these disorders, given their relevance to the treatment of smokers.


Measuring 3D growth plate shape: Methodology and application to cam morphology.
Physeal changes corresponding to cam morphology are currently measured using two-dimensional (2D) methods. These methods are limited by definitions of the femoral neck axis and head center that are dependent on the radiographic plane of view. To address these limitations, we developed three-dimensional (3D) methods for analyzing continuous growth plate shape using magnetic resonance imaging scans. These new methods rely on a single definition of the femoral neck axis and head center that are both nondependent on the radiographic plane of view and allow for analysis of growth plate shape across the growth plate surface (performed using statistical parametric mapping). Using our 3D method, we analyzed the position of the growth plate in the femoral head (relative to a plane tangent to the femoral head) and the curvature of the growth plate (relative to a plane through the center of the growth plate) in 9-16-year-old males at risk for cam morphology and their recreationally active peers (n = 17/cohort). These two measurements provide an avenue to separately analyze the effects of these variables in the overall growth plate shape. We detected differences in growth plate shape with age in recreationally active adolescents but did not detect differences between at risk and recreationally adolescents.


G-proteins of fat-cells. Role in hormonal regulation of intracellular inositol 1,4,5-trisphosphate.
Pertussis toxin abolishes hormonal inhibition of adenylate cyclase, hormonal stimulation of inositol 1,4,5-trisphosphate accumulation in rat fat-cells, and catalyses the ADP-ribosylation of two peptides, of Mr 39,000 and 41,000 [Malbon, Rapiejko & Mangano (1985) J. Biol. Chem. 260, 2558-2564]. The 41,000-Mr peptide is the alpha-subunit of the G-protein, referred to as Gi, that is believed to mediate inhibitory control of adenylate cyclase by hormones. The nature of the 39,000-Mr substrate for pertussis toxin was investigated. The fat-cell 39,000-Mr peptide was compared structurally and immunologically with the alpha-subunits of two other G-proteins, Gt isolated from the rod outer segments of bovine retina and Go isolated from bovine brain. After radiolabelling in the presence of pertussis toxin and [32P]NAD+, the electrophoretic mobilities of the fat-cell 39,000-Mr peptide and the alpha-subunits of Go and Gt were nearly identical. Partial proteolysis of these ADP-ribosylated proteins generates peptide patterns that suggest the existence of a high degree of homology between the fat-cell 39,000-Mr peptide and the alpha-subunit of Go. Antisera raised against purified G-proteins and their subunits were used to probe immunoblots of purified Gt, Gi, Go, and fat-cell membrane proteins. Although recognizing the 36,000-Mr beta-subunit band of Gt, Gi, Go and a 36,000-Mr fat-cell peptide, antisera raised against Gt failed to recognize either the 39,000- or the 41,000-Mr peptides of fat-cells or the alpha-subunits of Go and Gi. Antisera raised against the alpha-subunit of Go, in contrast, recognized the 39,000-Mr peptide of rat fat-cells, but not the alpha-subunit of either Gi or Gt. These data establish the identity of Go, in addition to Gi, in fat-cell membranes and suggest the possibility that either Go or Gi alone, or both, may mediate hormonal regulation of adenylate cyclase and phospholipase C.


Geochronological and taxonomic revisions of the middle Eocene Whistler Squat Quarry (Devil's Graveyard Formation, Texas) and implications for the early Uintan in Trans-Pecos Texas.
The Whistler Squat Quarry (TMM 41372) of the lower Devil's Graveyard Formation in Trans-Pecos Texas is a middle Eocene fossil locality attributed to Uintan biochronological zone Ui1b. Specimens from the Whistler Squat Quarry were collected immediately above a volcanic tuff with prior K/Ar ages ranging from ∼47-50 Ma and below a tuff previously dated to ∼44 Ma. New 40Ar/39Ar analyses of both of the original tuff samples provide statistically indistinguishable ages of 44.88±0.04 Ma for the lower tuff and 45.04±0.10 Ma for the upper tuff. These dates are compatible with magnetically reversed sediments at the site attributable to C20r (43.505-45.942 Ma) and a stratigraphic position above a basalt dated to 46.80 Ma. Our reanalysis of mammalian specimens from the Whistler Squat Quarry and a stratigraphically equivalent locality significantly revises their faunal lists, confirms the early Uintan designation for the sites, and highlights several biogeographic and biochronological differences when compared to stratotypes in the Bridger and Uinta Formations. Previous suggestions of regional endemism in the early Uintan are supported by the recognition of six endemic taxa (26% of mammalian taxa) from the Whistler Squat Quarry alone, including three new taxa. The revised faunal list for the Whistler Squat Quarry also extends the biostratigraphic ranges of nine non-endemic mammalian taxa to Ui1b.


Pilot study of Panax quinquefolius (American ginseng) to improve cancer-related fatigue: a randomized, double-blind, dose-finding evaluation: NCCTG trial N03CA.
This pilot trial sought to investigate whether any of three doses of American ginseng (Panax quinquefolius) might help cancer-related fatigue. A secondary aim was to evaluate toxicity. Eligible adults with cancer were randomized in a double-blind manner, to receive American ginseng in doses of 750, 1,000, or 2,000 mg/day or placebo given in twice daily dosing over 8 weeks. Outcome measures included the Brief Fatigue Inventory, vitality subscale of the Medical Outcome Scale Short Form-36 (SF-36), and the Global Impression of Benefit Scale at 4 and 8 weeks. Two hundred ninety patients were accrued to this trial. Nonsignificant trends for all outcomes were seen in favor of the 1,000- and 2,000-mg/day doses of American ginseng. Area under the curve analysis of activity interference from the Brief Fatigue Inventory was 460-467 in the placebo group and 750 mg/day group versus 480-551 in the 1,000- and 2,000-mg/day arms, respectively. Change from baseline in the vitality subscale of the SF-36 was 7.3-7.8 in the placebo and the 750-mg/day arm, versus 10.5-14.6 in the 1,000- and 2,000-mg/day arms. Over twice as many patients on ginseng perceived a benefit and were satisfied with treatment over those on placebo. There were no significant differences in any measured toxicities between any of the arms. There appears to be some activity and tolerable toxicity at 1,000-2,000 mg/day doses of American ginseng with regard to cancer-related fatigue. Thus, further study of American ginseng is warranted.


Evaluation of transforming growth factor beta 1 in dogs with osteoarthritis.
Osteoarthritis is a common problem in daily veterinary practice with insufficient knowledge about disease mechanism. Because fibrosis is a part of the alteration in the effected joints, we investigated the transforming growth factor beta 1 (TGF-β1) as an important regulation molecule of tissue fibrosis. We chose Osteoarthritis following a cruciate ligament rupture (CLR) because it is a common model of osteoarthritis. A total of 13 healthy dogs and 38 dogs suffering from CLR were included in this prospective study. The concentration of TGF-β1 was measured in synovial fluid using the "Quantikine Human TGF-β1 Immunoassay" (Fa. R & D Systems, Minneapolis, USA). There was a significant difference in the TGF-β1 concentration of the synovial fluid of healthy compared to affected patients (p < 0.001). The synovial TGF-β1 concentration also correlated significantly (p = 0.0147) with the synovial viscosity of the affected patients. No significant correlations could be observed to duration of disease, severity of lameness and degree of joint swelling, but also to joint capsule sickness, osteophyte development and degeneration of joint cartilage. The results of our study show an increased TGF-β1 concentration in knees affected with osteoarthritis as consequence of CLR. So we can conclude that TGF-β1 takes part at the osteoarthritic remodelling process, but different phases of the remodelling process cannot be distinguished by the measurement of TGF-β1.


A beta-hairpin comprising the nuclear localization sequence sustains the self-associated states of nucleosome assembly protein 1.
The histone chaperone nucleosome assembly protein 1 (NAP1) is implicated in histone shuttling as well as nucleosome assembly and disassembly. Under physiological conditions, NAP1 dimers exist in a mixture of various high-molecular-weight oligomers whose size may be regulated by the cell cycle-dependent concentration of NAP1. Both the functional and structural significance of the observed oligomers are unknown. We have resolved the molecular mechanism by which yeast NAP1 (yNAP1) dimers oligomerize by applying x-ray crystallographic, hydrodynamic, and functional approaches. We found that an extended beta-hairpin that protrudes from the compact core of the yNAP1 dimer forms a stable beta-sheet with beta-hairpins of neighboring yNAP1 dimers. Disruption of the beta-hairpin (whose sequence is conserved among NAP1 proteins in various species) by the replacement of one or more amino acids with proline results in complete loss of yNAP1 dimer oligomerization. The in vitro functions of yNAP1 remain unaffected by the mutations. We have thus identified a conserved structural feature of NAP1 whose function, in addition to presenting the nuclear localization sequence, appears to be the formation of higher-order oligomers.


Improving outcomes for children with cancer in low-income countries in Latin America: a report on the recent meetings of the Monza International School of Pediatric Hematology/Oncology (MISPHO)-Part I.
The difference in survival for children diagnosed with cancer between high- and low-income countries (LIC) continues to widen as curative therapies are developed in the former but not implemented in the latter. In 1996, the Monza International School of Pediatric Hematology/Oncology (MISPHO) was founded in an attempt to narrow this survival gap. During its sixth and seventh meetings, members recognized the problem of lack of affordability of essential drugs to treat childhood cancer in many LIC, and initiated an advocacy program. In 1998, MISPHO spawned a collaboration of Central American pediatric oncology centers: the Asociación de Hemato-Oncología Pediátrica Centroamericana (AHOPCA). AHOPCA members reported preliminary findings from several of the 10 cooperative protocols that are currently in progress. In 2003, a second regional collaborative group was formed that includes seven centers in South America. Twinning programs between MISPHO centers and centers in high-income countries (HIC) have proven invaluable to harness the resources of these centers to improve pediatric oncology care in LIC. MISPHO educational efforts include oncology nursing, supportive care, cancer-specific updates, epidemiology, and clinical research methods. Educational efforts are facilitated by educational content and online conferencing via www.cure4kids.org. Identifying preventable causes of abandonment of therapy and documenting the nutritional status of patients treated at MISPHO centers are areas of active research.


Long-term colonization with single and multiple strains of Helicobacter pylori assessed by DNA fingerprinting.
The gastric pathogen Helicobacter pylori establishes long-term chronic infections that can lead to gastritis, peptic ulcers, and cancer. The species is so diverse that distinctly different strains are generally recovered from each patient. To better understand the dynamics of long-term carriage, we characterized H. pylori isolates from initial and follow-up biopsy specimens from a patient population at high risk of H. pylori infection and gastric cancer. Eighty-five isolates were obtained from 23 patients and were analyzed by genomic restriction enzyme analysis, arbitrarily primed PCR fingerprinting, (random amplified polymorphic DNA analysis), and/or restriction of specific PCR-amplified genes (restriction fragment length polymorphism analysis). A single strain was found in sequential biopsy specimens from 12 of 15 patients (80%) receiving sucralfate. In the remaining three patients treated with sucralfate, two strains were identified in two patients and three strains were identified in the third patient. In contrast, a single strain was found in sequential biopsy specimens from only three of eight patients (37%) receiving bismuth, metronidazole, and nitrofurantoin. Two strains were identified in five other patients receiving bismuth-antibiotic (63%). Immunoglobulin G antibodies to H. pylori were present in the sera of all patients. Thus, H. pylori colonization can persist for long periods (up to at least 4 years), despite high titers of immunoglobulin G antibodies in serum. Resistance to metronidazole was noted in some strains before and/or after treatment, but all strains remained susceptible to amoxicillin, tetracycline, and nitrofurantoin. We conclude that H. pylori genotypes, as measured by several sensitive DNA fingerprinting methods, can remain stable for years in vivo, despite the acquisition or loss of drug resistance, circulating antibody, or exposure to antibiotics or sucralfate.


The prevention of emesis in plastic surgery: a randomized, prospective study.
Perhaps the most unpleasant experience following outpatient plastic surgery procedures is postoperative nausea and vomiting. Postoperative nausea and vomiting often results in delayed recovery time and unintended admission, and it can be a contributing factor to the formation of hematoma following rhytidectomy. Ondansetron (Zofran) has proven benefit in preventing postoperative nausea and vomiting if given before general anesthesia in a variety of surgical procedures. Its utility in cases performed under conscious sedation has not been determined. The purpose of this study was (1) to test the ability of prophylactic ondansetron to prevent postoperative nausea and vomiting in plastic surgery cases performed under conscious sedation, and (2) to determine relative risk factors for postoperative nausea and vomiting and a selection policy for the administration of antiemetic prophylaxis. This was a prospective, randomized, double-blind study. One hundred twenty patients were enrolled after giving informed consent. Patients received a single dose of either placebo or ondansetron (4 mg intravenously) before administration of sedation. Sedation administration followed a standardized institutional protocol, using midazolam and fentanyl. Data were recorded from a series of three questionnaires: preoperatively, immediately postoperatively, and at the time of the first office return. Data were confirmed by means of telephone interview, chart analysis, and nursing documentation. Multivariate analysis was conducted. Nausea and emesis occurred with an overall frequency of 33 percent and 22 percent, respectively. Postoperative nausea and vomiting was associated with statistically longer recovery periods. The incidence of emesis was statistically higher among women, among those undergoing facial rejuvenation, and among those with a history of opioid-induced emesis or postoperative nausea and vomiting following a previous operation (p < 0.05). The incidence of postoperative nausea and vomiting paralleled increases in case duration; the incidence of emesis was zero in cases less than 90 minutes in duration. Ondansetron significantly reduced the incidence of emesis overall (placebo, 30 percent; ondansetron, 13 percent; p < 0.05). Postoperative perception of nausea was significantly lower among those who had received ondansetron (p < 0.05). These results confirm the efficacy of ondansetron for the prevention of postoperative nausea and vomiting in plastic surgery cases under conscious sedation. In those who are at increased risk, prophylaxis should be considered. Such risks include female gender, facial rejuvenation procedures, and a patient history of opioid-induced emesis or postoperative nausea and vomiting following a prior operation. The zero incidence of emesis in cases less than 90 minutes does not support the routine use of prophylaxis in such cases. Patient satisfaction in plastic surgery is derived from the overall subjective experience of the event as much as by the final result. By remaining attentive to patient concerns and optimizing perioperative care, we can improve the subjective experience for our patients.


[Comparison of Doppler ultrasonography and hepatic venous pressure gradient in assessing portal hypertension in liver cirrhosis].
This prospective study aimed to determine if Doppler ultrasonography can be representative of hepatic venous pressure gradient (HVPG) in assessing the severity of portal hypertension and response to drug reducing portal pressure. The HVPG and the parameters of Doppler ultrasonography including portal venous velocity (PVV) and splenic venous velocity, the pulsatility and resistive index of hepatic, splenic and renal arteries were measured in 105 patients with liver cirrhosis. In 31 patients the changes of hepatic venous pressure gradient and portal venous velocity after administration of terlipressin were evaluated. The patients who showed a reduction in HVPG of more than 20% of the baseline were defined as responders to terlipressin. Any Doppler ultrasonographic parameters did not correlate with HVPG. Both HVPG and PVV showed a highly significant reduction after the administration of terlipressin(-28.3 +/- 3.9%, -31.2 +/- 2.2% respectively). However, PVV decreased significantly not only in responders(31.7 +/- 2.4%) but also in nonresponders(29.5 +/- 6.1%). Doppler ultrasonography can not be representative of HVPG in assessing the severity of portal hypertension and response to drug reducing portal pressure in liver cirrhosis.


[Pain management].
Analgesia and hypnosis are two separate entities and should result in distinct assessment and management for patients admitted to an intensive care unit (ICU). Those patients are exposed to moderate-severe pain and they are likely to remember pain as one bothersome experience. Any cause of patient discomfort is sought with the priority given to pain and adequate analgesia. Assessing pain must rely upon the use of clinical scoring systems, although these instruments are still underused in ICU. Satisfactory levels of analgesia by continuous infusion of opioids during times without stimulation do not guarantee against pain reactions during procedures (endotracheal suctioning, mobilization, wound care and dressing change, removal of chest tube). The concept of multimodal analgesia should be extended to the ICU since it may reduce the opioids requirements. In order to facilitate systematic pain and sedation assessment and to adjust daily drug dosages accordingly, it appears crucial to promote educational programs and elaboration of protocols/guidelines in ICU. Protocols/guidelines may help caregivers to rationally use sedatives and opioids and possibly reduce mechanical ventilation and ICU length of stay.


A light tunable differentiation system for the creation and control of consortia in yeast.
Artificial microbial consortia seek to leverage division-of-labour to optimize function and possess immense potential for bioproduction. Co-culturing approaches, the preferred mode of generating a consortium, remain limited in their ability to give rise to stable consortia having finely tuned compositions. Here, we present an artificial differentiation system in budding yeast capable of generating stable microbial consortia with custom functionalities from a single strain at user-defined composition in space and in time based on optogenetically-driven genetic rewiring. Owing to fast, reproducible, and light-tunable dynamics, our system enables dynamic control of consortia composition in continuous cultures for extended periods. We further demonstrate that our system can be extended in a straightforward manner to give rise to consortia with multiple subpopulations. Our artificial differentiation strategy establishes a novel paradigm for the creation of complex microbial consortia that are simple to implement, precisely controllable, and versatile to use.


The Escherichia coli K-12 gntP gene allows E. coli F-18 to occupy a distinct nutritional niche in the streptomycin-treated mouse large intestine.
Escherichia coli F-18 is a human fecal isolate that makes type 1 fimbriae, encoded by the fim gene cluster, and is an excellent colonizer of the streptomycin-treated mouse intestine. E. coli F-18 fimA::tet, lacking type 1 fimbriae, was constructed by bacteriophage P1 transduction of the fim region of the E. coli K-12 strain ORN151, containing the tetracycline resistance gene from Tn10 inserted in the fimA gene, into E. coli F-18. E. coli F-18 fimA::tet was found to occupy a distinct niche in the streptomycin-treated mouse intestine when fed in small numbers (10(4) CFU) to mice, along with large numbers (10(10) CFU) of E. coli F-18, as defined by the ability of the E. coli F-18 fimA::tet strain to grow and colonize only 1 order of magnitude below E. coli F-18. The same effect was observed when mice already colonized with E. coli F-18 were fed small numbers of E. coli F-18 fimA::tet. Experiments which show that the E. coli K-12 gene responsible for this effect is not fim::tet but gntP, which maps immediately downstream of the fim gene cluster, are presented. gntP encodes a high-affinity gluconate permease, suggesting that the distinct niche in the mouse large intestine is defined by the presence of gluconate. The data presented here support the idea that small numbers of an ingested microorganism can colonize the intestine as long as it can utilize an available nutrient better than any of the other resident species can.


Identification and initial response to children's exposure to intimate partner violence: a qualitative synthesis of the perspectives of children, mothers and professionals.
To synthesise evidence on the acceptable identification and initial response to children's exposure to intimate partner violence (IPV) from the perspectives of providers and recipients of healthcare and social services. We conducted a thematic synthesis of qualitative research, appraised the included studies with the modified Critical Appraisal Skills Programme checklist and undertook a sensitivity analysis of the studies scored above 15. We searched eight electronic databases, checked references and citations and contacted authors of the included studies. We included qualitative studies with children, parents and providers of healthcare or social services about their experiences of identification or initial responses to children's exposure to IPV. Papers that have not been peer-reviewed were excluded as well as non-English papers. Searches identified 2039 records; 11 studies met inclusion criteria. Integrated perspectives of 42 children, 212 mothers and 251 professionals showed that sufficient training and support for professionals, good patient-professional relationship and supportive environment for patient/clients need to be in place before enquiry/disclosure of children's exposure to IPV should occur. Providers and recipients of care favour a phased enquiry about IPV initiated by healthcare professionals, which focuses on 'safety at home' and is integrated into the context of the consultation or visit. Participants agreed that an acceptable initial response prioritises child safety and includes emotional support, education about IPV and signposting to IPV services. Participants had conflicting perspectives on what constitutes acceptable engagement with children and management of safety. Sensitivity analysis produced similar results. Healthcare and social service professionals should receive sufficient training and ongoing individual and system-level support to provide acceptable identification of and initial response to children's exposure to IPV. Ideal identification and responses should use a phased approach to enquiry and the WHO Listen, Inquire about needs and concerns, Validate, Enhance safety and Support principles integrated into a trauma-informed and violence-informed model of care.


Chromophobe Renal Cell Carcinoma Presenting as a Cystic Renal Mass: Case Report and Review of the Literature.
BACKGROUND Chromophobe renal cell carcinoma is a rare subtype of renal cell carcinoma which accounts for 4% to 6% of renal cell carcinoma subtypes. These tumors generally have a good prognosis. Typical radiological features include a well-defined homogeneous mass and a hypo-vascular soft tissue enhancement. Cystic chromophobe renal cell carcinoma is a very rare entity in which the overall estimated cystic renal cell carcinoma of all subtypes account for 4%. We describe a rare presentation of chromophobe renal cell carcinoma presenting as a unilocular cystic mass with mural nodules. CASE REPORT A 28-year-old healthy male presented with a history of right upper quadrant fullness and discomfort. Hepatomegaly was expected on examination. Ultrasonographic assessment revealed a normal liver size and echo texture. However, a large cystic mass measuring 15.7×12.8 cm was found arising from the right kidney which showed no internal vascularity but turbid fluid and debris within the dependent regions of the mass. Multiphase computed tomography scan showed a large unilocular cystic mass with enhancing mural nodules. There was no lymphovascular or peri-renal invasion. The patient underwent uneventful total right renal nephrectomy. Histological examination revealed a chromophobe renal cell carcinoma of grade II. CONCLUSIONS Cystic renal neoplasm differential diagnosis varies from benign entities to malignant neoplasms. Knowledge of the variable common features for each renal cell carcinoma subtypes can aid in narrowing the differential diagnosis and prompt appropriate surgical management since a possibility of nephron sparing technique might still take place in suspected cystic chromophobe renal cell carcinoma but never for cystic clear cell renal cell carcinoma.


Abundance of ruminal bacteria, epithelial gene expression, and systemic biomarkers of metabolism and inflammation are altered during the peripartal period in dairy cows.
Seven multiparous Holstein cows with a ruminal fistula were used to investigate the changes in rumen microbiota, gene expression of the ruminal epithelium, and blood biomarkers of metabolism and inflammation during the transition period. Samples of ruminal digesta, biopsies of ruminal epithelium, and blood were obtained during -14 through 28d in milk (DIM). A total of 35 genes associated with metabolism, transport, inflammation, and signaling were evaluated by quantitative reverse transcription-PCR. Among metabolic-related genes, expression of HMGCS2 increased gradually from -14 to a peak at 28 DIM, underscoring its central role in epithelial ketogenesis. The decrease of glucose and the increase of nonesterified fatty acids and β-hydroxybutyrate in the blood after calving confirmed the state of negative energy balance. Similarly, increases in bilirubin and decreases in albumin concentrations after calving were indicative of alterations in liver function and inflammation. Despite those systemic signs, lower postpartal expression of TLR2, TLR4, CD45, and NFKB1 indicated the absence of inflammation within the epithelium. Alternatively, these could reflect an adaptation to react against inducers of the immune system arising in the rumen (e.g., bacterial endotoxins). The downregulation of RXRA, INSR, and RPS6KB1 between -14 and 10 DIM indicated a possible increase in insulin resistance. However, the upregulation of IRS1 during the same time frame could serve to restore sensitivity to insulin of the epithelium as a way to preserve its proliferative capacity. The upregulation of TGFB1 from -14 and 10 DIM coupled with upregulation of both EGFR and EREG from 10 to 28 DIM indicated the existence of 2 waves of epithelial proliferation. However, the downregulation of TGFBR1 from -14 through 28 DIM indicated some degree of cell proliferation arrest. The downregulation of OCLN and TJP1 from -14 to 10 DIM indicated a loss of tight-junction integrity. The gradual upregulation of membrane transporters MCT1 and UTB to peak levels at 28 DIM reflected the higher intake and fermentability of the lactation diet. In addition, those changes in the diet after calving resulted in an increase of butyrate and a decrease of ruminal pH and acetate, which partly explain the increase of Anaerovibrio lipolytica, Prevotella bryantii, and Megasphaera elsdenii and the decrease of fibrolytic bacteria (Fibrobacter succinogenes, Butyrivibrio proteoclasticus). Overall, these multitier changes revealed important features associated with the transition into lactation. Alterations in ruminal epithelium gene expression could be driven by nutrient intake-induced changes in microbes; microbial metabolism; and the systemic metabolic, hormonal, and immune changes. Understanding causes and mechanisms driving the interaction among ruminal bacteria and host immunometabolic responses merits further study.


Fiscal Stewardship, Choice, and Control: The Context of Self-Directed Services for People With Intellectual and Developmental Disabilities (IDD) in the United States.
Self-directed home and community based services (HCBS) waiver services and supports for people with intellectual and developmental disabilities (IDD) have become a viable and widely used method of service provision in the United States. Grounded in theories of self-determination, previous literature on self-direction has suggested high satisfaction and positive outcomes for people who use self-directed programs as well as cost savings for state IDD service systems. This study explored the ways in which state IDD service administrators think about how self-direction may be used as a method of achieving cost savings while providing opportunities for people with IDD and their families to exercise choice and control. Informed by 54 high-level IDD service administrators in 34 states, and guided by a thematic analysis approach to data interpretation, the study found evidence that administrators typically see strong potential for self-direction to have cost-savings benefits, while also fostering choice. In the current political climate, the need for cautious fiscal stewardship may become a stronger driving force behind self-direction for people with IDD in the United States.


Experiences of Self-Criticism and Self-Compassion in People Diagnosed With Cancer: A Multimethod Qualitative Study.
Objective: Self-criticism is a self-condemning and self-compassion a supportive style of self-to-self relating. These concepts have increasingly been studied in people with cancer, but mainly with quantitative studies. This study is the first to explore how adult cancer patients experience self-criticism and self-compassion in the context of their illness. Design: A multimethod qualitative study design was used, combining individual and group semi-structured interviews. Participants were 26 people with cancer who familiarized themselves with the topic by doing various self-compassion exercises for 2 weeks prior to the interview. Individual and group interviews were analyzed together using thematic analysis. Results: Four themes regarding self-criticism were identified: (1) being harsh or strict with yourself, (2) feeling guilty or angry, (3) feeling useless or like a burden, (4) feeling ashamed and not wanting to show weakness. Six themes regarding self-compassion were identified: (1) being mild to yourself, (2) guarding your boundaries, (3) accepting the illness and limitations, (4) maintaining a positive perspective, (5) connecting to others, and (6) taking responsibility for your health. Conclusion: Our findings offer insights into practical and daily life experiences of self-criticism and self-compassion of people with cancer, which can aid the further development of theory, scales and interventions.


Catalase activity is regulated by c-Abl and Arg in the oxidative stress response.
The Abl family of mammalian non-receptor tyrosine kinases includes c-Abl and Arg. Recent studies have demonstrated that c-Abl and Arg are activated in the response of cells to oxidative stress. This work demonstrates that catalase, a major effector of the cellular defense against H2O2, interacts with c-Abl and Arg. The results show that H2O2 induced binding of c-Abl and Arg to catalase. The SH3 domains of c-Abl and Arg bound directly to catalase at a P293FNP site. c-Abl and Arg phosphorylated catalase at Tyr231 and Tyr386 in vitro and in the response of cells to H2O2. The functional significance of the interaction is supported by the demonstration that cells deficient in both c-Abl and Arg exhibit substantial increases in H2O2 levels. In addition, c-abl-/- arg-/- cells exhibited a marked increase in H2O2-induced apoptosis compared with that found in the absence of either kinase. These findings indicate that c-Abl and Arg regulate catalase and that this signaling pathway is of importance to apoptosis in the oxidative stress response.


On-field predictors of neuropsychological and symptom deficit following sports-related concussion.
Investigate the relationship between on-field markers of concussion severity and postinjury neuropsychological and symptom presentation in an athlete-specific population. Case control study. Multicenter analysis of high school and college athletes. A total of 78 athletes sustaining sports-related concussion were selected from a larger sample of 139 concussed athletes. ASSESSMENT OF PREDICTOR VARIABLES: On-field presence of disorientation, posttraumatic amnesia, retrograde amnesia, and loss of consciousness. ImPACT, a computerized neuropsychological test battery, was administered pre-season and, on average, 2 days postinjury. Good postinjury presentation (n = 44) was defined as no measurable change, relative to baseline, in terms of both ImPACT memory and symptom composite scores. Poor presentation (n = 34) was defined as a 10-point increase in symptom reporting and 10-point decrease in memory functioning (exceeding the 80% confidence interval for measurement error on ImPACT). Athletes failing to meet good or poor selection criteria (n = 61) were not included in the analysis. Odds ratios revealed that athletes demonstrating poor presentation at 2 days postinjury were over 10 times more likely (P < 0.001) to have exhibited retrograde amnesia following concussive injury when compared with athletes exhibiting good presentation. Similarly, athletes with poor presentation were over 4 times more likely (P < 0.013) to have exhibited posttraumatic amnesia and at least 5 minutes of mental status change. There were no differences between good and poor presentation groups in terms of on-field loss of consciousness. The presence of amnesia, not loss of consciousness, appears predictive of symptom and neurocognitive deficits following concussion in athletes. Athletes presenting with on-field amnesia should undergo comprehensive and individualized assessment prior to returning to sport participation. Continued refinement of sports concussion grading scales is warranted in lieu of consistent findings that brief loss of consciousness is not predictive of concussion injury severity.


Diagnostic stewardship of C. difficile testing: a quasi-experimental antimicrobial stewardship study.
We evaluated whether a diagnostic stewardship initiative consisting of ASP preauthorization paired with education could reduce false-positive hospital-onset (HO) Clostridioides difficile infection (CDI). Single center, quasi-experimental study. Tertiary academic medical center in Chicago, Illinois. Adult inpatients were included in the intervention if they were admitted between October 1, 2016, and April 30, 2018, and were eligible for C. difficile preauthorization review. Patients admitted to the stem cell transplant (SCT) unit were not included in the intervention and were therefore considered a contemporaneous noninterventional control group. The intervention consisted of requiring prescriber attestation that diarrhea has met CDI clinical criteria, ASP preauthorization, and verbal clinician feedback. Data were compared 33 months before and 19 months after implementation. Facility-wide HO-CDI incidence rates (IR) per 10,000 patient days (PD) and standardized infection ratios (SIR) were extracted from hospital infection prevention reports. During the entire 52 month period, the mean facility-wide HO-CDI-IR was 7.8 per 10,000 PD and the SIR was 0.9 overall. The mean ± SD HO-CDI-IR (8.5 ± 2.0 vs 6.5 ± 2.3; P < .001) and SIR (0.97 ± 0.23 vs 0.78 ± 0.26; P = .015) decreased from baseline during the intervention. Segmented regression models identified significant decreases in HO-CDI-IR (Pstep = .06; Ptrend = .008) and SIR (Pstep = .1; Ptrend = .017) trends concurrent with decreases in oral vancomycin (Pstep < .001; Ptrend < .001). HO-CDI-IR within a noninterventional control unit did not change (Pstep = .125; Ptrend = .115). A multidisciplinary, multifaceted intervention leveraging clinician education and feedback reduced the HO-CDI-IR and the SIR in select populations. Institutions may consider interventions like ours to reduce false-positive C. difficile NAAT tests.


Observations of measles virus infection of cultured human cells. I. A study of development and spread of virus antigen by means of immunofluorescence.
The development of measles virus in cultures of both primary human amnion cells and H.Ep.-2 cells has been followed by means of the indirect fluorescent antibody technic and concurrent light and electron microscope observations. The immunofluorescence studies revealed that there is a latent period for development of demonstrable measles virus antigen. In amnion cells the latent period lasted for at least 3 days. In contrast, virus antigen could be detected in H.Ep.-2 cells as early as 12 hours following inoculation. In each cell system virus antigen was seen in either nucleus or cytoplasm of infected cells, or both. Early localization tended to be perinuclear. Intranuclear fluorescence was generally less bright and less widespread than cytoplasmic fluorescence. Giant cells and long cytoplasmic spindle-shaped processes appeared regularly in infected cultures. Infectious virus was liberated into the nutrient fluid but when extracellular virus was inhibited by antibody, spread of infection from cell to cell in the monolayer still continued. Results obtained in concurrent electron microscope studies will be presented separately. Correlation of the results of the immunofluorescence and electron microscope studies suggests the possibility that much of the immunofluorescence observed might be due to antigen in virus precursors or components.


Weight reduction by means of intragastric device: experience with the bioenterics intragastric balloon.
A new intragastric balloon is available for weight reduction. Patients consulting for a weight reduction plan and refusing any kind of actual surgery, or suffering from obesity but not meeting the IFSO standards for surgery, were offered the possibility of weight reduction by the BioEnterics intragastric balloon (BIB). A preoperative questionnaire was completed by all patients, inquiring about medical history, co-morbidity factors, dietary habits, previous treatments for weight and social, psychological, relational and economic impact of the obesity. BIB placement was done on an inpatient basis, under general anesthesia. Inflation was standardized at 500 ml saline. Intravenous antiemetic and spasmolytic drugs were given to control post-insertion nausea for 24 hours, and oral medication was administered on the patient's discharge. A standard 800 calorie diet was prescribed after dietitian's consultation. Extraction of the balloon was left to the patient's discretion at 3 or maximum 6 months after placement. Patients choosing for the maximal period received a formal invitation to extract the balloon. Evaluation of weight reduction was done at extraction and by questionnaire. 126 patients (5 M, 121 F) with mean age of 35.6 years (20-62) were included after preoperative evaluation. Mean preoperative BMI was 37.7 kg/m2 (26.7-57.7 kg/m2), with a mean initial excessive weight of 35.3 kg (8.8-96.4 kg) and mean initial % excess weight of 32.2 (6.3-102). 69 patients were eligible for review; mean excess weight loss after 3 months was 48.6% and after 6 months 50.8%. Mean weight loss was 15.4 kg (0-35 kg). 76.8% of the patients (41/69) complained of severe nausea and vomiting lasting an average of 1 week (1 day-6 months), resulting in 3 patients in early removal of the balloon (at 1 day, 1 week, 1 month after placement respectively). 2 patients suffered gastric perforation presenting as acute peritonitis 3 and 4 months after placement and were operated. Extraction of the balloon was performed in 3 patients after 3 months and in 66 patients after 6 months. In 11 patients (22%), esophagitis was present (8 grade 1, 2 grade II, 1 grade III), and one patient showed diffuse gastric erosion. One patient required removal of the balloon by rigid esphagoscopy following technical failure of the endoscopic extraction device. 45 patients replied to the mailed, questionnaire; 15% (7/45) were very satisfied, 13% (6/45) satisfied, 22% (10/45) reasonably satisfied, 8.8% (4/45) unsatisfied and 40% very unsatisfied. Degree of satisfaction correlated poorly with weight loss. Results may be better with close continuous guidance by a counselor. BIB as a means of weight reduction in the obese patient led to a 50.8% loss of excess weight after 6 months. Although severe morbidity can occur, the BIB provides a means for short-term weight reduction in conjunction with dietary measures.


Neural representation of spatial topology in the rodent hippocampus.
Pyramidal cells in the rodent hippocampus often exhibit clear spatial tuning in navigation. Although it has been long suggested that pyramidal cell activity may underlie a topological code rather than a topographic code, it remains unclear whether an abstract spatial topology can be encoded in the ensemble spiking activity of hippocampal place cells. Using a statistical approach developed previously, we investigate this question and related issues in greater detail. We recorded ensembles of hippocampal neurons as rodents freely foraged in one- and two-dimensional spatial environments and used a "decode-to-uncover" strategy to examine the temporally structured patterns embedded in the ensemble spiking activity in the absence of observed spatial correlates during periods of rodent navigation or awake immobility. Specifically, the spatial environment was represented by a finite discrete state space. Trajectories across spatial locations ("states") were associated with consistent hippocampal ensemble spiking patterns, which were characterized by a state transition matrix. From this state transition matrix, we inferred a topology graph that defined the connectivity in the state space. In both one- and two-dimensional environments, the extracted behavior patterns from the rodent hippocampal population codes were compared against randomly shuffled spike data. In contrast to a topographic code, our results support the efficiency of topological coding in the presence of sparse sample size and fuzzy space mapping. This computational approach allows us to quantify the variability of ensemble spiking activity, examine hippocampal population codes during off-line states, and quantify the topological complexity of the environment.


Coping with the New Era: Noise and Light Pollution, Hperactivity and Steroid Hormones. Towards an Evolutionary View of Bipolar Disorders.
Human population is increasing in immense cities with millions of inhabitants, in which life is expected to run 24 hours a day for seven days a week (24/7). Noise and light pollution are the most reported consequences, with a profound impact on sleep patterns and circadian biorhythms. Disruption of sleep and biorhythms has severe consequences on many metabolic pathways. Suppression of melatonin incretion at night and the subsequent effect on DNA methylation may increase the risk of prostate and breast cancer. A negative impact of light pollution on neurosteroids may also affect mood. People who carry the genetic risk of bipolar disorder may be at greater risk of full-blown bipolar disorder because of the impact of noise and light pollution on sleep patterns and circadian biorhythms. However, living in cities may also offers opportunities and might be selective for people with hyperthymic temperament, who may find themselves advantaged by increased energy prompted by increased stimulation produced by life in big cities. This might result in the spreading of the genetic risk of bipolar disorder in the coming decades. In this perspective the burden of poor quality of life, increased disability adjusted life years and premature mortality due to the increases of mood disorders is the negative side of a phenomenon that in its globality also shows adaptive aspects. The new lifestyle also influences those who adapt and show behaviors, reactions and responses that might resemble the disorder, but are on the adaptive side.


A comparison between the determination of free Pb(II) by two techniques: absence of gradients and Nernstian equilibrium stripping and resin titration.
Absence of gradients and Nernstian equilibrium stripping (AGNES) is an emerging electroanalytical technique designed to measure free metal ion concentration. The practical implementation of AGNES requires a critical selection of the deposition time, which can be drastically reduced if the contribution of the complexes is properly taken into account. The resin titration (RT) is a competition method based on the sorption of metal ions on a complexing resin. The competitor here considered is the resin Chelex 100 whose sorbing properties towards Pb(II) are well known. The RT is a consolidated technique especially suitable to perform an intercomparison with AGNES, due to its independent physicochemical nature. Two different ligands for Pb(II) complexation have been analyzed here: nitrilotriacetic acid (NTA) and pyridinedicarboxylic acid (PDCA). The complex PbNTA is practically inert in the diffusion layer, so, for ordinary deposition potentials, its contribution is almost negligible; however, at potentials more negative than -0.8 V vs. Ag/AgCl the complex dissociates on the electrodic surface giving rise to a second wave in techniques such as normal pulse polarography. The complex Pb-PDCA is partially labile, so that its contribution can be estimated from an expression of the lability degree of the complex. These new strategies allow us to reduce the deposition time. The free Pb(II) concentrations obtained by AGNES and by RT are in full agreement for both systems here considered. The main advantage of the use of AGNES in these systems lies in the reduction of the time of the experiment, while RT can be applied to non-amalgamating elements and offers the possibility of simultaneous determinations.


Determining the probability of cyanobacterial blooms: the application of Bayesian networks in multiple lake systems.
A Bayesian network model was developed to assess the combined influence of nutrient conditions and climate on the occurrence of cyanobacterial blooms within lakes of diverse hydrology and nutrient supply. Physicochemical, biological, and meteorological observations were collated from 20 lakes located at different latitudes and characterized by a range of sizes and trophic states. Using these data, we built a Bayesian network to (1) analyze the sensitivity of cyanobacterial bloom development to different environmental factors and (2) determine the probability that cyanobacterial blooms would occur. Blooms were classified in three categories of hazard (low, moderate, and high) based on cell abundances. The most important factors determining cyanobacterial bloom occurrence were water temperature, nutrient availability, and the ratio of mixing depth to euphotic depth. The probability of cyanobacterial blooms was evaluated under different combinations of total phosphorus and water temperature. The Bayesian network was then applied to quantify the probability of blooms under a future climate warming scenario. The probability of the "high hazardous" category of cyanobacterial blooms increased 5% in response to either an increase in water temperature of 0.8°C (initial water temperature above 24°C) or an increase in total phosphorus from 0.01 mg/L to 0.02 mg/L. Mesotrophic lakes were particularly vulnerable to warming. Reducing nutrient concentrations counteracts the increased cyanobacterial risk associated with higher temperatures.


Comparison of several atherogenicity indices by the analysis of serum lipoprotein composition in patients with chronic renal failure with or without haemodialysis, and in renal transplant patients.
We investigated serum lipoproteins in uraemic and kidney transplant patients, and compared the results with those from normal persons. In uraemic patients, with or without haemodialysis, and in kidney transplant patients, the ratio of HDL-cholesterol to total cholesterol minus HDL-cholesterol, the ratio of HDL-phospholipids to total phospholipids minus HDL-phospholipids, and the ratio of apolipoprotein A to apolipoprotein B are all decreased, though to different extents, when compared with healthy control subjects. Furthermore, important differences exist in the relative HDL composition between the 3 patient groups and healthy control subjects. Thus, the ratio of apolipoprotein A/HDL-cholesterol and that of HDL-cholesterol/HDL-phospholipids are significantly altered in uraemic haemodialyzed patients, while the ratio of apolipoprotein A/HDL-phospholipids is normal. By contrast, the 3 ratios are normal in uraemic undialyzed patients, and the 2 ratios, apolipoprotein A/HDL-cholesterol and apolipoprotein A/HDL-phospholipids are normal in renal transplant patients. The last ratio, HDL-cholesterol/HDL-phospholipids, is increased in this group of patients. Thus, it appears that HDL-cholesterol, HDL-phospholipids, and total apolipoprotein A represent different aspects of the same lipoprotein. The determination of all three parameters could lead to a different approach in the evaluation of the so-called cardiovascular risk, at least for uraemic patients and renal transplant patients.


Flaxseed reduces plasma cholesterol and atherosclerotic lesion formation in ovariectomized Golden Syrian hamsters.
Dietary modification contributes significantly in reducing cardiovascular disease (CVD) risk factors including lowering cholesterol and atherosclerosis. The purpose of this study was to investigate the effects of flaxseed, a rich source of lignans, alpha-linolenic acid and soluble fiber mucilage, on the prevention of ovariectomy-induced rise in total cholesterol and atherosclerotic lesions. Seventy-two 6-month-old female Golden Syrian hamsters were either ovariectomized (ovx) or sham-operated (sham) and randomly assigned to six groups (n = 12): sham, ovx, or ovx plus either 17beta-estradiol (E(2), 10 microg/kg body weight) or semi-purified diet adjusted for macronutrients and fiber to contain one of the three doses of flaxseed (7.5, 15, or 22.5%) for 120 days. Ovariectomy significantly elevated plasma total-, HDL-, and free-cholesterol concentrations. Similar to estrogen, all doses of flaxseed were effective in preventing the ovx-induced rise in plasma total cholesterol. Triglyceride concentrations were significantly higher in the flax-fed hamsters. There were no significant differences in plasma non-HDL- and esterified-cholesterol among the treatment groups. Ovariectomy also increased the number of hamsters with lesions and the aortic fatty streak area. All three doses of flaxseed reduced the fatty streak area and the incidence of lesions to levels similar to the sham group. The findings of this study show that flaxseed is beneficial in reducing plasma cholesterol and plaque formation induced by ovarian hormone deficiency.


Non-antibiotic agent ginsenoside 20(S)-Rh2 enhanced the antibacterial effects of ciprofloxacin in vitro and in vivo as a potential NorA inhibitor.
The aim of this study is to explore the potential enhancing effect of ginsenoside 20(S)-Rh2 (Rh2) towards ciprofloxacin (CIP) against Staphylococcus aureus (S. aureus) infection in vitro and in vivo, and analyze the possible mechanisms through NorA inhibition from a target cellular pharmacokinetic view. In combination with non-toxic dosage of Rh2, the susceptibilities of S. aureus strains to CIP were significantly augmented, and the antibacterial kinetics of CIP in the S. aureus strains were markedly promoted. This enhancing effect of Rh2 towards CIP was also observed in S. aureus infected peritonitis mice, with elevated survival rate and reduced bacteria counts in blood. However, Rh2 did not influence the plasma concentrations of CIP. Further analysis indicated that Rh2 significantly promoted the accumulations of CIP in S. aureus, and inhibited the NorA mediated efflux of pyronin Y. The expressions of NorA gene on S. aureus were positively correlated with the enhancing effect of Rh2 with CIP. This is the first report of the enhancing effect of Rh2 with CIP for S. aureus infection in vitro and in vivo, of which it is probably that Rh2 inhibited NorA-mediated efflux and promoted the accumulation of CIP in the bacteria.


Analysis of polyomavirus-infected renal transplant recipients' urine specimens: correlation of routine urine cytology, fluorescence in situ hybridization, and digital image analysis.
Urinary polyomavirus subtype BK (BKV)-infected urothelial and renal tubular cells, "decoy cells," have been shown to be aneuploid with digital image analysis (DIA). We wanted to determine whether decoy cells cause false-positive fluorescence in situ hybridization (FISH) results in BKV-infected urine samples. Urine samples from 38 renal transplant recipients were split 3 ways and evaluated for number of decoy cells per 10 high-power fields by routine cytology, total nuclear DNA content by DIA, and chromosomal abnormalities by FISH. For DIA, Feulgen-stained cells were quantified with a CAS 200 image analyzer (Bacus Laboratories, Lombard, IL), and for FISH, the UroVysion probe set (Vysis, Downers Grove, IL) consisting of chromosome enumeration probes 3, 7, 17, and locus-specific identifier probe 9p21 (p16 gene) was used. Of the 38 specimens, 32 (84.2%) had evidence of BKV infection by routine cytology. DIA and FISH results were aneuploid/aneusomic in 30 (93.8%) and 4 (12.5%) cases, respectively. All aneusomic FISH specimens occurred in men older than 52 years of age and who also had 4 of the 6 highest PCR-BKV titers. To date, no patient has clinical evidence of malignancy. The 6 specimens without decoy cells were DNA diploid/disomic by DIA and FISH. Abnormal FISH results of urinary decoy cells occur much less frequently than aneuploidy by DIA in renal transplant recipients and might merit close follow-up in some transplant recipients.


Prophylactic therapy with human amniotic fluid stem cells improved survival in a rat model of lipopolysaccharide-induced neonatal sepsis through immunomodulation via aggregates with peritoneal macrophages.
Despite recent advances in neonatal care, sepsis remains a leading cause of mortality in neonates. Mesenchymal stem cells derived from various tissues, such as bone marrow, umbilical cord, and adipose tissue, have beneficial effects on adult sepsis. Although human amniotic fluid stem cells (hAFSCs) have mesenchymal stem cell properties, the efficacy of hAFSCs on neonatal sepsis is yet to be elucidated. This study aimed to investigate the therapeutic potential of hAFSCs on neonatal sepsis using a rat model of lipopolysaccharide (LPS)-induced sepsis. hAFSCs were isolated as CD117-positive cells from human amniotic fluid. Three-day-old rat pups were intraperitoneally treated with LPS to mimic neonatal sepsis. hAFSCs were administered either 3 h before or at 0, 3, or 24 h after LPS exposure. Serum inflammatory cytokine levels, gene expression profiles from spleens, and multiple organ damage were analyzed. hAFSC localization was determined in vivo. In vitro LPS stimulation tests were performed using neonatal rat peritoneal macrophages co-cultured with hAFSCs in a cell-cell contact-dependent/independent manner. Immunoregulation in the spleen was determined using a DNA microarray analysis. Prophylactic therapy with hAFSCs improved survival in the LPS-treated rats while the hAFSCs transplantation after LPS exposure did not elicit a therapeutic response. Therefore, hAFSC pretreatment was used for all subsequent studies. Inflammatory cytokine levels were elevated after LPS injection, which was attenuated by hAFSC pretreatment. Subsequently, inflammation-induced damages in the brain, lungs, and liver were ameliorated. hAFSCs aggregated with peritoneal macrophages and/or transiently accumulated in the liver, mesentery, and peritoneum. Paracrine factors released by hAFSCs induced M1-M2 macrophage polarization in a cell-cell contact-independent manner. Direct contact between hAFSCs and peritoneal macrophages further enhanced the polarization. Microarray analysis of the spleen showed that hAFSC pretreatment reduced the expression of genes involved in apoptosis and inflammation and subsequently suppressed toll-like receptor 4 signaling pathways. Prophylactic therapy with hAFSCs improved survival in a rat model of LPS-induced neonatal sepsis. These effects might be mediated by a phenotypic switch from M1 to M2 in peritoneal macrophages, triggered by hAFSCs in a cell-cell contact-dependent/independent manner and the subsequent immunomodulation of the spleen.


Percutaneous bladder catheterization in microgravity.
Urinary obstruction (UO) or failure to void has been observed during several episodes of short-duration spaceflight, necessitating bladder catheterization. It should be considered a possible medical condition in long-duration space missions as well. Antiemetics used early in space flight add to the risk and severity of voiding problems, along with the sensory and psychological peculiarities of voiding without gravity and in the unusual setting of a spacecraft. Urolithiasis due to the above-normal calcium excretion increases the risk of UO in long duration space missions. Finally, the individual risk of UO is higher against the background of preexisting conditions such as benign prostatic hyperplasia (BPH) or urethral stricture. Both acute retention and ureteral obstruction are associated with substantial patient distress, and carry a risk of urosepsis and/or acute renal failure. If UO in orbital flight is unresolved or complicated, it would likely result in crew emergency return from orbit. Exploration missions, however, may require means for definitive treatment of urinary tract obstruction. This study documents successful ultrasound-guided percutaneous catheterization of the urinary bladder in microgravity. A porcine model of urethral occlusion was used. The results demonstrate an additional capability from our previous investigations describing endoscopic catheterization and stenting of the ureters in microgravity conditions. In an anesthetized porcine model, a Foley catheter was placed in the bladder and clamped after instillation of 200 ml of colored liquid. The bladder was visualized and then drained under ultrasound guidance through suprapubic puncture, employing a 10.3 F pigtail catheter with introducer. The procedural elements were conducted only during microgravity portions of the parabolic flight. Ultrasound imaging was used to successfully perform image-guided percutaneous puncture through the anterior bladder wall with the catheter, without injury to adjacent organs. The percutaneous catheter was able to successfully drain the bladder in microgravity conditions. Percutaneous bladder catheterization and drainage can be successfully performed in weightless conditions under ultrasound guidance. Ultrasound provides a low-power, portable means to safely conduct minimally invasive procedures in pertinent organs and tissues. Percutaneous bladder catheterization is a standard procedure when luminal bladder catheterization is not possible; this technique can be successfully modified for use in space medicine applications.


Transporter-Mediated Delivery of Small Molecule Drugs to the Brain: A Critical Mechanism That Can Advance Therapeutic Development for Ischemic Stroke.
Ischemic stroke is the 5th leading cause of death in the United States. Despite significant improvements in reperfusion therapies, stroke patients still suffer from debilitating neurocognitive deficits. This indicates an essential need to develop novel stroke treatment paradigms. Endogenous uptake transporters expressed at the blood-brain barrier (BBB) provide an excellent opportunity to advance stroke therapy via optimization of small molecule neuroprotective drug delivery to the brain. Examples of such uptake transporters include organic anion transporting polypeptides (OATPs in humans; Oatps in rodents) and organic cation transporters (OCTs in humans; Octs in rodents). Of particular note, small molecule drugs that have neuroprotective properties are known substrates for these transporters and include 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (i.e., statins) for OATPs/Oatps and 1-amino-3,5-dimethyladamantane (i.e., memantine) for OCTs/Octs. Here, we review current knowledge on specific BBB transporters that can be targeted for improvement of ischemic stroke treatment and provide state-of-the-art perspectives on the rationale for considering BBB transport properties during discovery/development of stroke therapeutics.


First Report of Bacterial Wilt Caused by Ralstonia solanacearum on Roselle in Taiwan.
Roselle (Hibiscus sabdariffa L.) is a herbaceous plant belonging to the Malvaceae family. Its calyxes are rich in vitamin C and anthocyanins and are used to make roselle drink and hibiscus tea. Roselles are grown in counties of Taitung, Pingtung, and Chiayi in Taiwan. In addition to a few local cultivars, the major cultivar currently grown in Taiwan is Roselle cv. Victor. In April of 2012, a wilt disease appeared on seedlings of a cultivar, Chiada 1, at the Chungpu Township of Chiayi County. Mature plants were free from this disease. Leaves appeared weak and drooping when they were still green, followed by collapse of the whole plant a few days later. Browning of vascular and pith tissues was evident, especially at the base of the stem. A whitish mass of bacteria oozed from the cut end of diseased stems, suggesting that bacteria might be the cause of this disease. A total of 15 bacterial strains were collected. Colonies on tetrazolium chloride medium (3) were round to oval and fluidal, each with a pink or red center after incubation at 30°C for 48 h. When tobacco leaves were infiltrated with these strains, a hypersensitive reaction (HR) typical of phytopathogenic bacteria was induced. All strains produced the expected amplicon (282 bp) after PCR with the Ralstonia solanacearum-specific primer pair, AU759f and AU760r (4). Three hexose alcohols (mannitol, sorbitol, and dulcitol), rather than three disaccharides (lactose, maltose, and cellobiose), were utilized, which suggests R. solanacearum biovar 4 (2). R. solanacearum phylotype I was determined by phylotype-specific multiplex PCR (1). Pathogenicity of the strains was tested on roselle, tomato, pepper, and eggplant. Young plants of the various species were inoculated at the four- to six-leaf stage by soil drenching with 30 ml of bacterial suspension (about 108 CFU/ml). Control plants were inoculated with sterile water. Each treatment comprised eight plants with a single plant in each pot. Plants were incubated in a greenhouse at 25 to 31°C and 56 to 93% humidity. Wilting was observed 4 to 6 days after inoculation, while the control did not wilt. To find the correlation between plant growth stage and resistance to the pathogen, 2-, 3-, 4-, and 5-week-old roselle plants cv. Chiada 1 were transplanted into artificially infested soil. Eight plants in each treatment were planted with a single plant in each pot. The disease incidences for plants of different ages were 75%, 62.5%, 50%, and 12.5%, respectively. This study showed that resistance increases with plant age. Hence, if older seedlings are transplanted, the risk of bacterial wilt of roselle can be reduced. To our knowledge, this is the first report of R. solanacearum on roselle in Taiwan. References: (1) M. Fegan and P. Prior. Bacterial Wilt Disease and the Ralstonia solanacearum Species Complex, page 449. C. Allen et al., eds. The American Phytopathological Society. St. Paul, MN, 2005. (2) A. C. Hayward. J. Appl. Bacteriol. 27:265, 1964. (3) A. Kelman. Phytopathology 44:693, 1954. (4) N. Opina et al. Asia Pac. J. Mol. Biol. Biotechnol. 5:19, 1997.


A comparative analysis of island floras challenges taxonomy-based biogeographical models of speciation.
Speciation on islands, and particularly the divergence of species in situ, has long been debated. Here, we present one of the first, complete assessments of the geographic modes of speciation for the flora of a small oceanic island. Cocos Island (Costa Rica) is pristine; it is located 550 km off the Pacific coast of Central America. It harbors 189 native plant species, 33 of which are endemic. Using phylogenetic data from insular and mainland congeneric species, we show that all of the endemic species are derived from independent colonization events rather than in situ speciation. This is in sharp contrast to the results of a study carried out in a comparable system, Lord Howe Island (Australia), where as much as 8.2% of the plant species were the product of sympatric speciation. Differences in physiography and age between the islands may be responsible for the contrasting patterns of speciation observed. Importantly, comparing phylogenetic assessments of the modes of speciation with taxonomy-based measures shows that widely used island biogeography approaches overestimate rates of in situ speciation.


Effect of ultrasound on the extraction of total anthocyanins from Purple Majesty potato.
This study examined anthocyanin extraction using the application of ultrasound to raw freeze dried, microwaved and raw sliced Purple Majesty potato, a new pigmented potato variety rich in anthocyanins. A 20 kHz probe was used for the sonication at 3 different amplitudes (30%, 50% and 70%) and ethanol in water at different ratios (50:50 and 70:30 v/v) was used for the extraction. Anthocyanin extraction from raw freeze dried purple potato was optimal at an ethanol:water ratio (70:30; v/v) after 5 min of ultrasonication, while the least amount of anthocyanins was extracted from raw sliced potatoes. The application of microwaves (as a pre-treatment) before the UAE resulted in an increase in the amount of anthocyanins extracted and a decrease in the amount of solvent used. Analysis of variance showed that potato form, ultrasonication time, ultrasonication amplitude and solvent ratio as well as two and three way interactions between some of these factors had a very significant effect (p<0.000) on the amount of anthocyanins extracted.


Cost-effectiveness of early versus standard antiretroviral therapy in HIV-infected adults in Haiti.
In a randomized clinical trial of early versus standard antiretroviral therapy (ART) in HIV-infected adults with a CD4 cell count between 200 and 350 cells/mm³ in Haiti, early ART decreased mortality by 75%. We assessed the cost-effectiveness of early versus standard ART in this trial. Trial data included use of ART and other medications, laboratory tests, outpatient visits, radiographic studies, procedures, and hospital services. Medication, laboratory, radiograph, labor, and overhead costs were from the study clinic, and hospital and procedure costs were from local providers. We evaluated cost per year of life saved (YLS), including patient and caregiver costs, with a median of 21 months and maximum of 36 months of follow-up, and with costs and life expectancy discounted at 3% per annum. Between 2005 and 2008, 816 participants were enrolled and followed for a median of 21 months. Mean total costs per patient during the trial were US$1,381 for early ART and US$1,033 for standard ART. After excluding research-related laboratory tests without clinical benefit, costs were US$1,158 (early ART) and US$979 (standard ART). Early ART patients had higher mean costs for ART (US$398 versus US$81) but lower costs for non-ART medications, CD4 cell counts, clinically indicated tests, and radiographs (US$275 versus US$384). The cost-effectiveness ratio after a maximum of 3 years for early versus standard ART was US$3,975/YLS (95% CI US$2,129/YLS-US$9,979/YLS) including research-related tests, and US$2,050/YLS excluding research-related tests (95% CI US$722/YLS-US$5,537/YLS). Initiating ART in HIV-infected adults with a CD4 cell count between 200 and 350 cells/mm³ in Haiti, consistent with World Health Organization advice, was cost-effective (US$/YLS <3 times gross domestic product per capita) after a maximum of 3 years, after excluding research-related laboratory tests. ClinicalTrials.gov NCT00120510.


Association between Serum Cholesterol Level and Bone Mineral Density at Lumbar Spine and Femur Neck in Postmenopausal Korean Women.
Blood lipid profiles have been suggested to be a risk factor for osteoporosis. However, the association between lipid profiles and bone mineral density (BMD) is still unclear. This study aimed to evaluate an association between blood lipid profiles and BMD through both a cross-sectional and a longitudinal study. Study subjects were 958 postmenopausal Korean women who have repeatedly undertaken laboratory tests and BMD measurements at lumbar spine and femur neck with an interval of 7.1 years. The associations between lipid profiles and BMD were examined using Spearman correlation analysis with an adjustment for age, smoking, alcohol drinking, physical activity, body mass index, and follow-up duration. Lumbar spine BMD was not associated with total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HLD-C) regardless of when the measurement was performed. In an analysis using data measured at the beginning of the study, femur neck BMD was not associated with TC and LDL-C. However, femur neck BMD showed weak but significantly positive correlation with HDL-C (correlation coefficient, 0.077; 95% confidence interval, 0.005 to 0.149). When the analysis was repeated with data measured at the end of the follow-up, there was no significant correlation between femur neck BMD and any lipid profile. In addition, change in femur neck BMD during follow-up was not associated with the change in lipid profiles. Although further study with a consideration of calcium intake and osteoporosis medication seems necessary, this study found no association between serum lipid profiles and BMD in postmenopausal Korean women.