Genotype-phenotype correlations in SCN8A-related disorders reveal prognostic and therapeutic implications.
We report detailed functional analyses and genotype-phenotype correlations in 392 individuals carrying disease-causing variants in SCN8A, encoding the voltage-gated Na+ channel Nav1.6, with the aim of describing clinical phenotypes related to functional effects. Six different clinical subgroups were identified: Group 1, benign familial infantile epilepsy (n = 15, normal cognition, treatable seizures); Group 2, intermediate epilepsy (n = 33, mild intellectual disability, partially pharmaco-responsive); Group 3, developmental and epileptic encephalopathy (n = 177, severe intellectual disability, majority pharmaco-resistant); Group 4, generalized epilepsy (n = 20, mild to moderate intellectual disability, frequently with absence seizures); Group 5, unclassifiable epilepsy (n = 127); and Group 6, neurodevelopmental disorder without epilepsy (n = 20, mild to moderate intellectual disability). Those in Groups 1-3 presented with focal or multifocal seizures (median age of onset: 4 months) and focal epileptiform discharges, whereas the onset of seizures in patients with generalized epilepsy was later (median: 42 months) with generalized epileptiform discharges. We performed functional studies expressing missense variants in ND7/23 neuroblastoma cells and primary neuronal cultures using recombinant tetrodotoxin-insensitive human Nav1.6 channels and whole-cell patch-clamping. Two variants causing developmental and epileptic encephalopathy showed a strong gain-of-function (hyperpolarizing shift of steady-state activation, strongly increased neuronal firing rate) and one variant causing benign familial infantile epilepsy or intermediate epilepsy showed a mild gain-of-function (defective fast inactivation, less increased firing). In contrast, all three variants causing generalized epilepsy induced a loss-of-function (reduced current amplitudes, depolarizing shift of steady-state activation, reduced neuronal firing). Functional effects were known for 170 individuals. All 136 individuals carrying a functionally tested gain-of-function variant had either focal (n = 97, Groups 1-3) or unclassifiable (n = 39) epilepsy, whereas 34 individuals with a loss-of-function variant had either generalized (n = 14), no (n = 11) or unclassifiable (n = 6) epilepsy; only three had developmental and epileptic encephalopathy. Computational modelling in the gain-of-function group revealed a significant correlation between the severity of the electrophysiological and clinical phenotypes. Gain-of-function variant carriers responded significantly better to sodium channel blockers than to other anti-seizure medications, and the same applied for all individuals in Groups 1-3. In conclusion, our data reveal clear genotype-phenotype correlations between age at seizure onset, type of epilepsy and gain- or loss-of-function effects of SCN8A variants. Generalized epilepsy with absence seizures is the main epilepsy phenotype of loss-of-function variant carriers and the extent of the electrophysiological dysfunction of the gain-of-function variants is a main determinant of the severity of the clinical phenotype in focal epilepsies. Our pharmacological data indicate that sodium channel blockers present a treatment option in SCN8A-related focal epilepsy with onset in the first year of life.


Reliability and Validity of the Pediatric Palliative Care Questionnaire for Measuring Self-Efficacy, Knowledge, and Adequacy of Prior Medical Education among Pediatric Fellows.
Interventions to improve pediatric trainee education in palliative care have been limited by a lack of reliable and valid tools for measuring effectiveness. We developed a questionnaire to measure pediatric fellows' self-efficacy (comfort), knowledge, and perceived adequacy of prior medical education. We measured the questionnaire's reliability and validity. The questionnaire contains questions regarding self-efficacy (23), knowledge (10), fellow's perceived adequacy of prior medical education (6), and demographics. The survey was developed with palliative care experts, and sent to fellows in U.S. pediatric cardiology, critical care, hematology/ oncology, and neonatal-perinatal medicine programs. Measures of reliability, internal consistency, and validity were calculated. One hundred forty-seven fellows completed the survey at test and retest. The self-efficacy and medical education questionnaires showed high internal consistency of 0.95 and 0.84. The test-retest reliability for the Self-Efficacy Summary Score, measured by intraclass correlation coefficient (ICC) and weighted kappa, was 0.78 (item range 0.44-0.81) and 0.61 (item range 0.36-0.70), respectively. For the Adequacy of Medical Education Summary Score, ICC was 0.85 (item range 0.6-0.78) and weighted kappa was 0.63 (item range 0.47-0.62). Validity coefficients for these two questionnaires were 0.88 and 0.92. Fellows answered a mean of 8.8/10 knowledge questions correctly; percentage agreement ranged from 65% to 99%. This questionnaire is capable of assessing self-efficacy and fellow-perceived adequacy of their prior palliative care training. We recommend use of this tool for fellowship programs seeking to evaluate fellow education in palliative care, or for research studies assessing the effectiveness of a palliative care educational intervention.


Intentions of older homebound women to reduce the risk of falling again.
Researchers have tested interventions to prevent recurrent falls for older people without exploring their intentions to prevent another fall. Lack of knowledge about such intentions is an impediment to intervention effectiveness. The purpose was to describe intentions to prevent another fall as discerned during a study with older homebound women. Data were obtained during a series of four in-home interviews over 18 months with monthly telephone contacts between interviews; fall history was updated at each contact. A descriptive phenomenological method was used to analyze data. Of the 40 women, 36 (aged 85 to 98 years) had fallen at home at least once before enrolling in the study, or had a subsequent or initial fall during the study. The overall intention was reducing my risk of falling again at home, with its components figuring out the reason that I fell and changing my ways to reduce my risk of falling again at home. Most women explained reasons for a fall and voiced intentions to prevent a similar fall. Women who viewed falls as unexpected events were uncertain that they could prevent a fall or felt unable to do so and voiced few preventive intentions. A few women voiced generalized preventive intentions to change health-related behaviors or habits. Most intentions were tied directly to the situation in which a fall had occurred. Such specificity could limit effectiveness of personal efforts to prevent falls in other situations. Practitioners should converse with older women who have fallen about their intentions to prevent another fall and weigh the need to help them generalize both the risk of falling again and their risk reduction intentions.


A polymorphism in IL28B distinguishes exposed, uninfected individuals from spontaneous resolvers of HCV infection.
Polymorphisms in the interleukin-28B (IL28B) gene are associated with outcomes from infection with hepatitis C virus (HCV). However, the role of these polymorphisms in protecting injection drug users who are at high risk for HCV infection but do not have detectable antibodies against HCV or HCV RNA (exposed uninfected) has not been demonstrated. We investigated whether these individuals have the IL28B genotype rs12979860-CC, which protects some individuals against HCV infection. Seventy-four exposed uninfected individuals, 89 spontaneous resolvers, and 234 chronically infected individuals were genotyped to determine single nucleotide polymorphisms at IL28B.rs12979860. Exposed, uninfected individuals had a significantly lower frequency of the protective genotype (rs12979860-CC) than anti-HCV-positive spontaneous resolvers (41.9% vs 69.7%, respectively; P=.0005; odds ratio [OR], 0.31; 95% confidence interval [CI]: 0.16-0.60) but a similar frequency to patients who were chronically infected (41.9% vs 43.6%, respectively; P=ns). However, exposed, uninfected individuals had a significantly higher frequency of homozygosity for killer cell immunoglobulin-like receptor 2DL3:group 1 HLA-C (KIR2DL3:HLA-C1) than those with chronic infection (31.1% vs 13.3%, respectively; P=.0008; OR, 2.95; 95% CI: 1.59-5.49). For patients who spontaneously resolved infection, IL28B and KIR:HLA protected, independently, against chronic HCV infection, based on logistic regression and synergy analyses (synergy factor, 1.3; 95% CI: 0.37-4.75; P synergy=.6). IL28B and KIR2DL3:HLA-C1 are independently associated with spontaneous resolution of viremia following HCV exposure. Resistance to HCV infection in exposed uninfected cases is associated with homozygosity for KIR2DL3:HLA-C1 but not the single nucleotide polymorphism IL28B.rs12979860. Uninfected individuals are therefore a distinct population from patients who spontaneously resolve HCV infection. Distinct, nonsynergistic innate immune mechanisms can determine outcomes of HCV exposure.


Technology evaluation: electroporation therapy, Genetronics Inc.
Genetronics Inc has developed a MedPulser and a needle array applicator that delivers electric pulses to tumors and induces a transient permeabilization. Used in conjunction with intratumoral injections of anticancer drugs, this form of therapy, termed electroporation therapy, allows for intracellular accumulation of cytotoxic drugs without the toxic side effects associated with systemic administration. Accrual of preclinical data of electroporation therapy with bleomycin has led to clinical studies in patients with cutaneous and subcutaneous tumors. In 1999, objective responses were reported in a phase II study involving 30 patients with head and neck cancer with minimal side effects [328332]. Other indications for this form of treatment include liver and pancreatic cancers, Kaposi's sarcoma and melanoma [273388,290144]. Genetronics Inc and Ethicon Endo-Surgery (Johnson & Johnson Development Corp) are reviewing all clinical data from phase II studies prior to the initiation of a pivotal clinical trial in head and neck cancer.


Impact of The Real Cost Media Campaign on Youth Smoking Initiation.
The purpose of this study was to assess the relationship between youth exposure to the U.S. Food and Drug Administration's national tobacco public education campaign, The Real Cost, and changes in smoking initiation. From November 2013 to November 2016, a longitudinal study of youth was conducted with a baseline and 4 post-campaign follow-up surveys. The sample consisted of nonsmoking youths from 75 U.S. media markets (n=5,103) who completed a baseline and at least 1 follow-up survey. Exposure was measured by media market-level target rating points and self-reported ad exposure frequency. Smoking initiation was examined among youths who had never smoked at baseline and defined as first trial of a cigarette. Discrete-time survival models using logistic regression and controlling for confounding influences were estimated. Analyses were conducted in 2018. The odds of reporting smoking initiation at follow-up was lower among youths in media markets with higher levels of campaign advertisements than among those with less. Both between-wave and cumulative target rating points were associated with decreased risk of smoking initiation (AOR=0.69 [p<0.01] and AOR=0.89 [p<0.05], respectively); for every 3,500 between-wave target rating points on air, there was an associated 30% reduction in the hazard of smoking initiation among youths. Results from self-reported recall of the campaign advertisements found similar dose-response effects. The campaign is associated with an estimated 380,000-587,000 youths aged 11-19 years being prevented from initiating smoking nationwide. Sustained national tobacco public education campaigns like The Real Cost can change population-level smoking initiation among youths, preventing future generations from tobacco-related harms.


Iloperidone, asenapine, and lurasidone: a brief overview of 3 new second-generation antipsychotics.
Three new second-generation antipsychotics were approved by the US Food and Drug Administration in 2009 and 2010: iloperidone, asenapine, and lurasidone. All 3 agents are approved for the treatment of acute schizophrenia in adults, and asenapine is also approved for the maintenance treatment of schizophrenia and as a monotherapy or as an adjunct to lithium or valproate for the treatment of bipolar manic or mixed episodes. The expectation is that these new agents will be less problematic regarding treatment-emergent weight gain and metabolic disturbances, which unfortunately can occur with several other second-generation antipsychotics. Asenapine is a sublingual preparation, in contrast to iloperidone and lurasidone, which are swallowed. Iloperidone and asenapine are dosed twice daily, in contrast to lurasidone, which is dosed once daily with food. Both asenapine and lurasidone can be initiated at a dose that is possibly therapeutic, but iloperidone requires 4 days of titration to reach its recommended target dose range. Although both asenapine and lurasidone can be associated with dose-related treatment-emergent akathisia, iloperidone is essentially free of extrapyramidal adverse effects or akathisia throughout its recommended dose range. Sedation and/or somnolence have been reported with each medication. They are the most common adverse events associated with asenapine treatment, and are clearly dose-related for lurasidone. In contrast, no therapeutic dose response for iloperidone, asenapine, or lurasidone is clearly evident from short-term clinical trials. Longer-term and naturalistic studies will be helpful in evaluating these agents and their role in the psychiatric armamentarium.


Sector hamartoma of the iris.
A 15-year old girl with a jet black-pigmented lesion on the anterior surface of the upper nasal quadrant of her iris was followed up for 12 years, during which interval growth of the lesion was documented. Histopathological evaluation of the excised segment of the iris revealed a pigment epithelial proliferation originating from posterior pigment epithelium. Additionally, there was a band of spindle shaped smooth muscle cells in the posterior stroma and a zone of plump melanocytes in the anterior stroma beneath the proliferating pigment epithelial cells. This combination of abnormalities is thought to represent a benign hamartomatous maldevelopment of the involved sector of the iris rather than a true neoplasm. Such an elaborate dysgenesis is unlike the more modest melanocytic hamartomas of the iris associated with the phakomatoses.


New model for the study of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane: the gelatin sponge/chorioallantoic membrane assay.
Several methods for the in vivo study of angiogenesis are available, and each angiogenic assay presents distinct advantages and disadvantages. In this study, we present a new method for the quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM), based on the implantation of gelatin sponges on the top of growing CAM, on day 8 of incubation. After implantation, the sponges were treated with a stimulator (recombinant human basic fibroblast growth factor, FGF2) or an inhibitor (a rabbit polyclonal anti-FGF2 antibody) of blood vessel formation. Blood vessels growing vertically into the sponge and at the boundary between sponge and surrounding CAM mesenchyme were counted by a morphometric method on day 12. In addition, to assess whether the gelatin sponge is an appropriate vehicle to deliver cultured cells and evaluate their angiogenic potential, mouse aortic endothelial cells were cotransfected with human FGF2 and the Escherichia coli beta-galactosidase (beta-GAL) reporter gene. Stable transfectants were absorbed by the sponge, and evaluation of the angiogenic response was paralleled by beta-GAL staining to visualize implanted cells. This technique may facilitate the discovery and development of agonists or antagonists of angiogenesis.


Sleep disorders in children with blindness.
To evaluate the frequency and type of sleep disorders seen in blind children compared with matched controls, a 42-item questionnaire was used on 156 children (77 blind children) ranging from 3 to 18 years of age. A total of 17.4% of blind children reported sleeping less than 7 hours per night on weekdays compared with 2.6% of controls, with blind children awakening much earlier. Blind children had more sleep complaints, and 13.4% of blind subjects had daily episodes of involuntary sleepiness compared with 1.3% of controls. Blindness has an impact on sleep and alertness that adds to the primary disability.


[The functional status of the cerebral cortex after replacement of skull base bone defects].
The functional status of the cerebral cortex was evaluated by EEG before and different periods after prolonged cranial osteoplasty made by the type of replacement of skull base bone defects in patients with dyscirculatory encephalopathies of various genesis. Seven patients aged 20 to 50 years undergone repair of skull base bone defects (both posttraumatic and simulated) by the procedures developed at the Russian Research Center "VTO" were examined. The bioelectrical activity of the brain was recorded and analyzed on a PEGASUS digital 16-channel EEG system (EMS, Austria) by using the Fourie prompt conversion test (spectral analysis) and amplitude-frequency mapping. After multimodality treatment, all the patients showed a positive changes in the whole range of EEG parameters, which appeared as an increase in basic rhythm power, a decrease in the focal area of slow-wave activity and in the interhemispheral asymmetry coefficient, which in combination of neurological regression is regarded as indirect evidence for the vasoactivity of prolonged cranial osteoplasty.


Antibiotic management of lung infections in cystic fibrosis. II. Nontuberculous mycobacteria, anaerobic bacteria, and fungi.
Airway infections are a key component of cystic fibrosis (CF) lung disease. Whereas the approach to common pathogens such as Pseudomonas aeruginosa is guided by a significant body of evidence, other infections often pose a considerable challenge to treating physicians. In Part I of this series on the antibiotic management of difficult lung infections, we discussed bacterial organisms including methicillin-resistant Staphylococcus aureus, gram-negative bacterial infections, and treatment of multiple bacterial pathogens. Here, we summarize the approach to infections with nontuberculous mycobacteria, anaerobic bacteria, and fungi. Nontuberculous mycobacteria can significantly impact the course of lung disease in patients with CF, but differentiation between colonization and infection is difficult clinically as coinfection with other micro-organisms is common. Treatment consists of different classes of antibiotics, varies in intensity, and is best guided by a team of specialized clinicians and microbiologists. The ability of anaerobic bacteria to contribute to CF lung disease is less clear, even though clinical relevance has been reported in individual patients. Anaerobes detected in CF sputum are often resistant to multiple drugs, and treatment has not yet been shown to positively affect patient outcome. Fungi have gained significant interest as potential CF pathogens. Although the role of Candida is largely unclear, there is mounting evidence that Scedosporium species and Aspergillus fumigatus, beyond the classical presentation of allergic bronchopulmonary aspergillosis, can be relevant in patients with CF and treatment should be considered. At present, however there remains limited information on how best to select patients who could benefit from antifungal therapy.


Review article: monitoring for drug side-effects in inflammatory bowel disease.
The side-effects suitable for monitoring in patients with inflammatory bowel disease being treated with the four main groups of drugs (5-aminosalicylic acid preparations, azathioprine and 6-mercaptopurine, methotrexate, and corticosteroids) are reviewed. On the basis of the reported frequency, severity and timing of side-effects, a practical scheme of monitoring is recommended. This includes a baseline measurement of full blood count, creatinine and liver function tests in all patients. In the absence of worrying symptoms, we recommend the following: (i) no monitoring for sulfasalazine; (ii) for other 5-aminosalicylic acid preparations, the measurement of creatinine at 6 and 12 months and then annually; (iii) for azathioprine/6-mercaptopurine, thiopurine methyltransferase genotype/phenotype determination has no role in treatment monitoring, but a full blood count at 2 weeks, 1 month, 3 months and then every 3 months should be performed; (iv) for methotrexate, a full blood count and liver function tests should be performed every 3 months; (v) for steroids, dual energy X-ray absorptiometry bone scanning should be performed at the start of therapy, every year in which steroids are used if the T score is < 0, and every 3-5 years if the T score is > 0.


Single time point diagnostic breath tests: a review.
The metabolism of ingested xenobiotics is clinically significant to minimize risk and optimize therapeutic benefits. A majority of the drugs approved by the FDA are metabolized by phase I enzymes. Stable isotope-labeled xenobiotics can be used to provide rapid in vivo phenotype assessment of phase I enzymes. In this paper, we describe three simple, noninvasive phenotype breath tests using [13C]-dextromethorphan and [13C]-pantoprazole for assessing polymorphic CYP2D6 and CYP2C19 enzyme activity and [13C]-uracil to assess the enzyme activity of DPD, DPHD and β-ureidopropionase for identifying pyrimidine metabolic disorder. The results of the [13C]-dextromethorphan, [13C]-pantoprazole and [13C]-uracil breath test studies suggest that they have great potential for evaluating CYP2D6, CYP2C19 and DPD enzyme activities in a relatively short time with a single time point breath collection in a clinic or hospital setting. This would enable physicians to prescribe personalized therapy for each individual by selecting the ideal medication at the right dose for optimal efficacy of xenobiotics metabolized by these enzymes.


Thermal environment and sudden infant death syndrome: case-control study.
To compare the thermal environment of infants who died of the sudden infant death syndrome with that of age matched control infants. Case-control study. Infants who died were matched with two controls, one for age and one for age and birth weight. Thermal measurements were conducted at the death scene for cases and at the scene of last sleep for control infants, who were visited unexpectedly within four weeks of the index infant's death on a day of similar climatic conditions. A follow up questionnaire was administered to parents of cases and controls. The geographical area served by the professional Tasmanian state ambulance service, which includes 94% of the Tasmanian population. 41 infants died of the sudden infant death syndrome at home; thermal observations at death scene were available for 28 (68%), parental questionnaire data were available for 40 (96%). 38 controls matched for age and 41 matched for age and birth weight. Cases had more excess thermal insulation for their given room temperature (2.3 togs) than matched controls (0.6 togs) (p = 0.009). For every excess thermal insulation unit (tog) the relative risk of the sudden infant death syndrome was 1.26 (95% confidence interval 1.05 to 1.52). The average thermal bedding value calculated from parental recall was similar to that observed by attendant ambulance officers (mean difference = 0.4 tog, p = 0.39). Cases were more likely to have been found prone (odds ratio 4.58; 1.48 to 14.11). Prone sleeping position was not a confounder or effect modifier of the relation between excess thermal insulation and the syndrome. Overheating and the prone sleeping position are independently associated with an increased risk of the sudden infant death syndrome. Further work on infant thermal balance and sudden infant death is required and guidelines for appropriate infant thermal care need to be developed.


Quantification of Neoantigen-Mediated Immunoediting in Cancer Evolution.
Immunoediting includes three temporally distinct stages, termed elimination, equilibrium, and escape, and has been proposed to explain the interactions between cancer cells and the immune system during the evolution of cancer. However, the status of immunoediting in cancer remains unclear, and the existence of neoantigen depletion in untreated cancer has been debated. Here we developed a distribution pattern-based method for quantifying neoantigen-mediated negative selection in cancer evolution. The method can provide a robust and reliable quantification for immunoediting signal in individual patients with cancer. Moreover, this method demonstrated the prevalence of immunoediting in the immunotherapy-naive cancer genome. The elimination and escape stages of immunoediting can be quantified separately, where tumor types with strong immunoediting-elimination exhibit a weak immunoediting-escape signal, and vice versa. The quantified immunoediting-elimination signal was predictive of clinical response to cancer immunotherapy. Collectively, immunoediting quantification provides an evolutionary perspective for evaluating the antigenicity of neoantigens and reveals a potential biomarker for precision immunotherapy in cancer. Quantification of neoantigen-mediated negative selection in cancer progression reveals distinct features of cancer immunoediting and can serve as a potential biomarker to predict immunotherapy response.


Antinuclear antibody positivity in patients with chronic hepatitis C: clinically relevant or an epiphenomenon?
Serum autoantibodies such as antinuclear antibody (ANA) are frequently detected in patients with chronic hepatitis C virus (HCV) infection, but its relevance is a matter of discussion. To assess the association of ANA positivity with clinical and histological features, and with the outcome of antiviral therapy in patients with HCV infection. Baseline samples from patients with hepatitis C treated with interferon and ribavirin were tested for ANA positivity by indirect immunofluorescence. The mean age was 48.3+/-11.1 years and 56% were men. Among 234 included patients, 22 patients (9.4%) were positive for ANA. These patients showed significantly higher median alanine aminotransferase level (3.52 vs. 2.39 x upper limit of normal, P=0.009) when compared with ANA-negative patients. Fibrosis stage and necroinflammatory grading were not influenced by ANA positivity. Sustained virological response (SVR) rates were similar between ANA-positive and ANA-negative patients (27 vs. 29%, P=0.882). Alanine aminotransferase flares (> or = 1.5-fold the baseline) during treatment were observed in 28 patients (12%), irrespective of the presence of ANA and without any clinical significance. Among HCV patients, ANA positivity seems to represent an immunological epiphenomenon. It neither influences clinical, biochemical, and histological features of chronic hepatitis C nor predicts response to antiviral treatment.


Continuing versus withdrawing ixekizumab treatment in patients with axial spondyloarthritis who achieved remission: efficacy and safety results from a placebo-controlled, randomised withdrawal study (COAST-Y).
The objective of COAST-Y was to evaluate the effect of continuing versus withdrawing ixekizumab (IXE) in patients with axial spondyloarthritis (axSpA) who had achieved remission. COAST-Y is an ongoing, phase III, long-term extension study that included a double-blind, placebo (PBO)-controlled, randomised withdrawal-retreatment period (RWRP). Patients who completed the originating 52-week COAST-V, COAST-W or COAST-X studies entered a 24-week lead-in period and continued either 80 mg IXE every 2 (Q2W) or 4 weeks (Q4W). Patients who achieved remission (an Ankylosing Spondylitis Disease Activity Score (ASDAS)<1.3 at least once at week 16 or week 20, and <2.1 at both visits) were randomly assigned equally at week 24 to continue IXE Q4W, IXE Q2W or withdraw to PBO in a blinded fashion. The primary endpoint was the proportion of flare-free patients (flare: ASDAS≥2.1 at two consecutive visits or ASDAS>3.5 at any visit) after the 40-week RWRP, with time-to-flare as a major secondary endpoint. Of 773 enrolled patients, 741 completed the 24-week lead-in period and 155 entered the RWRP. Forty weeks after randomised withdrawal, 83.3% of patients in the combined IXE (85/102, p<0.001), IXE Q4W (40/48, p=0.003) and IXE Q2W (45/54, p=0.001) groups remained flare-free versus 54.7% in the PBO group (29/53). Continuing IXE significantly delayed time-to-flare versus PBO, with most patients remaining flare-free for up to 20 weeks after IXE withdrawal. Patients with axSpA who continued treatment with IXE were significantly less likely to flare and had significantly delayed time-to-flare compared with patients who withdrew to PBO.


Immunologic analysis of blood samples obtained from horses and stored for twenty-four hours.
To determine whether immune function can be accurately assessed in blood samples obtained from horses and refrigerated overnight and whether a nonradioactive lymphocyte proliferation assay can be used to evaluate samples obtained from horses. 224 blood samples from 28 clinically normal adult horses. Heparinized blood samples were collected. Each sample was divided into 2 equal aliquots. One aliquot was refrigerated overnight to simulate overnight shipping of blood samples, and the other aliquot was evaluated on the day of blood collection. Lymphocytes were isolated and enumerated by use of a modified single-gradient procedure. Cell viability and function were assessed by use of cytologic examination, flow cytometry, and mitogen-induced proliferation assays. Lymphocyte proliferation in response to T- and B-cell mitogens was measured by use of [3H]-thymidine incorporation and a nonradioactive lymphocyte proliferation assay. Lymphocytes refrigerated for up to 24 hours continued to be acceptable for use in immunologic analysis on the basis that they maintained viability and did not have significant alterations in lymphocyte subsets, except for CD8, when compared with freshly isolated lymphocytes. Furthermore, results for mitogen-induced lymphocyte proliferation assays were also comparable between fresh and refrigerated aliquots. CONDUSIONS AND CLINICAL RELEVANCE: The nonradioactive lymphocyte proliferation assay is a reliable alternative to [3H]-thymidine assay for assessing proliferation of equine lymphocytes. Collectively, our results imply that blood samples refrigerated and shipped ovenight to a laboratory can be used to perform cellular-immune assays; results of those assays would enhance a clinician's diagnostic abilities to monitor the efficacy of treatment.


The Influence of Motivational Interviewing on Patients With Inflammatory Bowel Disease: A Systematic Review of the Literature.
Inflammatory bowel disease (IBD) is a condition accompanied by several physical and often psychological symptoms (e.g., depression). Treatments generally involve dietary modifications and prescription medications. Of concern, non-adherence rates with prescription medications for this population have been reported to be between 30% and 45%. In order to examine an intervention that has shown promise in improving adherence, researchers systematically reviewed the literature in order to determine the impact of a motivational interviewing (MI) intervention on outcomes for individuals diagnosed with IBD. The outcomes assessed were broad and included, among others, the target behaviors of medication adherence and advice-seeking, and also patient-perceived provider empathy. Results suggest that MI can be effective in improving outcomes for individuals with IBD since patients experienced improved adherence rates, displayed greater advice-seeking behavior, and perceived providers as having more empathy and better communication skills. Further research is required since the pool of retained studies is small, evidencing a paucity of literature focusing on this evidence-based health behavior intervention for the behaviors needed to optimally manage IBD. Further, only adults were examined in these studies, so generalizations to children and adolescents are limited.


Inline amplification of mid-infrared intrapulse difference frequency generation.
We demonstrate an ultrafast mid-infrared source architecture that implements both intrapulse difference frequency generation (iDFG) and further optical parametric amplification (OPA), in an all-inline configuration. The source is driven by a nonlinearly compressed high-energy Yb-doped-fiber amplifier delivering 7.4 fs pulses at a central wavelength of 1030 nm, at a repetition rate of 250 kHz. It delivers 1 µJ, 73 fs pulses at a central wavelength of 8 µm, tunable over more than one octave. By enrolling all the pump photons in the iDFG process and recycling the long wavelength pump photons amplified in the iDFG in the subsequent OPA, we obtain an unprecedented overall optical efficiency of 2%. These performances, combining high energy and repetition rate in a very simple all-inline setup, make this technique ideally suited for a growing number of applications, such as high harmonic generation in solids or two-dimensional infrared spectroscopy experiments.


The Use of Ultrasound in Breast Cancer Screening of Asymptomatic Women with Dense Breast Tissue: A Narrative Review.
Mammography is the standard screening modality for breast cancer; however, sensitivity reduces with increasing breast density, resulting in the potential for masking of cancer. Ultrasound is a potential supplemental screening tool, but its routine use is controversial. A database search was performed with keywords "ultrasound" and "breast density and screening", including variations. Articles were included if they assessed the use of hand-held ultrasound as a supplemental screening modality in women with dense breasts. Twelve articles were identified. No high-level evidence articles were identified. Cancer detection rates increased with the addition of ultrasound-to-mammography screening protocols. However, this was associated with increased costs per cancer detected, an increased biopsy rate, and a low positive predictive value. The survival benefit, cost versus benefit, and psychological impact of the addition of ultrasound is unknown. The addition of ultrasound to a screening program in an asymptomatic population of women with dense breast tissue detects additional cancers compared with mammography alone. Knowledge regarding a survival or cost benefit associated with increased cancer detection, and the psychological impact of the addition of ultrasound is unknown. Further research is needed to assess whether the addition of ultrasound is cost-effective with respect to clinical outcome and survival.


Compact and Efficient 3.2-W Diode-Pumped Nd:YVO(4)/KTP Green Laser.
We demonstrate a simple way to achieve single-frequency operation by using fiber-coupled diode-pumped Nd:YVO(4)/KTP green lasers in a short standing-wave linear cavity. A single-mode output with 3.2-W green power was generated with a 12.6-W pump power corresponding to a conversion efficiency of 25.4%. The single-mode operation was obtained through the combined action of the anisotropic emission cross section of Nd:YVO(4) and the KTP crystal acting as a birefringent filter.


[The development of intestinal dysbacteriosis in sexually mature rats after thymectomy].
Thymectomy in puberal rats resulted in the development of pronounced enteric dysbacteriosis, correlating with the degree of T- and B-lymphocyte deficiency in the lymphoid organs and the blood stream during the first 2 months. Microbiocenosis restores probably due to more intensive functioning of alternative regulating mechanisms. A simple and informative characteristic indicating the degree of dysbacteriosis developing in the intestine is proposed: the index of dysbacteriosis.


[Allergy to Anisakis simplex].
Anisakis simplex is a parasite, belonging to the Anisakidae family. The life cycle of the parasite can include one or more intermediary hosts, their final hosts being marine mammals or large fish, in which the larvae develop until the adult stage is reached. Man is an accidental host who acquires the larvae by eating raw or undercooked fish. Since the mid-50s, when the first case studies were published in Holland and Japan, new cases have been emerging in different countries including Spain. Parasitization of man by the living larva is known as anisakiasis, principally giving rise to digestive symptomology, with other rare cases described of invasion of other organs such as the lung, the liver, the spleen, the pancreas, etc. Clinical pictures of allergy to IgE mediated anisakis simplex have also been described: reactions by thermostable antigens of the parasite that develop in spite of the fish being cooked or frozen, and an acute digestive parasitization with allergic symptoms called gastro-allergic anisakiasis. In the diagnosis of anisakiasis and/or allergy to Anisakis, the antecedent of the prior ingestion of fish as well as the clinical accompaniment can form basic data of considerable orientational value, and endoscopy can reveal the presence of the larvae and make possible their extraction. Besides, in cases of allergy the detection test for specific IgE facing Anisakis simplex, and cutaneous tests with fish should be carried out. The best treatment for avoiding this parasitization is prophylactic, avoiding the consumption of raw or undercooked fish, while a fish free diet is necessary in cases of true allergy to the thermostable proteins of the parasite.


Occurrence and Risk Factors of Uveitis in Juvenile Psoriatic Arthritis: Data From a Population-based Nationwide Study in Germany.
Data on uveitis in juvenile psoriatic arthritis (JPsA), a category of juvenile idiopathic arthritis (JIA), are scarce. We describe prevalence and risk factors for JPsA-associated uveitis (JPsA-U). Cross-sectional data from the German National Pediatric Rheumatological Database (2002-2014) were used to characterize JPsA-U and assess risk factors for the development of uveitis. Uveitis developed in 6.6% of 1862 patients with JPsA. Patients with JPsA-U were more frequently female (73.0 vs 62.9%, P = 0.03), antinuclear antibody (ANA) positive (60.3 vs 37.0%, P < 0.001), younger at JPsA onset (5.3 ± 4.1 vs 9.3 ± 4.4 yrs, P < 0.001), and treated with disease-modifying antirheumatic drugs (DMARDs) significantly more frequently compared with JPsA patients without uveitis. On a multivariable analysis of a subgroup of 655 patients enrolled in the study ≤ 1 year after arthritis onset, mean clinical Juvenile Arthritis Disease Activity Score for 10 joints during study documentation was significantly associated with uveitis development. Children with early onset of JPsA (aged < 5 yrs vs ≥ 5 yrs) were significantly more frequently ANA positive (48.4% vs 35.7%, P < 0.001), affected by uveitis (17.3% vs 3.8%, P < 0.001), and treated with DMARDs (52.9% vs 43.8%, P < 0.001), but less often affected by skin disease (55.3% vs 61.0%, P = 0.03). The characteristics of patients with JPsA developing uveitis are similar to those of patients with uveitis in other JIA categories, such as oligoarticular JIA. Children with early-onset JPsA are at a higher risk for ocular involvement. Our data support the notion of a major clinical difference between those patients with early vs late onset of JPsA.


Full-color illumination that needs no electric power.
Red, green, and blue phosphorescent resins were developed to realize full-color illuminators that shine without the need for electric power. Red phosphorescence, which used to be weak with conventional red phosphors, was enhanced notably with dye-doped resins in which green or blue phosphors were dispersed uniformly; i.e., brightness increased by seven times and afterglow duration extended by two times. Full-color phosphorescence from violet to red was attained by selecting suitable mixing ratios of the phosphors and dyes.


Merozoite proteins from Babesia sp. BQ1 (Lintan) as potential antigens for serodiagnosis by ELISA.
Babesia sp. BQ1 (Lintan) is a Babesia isolated from sheep infested with Haemaphysalis qinghaiensis in China, and is closely related to B. motasi based on the 18S rRNA gene sequence. In the present study, an ELISA was developed with merozoite antigens of Babesia sp. BQ1 (Lintan) (BQMA) purified from in vitro culture. When the positive threshold was chosen as 30% of the antibodies rate, evaluated with 198 negative sera, the specificity was 95.5%. Except for Babesia sp. Tianzhu, there was no cross-reaction between BQMA and positive sera from Babesia sp. BQ1 (Ningxian)-, Babesia sp. Hebei-, Babesia sp. Xinjiang-, Theileria luwenshuni-, T. uilenbergi-, or Anaplasma ovis-infected sheep, which are the dominant haemoparasites of small ruminants in China. Specific antibodies against Babesia sp. BQ1 (Lintan) were produced 1 or 2 weeks post-infection and a high level of antibodies persisted for more than 8 months in experimentally infected sheep. This ELISA was tested on 974 sera collected from field-grazing sheep in 3 counties of Gansu province, northwestern China to evaluate the seroprevalence of Babesia sp. BQ1 (Lintan) infection and the average positive rate was 66.84%. The feasibility of increasing the specificity of this BQMA-based ELISA, by using some BQMA antigens for serodiagnosis is discussed.


Phase ii comparator study of oral versus intravenous topotecan in patients with chemosensitive small-cell lung cancer.
Topotecan, administered intravenously, is active in small-cell lung cancer (SCLC). In this study, the comparability of oral topotecan to IV topotecan was investigated. Patients with SCLC that had relapsed 90 days or more after cessation of initial chemotherapy were randomized to receive either oral topotecan (Hycamtin) 2.3 mg/m(2)/d x 5 (52 patients) or IV topotecan 1.5 mg/m(2)/d x 5 (54 patients), every 21 days. Response rates in this phase II randomized study were 23% (12/52) in the oral topotecan arm and 15% (8/54) in the IV topotecan arm. All radiological responses were confirmed by an independent radiologist. Median survival was 32 weeks (oral) and 25 weeks (IV). Good symptom control, defined as sustained improvement or no deterioration, was evident in both treatment groups. Topotecan was generally well tolerated, with myelosuppression being the major toxicity. Grade 4 neutropenia occurred in 35.3% of patients on oral topotecan and in 67.3% of patients on IV topotecan, which was statistically significant (P =.001). Fever/infection more than or equal to grade 2 associated with grade 4 neutropenia, together with sepsis, occurred in only 5.1% of courses (oral) and 3.3% of courses (IV). Non-hematological toxicity consisted mainly of vomiting (oral: 36.5% of patients; IV: 31.5% of patients) and nausea (oral: 26.9% of patients; IV: 40.7% of patients). This study found oral topotecan to be similar in efficacy to IV topotecan in the treatment of patients with relapsed SCLC, sensitive to first-line chemotherapy, with less grade 4 neutropenia and greater convenience of administration.


Learning Abroad: Residents' Narratives of Clinical Experiences From a Global Health Elective.
While resident participation in global health (GH) rotations has grown, little is known about trainee perceptions of the personal value of these international clinical experiences and their importance to the objectives of GH training. We sought to better understand the clinical scenarios experienced during international rotations that residents perceived as most meaningful and the frequency of these experiences across scenarios and participating residents. Using the conceptual framework of Schön's reflection on action, we asked University of Minnesota GH track pediatric and internal medicine-pediatric residents to describe 10 clinical scenarios they found interesting or impactful during their 2016-2017 GH elective. We conducted a qualitative analysis of the deidentified resident narratives and mapped themes to the Accreditation Council for Graduate Medical Education (ACGME) competencies. All eligible residents (n = 13) participated, yielding 129 unique clinical scenarios from 7 countries. We identified 5 thematic groups: (1) addressing challenges in making diagnoses in resource-limited settings; (2) dealing with patient outcomes different from those expected in the United States; (3) encountering and managing diseases in a different clinical context; (4) encountering and managing diseases in a different cultural context; and (5) reflecting on learning and self-growth. Of the 129 unique clinical scenarios, 30% (n = 39) had not been previously experienced by participants. Across the 5 themes, all ACGME core competencies were addressed. Residents identified meaningful scenarios of their GH experiences that are relevant to the educational and clinical objectives of GH training.


Maternal dietary resveratrol alleviates weaning-associated diarrhea and intestinal inflammation in pig offspring by changing intestinal gene expression and microbiota.
Early weaning commonly results in gastrointestinal disorders, inflammation and diarrhea in infants and young animals. Resveratrol, a plant phenol, affords protection against inflammation and cancer. A porcine model was used to investigate the effects of maternal dietary resveratrol on diarrhea, intestinal inflammation and the intestinal morphology in offspring during weaning. The results showed that maternal dietary resveratrol alleviated weaning-associated intestinal inflammation and diarrhea and improved the intestinal morphology in offspring. In weaning piglets, maternal dietary resveratrol increased the proportion of butyrate-producing bacteria, such as Flavonifractor, Odoribacter and Oscillibacter, as determined by 16S rRNA sequencing. RNA-seq analysis identified 189 and 139 differentially expressed genes (DEGs) in weaning and post-weaning piglets, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that DEGs were enriched for the T cell receptor, primary immunodeficiency, mitogen-activated protein kinase (MAPK) and Ras signaling pathways in weaning piglets and for the cytokine-cytokine receptor interaction pathway and metabolism-related pathways in post-weaning piglets.


Low serum adiponectin levels in Korean children with a family history of type 2 diabetes mellitus.
The current worldwide increases of type 2 diabetes mellitus (T2DM) in children coincide with increases in the prevalence of obesity. We investigated the insulin resistance and adiponectin levels of children and adolescents with a family history of T2DM (FHD). Our sample included 131 children and adolescents aged 8-15 years. Fasting plasma glucose, lipids, fasting insulin, adiponectin levels and HOMA-IR were analyzed according to FHD and obesity. Oral glucose tolerance tests were performed in all subjects except non-obese subjects without FHD. Adiponectin levels of subjects with FHD were significantly lower than those of subjects without FHD in both the obese and nonobese groups. HOMA-IR was significantly higher in obese subjects with FHD than in those without FHD. Adiponectin levels were found to be independently associated with FHD and Matsuda index. The frequency of impaired glucose tolerance in obese subjects with FHD was more than four times higher compared to obese subjects without FHD. Our results suggest that FHD could be a risk factor of T2DM in obese Korean children, especially with low serum levels of adiponectin.


Immunomodulatory Effect of SINA 1.2 Therapy Protocol in Asthmatic Mice Model: A Combination of Oxymel and Sauna.
Alternative medicine, has become popular in asthmatic patients. We evaluated the immunomodulatory effects of SINA 1.2 therapy protocol derived from Persian medicine in an asthmatic mice model. Forty-two male BALB/c mice divided into six groups: one control (sham) and five sensitized groups (by parenteral injection of 20 μg ovalbumin in 100 μL normal saline plus 50 μL alum on days 1 and 14). Sensitized groups were as: untreated, budesonide (1 mg nebulized budesonide: 200 μg/puff every 5 min for 25 min), dry sauna (30 min, 37°C), oral oxymel (gavaged: 0.2 mL of the syrup plus 0.8 mL of water), and SINA protocol No.1.2 (oxymel followed by sauna) groups. Treatments were given for 10 days from day 23 to 33 then sacrificed. Significant gene expression reduction of interleukin(IL)-4, IL-5, and MUC5AC and increase of interferon(IFN)-γ and IFN-γ/IL-4 ratio and decreased perivascular and peribronchial inflammation, goblet cell hyperplasia, and subsequent mucus hypersecretion in SINA group were seen compared to untreated group. SINA lowered IL-5 and MUC5AC gene expression levels similar to the budesonide and acted better than budesonide in increasing IFN-γ gene expression up to normal level. Compared with the asthma group, sauna alone only affected MUC5AC and IFN-γ gene expressions and oxymel alone, only reduced IL-4 gene expression, perivascular and peribronchial inflammation, and mucus hypersecretion. It seems that SINA therapy alleviates asthma via immune modulation of pro-inflammatory cytokines and improvement of pathological changes in ovalbumin-induced asthma in mice, supporting the notion of innate healing power mentioned in Persian medicine literature.


Panfungal Polymerase Chain Reaction for Identification of Fungal Pathogens in Formalin-Fixed Animal Tissues.
Identification of fungal organisms often poses a problem for pathologists because the histomorphology of some fungal organisms is not specific, fresh tissues may not be available, and isolation and identification in culture may take a long time. The purpose of this study was to validate the use of panfungal polymerase chain reaction (PCR) to identify fungal organisms from formalin-fixed paraffin-embedded (FFPE) tissues. Formalin-fixed paraffin-embedded curls were tested from 128 blocks containing canine, feline, equine, and bovine tissues with cutaneous, nasal, pulmonary, and systemic fungal infections, identified by the presence of fungi in histologic sections. Quantitative scoring of histologic sections identified rare (11.9%), occasional (17.5%), moderate (17.5%), or abundant (53.1%) fungal organisms. DNA was isolated from FFPE tissues and PCR was performed targeting the internal transcribed spacer 2 (ITS-2) region, a segment of noncoding DNA found in all eukaryotes. Polymerase chain reaction products were sequenced and identified at ≥97% identity match using the Basic Local Alignment Search Tool and the NCBI database of ITS sequences. Of the 128 blocks, 117 (91.4%) yielded PCR products and high-quality sequences were derived from 89 (69.5%). Sequence and histologic identifications matched in 79 blocks (61.7%). This assay was capable of providing genus- and species-level identification when histopathology could not and, thus, is a beneficial complementary tool for diagnosis of fungal diseases.


Ammonia modeling for assessing potential toxicity to fish species in the Rio Grande, 1989-2002.
Increasing volumes of treated and untreated human sewage discharged into rivers around the world are likely to be leading to high aquatic concentrations of toxic, unionized ammonia (NH3), with negative impacts on species and ecosystems. Tools and approaches are needed for assessing the dynamics of NH3. This paper describes a modeling approach for first-order assessment of potential NH3 toxicity in urban rivers. In this study daily dissolved NH3 concentrations in the Rio Grande of central New Mexico, USA, at the city of Albuquerque's treated sewage outfall were modeled for 1989-2002. Data for ammonium (NH4+) concentrations in the sewage and data for discharge, temperature, and pH for both sewage effluent and the river were used. We used State of New Mexico acute and chronic NH3- N concentration values (0.30 and 0.05 mg/L NH3-N, respectively) and other reported standards as benchmarks for determining NH3 toxicity in the river and for assessing potential impact on population dynamics for fish species. A critical species of concern is the Rio Grande silvery minnow (Hybognathus amarus), an endangered species in the river near Albuquerque. Results show that NH3 concentrations matched or exceeded acute levels 13%, 3%, and 4% of the time in 1989, 1991, and 1992, respectively. Modeled NH3 concentrations matched or exceeded chronic values 97%, 74%, 78%, and 11% of the time in 1989, 1991, 1992, and 1997, respectively. Exceedences ranged from 0% to 1% in later years after enhancements to the wastewater treatment plant. Modeled NH3 concentrations may differ from actual concentrations because of NH3 and NH4+ loss terms and additive terms such as mixing processes, volatilization, nitrification, sorbtion, and NH4+ uptake. We conclude that NH3 toxicity must be considered seriously for its potential ecological impacts on the Rio Grande and as a mechanism contributing to the decline of the Rio Grande fish community in general and the Rio Grande silvery minnow specifically. Conclusions drawn for the Rio Grande suggest that NH3 concentrations may be high in rivers around the world where alkaline pH values are prevalent and sewage treatment capabilities are poorly developed or absent.


Anammox process for nitrogen removal from anaerobically digested fish canning effluents.
The Anammox process was used to treat the effluent generated in an anaerobic digester which treated the wastewater from a fish cannery once previously processed in a Sharon reactor. The effluents generated from the anaerobic digestion are characterised by their high ammonium content (700-1000 g NH4+ -Nm(-3)), organic carbon content (1000-1300 g TOCm(-3)) and salinity up to 8,000-10,000 g NaCl m(-3). In the Sharon reactor, approximately 50% of the NH4+ -N was oxidised to NO2- -N via partial nitrification. The effluent of the Sharon step was fed to the Anammox reactor which treated an averaged nitrogen loading rate of 500 g N m(-3) x d(-1). The system reached an averaged nitrogen removal efficiency of 68%, mainly limited due to the nonstoichiometric relation, for the Anammox process, between the ammonium and nitrite added in the feeding. The Anammox reactor bacterial population distribution, followed by FISH analysis and batch activity assays, did not change significantly despite the continuous entrance to the system of aerobic ammonium oxidisers coming from the Sharon reactor. Most of the bacteria corresponded to the Anammox population and the rest with slight variable shares to the ammonia oxidisers. The Anammox reactor showed an unexpected robustness despite the continuous variations in the influent composition regarding ammonium and nitrite concentrations. Only in the period when NO2- -N concentration was higher than the NH4+ -N concentration did the process destabilise and it took 14 days until the nitrogen removal percentage decreased to 34% with concentrations in the effluent of 340g NH4+ -N m(-3) and 440 g NO2- -N m(-3), respectively. Based on these results, it seems that the Sharon-Anammox system can be applied for the treatment of industrial wastewaters with high nitrogen load and salt concentration with an appropriate control of the NO2- -N/NH4+ -N ratio.


Sulfated Polysaccharides from Macroalgae Are Potent Dual Inhibitors of Human ATP-Hydrolyzing Ectonucleotidases NPP1 and CD39.
Extracellular ATP mediates proinflammatory and antiproliferative effects via activation of P2 nucleotide receptors. In contrast, its metabolite, the nucleoside adenosine, is strongly immunosuppressive and enhances tumor proliferation and metastasis. The conversion of ATP to adenosine is catalyzed by ectonucleotidases, which are expressed on immune cells and typically upregulated on tumor cells. In the present study, we identified sulfopolysaccharides from brown and red sea algae to act as potent dual inhibitors of the main ATP-hydrolyzing ectoenzymes, ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) and ecto-nucleoside triphosphate diphosphohydrolase-1 (NTPDase1, CD39), showing nano- to picomolar potency and displaying a non-competitive mechanism of inhibition. We showed that one of the sulfopolysaccharides tested as a representative example reduced adenosine formation at the surface of the human glioblastoma cell line U87 in a concentration-dependent manner. These natural products represent the most potent inhibitors of extracellular ATP hydrolysis known to date and have potential as novel therapeutics for the immunotherapy of cancer.


[Role of reflexotherapy in the complex treatment of neuroendocrine obesity].
The paper is concerned with comparative estimation of the effect of different therapeutic methods (hypocaloric diet, physiotherapeutic, drug and exercise therapy, reflexotherapy) on loss of body mass. The experience of reflexotherapy of 86 patients showed its positive effect on the psychoemotional activities of patients with obesity, a rise of adaptation capabilities of the body under physical exercise, improved external respiration function, an increase in oxygen saturation of tissues, the stimulation of metabolism (by the basal metabolism findings) by way of increasing the secretion of hypophyseal tropic hormones, triiodothyronine and thyroxin, and potentiation of the time course of loss of body mass.


Vegetable and Fruit Intake and Fracture-Related Hospitalisations: A Prospective Study of Older Women.
The importance of vegetable and fruit intakes for the prevention of fracture in older women is not well understood. Few studies have explored vegetable and fruit intakes separately, or the associations of specific types of vegetables and fruits with fracture hospitalisations. The objective of this study was to examine the associations of vegetable and fruit intakes, separately, and specific types of vegetables and fruits with fracture-related hospitalisations in a prospective cohort of women aged ≥70 years. Vegetable and fruit intakes were assessed at baseline (1998) in 1468 women using a food frequency questionnaire. The incidence of fracture-related hospitalisations over 14.5 years of follow-up was determined using the Hospital Morbidity Data Collection, linked via the Western Australian Data Linkage System. Fractures were identified in 415 (28.3%) women, of which 158 (10.8%) were hip fractures. Higher intakes of vegetables, but not fruits, were associated with lower fracture incidence. In multivariable-adjusted models for vegetable types, cruciferous and allium vegetables were inversely associated with all fractures, with a hazard ratio (HR) (95% confidence interval) of 0.72 (0.54, 0.95) and 0.66 (0.49, 0.88), respectively, for the highest vs. lowest quartiles. Increasing vegetable intake, with an emphasis on cruciferous and allium vegetables, may prevent fractures in older postmenopausal women.


Postoperative nonsteroidal antiinflammatory drugs and the prevention of heterotopic ossification after cervical arthroplasty: analysis using CT and a minimum 2-year follow-up.
OBJECT Heterotopic ossification (HO) after cervical arthroplasty is not uncommon and may cause immobility of the disc. To prevent HO formation, study protocols of clinical trials for cervical arthroplasty undertaken by the US FDA included perioperative use of nonsteroidal antiinflammatory drugs (NSAIDs). However, there are few data supporting the use of NSAIDs to prevent HO after cervical arthroplasty. Therefore, this study aimed to evaluate the efficacy of NSAIDs in HO formation and clinical outcomes. METHODS Consecutive patients who underwent 1- or 2-level cervical arthroplasty with a minimum follow-up of 24 months were retrospectively reviewed. All patients were grouped into 1 of 2 groups, an NSAID group (those patients who had used NSAIDs postoperatively) and a non-NSAID group (those patients who had not used NSAIDs postoperatively). The formation of HO was detected and classified using CT in every patient. The incidence of HO formation, disc mobility, and clinical outcomes, including visual analog scale (VAS) scores of neck and arm pain, neck disability index (NDI) scores, and complications were compared between the two groups. Furthermore, a subgroup analysis of the patients in the NSAID group, comparing the selective cyclooxygenase (COX)-2 to nonselective COX-2 NSAID users, was also conducted for each of the above-mentioned parameters. RESULTS A total of 75 patients (mean age [± SD] 46.71 ± 9.94 years) with 107 operated levels were analyzed. The mean follow-up duration was 38.71 ± 9.55 months. There were no significant differences in age, sex, and levels of arthroplasty between the NSAID and non-NSAID groups. There was a nonsignificantly lower rate of HO formation in the NSAID group than the non-NSAID group (47.2% vs. 68.2%, respectively; p = 0.129). During follow-up, most of the arthroplasty levels remained mobile, with similar rates of immobile discs in the NSAID and non-NSAID groups (13.2% and 22.7%, respectively; p = 0.318). Furthermore, there was a nonsignificantly lower rate of HO formation in the selective COX-2 group than the nonselective COX-2 group (30.8% vs 52.5%, respectively; p = 0.213). The clinical outcomes, including VAS neck, VAS arm, and NDI scores at 24 months postoperatively, were all similar in the NSAID and non-NSAID groups, as well as the selective and nonselective COX-2 groups (all p > 0.05). CONCLUSIONS In this study there was a trend toward less HO formation and fewer immobile discs in patients who used postoperative NSAIDs after cervical arthroplasty than those who did not, but this trend did not reach statistical significance. Patients who used selective COX-2 NSAIDs had nonsignificantly less HO than those who used nonselective COX-2 NSAIDs. The clinical outcomes were not affected by the use of NSAIDs or the kinds of NSAIDs used (selective vs nonselective COX-2). However, the study was limited by the number of patients included, and the efficacy of NSAIDs in the prevention of HO after cervical arthroplasty may need further investigation to confirm these results.


'Obligate' anaerobic Salmonella strain YB1 suppresses liver tumor growth and metastasis in nude mice.
The antitumor properties of bacteria have been demonstrated over the past decades. However, the efficacy is limited and unclear. Furthermore, systemic infection remains a serious concern in bacteria treatment. In this study, the effect of YB1, a rationally designed 'obligate' anaerobic Salmonella typhimurium strain, on liver tumor growth and metastasis in a nude mouse orthotopic liver tumor model was investigated. The orthotopic liver tumor model was established in nude mice using the hepatocellular carcinoma cell line MHCC-97L. Two weeks after orthotopic liver tumor implantation, YB1, SL7207 and saline were respectively administered through the tail vein of the mice. Longitudinal monitoring of tumor growth and metastasis was performed using Xenogen IVIS, and direct measurements of tumor volume were taken 3 weeks after treatment. In vitro, MHCC-97L and PLC cells were incubated with YB1 or SL7207 under anaerobic conditions. YB1 was observed to invade tumor cells and induce tumor cell apoptosis and death. The results revealed that all mice in the YB1 group were alive 3 weeks after YB1 injection while all mice in the SL7207 group died within 11 days of the SL7207 injection. The body weight decreased by ~9% on day 1 after YB1 injection and but subsequently recovered. Liver tumor growth and metastases were significantly inhibited following YB1 treatment. By contrast to the control group, a large number of Gr1-positive cells were detected on days 1 to 21 following YB1 treatment. Furthermore, YB1 also effectively invaded tumor cells and induced tumor cell apoptosis and death. In conclusion, YB1 suppressed liver tumor growth and metastasis in a nude mice liver tumor model. The potential mechanism may be through enhancing innate immune response and inducing tumor cell apoptosis and cell death.


Ethanol alters the balance of Sox2, Oct4, and Nanog expression in distinct subpopulations during differentiation of embryonic stem cells.
The transcription factors Sox2, Oct4, and Nanog regulate within a narrow dose-range embryonic stem (ES) cell pluripotency and cell lineage commitment. Excess of Oct4 relative to Sox2 guides cells to mesoendoderm (ME), while abundance of Sox2 promotes neuroectoderm (NE) formation. Literature does not address whether ethanol interferes with these regulatory interactions during neural development. We hypothesized that ethanol exposure of ES cells in early differentiation causes an imbalance of Oct4 and Sox2 that diverts cells away from NE to ME lineage, consistent with the teratogenesis effects caused by prenatal alcohol exposure. Mouse ES cells were exposed to ethanol (0, 25, 50, and 100 mM) during retinoic acid (10 nM)-directed differentiation to NE for 0-6 days, and the expression of Sox2, Oct4, and Nanog was measured in single live cells by multiparametric flow cytometry, and the cellular phenotype was characterized by immunocytochemistry. Our data showed an ethanol dose- and time-dependent asymmetric modulation of Oct4 and Sox2 expression, as early as after 2 days of exposure. Single-cell analysis of the correlated expression of Sox2, Oct4, and Nanog revealed that ethanol promoted distinct subpopulations with a high Oct4/Sox2 ratio. Ethanol-exposed cells differentiated to fewer β-III tubulin-immunoreactive cells with an immature neuronal phenotype by 4 days. We interpret these data as suggesting that ethanol diverted cells in early differentiation from the NE fate toward the ME lineage. Our results provide a novel insight into the mode of ethanol action and opportunities for discovery of prenatal biomarkers at early stages.


Plasma fibroblast skin tightening treatment resulting in bilateral chemical eye injury secondary to EMLA cream: a case report.
Plasma fibroblast skin tightening treatment is a relatively novel and growing minimally invasive aesthetic skin procedure. The treatment claims to rejuvenate skin by improving facial lines, wrinkles and skin pigmentation associated with photo-ageing. The skin is often anaesthetised prior to the procedure with topical creams such as EMLA (Eutectic mixture of local anaesthetics). We present the first case of bilateral chemical eye injury following plasma fibroblast skin tightening treatment secondary to EMLA cream. EMLA cream was inadvertently administered to both eyes in preparation for the treatment. A patient was referred from the emergency department to a tertiary ophthalmology centre with bilateral exquisite eye pain, inability to open the eyes, photosensitivity and reduced vision. She underwent cosmetic plasma fibroblast skin tightening treatment at her local salon four hours earlier. She was found to have bilateral alkali chemical eye injuries with significant diffuse corneal epithelial loss. The injury was thought to be caused by inadvertent ocular exposure to EMLA cream, which was used in preparation for the plasma fibroblast skin tightening treatment. She was treated with topical antibiotics and achieved satisfactory recovery. This case report highlights a possible complication following plasma fibroblast skin tightening treatment. We lay emphasis on the importance identifying chemical injury and recommend that medication attention should be sought if there is any concern.


Support-based distributed optimisation: an approach to radiotherapy patient scheduling.
In the health system inefficiency leads to poor use of scarce expensive resources. Lengthy patient treatment waiting time can result from inefficiency in scheduling. The use of state-of-the art multi-agent and distributed computing technologies can provide a solution to address this problem. However, distributed optimisation in such a multi-agent setting poses an important challenge that requires protocols to enable agents to optimise shared objectives without necessarily revealing all of their private constraints. In this study we show that if the problem is expressed as a Dynamic Distributed Constraint Optimisation Problem a powerful algorithm such as SBDO can be deployed to solve it. As SBDO can be deployed in a grid all of the advantages of grid computing are also gained.


Expression and function of peroxisome proliferator-activated receptor-gamma in mesangial cells.
P:eroxisome proliferator-activated receptor-gamma (PPARgamma) is a novel nuclear receptor, which enhances insulin-mediated glucose uptake. Ligands to PPARgamma are currently used as therapy for type II diabetes. Using Western blot analysis, RNase protection assay, and immunostaining, we identified the presence of PPARgamma message and protein in cultured primary rat mesangial cells. Electrophoretic mobility of a labeled PPARgamma response element (PPRE) was retarded in the presence of mesangial cell nuclear extract, suggesting that PPARgamma is functional in these cells. The addition of unlabeled PPRE efficiently competed away the PPARgamma-PPRE protein complex, confirming specificity of binding of the PPARgamma to the PPRE. PPARgamma ligands rosiglitazone (1 to 10 micromol/L) and troglitazone (1 to 10 micromol/L) inhibited platelet-derived growth factor-induced DNA synthesis, measured as bromodeoxyuridine incorporation (P<0.01). This inhibition was dose dependent. When administered in antidiabetic doses to streptozotocin-induced diabetic rats, troglitazone substantially normalized albumin excretion at 3 months (from 687.1 to 137.6 microgram urinary albumin/mg creatinine, P:<0.05) but did not affect hyperglycemia or blood pressure in this model. This treatment also decreased glomerular plasminogen activator inhibitor-1 (PAI-1) expression. These data suggest that PPARgamma activation may directly attenuate diabetic glomerular disease, possibly by inhibiting mesangial growth, which occurs early in the process of diabetic nephropathy, or by inhibiting PAI-1 expression. PAI-1 inhibits the activation of plasmin and matrix metalloproteinase, which degrade extracellular matrix in the glomerulus. Excess glomerular PAI-1 allows the accumulation of extracellular matrix, leading to glomerulosclerosis. These results have therapeutic implications for diabetic nephropathy as well as for proliferative mesangial diseases of the kidney.


A stair-climb test of cardiorespiratory fitness for Singapore.
The purpose of this study was to develop a stair-climb test of cardiorespiratory fitness for adult Singaporeans, particularly those staying in Singapore Housing and Development Board (HDB) flats. 103 subjects [56 males, of mean (SD) age 44.8 (13.9) years and Body Mass Index, or BMI, 23.3 (3.2); and 47 females, of age 43.2 (12.9) years and BMI 21.9 (2.8)] were first assessed for their cardiorespiratory fitness, measured using maximal oxygen uptake (or VO2max) on a treadmill, before undergoing a stair-climb test up to the 12th storey (11 floors) of a typical HDB flat (180 steps, vertical height 27.0 metres). The mean (SD) time taken for the climb (CT) and heart rate at the end of the climb (HRend) averaged 111.3 (16) seconds and 154.4 (13.4) beats.min-1 respectively for males, and 121.0 (18.1) seconds and 164.6 (15.7) beats.min-1 respectively for females. Regression equations were developed to predict VO2max from age (years), BMI, CT (seconds), HRend (beats.min-1): For males: VO2max (ml.kg-1.min-1) = 133 - 0.273 (Age) - 0.672 (BMI) - 0.236 (CT) - 0.232 (HRend). For females: VO2max (ml.kg-1.min-1) = 66.69 - 0.135 (Age) - 0.249 (BMI) - 0.128 (CT) - 0.021 (HRend). Validation of the regression equations conducted on a different sample consisting of 18 subjects (11 male and 7 female) showed significant correlations between the predicted and directly measured VO2max (males, r = 0.81 and females, r = 0.90; p < 0.01). There were no significant differences between the means of predicted and directly measured VO2max. A stair-climb test using HDB stairs was developed which was able to estimate cardiorespiratory fitness with reasonable accuracy.


The gene Sr33, an ortholog of barley Mla genes, encodes resistance to wheat stem rust race Ug99.
Wheat stem rust, caused by the fungus Puccinia graminis f. sp. tritici, afflicts bread wheat (Triticum aestivum). New virulent races collectively referred to as "Ug99" have emerged, which threaten global wheat production. The wheat gene Sr33, introgressed from the wild relative Aegilops tauschii into bread wheat, confers resistance to diverse stem rust races, including the Ug99 race group. We cloned Sr33, which encodes a coiled-coil, nucleotide-binding, leucine-rich repeat protein. Sr33 is orthologous to the barley (Hordeum vulgare) Mla mildew resistance genes that confer resistance to Blumeria graminis f. sp. hordei. The wheat Sr33 gene functions independently of RAR1, SGT1, and HSP90 chaperones. Haplotype analysis from diverse collections of Ae. tauschii placed the origin of Sr33 resistance near the southern coast of the Caspian Sea.


Transjugular intrahepatic portosystemic shunt placement before abdominal intervention in cirrhotic patients with portal hypertension: lessons from a pilot study.
Abdominal interventions are usually contraindicated in patients with cirrhosis and portal hypertension because of increased morbidity and mortality. Decreasing portal pressure with transjugular intrahepatic portosystemic shunt (TIPS) may improve patient outcomes. We report our experience with patients treated by neoadjuvant TIPS to identify those who would most benefit from this two-step procedure. All patients treated by dedicated neoadjuvant TIPS between 2005 and March 2013 in two tertiary referral hospitals were included. The primary endpoint was the rate of failure, defined by the inability to proceed to the planned intervention after TIPS placement or persistent liver decompensation 3 months after intervention. The secondary endpoints were the rate of complications, parameters associated with failure, and 1-year survival. Twenty-eight consecutive patients were included, with a mean age of 61.2±6.6 years, mean Child-Pugh score of 6.6±1.5, and mean model for end-stage liver disease score of 10.4±3.3. Procedures were digestive (43%) or liver (25%) resections, abdominal wall surgery (21%), or interventional gastrointestinal endoscopies (11%). The scheduled procedure was performed in 24 (86%) patients within a median of 25 days after TIPS. Procedure failures occurred in six (21%) patients: four did not undergo surgery and two experienced persistent liver decompensation. Seven (25%) patients had postoperative complications, mainly local. Viral origin of cirrhosis, history of encephalopathy, and hepatic surgery were found to be associated with failure. One-year survival in the whole cohort was 70%. In selected patients, extrahepatic surgery or interventional endoscopies can be safely performed after portal hypertension has been controlled by TIPS.


Benzodiazepines for treatment of patients with delirium excluding those who are cared for in an intensive care unit.
Delirium is a very common condition associated with significant morbidity, mortality, and costs. Current critical care guidelines recommend first and foremost the use of nonpharmacological strategies in both the prevention and treatment of delirium. Pharmacological interventions may augment these approaches and they are currently used widely in clinical practice to manage the symptoms of delirium. Benzodiazepines are currently used in clinical practice to treat behavioural disturbances associated with delirium but current guidelines do not recommend their use for this indication. The use of these medicines is controversial because there is uncertainty about whether they are effective for patients or have the potential to harm them. To assess the effectiveness and safety of benzodiazepines in the treatment of delirium (excluding delirium related to withdrawal from alcohol or benzodiazepines) in any healthcare settings other than intensive care units (ICU). We searched ALOIS: the Cochrane Dementia and Cognitive Improvement Group's Specialized Register up to 10 April 2019. ALOIS contains records of clinical trials identified from monthly searches of a number of major healthcare databases (including MEDLINE, Embase, PsycINFO, CINAHL, LILACS), numerous trial registries (including national, international and pharmaceutical registries), and grey literature sources. We included randomised controlled trials (RCTs) conducted in healthcare settings that ranged from nursing homes and long-term care facilities to any hospital setting except for ICUs, involving adult patients with delirium excluding those with delirium related to alcohol or benzodiazepine withdrawal. Included RCTs had to assess the effect of benzodiazepines, at any dose and given by any route, compared with placebo or another drug intended to treat delirium. Two review authors independently assessed study eligibility, extracted data, and assessed the risk of bias of included studies. We decided whether or not to pool data on the basis of clinical heterogeneity between studies. We used GRADE (Grades of Recommendation, Assessment, Development and Evaluation) methods to assess the quality of evidence. We identified only two trials that satisfied the selection criteria. We did not pool the data because of the substantial clinical differences between the trials. In one trial, participants (n = 58) were patients in an acute palliative care unit with advanced cancer who had a mean age of 64 years. All of the participants had delirium, were treated with haloperidol, and were randomised to receive either lorazepam or placebo in combination with it. Due to very serious imprecision, all evidence was of low certainty. We were unable to determine whether there were clinically important differences in the severity of delirium (mean difference (MD) 2.10, 95% CI -0.96 to 5.16; n = 50), length of hospital admission (MD 0.00, 95% CI -3.45 to 3.45; n = 58), mortality from all causes (risk ratio (RR) 0.33, 95% CI 0.04 to 3.02; participants = 58) or any of a number of adverse events. Important effects could not be confirmed or excluded. The study authors did not report the length of the delirium episode. In the other trial, participants (n = 30) were patients in general medical wards with acquired immune deficiency syndrome (AIDS) who had a mean age of 39.2 years. Investigators compared three drug treatments: all participants had delirium, and were randomised to receive lorazepam, chlorpromazine, or haloperidol. Very low-certainty evidence was identified, and we could not determine whether lorazepam differed from either of the other treatments in the effect on severity of delirium, any adverse event, or mortality from all causes. The study authors did not report the length of the delirium episode or the length of hospital admission. There is no enough evidence to determine whether benzodiazepines are effective when used to treat patients with delirium who are cared for in non-ICU settings. The available evidence does not support their routine use for this indication. Because of the scarcity of data from randomised controlled trials, further research is required to determine whether or not there is a role for benzodiazepines in the treatment of delirium in non-ICU settings.


A recurrent mutation, ala391glu, in the transmembrane region of FGFR3 causes Crouzon syndrome and acanthosis nigricans.
Mutations in the fibroblast growth factor receptor 2 (FGFR2) gene have previously been identified in Crouzon syndrome, an autosomal dominant condition involving premature fusion of the cranial sutures. Several different missense and other mutations have been identified in Crouzon syndrome patients, clustering around the third immunoglobulin-like domain. We report here the identification of a mutation in the transmembrane region of FGFR3, common to three unrelated patients with classical Crouzon syndrome and acanthosis nigricans, a dermatological condition associated with thickening and abnormal pigmentation of the skin. The mutation within the FGFR3 transcript was determined by direct sequencing as a specific gcg to gag transversion, resulting in an amino acid substitution ala391glu within the transmembrane region.


Localization of membrane proteins by the use of a photoreactive fatty acid incorporated in vivo into vesicular stomatitis virus.
Vesicular stomatitis virus grown in the presence of omega-[9-3H]diazirinophenoxy nonanoate resulted in the biosynthetic incorporation of this photoreactive fatty acid into viral phospholipids as well as into the membrane anchoring domain of the viral G glycoprotein. Photolysis of the isolated virus at 360 nm resulted in extensive labeling of the G protein but none of the other proteins by the viral phospholipids. In addition, a new product was obtained and was identified as a G-G dimer by its molecular weight and its reaction with anti-G antibody. These results demonstrate the use of photoreactive fatty acid to identify integral membrane proteins and to make photoaffinity probes of fatty acylated membrane proteins.


Rapidly exploring structural and dynamic properties of signaling networks using PathwayOracle.
In systems biology the experimentalist is presented with a selection of software for analyzing dynamic properties of signaling networks. These tools either assume that the network is in steady-state or require highly parameterized models of the network of interest. For biologists interested in assessing how signal propagates through a network under specific conditions, the first class of methods does not provide sufficiently detailed results and the second class requires models which may not be easily and accurately constructed. A tool that is able to characterize the dynamics of a signaling network using an unparameterized model of the network would allow biologists to quickly obtain insights into a signaling network's behavior. We introduce PathwayOracle, an integrated suite of software tools for computationally inferring and analyzing structural and dynamic properties of a signaling network. The feature which differentiates PathwayOracle from other tools is a method that can predict the response of a signaling network to various experimental conditions and stimuli using only the connectivity of the signaling network. Thus signaling models are relatively easy to build. The method allows for tracking signal flow in a network and comparison of signal flows under different experimental conditions. In addition, PathwayOracle includes tools for the enumeration and visualization of coherent and incoherent signaling paths between proteins, and for experimental analysis - loading and superimposing experimental data, such as microarray intensities, on the network model. PathwayOracle provides an integrated environment in which both structural and dynamic analysis of a signaling network can be quickly conducted and visualized along side experimental results. By using the signaling network connectivity, analyses and predictions can be performed quickly using relatively easily constructed signaling network models. The application has been developed in Python and is designed to be easily extensible by groups interested in adding new or extending existing features. PathwayOracle is freely available for download and use.


Insufficient compensation for self-motion during perception of object speed: The vestibular Aubert-Fleischl phenomenon.
To estimate object speed with respect to the self, retinal signals must be summed with extraretinal signals that encode the speed of eye and head movement. Prior work has shown that differences in perceptual estimates of object speed based on retinal and oculomotor signals lead to biased percepts such as the Aubert-Fleischl phenomenon (AF), in which moving targets appear slower when pursued. During whole-body movement, additional extraretinal signals, such as those from the vestibular system, may be used to transform object speed estimates from a head-centered to a world-centered reference frame. Here we demonstrate that whole-body pursuit in the form of passive yaw rotation, which stimulates the semicircular canals of the vestibular system, leads to a slowing of perceived object speed similar to the classic oculomotor AF. We find that the magnitude of the vestibular and oculomotor AF is comparable across a range of speeds, despite the different types of input signal involved. This covariation might hint at a common modality-independent mechanism underlying the AF in both cases.


The potential impact of country-level migration networks on HIV epidemics in sub-Saharan Africa: the case of Botswana.
Generalised HIV epidemics in sub-Saharan Africa show substantial geographical variation in prevalence, which is considered when designing epidemic control strategies. We hypothesise that the migratory behaviour of the general population of countries in sub-Saharan Africa could have a substantial effect on HIV epidemics and challenge the elimination effort. To test this hypothesis, we used census data from 2017 to identify, construct, and visualise the migration network of the population of Botswana, which has one of the most severe HIV epidemics worldwide. We found that, over 12 months, approximately 14% of the population moved their residency from one district to another. Four types of migration occurred: urban-to-urban, rural-to-urban, urban-to-rural, and rural-to-rural. Migration is leading to a marked geographical redistribution of the population, causing high rates of population turnover in some areas, and further concentrating the population in urban areas. The migration network could potentially be having a substantial effect on the HIV epidemic of Botswana: changing the location of high-transmission areas, generating cross-country transmission corridors, creating source-sink dynamics, and undermining control strategies. Large-scale migration networks could present a considerable challenge to eliminating HIV in Botswana and in other countries in sub-Saharan Africa, and should be considered when designing epidemic control strategies.


Alzheimer's disease in humans and other animals: A consequence of postreproductive life span and longevity rather than aging.
Alzheimer's disease and diabetes mellitus are linked by epidemiology, genetics, and molecular pathogenesis. They may also be linked by the remarkable observation that insulin signaling sets the limits on longevity. In worms, flies, and mice, disrupting insulin signaling increases life span leading to speculation that caloric restriction might extend life span in man. It is our contention that man is already a long-lived organism, specifically with a remarkably high postfertility life span, and that it is this that results in the prevalence of Alzheimer's disease and diabetes. We review evidence for this hypothesis that carries specific predictions including that other animals with exceptionally long postreproductive life span will have increased risk of both diabetes and Alzheimer's disease. We present novel evidence that Dolphin, like man, an animal with exceptional longevity, might be one of the very few natural models of Alzheimer's disease.


Redundancy and synergy of neuronal ensembles in motor cortex.
We examined the ability of neuronal ensembles from rat motor cortex to predict behavioral performance during a reaction time task. We found that neurons that were the best individual predictors of task performance were not necessarily the neurons that contributed the most predictive information to an ensemble of neurons. To understand this result, we applied a framework for quantifying statistical relationships between neurons (Schneidman et al., 2003) to all possible combinations of neurons within our ensembles. We found that almost all neurons (96%) contributed redundant predictive information to the ensembles. This redundancy resulted in the maintenance of predictive information despite the removal of many neurons from each ensemble. Moreover, the balance of synergistic and redundant interactions depended on the number of neurons in the ensemble. Small ensembles could exhibit synergistic interactions (e.g., 23 +/- 9% of ensembles with two neurons were synergistic). In contrast, larger ensembles exhibited mostly redundant interactions (e.g., 99 +/- 0.1% of ensembles with eight neurons were redundant). We discuss these results with regard to constraints on interpreting neuronal ensemble data and with respect to motor cortex involvement in reaction time performance.


To maximize or randomize? An experimental study of probability matching in financial decision making.
Probability matching, also known as the "matching law" or Herrnstein's Law, has long puzzled economists and psychologists because of its apparent inconsistency with basic self-interest. We conduct an experiment with real monetary payoffs in which each participant plays a computer game to guess the outcome of a binary lottery. In addition to finding strong evidence for probability matching, we document different tendencies towards randomization in different payoff environments-as predicted by models of the evolutionary origin of probability matching-after controlling for a wide range of demographic and socioeconomic variables. We also find several individual differences in the tendency to maximize or randomize, correlated with wealth and other socioeconomic factors. In particular, subjects who have taken probability and statistics classes and those who self-reported finding a pattern in the game are found to have randomized more, contrary to the common wisdom that those with better understanding of probabilistic reasoning are more likely to be rational economic maximizers. Our results provide experimental evidence that individuals-even those with experience in probability and investing-engage in randomized behavior and probability matching, underscoring the role of the environment as a driver of behavioral anomalies.


Molecular entomology and prospects for malaria control.
During the past decade, the techniques of molecular and cell biology have been embraced by many scientists doing research on anopheline vectors of malaria parasites. Some of the most important research advances in molecular entomology have concerned the development of sophisticated molecular tools for procedures such as genetic and physical mapping and germ line transformation. Major advances have also been made in the study of specific biological processes such as insect defence against pathogens and the manner in which malaria parasites and their anopheline hosts interact during sporogony. One of the most important highlights of this research trend has been the emergence during the past year of a formal international Anopheles gambiae genome project, which at present includes investigators in several laboratories in Europe and the USA. Although much of this molecular research is directed towards the development of malaria control strategies that are probably many years from implementation, there are some important areas of molecular entomology that may have a more near-term impact on malaria control. We highlight developments over the past decade in three such areas that we believe can make important contributions to the development of near-term malaria control strategies. These areas are anopheline species identification, the detection and monitoring of insecticide susceptibility/resistance in wild anopheline populations and the determination of the genetic structure of anopheline populations.


The Role and Mechanism of SIRT6 in Regulating Phenotype Transformation of Vascular Smooth Muscle Cells in Abdominal Aortic Aneurysm.
Data mining of current gene expression databases has not been previously performed to determine whether sirtuin 6 (SIRT6) expression participates in the pathological process of abdominal aortic aneurysm (AAA). The present study was aimed at investigating the role and mechanism of SIRT6 in regulating phenotype transformation of vascular smooth muscle cells (VSMC) in AAA. Three gene expression microarray datasets of AAA patients in the Gene Expression Omnibus (GEO) database and one dataset of SIRT6-knockout (KO) mice were selected, and the differentially expressed genes (DEGs) were identified using GEO2R. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of both the AAA-related DEGs and the SIRT6-related DEGs were conducted. GEO2R analysis showed that the expression of SIRT6 was downregulated for three groups and upregulated for one group in the three datasets, and none of them satisfied statistical significance. There were top 5 DEGs (KYNU, NPTX2, SCRG1, GRK5, and RGS5) in both of the human AAA group and SIRT6-KO mouse group. Top 25 ontology of the SIRT6-KO-related DEGs showed that several pathways including tryptophan catabolic process to kynurenine and negative regulation of cell growth were enriched in the tissues of thickness aortic wall biopsies of AAA patients. Although SIRT6 mRNA level itself did not change among AAA patients, SIRT6 may play an important role in regulating several signaling pathways with significant association with AAA, suggesting that SIRT6 mRNA upregulation is a protective factor for VSMC against AAA.


Bleomycin induced sensitivity to TRAIL/Apo-2L-mediated apoptosis in human seminomatous testicular cancer cells is correlated with upregulation of death receptors.
The most common solid tumor is testicular cancer among young men. Bleomycin is an antitumor antibiotic used for the therapy of testicular cancer. TRAIL is a proapoptotic cytokine that qualified as an apoptosis inducer in cancer cells. Killing cancer cells selectively via apoptosis induction is an encouraging therapeutic strategy in clinical settings. Combination of TRAIL with chemotherapeutics has been reported to enhance TRAIL-mediated apoptosis of different kinds of cancer cell lines. The molecular ground for sensitization of tumour cells to TRAIL by chemotherapeutics might involve upregulation of TRAIL-R1 (TR/1, DR4) and/or TRAIL-R2 (TR/2, DR5) receptors or activation of proapoptotic proteins including caspases. The curative potential of TRAIL to eradicate cancer cells selectively in testicular cancer has not been studied before. In this study, we investigated apoptotic effects of bleomycin, TRAIL, and their combined application in NTera-2 and NCCIT testicular cancer cell lines. We measured caspase 3 levels as an apoptosis indicator, and TRAIL receptor expressions using flow cytometry. Both NTera-2 and NCCIT cells were fairly resistant to TRAIL's apoptotic effect. Incubation of bleomycin alone caused a significant increase in caspase 3 activity in NCCIT. Combined incubation with bleomycin and TRAIL lead to elevated caspase 3 activity in Ntera-2. Exposure to 72 h of bleomycin increased TR/1, TR/2, and TR/3 cell-surface expressions in NTera-2. Elevation in TR/1 cell-surface expression was evident only at 24 h of bleomycin application in NCCIT. It can be concluded that TRAIL death receptor expressions in particular are increased in testicular cancer cells via bleomycin treatment, and TRAIL-induced apoptosis is initiated.


Phosphate-Arsenic Interactions in Halophilic Microorganisms of the Microbial Mat from Laguna Tebenquiche: from the Microenvironment to the Genomes.
Arsenic (As) is a metalloid present in the earth's crust and widely distributed in the environment. Due to its high concentrations in the Andean valleys and its chemical similarity with phosphorus (P), its biological role in Andean Microbial Ecosystems (AMEs) has begun to be studied. The AMEs are home to extremophilic microbial communities that form microbial mats, evaporites, and microbialites inhabiting Andean lakes, puquios, or salt flats. In this work, we characterize the biological role of As and the effect of phosphate in AMEs from the Laguna Tebenquiche (Atacama Desert, Chile). Using micro X-ray fluorescence, the distribution of As in microbial mat samples was mapped. Taxonomic and inferred functional profiles were obtained from enriched cultures of microbial mats incubated under As stress and different phosphate conditions. Additionally, representative microorganisms highly resistant to As and able to grow under low phosphate concentration were isolated and studied physiologically. Finally, the genomes of the isolated Salicola sp. and Halorubrum sp. were sequenced to analyze genes related to both phosphate metabolism and As resistance. The results revealed As as a key component of the microbial mat ecosystem: (i) As was distributed across all sections of the microbial mat and represented a significant weight percentage of the mat (0.17 %) in comparison with P (0.40%); (ii) Low phosphate concentration drastically changed the microbial community in microbial mat samples incubated under high salinity and high As concentrations; (iii) Archaea and Bacteria isolated from the microbial mat were highly resistant to arsenate (up to 500 mM), even under low phosphate concentration; (iv) The genomes of the two isolates were predicted to contain key genes in As metabolism (aioAB and arsC/acr3) and the genes predicted to encode the phosphate-specific transport operon (pstSCAB-phoU) are next to the arsC gene, suggesting a functional relationship between these two elements.