The histone methyltransferase Suv39h1 increases class switch recombination specifically to IgA.
Ab class (isotype) switching allows the humoral immune system to adaptively respond to different infectious organisms. Isotype switching occurs by intrachromosomal DNA recombination between switch (S) region sequences associated with C(H) region genes. Although isotype-specific transcription of unrearranged (germline) C(H) genes is required for switching, recent results suggest that isotype specificity is also determined by the sequences of downstream (acceptor) S regions. In the current study, we identify the histone methyltransferase Suv39h1 as a novel Salpha-specific factor that specifically increases IgA switching (Smu-Salpha recombination) in a transiently transfected plasmid S substrate, and demonstrate that this effect requires the histone methyltransferase activity of Suv39h1. Additionally, B cells from Suv39h1-deficient mice have an isotype-specific reduction in IgA switching with no effect on the level of germline Ialpha-Calpha transcripts. Taken together, our results suggest that Suv39h1 activity inhibits the activity of a sequence-specific DNA-binding protein that represses switch recombination to IgA.


First Report of Myrothecium roridum Causing Myrothecium Leaf Spot on Dieffenbachia picta 'Camilla' in Taiwan.
Dumb cane (Dieffenbachia picta (Lodd.) Schott 'Camilla'), family Araceae, is a popular houseplant in Taiwan. During the winter of 2012, dumb canes with dark brown concentric spots on leaves and bright yellow borders were found in a protected ornamental nursery in Wandan township, Pingtung County, Taiwan. On diseased leaves, fungal fruiting bodies were sometimes observed in the concentric lesions and a fungal isolate was consistently isolated from the lesions. A single spore isolate, myr 2-2, was maintained on potato dextrose agar (PDA) for further tests. To fulfill Koch's postulates, the spores of myr 2-2 were suspended in sterilized distilled water containing 0.05% of Tween 20, 1 × 105 conidia ml-1, and then sprayed on leaves of D. picta 'Camilla' growing in polypropylene plant pots (about 7 cm in diameter), three plants per treatment. For the control, three plants were sprayed with sterilized distilled water containing 0.05% of Tween 20. Both inoculated and non-inoculated plants were covered with plastic bags and incubated in a growth chamber at 26 ± 1°C. Nine to 12 days after inoculation, symptoms described above were observed on inoculated plants whereas the plants in control remained healthy. The same fungus was reisolated from inoculated plants but not from the controls. Furthermore, the fungal pathogen was identified using its physiological, morphological, and molecular characteristics. In the mycelial growth test, the diameter of the fungal colony reaches 58.2 mm on PDA at 25°C after 14 days. The colonies were floccose, white to buff, and sporulate in concentric zones with olivaceous black to black sporodochia bearing viscid masses of conidia. Conidia were narrowly ellipsoid with rounded ends. The average size of 100 conidia was 6.25 ± 0.04 × 1.63 ± 0.02 μm. For molecular identification, the rDNA internal transcribed spacer (ITS) of isolate myr 2-2 was PCR amplified using ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'- TCCTCCGCTTATTGATATGC-3') primer pairs (3) and sequenced. The rDNA sequence was deposited in GenBank (KC469695) and showed 100% identity to the Myrothecium roridum isolates BBA 71015 (AJ302001) and BBA 67679 (AJ301995) (4). According to the physiological, morphological (1,2), and molecular characteristics, the fungal isolate was identified as M. roridum Tode ex Fr. To the best of our knowledge, this is the first report of Myrothecium leaf spot caused by M. roridum on D. picta 'Camilla' in Taiwan. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , January 31, 2013. (2) M. Tulloch. Mycol. Pap. 130: 1-42, 1972. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990. (4) Y. X. Zhang et al. Plant Dis. 95:1030, 2011.


Cell wall ultrastructure of flocculent and non-flocculent Schizosaccharomyces pombe strains. Effect of cell wall hydrolysing enzymes on flocculation and cell wall ultastructure.
Scanning and transmission electron microscopic studies revealed the presence of slime-like, amorphous material on the surface of Schizosaccahromyces pombe RIVE 4-2-1 cells, independently, whether they were in flocculated or in non-flocculated state. Close contact of the adjacent cells via the merging outermost cell wall layers was found, however, only in the case of floc formation, which was induced by cultivating the cells in the presence of 6% (v/v) ethanol. Irreversible loss of the flocculation ability of the cells by treatment with proteinases suggests that proteinaceous cell surface molecules as lectins contribute to the cell-to-cell interaction during flocculation. Both proteinase K and pronase treatments removed a distinct outer layer of the cell wall, which indicated that the protein moieties of the phosphogalactomannan outer surface layer has a crucial role in the maintenance of cell wall integrity. In the case of lysing enzyme treatment the removal of the outermost layer was also observed as the first step of the cell wall digestion, while driselase treatment resulted in almost complete digestion of the cell wall.


Body Mass-Related Predictors of the Female Athlete Triad Among Adolescent Athletes.
Early detection of the female athlete triad is essential for the long-term health of adolescent female athletes. The purpose of this study was to assess relationships between common anthropometric markers (ideal body weight [IBW] via the Hamwi formula, youth-percentile body mass index [BMI], adult BMI categories, and body fat percentage [BF%]) and triad components, (low energy availability [EA], measured by dietary restraint [DR], menstrual dysfunction [MD], low bone mineral density [BMD]). In the sample (n = 320) of adolescent female athletes (age 15.9± 1.2 y), Spearman's rho correlations and multiple logistic regression analyses evaluated associations between anthropometric clinical cutoffs and triad components. All underweight categories for the anthropometric measures predicted greater likelihood of MD and low BMD. Athletes with an IBW >85% were nearly 4 times more likely to report MD (OR = 3.7, 95% CI [1.8, 7.9]) and had low BMD (OR = 4.1, 95% CI [1.2, 14.2]). Those in <5th percentile for their age-specific BMI were 9 times more likely to report MD (OR 9.1, 95% CI [1.8, 46.9]) and had low BMD than those in the 50th to 85th percentile. Athletes with a high BF% were almost 3 times more likely to report DR (OR = 2.8, 95% CI [1.4, 6.1]). Our study indicates that low age-adjusted BMI and low IBW may serve as evidence-based clinical indicators that may be practically evaluated in the field, predicting MD and low BMD in adolescents. These measures should be tested for their ability as tools to minimize the risk for the triad.


Aggregation-induced enhanced green light emission from a simple donor-π-acceptor (D-π-A) material: a structure-property relationship study.
Organic D-π-A materials, possessing intramolecular charge transfer, have attracted much scientific attention in recent years because of their potential applications in the development of organic light emitting devices (OLEDs). Two new compounds, A1 and A2, having a D-π-A skeleton have been synthesized and single crystals were grown by the solution growth technique. Both compounds were characterized for crystallographic, thermal and photophysical properties. Upon photo-excitation in the solid state, A1 showed very strong green light emission while A2 gave sky-blue emission with much lower intensity. A single crystal X-ray diffraction study revealed that in the crystal lattice of A1, both the donor and acceptor groups are involved in the intermolecular interactions. This results in the restricted intramolecular rotation (RIR) of the D and A moieties, and enables A1 to emit more intensely in the solid state due to aggregation-induced emission (AIE). Intense green light emission, along with a good crystalline nature indicates that A1 might be a potential candidate for opto-electronic devices.


"Deletion of both Rab-GTPase–activating proteins TBC1D1 and TBC1D4 in mice eliminates insulin- and AICAR-stimulated glucose transport [corrected].
The Rab-GTPase–activating proteins TBC1D1 and TBC1D4 (AS160) were previously shown to regulate GLUT4 translocation in response to activation of AKT and AMP-dependent kinase [corrected]. However, knockout mice lacking either Tbc1d1 or Tbc1d4 displayed only partially impaired insulin-stimulated glucose uptake in fat and muscle tissue. The aim of this study was to determine the impact of the combined inactivation of Tbc1d1 and Tbc1d4 on glucose metabolism in double-deficient (D1/4KO) mice. D1/4KO mice displayed normal fasting glucose concentrations but had reduced tolerance to intraperitoneally administered glucose, insulin, and AICAR. D1/4KO mice showed reduced respiratory quotient, indicating increased use of lipids as fuel. These mice also consistently showed elevated fatty acid oxidation in isolated skeletal muscle, whereas insulin-stimulated glucose uptake in muscle and adipose cells was almost completely abolished. In skeletal muscle and white adipose tissue, the abundance of GLUT4 protein, but not GLUT4 mRNA, was substantially reduced. Cell surface labeling of GLUTs indicated that RabGAP deficiency impairs retention of GLUT4 in intracellular vesicles in the basal state. Our results show that TBC1D1 and TBC1D4 together play essential roles in insulin-stimulated glucose uptake and substrate preference in skeletal muscle and adipose cells.


Proof of principle for epitope-focused vaccine design.
Vaccines prevent infectious disease largely by inducing protective neutralizing antibodies against vulnerable epitopes. Several major pathogens have resisted traditional vaccine development, although vulnerable epitopes targeted by neutralizing antibodies have been identified for several such cases. Hence, new vaccine design methods to induce epitope-specific neutralizing antibodies are needed. Here we show, with a neutralization epitope from respiratory syncytial virus, that computational protein design can generate small, thermally and conformationally stable protein scaffolds that accurately mimic the viral epitope structure and induce potent neutralizing antibodies. These scaffolds represent promising leads for the research and development of a human respiratory syncytial virus vaccine needed to protect infants, young children and the elderly. More generally, the results provide proof of principle for epitope-focused and scaffold-based vaccine design, and encourage the evaluation and further development of these strategies for a variety of other vaccine targets, including antigenically highly variable pathogens such as human immunodeficiency virus and influenza.


Warfarin prevents venous obstruction after cardiac devices implantation in high-risk patients: partial analysis.
To evaluate the efficacy of prophylactic use of warfarin in patients with high risk of lead-associated thrombosis. Clinical, prospective, randomized and blinded study, in patients submitted to first transvenous leads implantation with LVEF <0.40 and/or previous ipsilateral temporary pacing. After device implantation, patients were randomly assigned to placebo or warfarin. Periodical clinical and laboratorial evaluations were performed to anticoagulant management. After a six-month period, every patient was submitted to a digital subtraction venography. From February 2004 to November 2006, 101 patients underwent randomization. Baseline characteristics were similar in both groups (P=NS). Venographic analysis showed 31.4% of venous obstructions in patients assigned to warfarin as compared with 57.1% in patients assigned to placebo (RR= 0.57 [95% CI, 0.33 to 0.98]; P=0.015). In the warfarin group, 72% of the PT/INR tests were in therapeutic INR range. Only one patient required warfarin discontinuation and cross-over to placebo group due to gastrointestinal bleeding. These preliminary results showed that the anticoagulation therapy has been safe and reduced the frequency of venous thrombosis after transvenous cardiac devices implantation in high risk patients.


Syndrome of Inappropriate Antidiuretic Hormone (SIADH) Induced by Long-Term Use of Citalopram and Short-Term Use of Naproxen.
BACKGROUND Use of selective serotonin reuptake inhibitors (SSRIs) has been reported to be associated with the syndrome of inappropriate antidiuretic hormone (SIADH), although it is uncommon. Nonsteroidal anti-inflammatory drugs (NSAIDs), as a sole agent, are an even rarer cause of SIADH. Despite being documented in the literature, the understanding of the mechanism of both agents is limited. Here, we report a case of a patient taking both of these medications, a dangerous combination that led to the development of SIADH. CASE REPORT An 88-year-old woman with a history of asymptomatic chronic hyponatremia presented to our facility with symptomatic acute-on-chronic hyponatremia after she started using naproxen in addition to her daily citalopram. Her hyponatremia symptoms resolved after discontinuing these 2 offending agents, along with administration of fluid restriction and oral sodium supplements. CONCLUSIONS Naproxen is commonly prescribed and is often taken by elderly patients to control long-term or short-term pain. SSRIs, on the other hand, are a first-line treatment for depression and are usually prescribe by a psychiatrist. Hyponatremia is a rare medication adverse effect that should be kept in mind when treating these patients with either of these medications, and should especially be considered when combining them. Medication reconciliation should be done carefully by the provider to avoid adverse effects and drug interactions. When hyponatremia is encountered, options for future medication prescriptions include rechallenging with the same medication, switching to a different medication with the same mechanism of action, or using a medication from another class altogether. Monitoring of serum sodium level is warranted when titrating the offending agent.


SAME-SEX PARTNERSHIPS AND THE HEALTH OF OLDER ADULTS.
While extensive research has examined associations between marriage, cohabitation, and the health of heterosexual adults, it remains unclear whether similar patterns of health are associated with same-sex partnerships for lesbian, gay, and bisexual (LGB) older adults. This article examines whether having a same-sex partner is associated with general self-reported health and depressive symptoms for LGB older adults. Based on survey data collected from LGB adults 50 years of age and older, having a same-sex partner was associated with better self-reported health and fewer depressive symptoms when compared with single LGB older adults, controlling for gender, age, education, income, sexuality, and relationship duration. Relationship duration did not significantly impact the association between partnership status and health. In light of recent public debates and changes in policies regarding same-sex partnerships, more socially integrated relationship statuses appear to play a role in better health for LGB older adults.


A pilot study of pulmonary rehabilitation in primary care.
Pulmonary rehabilitation is an effective intervention for patients with chronic obstructive pulmonary disease (COPD). It is usually available only through selected hospitals. A pilot study was undertaken to see if pulmonary rehabilitation performed by the primary health care team in one practice was feasible. Fourteen patients were recruited; 13 completed the programme and one year of follow-up. The programme was well received by patients and staff. There were not enough suitable patients among a practice list of 10,500 to justify the running of this programme for a single practice; one primary care group would suffice


Comparison of the gut microbiota from soldier and worker castes of the termite Reticulitermes grassei.
The bacterial microbiota from the whole gut of soldier and worker castes of the termite Reticulitermes grassei was isolated and studied. In addition, the 16S rDNA bacterial genes from gut DNA were PCR-amplified using Bacteria-selective primers, and the 16S rDNA amplicons subsequently cloned into Escherichia coli. Sequences of the cloned inserts were then used to determine closest relatives by comparison with published sequences and with sequences from our previous work. The clones were found to be affiliated with the phyla Spirochaetes, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Synergistetes, Verrucomicrobia, and candidate phyla Termite Group 1 (TG1) and Termite Group 2 (TG2). No significant differences were observed with respect to the relative bacterial abundances between soldier and worker phylotypes. The phylotypes obtained in this study were compared with reported sequences from other termites, especially those of phylotypes related to Spirochaetes, Wolbachia (an Alphaproteobacteria), Actinobacteria, and TG1. Many of the clone phylotypes detected in soldiers grouped with those of workers. Moreover, clones CRgS91 (soldiers) and CRgW68 (workers), both affiliated with 'Endomicrobia', were the same phylotype. Soldiers and workers also seemed to have similar relative protist abundances. Heterotrophic, poly-β-hydroxyalkanoate-accumulating bacteria were isolated from the gut of soldiers and shown to be affiliated with Actinobacteria and Gammaproteobacteria. We noted that Wolbachia was detected in soldiers but not in workers. Overall, the maintenance by soldiers and workers of comparable axial and radial redox gradients in the gut is consistent with the similarities in the prokaryotes and protists comprising their microbiota.


p53 prevents neurodegeneration by regulating synaptic genes.
DNA damage has been implicated in neurodegenerative disorders, including Alzheimer's disease and other tauopathies, but the consequences of genotoxic stress to postmitotic neurons are poorly understood. Here we demonstrate that p53, a key mediator of the DNA damage response, plays a neuroprotective role in a Drosophila model of tauopathy. Further, through a whole-genome ChIP-chip analysis, we identify genes controlled by p53 in postmitotic neurons. We genetically validate a specific pathway, synaptic function, in p53-mediated neuroprotection. We then demonstrate that the control of synaptic genes by p53 is conserved in mammals. Collectively, our results implicate synaptic function as a central target in p53-dependent protection from neurodegeneration.


Short-term adapted physical activity program improves bone quality in osteopenic/osteoporotic postmenopausal women.
It is known that people affected by osteopenia/osteoporosis can benefit from an adequate amount of physical activity, counteracting the progressive loss of bone and muscle mass caused by aging. Moreover, there is increasing evidence that exercise has positive effects on bone structure. The aim of our study was to evaluate the effects on bone tissue and muscular strength of a short-term exercise program in osteopenic/osteoporotic postmenopausal women. Forty-nine osteopenic/osteoporotic postmenopausal women were divided into 2 groups: exercise and control. All subjects underwent 2 evaluations: before and after a training period. Bone quality was assessed by phalangeal quantitative osteosonography, and maximal strength of leg extensor muscles was also evaluated. The experimental group participated in a specific supervised 20-week physical activity program that included aerobic, balance, and strength training. After the training period, all bone parameters and lower-limb maximal strength were significantly improved in the exercise group (P < .05), whereas no significant changes were observed in the control group. Our study showed that a broad-based training protocol, lasting 20 weeks, can improve leg strength and bone quality parameters-main determinants of fall and fracture risk, respectively.


Selenium and Selenoproteins in Adipose Tissue Physiology and Obesity.
Selenium (Se) homeostasis is tightly related to carbohydrate and lipid metabolism, but its possible roles in obesity development and in adipocyte metabolism are unclear. The objective of the present study is to review the current data on Se status in obesity and to discuss the interference between Se and selenoprotein metabolism in adipocyte physiology and obesity pathogenesis. The overview and meta-analysis of the studies on blood Se and selenoprotein P (SELENOP) levels, as well as glutathione peroxidase (GPX) activity in obese subjects, have yielded heterogenous and even conflicting results. Laboratory studies demonstrate that Se may modulate preadipocyte proliferation and adipogenic differentiation, and also interfere with insulin signaling, and regulate lipolysis. Knockout models have demonstrated that the selenoprotein machinery, including endoplasmic reticulum-resident selenoproteins together with GPXs and thioredoxin reductases (TXNRDs), are tightly related to adipocyte development and functioning. In conclusion, Se and selenoproteins appear to play an essential role in adipose tissue physiology, although human data are inconsistent. Taken together, these findings do not support the utility of Se supplementation to prevent or alleviate obesity in humans. Further human and laboratory studies are required to elucidate associations between Se metabolism and obesity.


Inflammatory potential of diet and risk of pancreatic cancer in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial.
Inflammation plays a central role in pancreatic cancer etiology and can be modulated by diet. We aimed to examine the association between the inflammatory potential of diet, assessed with the Dietary Inflammatory Index (DII®), and pancreatic cancer risk in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial prospective cohort. Our study included 101,449 participants aged 52-78 years at baseline who completed both baseline questionnaire and a diet history questionnaire. Energy-adjusted DII (E-DII) scores were computed based on food and supplement intake. Cox proportional hazards models and time dependent Cox models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) with participants in the lowest E-DII quintile (most anti-inflammatory scores) as referent. After a median 8.5 years of follow-up, 328 pancreatic cancer cases were identified. E-DII scores were not associated with pancreatic cancer risk in the multivariable model (HRQ5vsQ1 = 0.94; 95% CI = 0.66-1.35; p-trend = 0.43). Time significantly modified the association (p-interaction = 0.01). During follow up <4 years, there was suggestive evidence of an inverse association between E-DII and pancreatic cancer (HRQ5vsQ1 = 0.60; 95% CI = 0.35-1.02; p-trend = 0.20) while there was a significant positive trend in the follow up ≥4 years (HRQ5vsQ1 = 1.31; 95% CI = 0.83-2.08; p-trend = 0.03). Similar results were observed for E-DII from food only. Our study does not support an association between inflammatory potential of diet and pancreatic cancer risk; however, heterogeneous results were obtained with different follow-up times. These divergent associations may result from the influences of undetected disease in the short-term.


Novel dosing strategies increase exposures of the potent antituberculosis drug rifapentine but are poorly tolerated in healthy volunteers.
Rifapentine is a potent antituberculosis drug currently in phase III trials. Bioavailability decreases with increasing dose, yet high daily exposures are likely needed to improve efficacy and shorten the tuberculosis treatment duration. Further, the limits of tolerability are poorly defined. The phase I multicenter trial in healthy adults described here investigated two strategies to increase rifapentine exposures: dividing the dose or giving the drug with a high-fat meal. In arm 1, rifapentine was administered at 10 mg/kg of body weight twice daily and 20 mg/kg once daily, each for 14 days, separated by a 28-day washout; the dosing sequence was randomized. In arm 2, 15 mg/kg rifapentine once daily was given with a high-fat versus a low-fat breakfast. Sampling for pharmacokinetic analysis was performed on days 1 and 14. Population pharmacokinetic analyses were performed. This trial was stopped early for poor tolerability and because of safety concerns. Of 44 subjects, 20 discontinued prematurely; 11 of these discontinued for protocol-defined toxicity (a grade 3 or higher adverse event or grade 2 or higher rifamycin hypersensitivity). Taking rifapentine with a high-fat meal increased the median steady-state area under the concentration-time curve from time zero to 24 h (AUC0-24ss) by 31% (relative standard error, 6%) compared to that obtained when the drug was taken with a low-fat breakfast. Dividing the dose increased exposures substantially (e.g., 38% with 1,500 mg/day). AUC0-24ss was uniformly higher in our study than in recent tuberculosis treatment trials, in which toxicity was rare. In conclusion, two strategies to increase rifapentine exposures, dividing the dose or giving it with a high-fat breakfast, successfully increased exposures, but toxicity was common in healthy adults. The limits of tolerability in patients with tuberculosis remain to be defined. (AIDS Clinical Trials Group study A5311 has been registered at ClinicalTrials.gov under registration no. NCT01574638.).


p15Ink4b is a critical tumour suppressor in the absence of p16Ink4a.
The CDKN2b-CDKN2a locus on chromosome 9p21 in human (chromosome 4 in mouse) is frequently lost in cancer. The locus encodes three cell cycle inhibitory proteins: p15INK4b encoded by CDKN2b, p16INK4a encoded by CDKN2a and p14ARF (p19Arf in mice) encoded by an alternative reading frame of CDKN2a (ref. 1). Whereas the tumour suppressor functions for p16INK4a and p14ARF have been firmly established, the role of p15INK4b remains ambiguous. However, many 9p21 deletions also remove CDKN2b, so we hypothesized a synergistic effect of the combined deficiency for p15INK4b, p14ARF and p16INK4a. Here we report that mice deficient for all three open reading frames (Cdkn2ab-/-) are more tumour-prone and develop a wider spectrum of tumours than Cdkn2a mutant mice, with a preponderance of skin tumours and soft tissue sarcomas (for example, mesothelioma) frequently composed of mixed cell types and often showing biphasic differentiation. Cdkn2ab-/- mouse embryonic fibroblasts (MEFs) are substantially more sensitive to oncogenic transformation than Cdkn2a mutant MEFs. Under conditions of stress, p15Ink4b protein levels are significantly elevated in MEFs deficient for p16Ink4a. Our data indicate that p15Ink4b can fulfil a critical backup function for p16Ink4a and provide an explanation for the frequent loss of the complete CDKN2b-CDKN2a locus in human tumours.


Relevance of gingival crevice fluid components in assessment of periodontal disease - A critical analysis.
Monitoring infections which are episodic, site specific, localized or generalized, initiated by a number of microorganisms and assessed with measurement devices that have inherent shortcomings presents a difficult task. The lack of precise clinical criteria for assessment of periodontal disease has led to a search for alternative means of determining active disease sites, predicting future sites of breakdown and evaluating response to therapy. This paper highlights the potential array of biomarkers present in gingival crevice fluid (GCF) and provides an insight of which of these factors has the greatest diagnostic potential. The rationale behind using GCF as sampling source is also described. Finally it discusses the possible use of predictive chair side diagnostic tests in periodontitis.


Addressing Consent Issues in Donation After Circulatory Determination of Death.
Given the widening gap between the number of individuals on transplant waiting lists and the availability of donated organs, as well as the recent plateau in donations based on neurological criteria (i.e., brain death), there has been a growing interest in expanding donation after circulatory determination of death. While the prevalence of this form of organ donation continues to increase, many thorny ethical issues remain, often creating moral distress in both clinicians and families. In this article, we address one of these issues, namely, the challenges surrounding patient and surrogate informed consent for donation after circulatory determination of death. First we discuss several general concerns regarding consent related to this form of organ donation, and then we address additional issues that are unique to three different patient categories: adult patients with medical decision-making capacity or potential capacity, adult patients who lack capacity, and pediatric patients.


Symptomatic perioperative venous thromboembolism is a frequent complication in patients with a history of deep vein thrombosis.
Patients who undergo surgery are at risk for venous thromboembolism (VTE), and a history of prior deep vein thrombosis (DVT) increases that risk. This study determined the incidence and risk factors for symptomatic perioperative VTE in patients with a prior diagnosis of DVT. All lower extremity DVTs, diagnosed between January 2002 and December 2006, were identified through a vascular database. Patients who had subsequent surgery were reviewed. The following data were evaluated: location of DVT, time interval between DVT and surgery, type of surgery, common clinical VTE risk factors, postoperative venous duplex scans, computed tomography (CT) scans of the chest, and ventilation-perfusion scans. A total of 372 patients with prior DVT underwent 1081 subsequent surgical procedures. One hundred nine patients undergoing 211 procedures had a follow-up venous duplex scan within 30 days after surgery. Of them, 46% received an inferior vena caval (IVC) filter, and pulmonary emboli were diagnosed in 3 patients (<1%). Overall, 24% of the patients developed DVT extension or new-site DVT in the perioperative period. The median time interval between the original DVT and surgery was 1.5 weeks in patients with DVT recurrence and 4 weeks in patients without recurrence (P = .22, Mann-Whitney). High-risk surgeries were associated with a >three-fold increased risk for recurrence, when compared with low-risk procedures (34% vs 11%; P = .009, chi(2)). Perioperative VTE recurrence was not influenced by the location of the original thrombus or other VTE risk factors. In patients with prior DVT, perioperative symptomatic recurrence is common and is associated with high-risk procedures. A longer time interval between a DVT episode and subsequent surgery may decrease the risk of recurrence, but large clinical trials are needed to confirm this. Further prospective evaluations are needed to identify and treat patients at greatest risk for recurrence.


The effects of digoxin and dopamine on the oxygen consumption, lactate production and haemodynamic performance of an isolated, perfused, working guinea-pig heart.
We have adapted an established technique for perfusing the isolated, working rat heart to the guinea-pig heart and characterized its biochemical and haemodynamic performance. After 20 min of anoxia the heart recovers about 50% of its pre-anoxic performance ('post-anoxic cardiac failure') and after 90 min of continuous work total cardiac output falls to 50% of its initial, stable value ('spontaneous cardiac failure'). We have studied the effects of digoxin (10(-7) M and 2 x 10(-7) M) and dopamine (10(-5) M) on these two forms of cardiac failure and shown that digoxin improves the haemodynamic performance of the heart without altering its metabolism and therefore increases its efficiency. In contrast dopamine improves the haemodynamic performance of the heart at the expense of increased aerobic and anaerobic metabolism. The preparation is of value in studying the effects of cardioactive drugs on hearts subjected to constant pre-load and after-load.


Risk factors for clinical leptospirosis from Western Jamaica.
A retrospective, matched case-control study was conducted in Jamaica's Western Regional Health Authority (WRHA). Forty-three individuals developing clinical leptospirosis between January 2005 and December 2007 (i.e., cases) were age and neighborhood matched to 89 controls. Odds ratios (OR) and associated 95% confidence intervals (CIs) and the relative excess risk due to interaction (RERI) were calculated. Cases had increased odds of contact with rodents OR 3.52, goats OR 3.38, and being engaged in outdoor labor OR 5.30. Knowledge of leptospirosis and indoor work was protective, OR 0.39 and OR 0.16, respectively. Positive RERI values were noted for joint exposure to rodents and goats (RERI 5.54), outdoor labor and goats (RERI 6.97), and outdoor labor and rodents (RERI 30.59). Our results suggest a synergistic effect of occupational and environmental exposures on clinical human leptospirosis from the WRHA. Knowledge of the disease and its risk factors allows for protection from the disease.


Rey's Auditory Verbal Learning Test scores can be predicted from whole brain MRI in Alzheimer's disease.
Rey's Auditory Verbal Learning Test (RAVLT) is a powerful neuropsychological tool for testing episodic memory, which is widely used for the cognitive assessment in dementia and pre-dementia conditions. Several studies have shown that an impairment in RAVLT scores reflect well the underlying pathology caused by Alzheimer's disease (AD), thus making RAVLT an effective early marker to detect AD in persons with memory complaints. We investigated the association between RAVLT scores (RAVLT Immediate and RAVLT Percent Forgetting) and the structural brain atrophy caused by AD. The aim was to comprehensively study to what extent the RAVLT scores are predictable based on structural magnetic resonance imaging (MRI) data using machine learning approaches as well as to find the most important brain regions for the estimation of RAVLT scores. For this, we built a predictive model to estimate RAVLT scores from gray matter density via elastic net penalized linear regression model. The proposed approach provided highly significant cross-validated correlation between the estimated and observed RAVLT Immediate (R = 0.50) and RAVLT Percent Forgetting (R = 0.43) in a dataset consisting of 806 AD, mild cognitive impairment (MCI) or healthy subjects. In addition, the selected machine learning method provided more accurate estimates of RAVLT scores than the relevance vector regression used earlier for the estimation of RAVLT based on MRI data. The top predictors were medial temporal lobe structures and amygdala for the estimation of RAVLT Immediate and angular gyrus, hippocampus and amygdala for the estimation of RAVLT Percent Forgetting. Further, the conversion of MCI subjects to AD in 3-years could be predicted based on either observed or estimated RAVLT scores with an accuracy comparable to MRI-based biomarkers.


Mortality and hospital utilization for hepatocellular carcinoma in the United States.
The incidence of hepatocellular carcinoma (HCC) has been increasing in the United States. Although resource-intensive treatment modalities have been increasingly applied, these patients still have poor survival. We examined 2 nationally representative databases, the Multiple Cause of Death file and the Nationwide Inpatient Sample database, to examine trends in mortality and hospital service utilization related to HCC. In both databases, a priori criteria were used to identify cases of HCC. All other available diagnostic fields were examined to characterize coexistent liver disease. Age-, sex-, and race-specific mortality from HCC was calculated, and temporal changes in mortality rates were evaluated using the multivariable Poisson model. Hospital service utilization was estimated based on length of stay, total hospitalization charges, and principal procedures. The age-, sex-, and race-specific mortality from HCC increased from 1.54 to 2.58 per 100,000 per year between 1980 and 1998. Male sex, African and Asian race, and increasing age were also associated with higher mortality. The estimated total charge for HCC hospitalizations nationwide increased from 241 million US dollars in 1988 to 509 million US dollars in 2000 after inflation adjustment. Commonly employed procedures in 2000 included angiography/embolization, resection, local ablative therapy, and liver transplantation. In the recent past, mortality and hospital service utilization related to HCC increased substantially. Closer epidemiologic surveillance to understand causation of HCC at the population level and to help implement primary and secondary prevention is urgently warranted.


Urodynamic investigation of the ileal conduit: upper tract dilatation and the effects of revision of the conduit.
Urodynamic studies of the ileal conduit were performed in 36 patients with normal upper urinary tracts (controls) and in 32 who had progressive upper tract dilatation within 6 years preceding the study. In controls the conduit emptied mainly by means of low pressure, to-and-fro activity. In contrast, high frequency, high amplitude peristaltic activity was found commonly in patients with upper tract dilatation, such activity being consistent with obstruction of the distal conduit. Of the 32 patients with upper tract dilatation 17 underwent reoperation on the conduit as well as postoperative urodynamic studies. Among these 17 patients a decrease was observed in basal pressure in the conduit postoperatively compared to preoperatively (5 +/- 4 and 12 +/- 6 cm. water, p less than 0.001). A decrease also was found postoperatively in the frequency (4 +/- 6 and 36 +/- 28 per hour, p less than 0.001) and amplitude (34 +/- 47 and 61 +/- 28 cm. water, p less than 0.001) of peristaltic activity. Radiological improvement in the appearance of the upper urinary tracts was found in 9 patients (53 per cent) postoperatively. These findings support the hypothesis that upper tract dilatation in certain patients with an ileal conduit may be caused by high pressure activity in the presence of free ureteral reflux. Such abnormalities, which are probably the result of obstruction of the distal conduit, may be detected by means of urodynamic evaluation and may be corrected by revision of the conduit.


Physical activity, diabetes, and risk of thyroid cancer: a systematic review and meta-analysis.
Thyroid cancer incidence has been increasing more rapidly over time than the occurrence of cancers of other sites, and interest in potential adverse relations of diabetes and lack of physical activity to thyroid cancer risk is accumulating. We conducted a systematic review and meta-analysis of published epidemiologic studies on the relations of physical activity and diabetes to thyroid cancer according to the Meta-analysis of Observational Studies in Epidemiology guidelines. Published studies were identified through a search in MEDLINE and EMBASE. Random-effects models were used to summarize thyroid cancer risk estimates comparing high versus low levels of physical activity, and separately, comparing individuals with diabetes versus those without diabetes. Meta-regression analyses were performed to evaluate potential effect modification by study design and thyroid cancer risk factors. Information was extracted from seven studies of physical activity and thyroid cancer and from six studies of diabetes and thyroid cancer. The number of individuals from studies on physical activity was 939,305 (yielding 2,250 incident thyroid cancer cases) and from studies on diabetes it was 960,840 (yielding 1,230 cases). The summary relative risk (RR) estimate from cohort and case-control studies combined indicated no association between physical activity and thyroid cancer (summary RR 1.06, 95 % confidence interval (CI) 0.79-1.42). Subgroup-analyses revealed a significant positive association between physical activity and thyroid cancer in cohort studies (summary RR 1.28; 95 % CI 1.01-1.63), whereas the relation was suggestively inverse in case-control studies (summary RR 0.70; 95 % CI 0.48-1.03; p for heterogeneity = 0.005). Individuals with diabetes showed a borderline statistically significant increased risk of thyroid cancer compared with those without diabetes (summary RR 1.17; 95 % CI 0.99-1.39). The relations of physical activity and diabetes to thyroid cancer were not modified by sex, number of adjustment factors, and adjustments for adiposity, smoking, and study quality. In this comprehensive systematic review and meta-analysis, no significant association between physical activity and thyroid cancer was found. Diabetes showed a suggestive positive relation with risk of thyroid cancer.


Automated Radiotherapy Planning for Patient-Specific Exploration of the Trade-Off Between Tumor Dose Coverage and Predicted Radiation-Induced Toxicity-A Proof of Principle Study for Prostate Cancer.
Background: Currently, radiation-oncologists generally evaluate a single treatment plan for each patient that is possibly adapted by the planner prior to final approval. There is no systematic exploration of patient-specific trade-offs between planning aims, using a set of treatment plans with a-priori defined (slightly) different balances. To this purpose, we developed an automated workflow and explored its use for prostate cancer. Materials and Methods: For each of the 50 study patients, seven plans were generated, including the so-called clinical plan, with currently clinically desired ≥99% dose coverage for the low-dose planning target volume (PTVLow). The six other plans were generated with different, reduced levels of PTVLow coverage, aiming at reductions in rectum dose and consequently in predicted grade≥2 late gastro-intestinal (GI) normal tissue complication probabilities (NTCPs), while keeping other dosimetric differences small. The applied NTCP model included diabetes as a non-dosimetric predictor. All plans were generated with a clinically applied, in-house developed algorithm for automated multi-criterial plan generation. Results: With diabetes, the average NTCP reduced from 24.9 ± 4.5% for ≥99% PTVLow coverage to 17.3 ± 2.6% for 90%, approaching the NTCP (15.4 ± 3.0%) without diabetes and full PTVLow coverage. Apart from intended differences in PTVLow coverage and rectum dose, other differences between the clinical plan and the six alternatives were indeed minor. Obtained NTCP reductions were highly patient-specific (ranging from 14.4 to 0.1%), depending on patient anatomy. Even for patients with equal NTCPs in the clinical plan, large differences were found in NTCP reductions. Conclusions: A clinically feasible workflow has been proposed for systematic exploration of patient-specific trade-offs between various treatment aims. For each patient, automated planning is used to generate a limited set of treatment plans with well-defined variations in the balances between the aims. For prostate cancer, trade-offs between PTVLow coverage and predicted GI NTCP were explored. With relatively small coverage reductions, significant NTCP reductions could be obtained, strongly depending on patient anatomy. Coverage reductions could also make up for enhanced NTCPs related to diabetes as co-morbidity, again dependent on the patient. The proposed system can play an important role in further personalization of patient care.


Wearable cameras can reduce dietary under-reporting: doubly labelled water validation of a camera-assisted 24 h recall.
Preliminary research has suggested that wearable cameras may reduce under-reporting of energy intake (EI) in self-reported dietary assessment. The aim of the present study was to test the validity of a wearable camera-assisted 24 h dietary recall against the doubly labelled water (DLW) technique. Total energy expenditure (TEE) was assessed over 15 d using the DLW protocol among forty adults (n 20 males, age 35 (sd 17) years, BMI 27 (sd 4) kg/m2 and n 20 females, age 28 (sd 7) years, BMI 22 (sd 2) kg/m2). EI was assessed using three multiple-pass 24 h dietary recalls (MP24) on days 2-4, 8-10 and 13-15. On the days before each nutrition assessment, participants wore an automated wearable camera (SenseCam (SC)) in free-living conditions. The wearable camera images were viewed by the participants following the completion of the dietary recall, and their changes in self-reported intakes were recorded (MP24+SC). TEE and EI assessed by the MP24 and MP24+SC methods were compared. Among men, the MP24 and MP24+SC measures underestimated TEE by 17 and 9%, respectively (P< 0.001 and P= 0.02). Among women, these measures underestimated TEE by 13 and 7%, respectively (P< 0.001 and P= 0.004). The assistance of the wearable camera (MP24+SC) reduced the magnitude of under-reporting by 8% for men and 6% for women compared with the MP24 alone (P< 0.001 and P< 0.001). The increase in EI was predominantly from the addition of 265 unreported foods (often snacks) as revealed by the participants during the image review. Wearable cameras enhance the accuracy of self-report by providing passive and objective information regarding dietary intake. High-definition image sensors and increased imaging frequency may improve the accuracy further.


[Radiation therapy for bone metastasis from hepatocellular carcinoma].
Recent advances in both diagnosis and treatment have markedly improved the prognosis in patients with hepatocellular carcinoma (HCC). Bone metastasis has become a clinical problem in the treatment of HCC patients. The purpose of this study was to evaluate the palliative effect of radiotherapy for painful bone metastasis from HCC. From January 1991 to June 2000, 51 patients (77 sites) with painful bone metastasis from HCC were retrospectively analyzed. Ages ranged from 21 to 80 years (median 55 years). The male:female ratio was 7.5:1. Synchronous or metachronous bone metastasis was seen in 20 (39%) and 31 patients (61%), respectively. The most common symptom of bone metastasis was pain (45 patients, 88%). Twenty-one patients (41%) had a solitary bone metastasis while 30 (59%) had multiple ones. The sites of bone metastasis, in order of frequency, were the vertebra (38), rib (20), and pelvis (19). The total radiation dose ranged from 12.5 to 50 Gy (median 30 Gy). The Wisconsin Brief Pain Questionnaire was used to evaluate pain response. The overall 1 and 2 year survival rates from the time of bone metastasis were 15% and 4%, respectively. The median survival time was 5 months. Intrahepatic stage(p=0.014), and metastasis to other organs(p=0.019) were significant prognostic factor for survival by univariate analysis. There was, however, no independent prognostic factor on multivariate analysis. Pain relief after radiotherapy was achieved for 56 sites (73%). The expected life span (median 5 months) in this group of patients suggests a strong necessity for effective treatment for symptomatic palliation. Radiation therapy was effective in pain palliation for bone metastasis from HCC, and this could improve patients' quality of life.


Infection dynamics in frog populations with different histories of decline caused by a deadly disease.
Pathogens can drive host population dynamics. Chytridiomycosis is a fungal disease of amphibians that is caused by the fungus Batrachochytrium dendrobatidis (Bd). This pathogen has caused declines and extinctions in some host species whereas other host species coexist with Bd without suffering declines. In the early 1990s, Bd extirpated populations of the endangered common mistfrog, Litoria rheocola, at high-elevation sites, while populations of the species persisted at low-elevation sites. Today, populations have reappeared at many high-elevation sites where they presently co-exist with the fungus. We conducted a capture-mark-recapture (CMR) study of six populations of L. rheocola over 1 year, at high and low elevations. We used multistate CMR models to determine which factors (Bd infection status, site type, and season) influenced rates of frog survival, recapture, infection, and recovery from infection. We observed Bd-induced mortality of individual frogs, but did not find any significant effect of Bd infection on the survival rate of L. rheocola at the population level. Survival and recapture rates depended on site type and season. Infection rate was highest in winter when temperatures were favourable for pathogen growth, and differed among site types. The recovery rate was high (75.7-85.8%) across seasons, and did not differ among site types. The coexistence of L. rheocola with Bd suggests that (1) frog populations are becoming resistant to the fungus, (2) Bd may have evolved lower virulence, or (3) current environmental conditions may be inhibiting outbreaks of the fatal disease.


eps15, a novel tyrosine kinase substrate, exhibits transforming activity.
An expression cloning method which allows direct isolation of cDNAs encoding substrates for tyrosine kinases was applied to the study of the epidermal growth factor (EGF) receptor (EGFR) signaling pathway. A previously undescribed cDNA was isolated and designated eps15. The structural features of the predicted eps15 gene product allow its subdivision into three domains. Domain I contains signatures of a regulatory domain, including a candidate tyrosine phosphorylation site and EF-hand-type calcium-binding domains. Domain II presents the characteristic heptad repeats of coiled-coil rod-like proteins, and domain III displays a repeated aspartic acid-proline-phenylalanine motif similar to a consensus sequence of several methylases. Antibodies specific for the eps15 gene product recognize two proteins: a major species of 142 kDa and a minor component of 155 kDa, both of which are phosphorylated on tyrosine following EGFR activation by EGF in vivo. EGFR is also able to directly phosphorylate the eps15 product in vitro. In addition, phosphorylation of the eps15 gene product in vivo is relatively receptor specific, since the erbB-2 kinase phosphorylates it very inefficiently. Finally, overexpression of eps15 is sufficient to transform NIH 3T3 cells, thus suggesting that the eps15 gene product is involved in the regulation of mitogenic signals.


The role of the area postrema in the anorectic effects of amylin and salmon calcitonin: behavioral and neuronal phenotyping.
Amylin reduces meal size by activating noradrenergic neurons in the area postrema (AP). Neurons in the AP also mediate the eating-inhibitory effects of salmon calcitonin (sCT), a potent amylin agonist, but the phenotypes of the neurons mediating its effect are unknown. Here we investigated whether sCT activates similar neuronal populations to amylin, and if its anorectic properties also depend on AP function. Male rats underwent AP lesion (APX) or sham surgery. Meal patterns were analysed under ad libitum and post-deprivation conditions. The importance of the AP in mediating the anorectic action of sCT was examined in feeding experiments of dose-response effects of sCT in APX vs. sham rats. The effect of sCT to induce Fos expression was compared between surgery groups, and relative to amylin. The phenotype of Fos-expressing neurons in the brainstem was examined by testing for the co-expression of dopamine beta hydroxylase (DBH) or tryptophan hydroxylase (TPH). By measuring the apposition of vesicular glutamate transporter-2 (VGLUT2)-positive boutons, potential glutamatergic input to amylin- and sCT-activated AP neurons was compared. Similar to amylin, an intact AP was necessary for sCT to reduce eating. Further, co-expression between Fos activation and DBH after amylin or sCT did not differ markedly, while co-localization of Fos and TPH was minor. Approximately 95% of neurons expressing Fos and DBH after amylin or sCT treatment were closely apposed to VGLUT2-positive boutons. Our study suggests that the hindbrain pathways engaged by amylin and sCT share many similarities, including the mediation by AP neurons.


Role of fibroblast growth factor-2 isoforms in the effect of estradiol on endothelial cell migration and proliferation.
Both 17beta-estradiol (E2) and fibroblast growth factor-2 (FGF2) stimulate angiogenesis and endothelial cell migration and proliferation. The first goal of this study was to explore the potential link between this hormone and this growth factor. E2-stimulated angiogenesis in SC Matrigel plugs in Fgf2+/+ mice, but not in Fgf2-/- mice. Cell cultures from subcutaneous Matrigel plugs demonstrated that E2 increased both migration and proliferation in endothelial cells from Fgf2+/+ mice, but not from in Fgf2-/- mice. Several isoforms of fibroblast growth factor-2 (FGF2) are expressed: the low molecular weight 18-kDa protein (FGF2lmw) is secreted and activates tyrosine kinase receptors (FGFRs), whereas the high molecular weight (21 and 22 kDa) isoforms (FGF2hmw) remains intranuclear, but their role is mainly unknown. The second goal of this study was to explore the respective roles of FGF2 isoforms in the effects of E2. We thus generated mice deficient only in the FGF2lmw (Fgf2lmw-/-). E2 stimulated in vivo angiogenesis and in vitro migration in endothelial cells from Fgf2lmw-/- as it did in Fgf2+/+ mice. E2 increased FGF2hmw protein abundance in endothelial cell cultures from Fgf2+/+ and Fgf2lmw-/- mice. As shown using siRNA transfection, these effects were FGFR independent but involved FGF2-Interacting Factor, an intracellular FGF2hmw partner. This is the first report for a physiological role for the intracellular FGF2hmw found to mediate the effect of E2 on endothelial cell migration via an intracrine action.


Hepatic and biochemical repercussions of a polyunsaturated fat-rich hypercaloric and hyperlipidic diet in Wistar rats.
Non-alcoholic fatty liver disease is characterized by lipid deposits in the hepatocytes and has been associated with obesity, dyslipidemia and type-2 diabetes. It is considered a hepatic manifestation of the metabolic syndrome, of which the main component is insulin resistance leading to hyperinsulinemia and increased production of inflammatory cytokines. Saturated fat promotes hypertriglyceridemia and hyperinsulinemia, reduces levels of high-density cholesterol and increases levels of low-density cholesterol, while polyunsaturated fat is associated with hypolipidemic, antiinflammatory and imunoregulating action. To evaluate the hepatic and biochemical repercussions of a polyunsaturated fat-rich diet in Wistar rats. Twenty-two rats were distributed equally in two groups: GI - standard diet (Biobase Bio-tec Ratos e Camundongos®) providing 3.000 kcal/kg and GII - hypercaloric and hyperlipidic diet providing 4.250 kcal/kg (ω-6:ω-3 = 3:1). The animals were euthanized after 23 weeks of experiment. The weight, biochemical parameters and hepatohistological changes were registered. Findings were submitted to variance analysis with the level of statistical significance at 5%. The average weight did not differ significantly between the groups at baseline (P = 0.711), but was greater in Group II by the end of the experiment (P = 0.000). The levels of triglycerides (P = 0.039), total cholesterol (P = 0.015) and HDL (P = 0.005) were higher in Group I than in Group II. Macrovesicular steatosis was significantly more common in Group II than in Group I (P = 0.03). Hypercaloric and hyperlipidic diet rich in polyunsaturated fat promotes weight gain and favors the development of hepatic steatosis while reducing serum levels of triglycerides, total cholesterol and HDL.


Vitreous levels of interleukin-35 as a prognostic factor in B-cell vitreoretinal lymphoma.
Vitreoretinal lymphoma (VRL) is a rare disease of B-cell origin with poor prognosis. Regulatory cytokines promote tumor development by suppressing antitumor immunity in several cancer types, including B-cell malignancies. To identify the regulatory cytokines associated with poor prognosis in patients with B-cell VRL, we determined the regulatory cytokines profiles in the vitreous humor of patients with VRL. This retrospective study included 22 patients with VRL, 24 with non-infectious uveitis (NIU), and 20 with idiopathic epiretinal membrane (control). Vitreous concentrations of regulatory cytokines were assessed using a cytometric beads assay and association with clinical data was examined. IL-35 and soluble IL-2 receptor α levels were significantly higher in patients with VRL and NIU than those in the control group. The 5-year overall survival (OS) rates for the group with high intravitreal IL-35 was significantly poorer than those for the group with low intravitreal IL-35, who were diagnosed with VRL at the onset (P = 0.024, log-rank test). The 5-year OS rates with intravitreal IL-35 levels above and below the median were 40.0% and 83.3%, respectively. Our results suggest that high intravitreal IL-35 levels indicate poor prognosis for patients diagnosed with B-cell VRL at the onset.


Circulating interleukin-1 and tumor necrosis factor in septic shock and experimental endotoxin fever.
Interleukins (IL) -1 beta and -1 alpha and tumor necrosis factor (TNF-alpha) were measured by radioimmunoassay in plasma samples from 44 healthy individuals, 15 patients in septic shock, and 6 volunteers infused with endotoxin. Plasma IL-1 alpha levels were low (40 pg/ml) or undetectable in all situations. In 67% of the healthy subjects, plasma IL-1 beta levels were less than 70 pg/ml. Septic patients had higher plasma IL-1 beta levels (120 +/- 17 pg/ml, P = .001); those of surviving patients were higher than those of patients who died (P = .05). Plasma TNF-alpha concentrations in septic individuals were elevated (119 +/- 30 pg/ml) and correlated with severity of illness (r = .73, P = .003), but no correlation was observed between plasma IL-1 beta and TNF-alpha concentrations in individual samples. Infusion of endotoxin caused a twofold elevation of IL-1 beta, from a baseline of 35 +/- 5 pg/ml to a maximum of 69 +/- 27 pg/ml at 180 min (P less than .05). Peak TNF-alpha levels after endotoxin infusion were 15 times higher than IL-1 beta levels, were attained more rapidly (90 min), and as with the septic patients, did not correlate with IL-1 beta levels. These data support the concept that plasma IL-1 beta and TNF-alpha concentrations are regulated independently and are associated with different clinical outcomes.


The downstream regulation of chemokine receptor signalling: implications for atherosclerosis.
Heterotrimeric G-protein-coupled receptors (GPCRs) are key mediators of intracellular signalling, control numerous physiological processes, and are one of the largest class of proteins to be pharmacologically targeted. Chemokine-induced macrophage recruitment into the vascular wall is an early pathological event in the progression of atherosclerosis. Leukocyte activation and chemotaxis during cell recruitment are mediated by chemokine ligation of multiple GPCRs. Regulation of GPCR signalling is critical in limiting vascular inflammation and involves interaction with downstream proteins such as GPCR kinases (GRKs), arrestin proteins and regulator of G-protein signalling (RGS) proteins. These have emerged as new mediators of atherogenesis by functioning in internalisation, desensitisation, and signal termination of chemokine receptors. Targeting chemokine signalling through these proteins may provide new strategies to alter atherosclerotic plaque formation and plaque biology.


Impact of diabetes in patients with pulmonary hypertension.
Diabetes complicates management in a number of disease states and adversely impacts survival; how diabetes affects patients with pulmonary hypertension (PH) has not been well characterized. With insulin resistance having recently been demonstrated in PH, we sought to examine the impact of diabetes in these patients. Demographic characteristics, echo data, and invasive hemodynamic data were prospectively collected for 261 patients with PH referred for initial hemodynamic assessment. Diabetes was defined as documented insulin resistance or treatment with antidiabetic medications. Fifty-five patients (21%) had diabetes, and compared with nondiabetic patients, they were older (mean years ± SD, 61 ± 13 vs. 56 ± 16; [Formula: see text]), more likely to be black (29% vs. 14%; [Formula: see text]) and hypertensive (71% vs. 30%; [Formula: see text]), and had higher mean (±SD) serum creatinine levels (1.1 ± 0.5 vs. 1.0 ± 0.4; [Formula: see text]). Diabetic patients had similar World Health Organization functional class at presentation but were more likely to have pulmonary venous etiology of PH (24% vs. 10%; [Formula: see text]). Echo findings, including biventricular function, tricuspid regurgitation, and pressure estimates were similar. Invasive pulmonary pressures and cardiac output were similar, but right atrial pressure was appreciably higher (14 ± 8 mmHg vs. 10 ± 5 mmHg; [Formula: see text]). Despite similar management, survival was markedly worse and remained so after statistical adjustment. In summary, diabetic patients referred for assessment of PH were more likely to have pulmonary venous disease than nondiabetic patients with PH, with hemodynamics suggesting greater right-sided diastolic dysfunction. The markedly worse survival in these patients merits further study.


Cost-effectiveness analyses alongside randomised clinical trials.
Many health economists increasingly advocate the use of model-based evaluations rather than trial-based evaluations. The purpose of this paper is to review the merits and limitations of RCT-based evaluations of cost-effectiveness. The paper draws on the examples of large studies such as the United Kingdom Prospective Diabetes Study and the Heart Protection Study to suggest that large randomised trials offer a number of advantages to health economists wishing to estimate cost-effectiveness, including access to patient-level data, unbiased estimates of resource use as well as effects, the estimation of cost-effectiveness in sub-groups of patients, and an enhanced ability to build and validate extrapolation models. While many methodological issues remain to be resolved, the use of patient-level data derived from clinical trials as a basis for economic evaluations is likely to remain an important part of the health economics evidence base, and will also continue to provide the data required for methodological research.


A bench stable formal Cu(iii) N-heterocyclic carbene accessible from simple copper(ii) acetate.
For years, Cu(iii)NHCs have been proposed as active intermediates in Cu(i)NHC catalyzed reactions, yielding the desired products by reductive elimination, but until today, no one has ever reported the characterisation of such a compound. When working on the synthesis of biomimetic transition metal (NHC) complexes and their application in homogeneous catalysis, we recently found a highly unusual reactivity for Cu(ii) acetate in the presence of a particular cyclic tetra(NHC) ligand. Therein, the formation of the first stable CuNHC compound, displaying Cu in the formal oxidation state +III, by simple disproportionation of Cu(ii) acetate in dimethyl sulfoxide (DMSO) was observed. At elevated temperatures selective mono-oxidation of the NHC ligand occurs, even under anaerobic conditions. Acetate was identified as the origin of the oxygen atom by 18O-labelling experiments. The remarkably high stability of the title compound was furthermore proven electrochemically by cyclic voltammetry. An in-depth investigation of its reactivity revealed the involvement of four additional compounds. Three of them could be isolated and characterised by 1H/13C-NMR, single crystal XRD, mass spectrometry and elemental analysis. The fourth, a Cu(i)NHC intermediate, formed by formal reductive elimination from the Cu(NHC)3+ compound, was characterised in situ by 1H/13C-NMR and computational methods.


Increasing lifestyle physical activity in patients with depression or other serious mental illness.
People with severe and persistent mental illness are more likely to be overweight and to suffer from obesity-related illnesses such as diabetes and heart disease than healthy individuals. Lifestyle change interventions that emphasize integrating physical activity into daily life have not been studied extensively in people with mental illness. The authors present the results of an initial feasibility study of a lifestyle modification program for individuals with serious mental illness. Thirty-nine individuals with depression or other serious mental illness were recruited from three different mental health facilities to attend an 18-week lifestyle intervention program promoting physical activity and healthy eating. At each session, participants discussed topics related to healthy lifestyle changes and participated in group walks. Data were collected at baseline, 6 weeks, and 18 weeks. The results demonstrated that individuals who have depression and other serious mental illnesses can participate in a lifestyle intervention program. Participants who attended the final follow-up session had lost weight over the course of the intervention. Study retention was a problem. However, the cost of this type of group-based lifestyle intervention was relatively low, so that such an intervention for this high-risk group may still be cost-effective.


HORMONE-INDUCED SIGNALING DURING MOSS DEVELOPMENT.
Understanding how a cell responds to hormonal signals with a new program of cellular differentiation and organization is an important focus of research in developmental biology. In Funaria hygrometrica and Physcomitrella patens, two related species of moss, cytokinin induces the development of a bud during the transition from filamentous to meristematic growth. Within hours of cytokinin perception, a single-celled initial responds with changes in patterns of cell expansion, elongation, and division to begin the process of bud assembly. Bud assembly in moss provides an excellent model for the study of hormone-induced organogenesis because it is a relatively simple, well-defined process. Since buds form in a nonrandom pattern on cells that are not embedded in other tissues, it is possible to predict which cells will respond and where the ensuing changes will take place. In addition, bud assembly is amenable to biochemical, cellular, and molecular biological analyses. This review examines our current understanding of cytokinin-induced bud assembly and the potential underlying mechanisms, reviews the state of genetic analyses in moss, and sets goals for future research with this organism.


Cerebrospinal fluid markers for Alzheimer's disease over the lifespan: effects of age and the APOEε4 genotype.
In Alzheimer's disease (AD), the cerebral pathological changes begin many years before the clinical manifestation of the disease. Biomarkers for AD, such as the cerebrospinal fluid (CSF) concentrations of amyloid-β1-42 (Aβ1-42) and tau phosphorylated at threonine 181 (pTau181), may reflect these cerebral changes relatively early. Accordingly, cognitively healthy subjects at risk for AD often have altered CSF concentrations of Aβ1-42 and pTau181. In this study, we assessed the effects and interaction of two strong risk factors for AD, aging and the presence of the APOEε4 allele, on the CSF Aβ1-42 and pTau181 concentrations in 280 adults with normal cognition across the lifespan. For comparison, we further included 152 patients with probable AD. We found significant effects of age on the CSF Aβ1-42 and pTau181, and of the APOEε4 genotype on the Aβ1-42 levels in the cognitively normal participants. Carrying the APOEε4 allele was associated with a significant decrease of the Aβ1-42 concentrations in middle-aged and older participants. In the group of participants with AD, the Aβ1-42 levels were significantly lower in the APOEε4 carriers compared to the non-carriers. These findings demonstrate significant age effects on the CSF Aβ1-42 and pTau181 across lifespan. They also suggest that the decrease of Aβ1-42, but not the increase of pTau181 CSF levels is accelerated by the APOEε4 genotype in middle-aged and older adults with normal cognition.


Inhibition of ricin A-chain with pyrrolidine mimics of the oxacarbenium ion transition state.
Ricin A-chain (RTA) catalyzes the hydrolytic depurination of a specific adenosine at position 4324 of 28S rRNA. Kinetic isotope effects on the hydrolysis of a small 10mer stem-tetraloop oligonucleotide substrate established the mechanism of the reaction as D(N)*A(N), involving an oxacarbenium ion intermediate in a highly dissociative transition state. An inhibitor with a protonated 1,4-dideoxy-1,4-imino-D-ribitol moiety, a 4-azasugar mimic, at the depurination site in the tetraloop of a 14mer oligonucleotide with a 5 bp duplex stem structure had previously been shown to bind to RTA with a K(d) of 480 nM, which improved to 12 nM upon addition of adenine. Second-generation stem-tetraloop inhibitors have been synthesized that incorporate a methylene bridge between the nitrogen of a 1-azasugar mimic, namely, (3S,4R)-3-hydroxy-4-(hydroxymethyl)pyrrolidine, and substituents, including phenyl, 8-aza-9-deazaadenyl, and 9-deazaadenyl groups, that mimic the activated leaving group at the transition state. The values for the dissociation constants (K(i)) for these were 99 nM for the phenyl 10mer, 163 and 94 nM for the 8-aza-9-deazaadenyl 10- and 14mers, respectively, and 280 nM for the 9-deazaadenyl 14mer. All of these compounds are among the tightest binding molecules known for RTA. A related phenyl-substituted inhibitor with a deoxyguanosine on the 5'-side of the depurination site was also synthesized on the basis of stem-loop substrate specificity studies. This molecule binds with a K(i) of 26 nM and is the tightest binding "one-piece" inhibitor. 8-Aza-9-deaza- and 9-deazaadenyl substituents provide an increased pK(a) at N7, a protonation site en route to the transition state. The binding of these inhibitors is not improved relative to the binding of their phenyl counterpart, however, suggesting that RTA might also employ protonation at N1 and N3 of the adenine moiety to activate the substrate during catalysis. Studies with methylated adenines support this argument. That the various stem-loop inhibitors have similar potencies suggests that an optimal one-piece inhibitor remains to be identified. The second-generation inhibitors described here incorporate ribose mimics missing the 2-hydroxy group. On the basis of inhibition data and substrate specificity studies, the 2'-hydroxyl group at the depurination site seems to be critical for recruitment as well as catalysis by RTA.


Pharmacological interventions for clozapine-induced hypersalivation.
Clozapine is widely used for people with schizophrenia. Although agranulocytosis, weight gain, and cardiac problems are serious problems associated with its use, hypersalivation, sometimes of a gross and socially unacceptable quantity, is also common (30-80%). To determine the clinical effects of pharmacological interventions for clozapine-induced hypersalivation. We searched the Cochrane Schizophrenia Group Trials Register (March 2007), inspected references of all identified studies for further trials, contacted relevant pharmaceutical companies, drug approval agencies and authors of trials. We included randomised controlled trials comparing pharmacological interventions, at any dose and by any route of administration, for clozapine-induced hypersalivation. We extracted data independently. For dichotomous data (homogenous) we calculated relative risk (RR) with 95% confidence intervals (CI) and numbers needed to treat (NNT) on an intention-to-treat basis. We calculated weighted mean difference (WMD) for continuous data. Of the 15 trials identified, 14 were conducted in China and 14 in hospitals. The quality of reporting was poor with no studies clearly describing allocation concealment and much data were missing or unusable. All results are vulnerable to considerable bias. Most frequently the primary outcome was the diameter of the wet patch on the pillow. Antimuscarinics (astemizole, diphenhydramine, propantheline, doxepin) were the most commonly evaluated drugs. For the outcome of 'no clinically important improvement' astemizole and diphenhydramine were more effective than placebo (astemizole: n=97, 2 RCTs, RR 0.61 CI 0.47 to 0.81 NNT 3 CI 2 to 5; diphenhydramine: n=131, 2 RCTs, RR 0.43 CI 0.31 to 0.58, NNT 2 CI 1.5 to 2.5), but the doses of astemizole used were those that can cause toxicity. Data involving propantheline were heterogeneous (I2= 86.6%), but both studies showed benefit over placebo. Adverse effects were poorly recorded. Of the other interventions, oryzanol (rice bran oil and rice embryo oil extract) showed benefit over the antimuscarinic doxepin in terms of 'no clinically important change' (n=104, 1 RCT, RR 0.45 CI 0.27 to 0.75, NNT 4 CI 2 to 7). The Chinese medicine suo quo wan (comprises spicebush root, Chinese yam and bitter cardamom) showed benefit over doxepin (n=70, 1 RCT, RR 'no clinically important change' 0.31 CI 0.16 to 0.59, NNT 3 CI 1.5 to 3.7). There are currently insufficient data to confidently inform clinical practice. The limitations of these studies are plentiful and the risk of bias is high. These trials, however, are invaluable guides for current and future study design. Well conducted randomised trials are possible. Some may be underway. Current practice outside of well designed randomised trials should be clearly justified.


Mutations in CHD7, encoding a chromatin-remodeling protein, cause idiopathic hypogonadotropic hypogonadism and Kallmann syndrome.
CHARGE syndrome and Kallmann syndrome (KS) are two distinct developmental disorders sharing overlapping features of impaired olfaction and hypogonadism. KS is a genetically heterogeneous disorder consisting of idiopathic hypogonadotropic hypogonadism (IHH) and anosmia, and is most commonly due to KAL1 or FGFR1 mutations. CHARGE syndrome, a multisystem autosomal-dominant disorder, is caused by CHD7 mutations. We hypothesized that CHD7 would be involved in the pathogenesis of IHH and KS (IHH/KS) without the CHARGE phenotype and that IHH/KS represents a milder allelic variant of CHARGE syndrome. Mutation screening of the 37 protein-coding exons of CHD7 was performed in 101 IHH/KS patients without a CHARGE phenotype. In an additional 96 IHH/KS patients, exons 6-10, encoding the conserved chromodomains, were sequenced. RT-PCR, SIFT, protein-structure analysis, and in situ hybridization were performed for additional supportive evidence. Seven heterozygous mutations, two splice and five missense, which were absent in > or = 180 controls, were identified in three sporadic KS and four sporadic normosmic IHH patients. Three mutations affect chromodomains critical for proper CHD7 function in chromatin remodeling and transcriptional regulation, whereas the other four affect conserved residues, suggesting that they are deleterious. CHD7's role is further corroborated by specific expression in IHH/KS-relevant tissues and appropriate developmental expression. Sporadic CHD7 mutations occur in 6% of IHH/KS patients. CHD7 represents the first identified chromatin-remodeling protein with a role in human puberty and the second gene to cause both normosmic IHH and KS in humans. Our findings indicate that both normosmic IHH and KS are mild allelic variants of CHARGE syndrome and are caused by CHD7 mutations.


Design, Synthesis, and Characterization of the Macrocyclic Tetrapeptide cyclo[Pro-Sar-Phe-d-Phe]: A Mixed Opioid Receptor Agonist-Antagonist Following Oral Administration.
Substance abuse remains a serious public health crisis, affecting millions of people worldwide. Macrocyclic tetrapeptides like CJ-15,208 and [d-Trp]CJ-15,208 demonstrate opioid activity shown to attenuate the rewarding effects of cocaine in conditioned place preference assays in mice, making them promising lead compounds for treating substance abuse. In the present study, we report the rational design, synthesis, conformational analysis, and continued pharmacological evaluation of the novel macrocyclic tetrapeptide cyclo[Pro-Sar-Phe-d-Phe] to further explore this unique molecular scaffold. This peptide was rationally designed based on X-ray and NMR structures of related macrocyclic tetrapeptides. Following synthesis, its solution-phase conformations were determined by NMR and molecular modeling. The peptide adopted multiple conformations in polar solvents, but a single conformation in chloroform that is stabilized by intramolecular hydrogen bonding. The peptide is orally bioavailable, producing antinociception and antagonism of kappa opioid receptor (KOR) stimulation following oral administration in a mouse 55 °C warm-water tail-withdrawal assay. Notably, cyclo[Pro-Sar-Phe-d-Phe] blocked both stress- and drug-induced reinstatement of cocaine and morphine conditioned place preference in mice following oral administration, and displayed a decreased side-effect profile compared to morphine. Thus, cyclo[Pro-Sar-Phe-d-Phe] is a promising lead compound for the treatment of substance abuse.


On the design and analysis of competent selecto-recombinative GAs.
In this paper, we study two recent theoretical models--a population-sizing model and a convergence model--and examine their assumptions to gain insights into the conditions under which selecto-recombinative GAs work well. We use these insights to formulate several design rules to develop competent GAs for practical problems. To test the usefulness of the design rules, we consider as a case study the map-labeling problem, an NP-hard problem from cartography. We compare the predictions of the theoretical models with the actual performance of the GA for the map-labeling problem. Experiments show that the predictions match the observed scale-up behavior of the GA, thereby strengthening our claim that the design rules can guide the design of competent selecto-recombinative GAs for realistic problems.


Preclinical Development and Assessment of Viral Vectors Expressing a Fusion Antigen of Plasmodium falciparum LSA1 and LSAP2 for Efficacy against Liver-Stage Malaria.
Despite promising progress in malaria vaccine development in recent years, an efficacious subunit vaccine against Plasmodium falciparum remains to be licensed and deployed. Cell-mediated protection from liver-stage malaria relies on a sufficient number of antigen-specific T cells reaching the liver during the time that parasites are present. A single vaccine expressing two antigens could potentially increase both the size and breadth of the antigen-specific response while halving vaccine production costs. In this study, we investigated combining two liver-stage antigens, P. falciparum LSA1 (PfLSA1) and PfLSAP2, and investigated the induction of protective efficacy by coadministration of single-antigen vectors or vaccination with dual-antigen vectors, using simian adenovirus and modified vaccinia virus Ankara vectors. The efficacy of these vaccines was assessed in mouse malaria challenge models using chimeric P. berghei parasites expressing the relevant P. falciparum antigens and challenging mice at the peak of the T cell response. Vaccination with a combination of the single-antigen vectors expressing PfLSA1 or PfLSAP2 was shown to improve protective efficacy compared to vaccination with each single-antigen vector alone. Vaccination with dual-antigen vectors expressing both PfLSA1 and PfLSAP2 resulted in responses to both antigens, particularly in outbred mice, and most importantly, the efficacy was equivalent to that of vaccination with a mixture of single-antigen vectors. Based on these promising data, dual-antigen vectors expressing PfLSA1 and PfLSAP2 will now proceed to manufacturing and clinical assessment under good manufacturing practice (GMP) guidelines.


Risk factors for in-hospital post-hip fracture mortality.
Approximately 10% of hip fracture patients die during hospitalization; however, it is not clear what risk factors contribute to the excess mortality. This study sought to examine risk factors of, and to develop prognostic model for, predicting in-hospital mortality among hip fracture patients. We studied outcomes among 410 men and 1094 women with a hip fracture who were admitted to a major-teaching-hospital in Sydney (Australia) between 1997 and 2007. Clinical data, including concomitant illnesses, were obtained from inpatient data. The primary outcome of the study was in-hospital mortality regardless of length of stay. A Log-binomial regression model was used to identify risk factors for in-hospital mortality. Using the identified risk factors, prognostic nomograms were developed for predicting short term risk of mortality for an individual. The median duration of hospitalization was 9 days. During hospitalization, the risk of mortality was higher in men (9%) than in women (4%). After adjusting for multiple risk factors, increased risk of in-hospital mortality was associated with advancing age (rate ratio [RR] for each 10-year increase in age: 1.91 95% confidence interval [CI]: 1.47 to 2.49), in men (RR 2.13; 95% CI 1.41 to 3.22), and the presence of comorbid conditions on admission (RR for one or more comorbid conditions vs. none: 2.30; 95% CI 1.52 to 3.48). Specifically, the risk of mortality was increased in patients with a pre-existing congestive heart failure (RR 3.02; 95% CI: 1.65 to 5.54), and liver disease (RR 4.75; 95% CI: 1.87 to 12.1). These factors collectively accounted for 69% of the risk for in-hospital mortality. A nomogram was developed from these risk factors to individualize the risk of in-hospital death following a hip fracture. The area under the receiver operating characteristic curve of the final model containing age, sex and comorbid conditions was 0.76. These data suggest that among hip fracture patients, advancing age, gender (men), and pre-existing concomitant diseases such as congestive heart failure and liver disease were the main risk factors for in-hospital mortality. The nomogram developed from this study can be used to convey useful prognostic information to help guide treatment decisions.


Frequent visitors at the psychiatric emergency room - A literature review.
Frequent visitors at the psychiatric emergency room (PER) constitute a small subgroup of patients, yet they are responsible for a disproportionate number of visits and thus claim considerable resources. Their needs are often left unmet and their repetitive visits reflect their dissatisfaction as well as that of PERs' staff. Motivated by these dilemmas, this study systematically reviews the literature about frequent visitors at PER and seeks to answer two questions: What characterizes frequent visitors at PER in the literature? and What characterizes PER in the literature? Based on 29 studies, this paper offers answers to the two questions based on a strength weakness opportunities and threats (SWOT) analysis. The results of the review and subsequent analysis of the literature revealed the multiplicity and complexity of frequent visitors' characteristics and how they appear to converge. Commonalities were more difficult to identify in PER characteristics. In some cases, this happened because the characteristics were poorly described or were context specific. As a result, it was not easy to compare the studies on PER. Based on SWOT and the findings of the analysis, the paper proposes new venues of research and suggests how the field of mental health might develop by taking into account its opportunities and threats.


Posttranscriptional regulation of chicken ccn2 gene expression by nucleophosmin/B23 during chondrocyte differentiation.
CCN2/CTGF is a multifunctional factor that plays a crucial role in the growth and differentiation of chondrocytes. The chicken ccn2 gene is regulated not only at the transcriptional level but also by the interaction between a posttranscriptional element in the 3' untranslated region (3'-UTR) and a cofactor. In the present study, we identified a nucleophosmin (NPM) (also called B23) as this cofactor. Binding of NPM to the element was confirmed, and subsequent analysis revealed a significant correlation between the decrease in cytosolic NPM and the increased stability of the ccn2 mRNA during chondrocyte differentiation in vivo. Furthermore, recombinant chicken NPM enhanced the degradation of chimeric RNAs containing the posttranscriptional cis elements in a chicken embryonic fibroblast extract in vitro. It is noteworthy that the RNA destabilization effect by NPM was far more prominent in the cytosolic extract of chondrocytes than in that of fibroblasts, representing a chondrocyte-specific action of NPM. Stimulation by growth factors to promote differentiation changed the subcellular distribution of NPM in chondrocytes, which followed the expected patterns from the resultant change in the ccn2 mRNA stability. Therefore, the present study reveals a novel aspect of NPM as a key player in the posttranscriptional regulation of ccn2 mRNA during the differentiation of chondrocytes.


Relationship of atrial fibrillation and stroke after coronary artery bypass graft surgery: when is anticoagulation indicated?
Atrial fibrillation (AF) is considered as a risk factor for stroke after coronary artery bypass grafting operations. A retrospective search in our hospital's medical record database was done to identify patients with postoperative strokes who underwent coronary artery bypass grafting operations from January 1, 1993, until December 31, 2004. All cases were individually reviewed, and the temporal relationship between neurologic event and postoperative episodes of AF was determined. During the study period it was our consistent policy to use only Coumadin anticoagulation limited to patients who had persistent AF or were to be discharged in AF. Of the 2,964 coronary artery bypass grafting operations, 576 patients (19.4%) had AF and 32 patients (1.1%) suffered stroke. Seventeen stroke patients maintained normal sinus rhythm during their hospital stay. Of the remaining 15 patients, 9 presented with neurologic deficit before the first episode of AF, with 5 having intraoperative and 4 having postoperative stroke. Of the 6 patients with AF before neurologic event, three strokes occurred within 1 week after spontaneous conversion to normal sinus rhythm. One patient with preoperative and also with intraoperative AF who underwent emergency coronary artery bypass grafting woke up with stroke. In the remaining two cases, the AF or atrial flutter episodes lasted less than 6 hours each before the neurologic event. More aggressive anticoagulation as suggested in the published guidelines could not have prevented strokes in any of these 6 patients. This retrospective analysis does not support the use of aggressive anticoagulation, particularly full intravenous heparinization as a bridging therapy to decrease the already low incidence of postoperative strokes after routine coronary artery bypass grafting surgery.


Toxicity methods for CPPs.
CPPs have for numerous years been utilized as delivery vectors of various pharmaceutically interesting cargoes, both in vitro and in vivo. As CPPs are gradually approaching the bedsides, investigating toxicity associated with these highly interesting peptides becomes increasingly important and thorough initial assessment of cytotoxicity in vitro is a first step towards advancing these delivery vehicles in to the clinics. The present chapter describes protocols for four cytotoxicity assays in order to provide a toolbox for toxicity assessment of CPPs. The foci lie on membrane integrity (deoxyglucose leakage and propidium iodide assays) and cell viability (the MTT assay), but the chapter also provides a protocol for assessing an important parameter for future clinical applications, namely the hemolytic properties of CPPs.


Neurostimulation in People with Oropharyngeal Dysphagia: A Systematic Review and Meta-Analysis of Randomised Controlled Trials-Part II: Brain Neurostimulation.
Objective. To assess the effects of brain neurostimulation (i.e., repetitive transcranial magnetic stimulation [rTMS] and transcranial direct current stimulation [tDCS]) in people with oropharyngeal dysphagia (OD). Methods. Systematic literature searches were conducted in four electronic databases (CINAHL, Embase, PsycINFO, and PubMed) to retrieve randomised controlled trials (RCTs) only. Using the Revised Cochrane risk-of-bias tool for randomised trials (RoB 2), the methodological quality of included studies was evaluated, after which meta-analysis was conducted using a random-effects model. Results. In total, 24 studies reporting on brain neurostimulation were included: 11 studies on rTMS, 9 studies on tDCS, and 4 studies on combined neurostimulation interventions. Overall, within-group meta-analysis and between-group analysis for rTMS identified significant large and small effects in favour of stimulation, respectively. For tDCS, overall within-group analysis and between-group analysis identified significant large and moderate effects in favour of stimulation, respectively. Conclusion. Both rTMS and tDCS show promising effects in people with oropharyngeal dysphagia. However, comparisons between studies were challenging due to high heterogeneity in stimulation protocols and experimental parameters, potential moderators, and inconsistent methodological reporting. Generalisations of meta-analyses need to be interpreted with care. Future research should include large RCTs using standard protocols and reporting guidelines as achieved by international consensus.


TRPC Channels and Mental Disorders.
Transient receptor potential canonical (TRPC) channels mediate the influx of different types of cations through the cell membrane and are involved in many functions of the organism. Evidences of involvement of TRPC channels in neuronal development suggest that this family of proteins might play a role in certain neurological disorders. As reported, knockout mice for different TRPC channels show alterations in neuronal morphological and functional parameters, with behavioral abnormalities, such as in exploratory and social behaviors. Although mutations in TRPC channels could be related to mental/neurological disorders, there are only a few cases reported in literature, indicating that this correlation should be further explored. Nonetheless, other functional evidences support the implication of these channels in neurological diseases. In this chapter, we summarize the main findings relating TRPC channels to neurological disorders, such as autism spectrum disorders, bipolar disorder, and intellectual disability among others.


Recovery of both vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus from culture of a single clinical specimen from colonized or infected patients.
To describe the incidence of recovery of both vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) from culture of a single clinical specimen, to describe the clinical characteristics of patients from whom these specimens were recovered, and to identify the risk factors of these patients. A retrospective cohort and case-control study. A tertiary care university hospital and referral center in Seoul, Korea. We identified 61 case patients for whom a single clinical specimen yielded both VRE and MRSA on culture, and 122 control patients for whom any clinical specimen yielded only VRE on culture. The control patients were selected by matching 2:1 with the case patients for age, sex, and first date of sampling that led to isolation of VRE or both VRE and MRSA among 1,536 VRE-colonized patients from January 1, 2003, through December 31, 2006. To identify patient risk factors for the recovery of both VRE and MRSA in a single clinical specimen, we performed univariate comparisons between the 2 groups and then multivariate logistic regression analysis. The incidence of recovery of both VRE and MRSA from culture of a single clinical specimen was 3.97% (for 61 of 1,536 VRE-colonized patients) over 4 years. Among these 82 single clinical specimens, the most common type was wound specimens (26.8%), followed by lower respiratory tract specimens (18.3%), urine specimens (17.1%), and catheter tips (15.9%). Of the 61 case patients, 14 (23.0%) had 2 or more single clinical specimens that yielded both VRE and MRSA on culture, and the longest interval from the first sampling that yielded both organisms to the last sampling that yielded both was 174 days. Independent patient risk factors for the presence of both VRE and MRSA in a single clinical specimen were chronic renal disease (odds ratio [OR], 7.00; P=.012 ), urinary catheterization (OR, 3.36; P=.026), and longer total cumulative duration of hospital stay within the previous year (OR, 1.03; P < .001). We confirmed that the recovery of VRE and MRSA from a single clinical specimen occurs continually. Because prolonged cell-to-cell contact can facilitate transfer of vanA, close observation and surveillance for vancomycin-resistant S. aureus, especially among patients with risk factors for the recovery of both VRE and MRSA from a single clinical specimen, should be continued.


A catalytic reactor for the organocatalyzed enantioselective continuous flow alkylation of aldehydes.
The use of immobilized metal-free catalysts offers the unique possibility to develop sustainable processes in flow mode. The challenging intermolecular organocatalyzed enantioselective alkylation of aldehydes was performed for the first time under continuous flow conditions. By using a packed-bed reactor filled with readily available supported enantiopure imidazolidinone, different aldehydes were treated with three distinct cationic electrophiles. In the organocatalyzed α-alkylation of aldehydes with 1,3-benzodithiolylium tetrafluoroborate, excellent enantioselectivities, in some cases even better than those obtained in the flask process (up to 95% ee at 25 °C), and high productivity (more than 3800 h(-1) ) were obtained, which thus shows that a catalytic reactor may continuously produce enantiomerically enriched compounds. Treatment of the alkylated products with Raney-nickel furnished enantiomerically enriched α-methyl derivatives, key intermediates for active pharmaceutical ingredients and natural products.


Refined Ordovician timescale reveals no link between asteroid breakup and biodiversification.
The catastrophic disruption of the L chondrite parent body in the asteroid belt c. 470 Ma initiated a prolonged meteorite bombardment of Earth that started in the Ordovician and continues today. Abundant L chondrite meteorites in Middle Ordovician strata have been interpreted to be the consequence of the asteroid breakup event. Here we report a zircon U-Pb date of 467.50±0.28 Ma from a distinct bed within the meteorite-bearing interval of southern Sweden that, combined with published cosmic-ray exposure ages of co-occurring meteoritic material, provides a precise age for the L chondrite breakup at 468.0±0.3 Ma. The new zircon date requires significant revision of the Ordovician timescale that has implications for the understanding of the astrogeobiologic development during this period. It has been suggested that the Middle Ordovician meteorite bombardment played a crucial role in the Great Ordovician Biodiversification Event, but this study shows that the two phenomena were unrelated.


X-ray Scattering and Coarse-Grained Simulations for Clustering and Interactions of Monoclonal Antibodies at High Concentrations.
Attractive protein?protein interactions (PPI) in concentrated monoclonal antibody (mAb) solutions may lead to reversible oligomers (clusters) that impact colloidal stability and viscosity. Herein, the PPI are tuned for two mAbs via the addition of arginine (Arg), NaCl, or ZnSO4 as characterized by the structure factor ( Seff( q)) with small-angle X-ray scattering (SAXS). The SAXS data are fit with molecular dynamics simulations by placing a physically relevant short-range attractive interaction on selected beads in coarse-grained 12-bead models of the mAb shape. The optimized 12-bead models are then used to differentiate key microstructural properties, including center of mass radial distribution functions ( gCOM( r)), coordination numbers, and cluster size distributions (CSD). The addition of cosolutes results in more attractive Seff( q) relative to the no cosolute control for all systems tested, with the most attractive systems showing an upturn at low q. Only the All1 model with an attractive site in each Fab and Fc region (possessing Fab?Fab, Fab?Fc, and Fc?Fc interactions) can reproduce this upturn, and the corresponding CSDs show the presence of larger clusters compared to the control. In general, for models with similar net attractions, i.e., second osmotic virial coefficients, the size of the clusters increases as the attraction is concentrated on a smaller number of evenly distributed beads. The cluster size distributions from simulations are used to improve the understanding and prediction of experimental viscosities. The ability to discriminate between models with bead interactions at particular Fab and Fc bead sites from SAXS simulations, and to provide real-space properties (CSD and gCOM( r)), will be of interest in engineering protein sequence and formulating protein solutions for weak PPI to minimize aggregation and viscosities.


Nonlinear transmission properties of hydrogenated amorphous silicon core optical fibers.
The nonlinear properties of a low loss hydrogenated amorphous silicon core fiber have been characterized for transmission of high power pulses at 1540 nm. Numerical modelling of the pulse propagation in the amorphous core material was used to establish the two-photon absorption, free-carrier absorption and the nonlinear refractive index, which were found to be larger than the values typical for crystalline silicon. Calculation of a nonlinear figure of merit demonstrates the potential for these hydrogenated amorphous silicon core fibers to be used in nonlinear silicon photonics applications.