Comparative physical, mechanical and crystallographic properties of a series of gemfibrozil salts.
Understanding the impact of the counterion on the properties of an acidic or basic drug may influence the choice of salt form, especially for less potent drugs with a high drug load per unit dose. The aim of this work was to determine the influence of the hydrogen bonding potential of the counterion on the crystal structure of salts of the poorly soluble, poorly compressible, acidic drug gemfibrozil and to correlate these with mechanical properties. Compacts of the parent drug and the salts were used to determine Young's modulus of elasticity using beam bending tests. Crystal structures were determined previously from X-ray powder diffraction data. The free acid, tert-butylamine, 2-amino-2-methylpropan-1-ol and 2-amino-2-methylpropan-1,3-diol salts had a common crystal packing motif of infinite hydrogen-bonded chains with cross-linking between pairs of adjacent chains. The tromethamine (trsi) salt, with different mechanical properties, had a two-dimensional sheet-like network of hydrogen bonds, with slip planes, forming a stiffer compact. The type of counter ion is important in determining mechanical properties and could be selected to afford slip and plastic deformation.


A role for the human DNA repair enzyme HAP1 in cellular protection against DNA damaging agents and hypoxic stress.
The HAP1 protein (also known as APE/Ref-1) is a bifunctional human nuclear enzyme required for repair of apurinic/apyrimidinic sites in DNA and reactivation of oxidized proto-oncogene products. To gain insight into the biological roles of HAP1, the effect of expressing antisense HAP1 RNA in HeLa cells was determined. The constructs for antisense RNA expression consisted of either a full-length HAP1 cDNA or a genomic DNA fragment cloned downstream of the CMV promoter in pcDNAneo. Stable HeLa cell transfectants expressing HAP1 antisense RNA were found to express greatly reduced levels of the HAP1 protein compared to equivalent sense orientation and vector-only control transfectants. The antisense HAP1 transfectants exhibited a normal growth rate, cell morphology and plating efficiency, but were hypersensitive to killing by a wide range of DNA damaging agents, including methyl methanesulphonate, hydrogen peroxide, menadione, and paraquat. However, survival after UV irradiation was unchanged. The antisense transfectants were strikingly sensitive to changes in oxygen tension, exhibiting increased killing compared to controls following exposure to both hypoxia (1% oxygen) and hyperoxia (100% oxygen). Consistent with a requirement for HAP1 in protection against hypoxic stress, expression of the HAP1 protein was found to be induced in a time-dependent manner in human cells during growth under 1% oxygen. The possible involvement of a depletion of cellular glutathione being linked to the hypoxic stress-sensitive phenotype of the antisense HAP1 transfectants came from the finding that they also exhibited hypersensitivity to buthionine sulphoximine, an inhibitor of glutathione biosynthesis. We conclude that the HAP1 protein is a key factor in cellular protection against a wide variety of cellular stresses, including DNA damage and a change in oxygen tension.


Design, construction, and characterization of a second-generation DARP in library with reduced hydrophobicity.
Designed ankyrin repeat proteins (DARPins) are well-established binding molecules based on a highly stable nonantibody scaffold. Building on 13 crystal structures of DARPin-target complexes and stability measurements of DARPin mutants, we have generated a new DARPin library containing an extended randomized surface. To counteract the enrichment of unspecific hydrophobic binders during selections against difficult targets containing hydrophobic surfaces such as membrane proteins, the frequency of apolar residues at diversified positions was drastically reduced and substituted by an increased number of tyrosines. Ribosome display selections against two human caspases and membrane transporter AcrB yielded highly enriched pools of unique and strong DARPin binders which were mainly monomeric. We noted a prominent enrichment of tryptophan residues during binder selections. A crystal structure of a representative of this library in complex with caspase-7 visualizes the key roles of both tryptophans and tyrosines in providing target contacts. These aromatic and polar side chains thus substitute the apolar residues valine, leucine, isoleucine, methionine, and phenylalanine of the original DARPins. Our work describes biophysical and structural analyses required to extend existing binder scaffolds and simplifies an existing protocol for the assembly of highly diverse synthetic binder libraries.


Relationship between the Millon Clinical Multiaxial Inventory and the Millon-II: value of scales for Aggressive and Self-defeating personalities in Posttraumatic Stress Disorder.
This study addressed two issues, the interrelationship between the Millon Clinical Multiaxial Inventory (MCMI) and the Millon II (MCMI-II) and the value of the new personality scales, Aggressive and Self-defeating, in a sample with diagnoses of combat-related Posttraumatic Stress Disorder. 100 confirmed cases of combat-related Posttraumatic Stress Disorder were given a battery of measures including both Millon inventories and the Minnesota Multiphasic Personality Inventory (basic scales and selected subscales). They were rated on discharge status during a structured treatment program. Basic treatment and background information were also obtained. Analysis showed scores on the MCMI-II scales were higher but generally reflective of MCMI scales and that the Self-defeating personality style tends to be reflective of greater psychopathology, suicidal problems, treatment/disposition difficulties, overreporting of symptoms, and intensity of problems. Discussion encouraged the use of the MCMI-II with special emphasis given to the Self-defeating style in this group with Posttraumatic Stress Disorder.


[Improvements in the management of rheumatic patients from vertebral image obtained through dual-energy X-ray absorptiometry].
The diagnosis of asymptomatic vertebral fracture is clinically useful and the identification of new fractures may influences the choice of appropriate therapeutic measures. In order to identify moderate and asymptomatic vertebral deformities in an objective and reproducible manner, vertebral morphometry is performed. This method measures the vertebral body's anterior, middle and posterior heights at the dorsal and lumbar level. Currently this technique is performed on lateral images of the spine obtained through the traditional X-ray method (radiological morphometry or morphometric X-ray radiography, MRX) and, more recently from images obtained through dual-energy X-ray absorptiometry (DXA) machines (visual assessment of X-ray absorptiometry scans or morphometric X-ray absorptiometry, MXA), commonly used to measure bone mineral density. The main advantage of MXA relative to MRX is the lower radiation dose to which the patient is exposed during the exam. In addition, MXA scans offers the advantage of acquiring a single image of thoracic and lumbar spine, without any distortion (e.g.: coning). The most obvious advantage of MXA is the opportunity of obtaining during the same session a bone mineral density evaluation, and digital images that are easily processable, manageable, recordable and comparable for the patient's follow up. A limitation of the MXA technique is the inferior quality of the images, that make often impossible the detection of the vertebral edges, and the impossibility to visualize the upper thoracic vertebral bodies. MXA, despite its intrinsic limitations, when carried out by trained personnel may provide substantial improvements in the management (diagnosis and follow-up) of rheumatic patients.


"I Must Try Harder": Design Implications for Mobile Apps and Wearables Contributing to Self-Efficacy of Patients With Chronic Conditions.
Diverse wellness-promoting mobile health technologies, including mobile apps and wearable trackers, became increasingly popular due to their ability to support patients' self-management of health conditions. However, the patient's acceptance and use depend on the perceived experience and the app appropriateness to the patient's context and needs. We have some understating of the experience and factors influencing the use of these technologies in the general public, but we have a limited understanding of these issues in patients. By presenting results from an explorative study, this paper aims to identify implications for the design of mobile apps and wearables to effectively support patients' efforts in self-management of health with a special emphasis on support for self-efficacy of activities contributing to health. An explorative mixed-method study involving 200 chronically ill patients of Stanford Medical Center (Stanford, CA, United States) was conducted between mid-2016 and end of 2018. Amongst these, 20 patients were involved in a 4-weeks study, in which we collected the underlying wearable device use logs (e.g., Fitbit) and subjective use experience [via an Ecological Momentary Assessment (EMA)], as well as patients' momentary perception of general self-efficacy in their natural environments and different daily contexts. The results indicate that mobile apps for health and wearables have the potential to enable better self-management and improve patients' wellbeing but must be further refined to address different human aspects of their use. Specifically, the apps/wearables should be easier to use, more personalized and context-aware for the patient's overall routine and lifestyle choices, as well as with respect to the momentary patient state (e.g., location, type of people around) and health(care) needs. Additionally, apps and devices should be more battery efficient and accurate; providing timely, non-judgmental feedback and personalized advice to the patients anywhere-anytime-anyhow. These results are mapped on major sources of the individuals' self-efficacy. Our results show how the apps/wearables that are aimed at supporting the patients' self-management should be designed to leverage and further improve the patients' general self-efficacy and self-efficacy of activities contributing to chronic disease management.


Circumferential esophageal replacement using a tube-shaped tissue-engineered substitute: An experimental study in minipigs.
Esophageal replacement by the colon or the stomach for malignant and nonmalignant esophageal diseases exposes to significant morbidity and mortality. In this setting, tissue engineering seems to be a seductive alternative. In a porcine model, we performed a 5-cm-long circumferential replacement of the cervical esophagus by a tubulized acellular matrix (small intestinal submucosa) cellularized with autologous skeletal myoblasts and covered by a human amniotic membrane seeded with autologous oral epithelial cells. The substitute was grown for 2 weeks in the great omentum before esophageal replacement. Eighteen minipigs (divided into 3 groups: group A [substitute with esophageal endoprothesis; n = 6], group B [substitute alone; n = 6], and group C [endoprothesis alone; n = 6]) were included. The esophageal endoprothesis was removed at 6 months. Animals were killed sequentially over a 12 month-period. Clinical, endoscopic, radiologic and histologic outcomes were analyzed. All animals except 1 of in groups B and C died during the first 2 months owing to refractory esophageal stenosis or endoprothesis extrusion. Nutritional autonomy without endoprothesis was observed in all animals of group A with a follow-up of >6 months (n = 3). A phenotype similar to that of native esophagus, consisting of a mature epithelium, submucosal glands, and a circular muscular layer, was observed after 9 months. In this model, the circumferential replacement of the cervical esophagus by a tube-shaped tissue-engineered substitute under the temporary cover of an esophageal endoprothesis allowed nutritional autonomy and tissue remodeling toward an esophageal phenotype.


A molecular test based on RT-LAMP for rapid, sensitive and inexpensive colorimetric detection of SARS-CoV-2 in clinical samples.
Until there is an effective implementation of COVID-19 vaccination program, a robust testing strategy, along with prevention measures, will continue to be the most viable way to control disease spread. Such a strategy should rely on disparate diagnostic tests to prevent a slowdown in testing due to lack of materials and reagents imposed by supply chain problems, which happened at the beginning of the pandemic. In this study, we have established a single-tube test based on RT-LAMP that enables the visual detection of less than 100 viral genome copies of SARS-CoV-2 within 30 min. We benchmarked the assay against the gold standard test for COVID-19 diagnosis, RT-PCR, using 177 nasopharyngeal RNA samples. For viral loads above 100 copies, the RT-LAMP assay had a sensitivity of 100% and a specificity of 96.1%. Additionally, we set up a RNA extraction-free RT-LAMP test capable of detecting SARS-CoV-2 directly from saliva samples, albeit with lower sensitivity. The saliva was self-collected and the collection tube remained closed until inactivation, thereby ensuring the protection of the testing personnel. As expected, RNA extraction from saliva samples increased the sensitivity of the test. To lower the costs associated with RNA extraction, we performed this step using an alternative protocol that uses plasmid DNA extraction columns. We also produced the enzymes needed for the assay and established an in-house-made RT-LAMP test independent of specific distribution channels. Finally, we developed a new colorimetric method that allowed the detection of LAMP products by the visualization of an evident color shift, regardless of the reaction pH.


Pregabalin add-on for drug-resistant focal epilepsy.
Epilepsy is a common neurological disease that affects approximately 1% of the UK population. Approximately one-third of these people continue to have seizures despite drug treatment. Pregabalin is one of the newer antiepileptic drugs which have been developed to improve outcomes.This is an updated version of the Cochrane Review published in Issue 3, 2014, and includes three new studies. To assess the efficacy and tolerability of pregabalin when used as an add-on treatment for drug-resistant focal epilepsy. For the latest update we searched the Cochrane Register of Studies (CRS Web), which includes the Cochrane Epilepsy Group Specialized Register and the Cochrane Central Register of Controlled Trials (CENTRAL), on 5 July 2018, MEDLINE (Ovid, 1946 to 5 July 2018), ClinicalTrials.gov (5 July 2018), and the World Health Organization International Clinical Trials Registry Platform (ICTRP, 5 July 2018), and contacted Pfizer Ltd, manufacturer of pregabalin, to identify published, unpublished, and ongoing trials. We included randomised controlled trials comparing pregabalin with placebo or an alternative antiepileptic drug as an add-on for people of any age with drug-resistant focal epilepsy. Double-blind and single-blind trials were eligible for inclusion. The primary outcome was 50% or greater reduction in seizure frequency; secondary outcomes were seizure freedom, treatment withdrawal for any reason, treatment withdrawal due to adverse effects, and proportion of individuals experiencing adverse effects. Two review authors independently selected and assessed trials for eligibility and extracted data. Analyses were by intention-to-treat. We presented results as risk ratios (RR) and odds ratios (OR) with 95% confidence intervals (CIs). Two review authors assessed the included studies for risk of bias using the Cochrane 'Risk of bias' tool. We included nine industry-sponsored randomised controlled trials (3327 participants) in the review. Seven trials compared pregabalin to placebo. For the primary outcome, participants randomised to pregabalin were significantly more likely to attain a 50% or greater reduction in seizure frequency compared to placebo (RR 2.28, 95% CI 1.52 to 3.42, 7 trials, 2193 participants, low-certainty evidence). The odds of response doubled with an increase in dose from 300 mg/day to 600 mg/day (OR 1.99, 95% CI 1.74 to 2.28), indicating a dose-response relationship. Pregabalin was significantly associated with seizure freedom (RR 3.94, 95% CI 1.50 to 10.37, 4 trials, 1125 participants, moderate-certainty evidence). Participants were significantly more likely to withdraw from pregabalin treatment than placebo for any reason (RR 1.35, 95% CI 1.11 to 1.65, 7 trials, 2193 participants, moderate-certainty evidence) and for adverse effects (RR 2.65, 95% CI 1.88 to 3.74, 7 trials, 2193 participants, moderate-certainty evidence).Three trials compared pregabalin to three active-control drugs: lamotrigine, levetiracetam, and gabapentin. Participants allocated to pregabalin were significantly more likely to achieve a 50% or greater reduction in seizure frequency than those allocated to lamotrigine (RR 1.47, 95% CI 1.03 to 2.12, 1 trial, 293 participants) but not those allocated to levetiracetam (RR 0.94, 95% CI 0.80 to 1.11, 1 trial, 509 participants) or gabapentin (RR 0.96, 95% CI 0.82 to 1.12, 1 trial, 484 participants). We found no significant differences between pregabalin and lamotrigine (RR 1.39, 95% CI 0.40 to 4.83) for seizure freedom, however, significantly fewer participants achieved seizure freedom with add-on pregabalin compared to levetiracetam (RR 0.50, 95% CI 0.30 to 0.85). No data were reported for this outcome for pregabalin versus gabapentin. We found no significant differences between pregabalin and lamotrigine (RR 1.07, 95% CI 0.75 to 1.52), levetiracetam (RR 1.03, 95% CI 0.71 to 1.49), or gabapentin (RR 0.78, 95% CI 0.57 to 1.07) for treatment withdrawal due to any reason or due to adverse effects (pregabalin versus lamotrigine: RR 0.89, 95% CI 0.53 to 1.48; versus levetiracetam: RR 1.29, 95% CI 0.66 to 2.54; versus gabapentin: RR 1.07, 95% CI 0.54 to 2.11). Ataxia, dizziness, somnolence, weight gain, and fatigue were significantly associated with pregabalin.We rated the overall risk of bias in the included studies as low or unclear due to the possibility of publication bias and lack of methodological details provided. We rated the certainty of the evidence as very low to moderate using the GRADE approach. Pregabalin, when used as an add-on drug for treatment-resistant focal epilepsy, is significantly more effective than placebo at producing a 50% or greater seizure reduction and seizure freedom. Results demonstrated efficacy for doses from 150 mg/day to 600 mg/day, with increasing effectiveness at 600 mg doses, however issues with tolerability were noted at higher doses. The trials included in this review were of short duration, and longer-term trials are needed to inform clinical decision making.


Epstein-Barr virus LMP1 modulates the malignant potential of gastric carcinoma cells involving apoptosis.
About 10% of gastric carcinomas including lymphoepithelioma-like carcinoma and adenocarcinoma are associated with Epstein-Barr virus (EBV) infection. In EBV-associated gastric carcinomas, the tumor cells express Epstein-Barr nuclear antigen 1 (EBNA-1) but not EBNA-2, -3A, -3B, or -3C, leader protein, or latent membrane proteins (LMPs) because of gene methylation. Only a few exceptional cases have LMP1 expression in tumor cells as demonstrated by immunohistochemical studies. To elucidate the biological effects of LMP1 and the significance of its restricted expression in EBV-associated gastric carcinomas, the LMP1 gene was transferred into EBV-negative gastric carcinoma cell lines (SCM1 and TMC1) and into EBV-negative nasopharyngeal carcinoma (NPC) cells (HONE-1) as a control. The biological effects of LMP1 in gastric carcinoma cells were monitored in vitro and in vivo. These results showed that the consequence of LMP1 expression is a growth enhancement in NPC cells, but it is a growth suppression in gastric carcinoma cells. The LMP1-expressing gastric carcinoma cells had a reduced growth rate, colony-forming efficiency, mean colony size, and tumorigenicity and a lower malignant cytological grade. The reduced growth rate, colony-forming efficiency, and mean colony size were partially reversible in vitro with treatment with LMP1 antisense oligonucleotide. In addition, enhanced apoptosis was found in the LMP1-expressing gastric carcinoma cells. This suggests that LMP1 may negatively modulate the malignant potential of gastric carcinoma cells via an enhancement of apoptosis. We concluded that the restriction of LMP1 expression in EBV-associated gastric carcinomas may lead to a growth advantage for tumor cells by avoiding LMP1 apoptotic effects and immunologically mediated elimination.


Tumor margin histology predicts tumor aggressiveness in papillary thyroid carcinoma: a study of 514 consecutive patients.
Histologic patterns at tumor margins may be related to prognosis in several malignancies. We investigated tumor aggressiveness with respect to tumor margin histology in patients with papillary thyroid carcinoma (PTC). Five hundred fourteen consecutive patients who underwent surgery for primary PTC between January and July 2009 were assigned to two groups, one with an infiltrative pattern (I-type, n = 347) at tumor margins and one with an expanding pattern (E-type, n = 167). Tumor aggressiveness was assessed by analyzing relationships between these patterns and known prognostic factors. The analysis showed that unfavorable prognostic factors such as tumor multiplicity (P = 0.002), extrathyroidal extension (P < 0.001), lateral neck lymph node metastasis (P < 0.001) and advanced TNM stage (P = 0.001) were significantly more prevalent in patients with I-type PTC than in those with the E-type. Central neck node metastases were more prevalent without statistical significance in the I-type patients (P = 0.376). Tumor margin histology was not related to gender or tumor size. These results suggest that histologic patterns at tumor margins predict aggressiveness in PTC.


Protein kinase C dependent inhibition of the heteromeric Kir4.1-Kir5.1 channel.
Heteromultimerization of Kir4.1 and Kir5.1 leads to a channel with distinct functional properties. The heteromeric Kir4.1-Kir5.1 channel is expressed in the eye, kidney and brainstem and has CO(2)/pH sensitivity in the physiological range, suggesting a candidate molecule for the regulation of K(+) homeostasis and central CO(2) chemoreception. It is known that K(+) transport in renal epithelium and brainstem CO(2) chemosensitivity are subject to modulation by hormones and neurotransmitters that activate distinct intracellular signaling pathways. If the Kir4.1-Kir5.1 channel is involved in pH-dependent regulation of cellular functions, it may also be regulated by some of the intracellular signaling systems. Therefore, we undertook studies to determine whether PKC modulates the heteromeric Kir4.1-Kir5.1 channel. The channel expressed using a Kir4.1-Kir5.1 tandem dimer construct was inhibited by the PKC activator PMA in a dose-dependent manner. The channel inhibition was produced via reduction of the P(open). The effect of PMA was abolished by specific PKC inhibitors. In contrast, exposure of oocytes to forskolin (a PKA activator) had no significant effect on Kir4.1-Kir5.1 currents. The channel inhibition appeared to be independent of PIP(2) depletion and PKC-dependent internalization. Several consensus sequences of potential PKC phosphorylation sites were identified in the Kir4.1 and Kir5.1 subunits by sequence scan. Although the C-terminal peptides of both Kir4.1 and Kir5.1 were phosphorylated in vitro, site-directed mutagenesis of individual residues failed to reveal the PKC phosphorylation sites suggesting that the channel may have multiple phosphorylation sites. Taken together, these results suggest that the Kir4.1-Kir5.1 but not the homomeric Kir4.1 channel is strongly inhibited by PKC activation.


ITS2 Database V: Twice as Much.
The internal transcribed spacer 2 (ITS2) is a well-established marker for phylogenetic analyses in eukaryotes. A reliable resource for reference sequences and their secondary structures is the ITS2 database (http://its2.bioapps.biozentrum.uni-wuerzburg.de/). However, the database was last updated in 2011. Here, we present a major update of the underlying data almost doubling the number of entities. This increases the number of taxa represented within all major eukaryotic clades. Moreover, additional data has been added to underrepresented groups and some new groups have been added. The broader coverage across the tree of life improves phylogenetic analyses and the capability of ITS2 as a DNA barcode.


Surgical site infection prevention: time to move beyond the surgical care improvement program.
The objective of this study was to evaluate whether the Surgical Care Improvement Project (SCIP) improved surgical site infection (SSI) rates using national data at the patient level for both SCIP adherence and SSI occurrence. The SCIP was established in 2006 with the goal of reducing surgical complications by 25% in 2010. National Veterans' Affairs (VA) data from 2005 to 2009 on adherence to 5 SCIP SSI prevention measures were linked to Veterans' Affairs Surgical Quality Improvement Program SSI outcome data. Effect of SCIP adherence and year of surgery on SSI outcome were assessed with logistic regression using generalized estimating equations, adjusting for procedure type and variables known to predict SSI. Correlation between hospital SCIP adherence and SSI rate was assessed using linear regression. There were 60,853 surgeries at 112 VA hospitals analyzed. SCIP adherence ranged from 75% for normothermia to 99% for hair removal and all significantly improved over the study period (P < 0.001). Surgical site infection occurred after 6.2% of surgeries (1.6% for orthopedic surgeries to 11.3% for colorectal surgeries). None of the 5 SCIP measures were significantly associated with lower odds of SSI after adjusting for variables known to predict SSI and procedure type. Year was not associated with SSI (P = 0.71). Hospital SCIP performance was not correlated with hospital SSI rates (r = -0.06, P = 0.54). Adherence to SCIP measures improved whereas risk-adjusted SSI rates remained stable. SCIP adherence was neither associated with a lower SSI rate at the patient level, nor associated with hospital SSI rates. Policies regarding continued SCIP measurement and reporting should be reassessed.


[Autism spectrum disorder and substance use disorder: an unknown comorbidity?].
We describe the diagnosis and treatment of a patient with autism spectrum disorder (ASD) and substance use disorder (SUD). The patient had been given the dual diagnosis of Asperger's disorder and SUD. On that basis the approach adopted and the treatment provided seemed appropriate. Little is known about the comorbidity of ASD and SUD and it has hardly been reported in the scientific literature or in clinical practice. Epidemiological research shows that this dual diagnosis occurs in clinical populations. A careful psychiatric diagnosis is important in order to ensure that the comorbidity is recognised at an early stage and treated in an appropriate manner.


Posttraumatic stress in intensive care unit survivors - a prospective study.
Aims: This study aimed to estimate the prevalence of severe Posttraumatic Stress Disorder (PTSD) symptoms and to identify factors associated with PTSD in survivors of intensive care unit (ICU) treatment following traumatic injury. Methods: Fifty-two patients who were admitted to an ICU through the emergency ward following traumatic injury were prospectively followed. Information on injury severity and ICU treatment were obtained through medical records. Demographic information and measures of acute stress symptoms, experienced social support, coping style, sense of coherence (SOC) and locus of control were assessed within one-month post-accident (T1). At the six months follow-up (T2), PTSD was assessed with the Harvard Trauma Questionnaire (HTQ). Results: In the six months follow-up, 10 respondents (19.2%) had HTQ total scores reaching a level suggestive of PTSD (N = 52), and 11 respondents (21%) had symptom levels indicating subclinical PTSD. Female, five illness factors: coma time, mechanical ventilation, sedation, benzodiazepine, pain relieving medication, and four psychological factors: symptoms of acute stress (T1), fear of death and/or feeling completely helpless and powerless in relation to the accident and/or ICU (T1), SOC (T1) and more external locus of control (T1) correlated significantly with PTSD symptoms at T2. In the linear regression analysis, female, length of sedation, dissociation (T1), hypervigilance (T1), and external locus of control predicted 58% of the variation of PTSD. Conclusions: High levels of PTSD symptoms occurred in 19.2% of respondents in six months following traumatic injury requiring ICU admission. Screening for the variables gender, length of sedation, dissociation, hypervigilance, and locus of control after ICU admission following traumatic injuries may help to predict who will develop PTSD.


Phosphorylation of AMPA receptor subunits is differentially regulated by phospholipase A2 inhibitors.
Our laboratory recently discovered that the phosphorylation of subunits forming the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) subtype of glutamate receptors is regulated by constitutive phospholipase A2 (PLA2) activity in rat brain sections. In the present investigation, arachidonyl trifluoromethyl ketone (AACOCF3) and bromoenol lactone (BEL) were used to compare the influence of calcium-dependent (cPLA2) and calcium-independent (iPLA2) enzymes on phosphorylation of AMPA and N-methyl-D-aspartate (NMDA) subtypes of glutamate receptors. Incubation of rat brain sections with 3 microM BEL enhanced phosphorylation on the serine (Ser) 831 residue of the AMPA receptor GluR1 subunit in synaptosomal P2 fractions, whereas AACOCF3 at the same concentration resulted in increased phosphorylation on residues Ser880/891 of GluR2/3 subunits. These effects were restricted to the AMPA receptor subtype as no changes in phosphorylation were elicited on the NMDA receptor NR1 subunit. The effects of BEL and AACOCF3 were not occluded during blockade of protein phosphatases since AMPA receptor phosphorylation was still apparent in the presence of okadaic acid, indicating that the PLA2 inhibitor-induced increase in AMPA receptor phosphorylation does not rely on a decrease in dephosphorylation reactions. However, pretreatment of rat brain sections with a cell-permeable protein kinase C (PKC) inhibitor prevented BEL- and AACOCF3-induced phosphorylation on the Ser831 and Ser880/891 sites of GluR1 and GluR2/3 subunits, respectively. These results suggest that constitutive cPLA2 and iPLA2 systems may differentially influence AMPA receptor properties and function in the rat brain through mechanisms involving PKC activity.


An N-terminal partial sequence of the 13 kDa Pycnopodia helianthoides sperm chemoattractant 'startrak' possesses sperm-attracting activity.
Freshwater extracts of starfish ovaries were used to purify the sperm-attracting peptide 'startrak' from Pycnopodia helianthoides using hydrophobic interaction chromatography and DEAE-high-pressure liquid chromatography. Partially purified attractant had a molecular mass of 13 kDa, estimated from gel filtration and polyacrylamide gel electrophoresis results. The purified attractant was subjected to amino acid analysis and direct sequencing, and was found to consist largely of a single peptide composed of an estimated 127 residues based on a molecular mass of 13kDa. An N-terminal sequence of amino acids from positions 3 to 34 was obtained and synthesized as: NH2-Ala-Glu-Leu-Gly-Leu-Cys-Ile-Ala-Arg-Val-Arg-Gln-Gln-Asn-Gln-Gly-Gln- Asp-Asp-Val-Ser-Ile-Tyr-Gln-Ala-Ile-Met-Ser-Gln-Cys-Gln-Ser-COOH. The synthetic peptide possessed sperm-attracting activity 130 times greater than the activity of partially purified startrak and showed a pattern of species-specificity of sperm chemotaxis similar to that of startrak. Antibody prepared against synthetic peptide removed the sperm-attracting activity from crude and partially purified preparations of startrak. The partial sequence of startrak was not homologous with that of any of the known echinoid sperm motility-activating peptides.


Factors that facilitate or constrain the use of continuous sedation at the end of life by physicians and nurses in Belgium: results from a focus group study.
Continuous sedation at the end of life (CS) is the practice whereby a physician uses sedatives to reduce or take away a patient's consciousness until death. Although the incidence of CS is rising, as of yet little research has been conducted on how the administration of CS is experienced by medical practitioners. Existing research shows that many differences exist between medical practitioners regarding how and how often they perform CS. We conducted a focus group study to find out which factors may facilitate or constrain the use of continuous sedation by physicians and nurses. The participants often had clear ideas on what could affect the likelihood that sedation would be used. The physicians and nurses in the focus groups testified that the use of continuous sedation was facilitated in cases where a patient has a very limited life expectancy, suffers intensely, makes an explicit request and has family members who can cope with the stress that accompanies sedation. However, this 'paradigm case' was considered to occur only rarely. Furthermore, deviations from the paradigm case were said to be sometimes due to physicians initiating the discussion on CS too late or not initiating it at all for fear of inducing the patient. Deviations from the paradigm case may also occur when sedation proves to be too difficult for family members who are said to sometimes pressure the medical practitioners to increase dosages and speed up the sedation.


Transport of phagosomes in mouse peritoneal macrophages.
Mouse macrophages were elicited by the peritoneal injection of chondroitin sulfate solution, harvested and purified, and used as experimental materials. Small and large (diameter: 0.9 microns and 3.0 microns, respectively) polystyrene beads (PB) were used as ingested particles. When the macrophages were incubated with Hank's solution containing small or large PB for 30 min, the phagosomes containing small or large PB were usually randomly distributed. When the macrophages were further incubated for 45 min in PB-free medium, both small and large phagosomes containing PB accumulated at the perinuclear region. The transport of large phagosomes containing 3.0 microns PB was inhibited by cytochalasin B, but not by vinblastine or podophyllotoxin. Conversely, the transport of small phagosomes containing 0.9 microns PB was not inhibited by cytochalasin B but was inhibited by vinblastine or podophyllotoxin. Immunofluorescence microscopy showed that the small phagosomes appeared to accumulate at the central region of the microtubule network. The large phagosomes, on the other hand, appeared to be surrounded by actin-rich cytoplasm, and in some cells actin filament-like structures could be seen around large phagosomes. These results suggest that there are two different transport systems of phagosomes in macrophages. Phagosomes smaller than 0.9 microns in diameter are, probably, mainly transported to the perinuclear region by a microtubule-based motility system and those larger than 3.0 microns in diameter by an actin-based mechanism. It was observed electron-microscopically that accumulated phagosomes containing PB could fuse with each other and form larger phagosomes.


Epitope discovery with phylogenetic hidden Markov models.
Existing methods for the prediction of immunologically active T-cell epitopes are based on the amino acid sequence or structure of pathogen proteins. Additional information regarding the locations of epitopes may be acquired by considering the evolution of viruses in hosts with different immune backgrounds. In particular, immune-dependent evolutionary patterns at sites within or near T-cell epitopes can be used to enhance epitope identification. We have developed a mutation-selection model of T-cell epitope evolution that allows the human leukocyte antigen (HLA) genotype of the host to influence the evolutionary process. This is one of the first examples of the incorporation of environmental parameters into a phylogenetic model and has many other potential applications where the selection pressures exerted on an organism can be related directly to environmental factors. We combine this novel evolutionary model with a hidden Markov model to identify contiguous amino acid positions that appear to evolve under immune pressure in the presence of specific host immune alleles and that therefore represent potential epitopes. This phylogenetic hidden Markov model provides a rigorous probabilistic framework that can be combined with sequence or structural information to improve epitope prediction. As a demonstration, we apply the model to a data set of HIV-1 protein-coding sequences and host HLA genotypes.


Extent of implementation of food environment policies by the Malaysian Government: gaps and priority recommendations.
To determine the degree of food environment policies that have been implemented and supported by the Malaysian Government, in comparison to international best practice, and to establish prioritised recommendations for the government based on the identified implementation gaps. The Healthy Food-Environment Policy Index (Food-EPI) comprises forty-seven indicators of government policy practice. Local evidence of each indicator was compiled from government institutions and verified by related government stakeholders. The extent of implementation of the policies was rated by experts against international best practices. Rating results were used to identify and propose policy actions which were subsequently prioritised by the experts based on 'importance' and 'achievability' criteria. The policy actions with relatively higher 'achievability' and 'importance' were set as priority recommendations for government action. Malaysia. Twenty-six local experts. Majority (62 %) of indicators was rated 'low' implementation with no indicator rated as either 'high' or 'very little, if any' in terms of implementation. The top five recommendations were (i) restrict unhealthy food marketing in children's settings and (ii) on broadcast media; (iii) mandatory nutrition labelling for added sugars; (iv) designation of priority research areas related to obesity prevention and diet-related non-communicable diseases; and (v) introduce energy labelling on menu boards for fast-food outlets. This first policy study conducted in Malaysia identified a number of gaps in implementation of key policies to promote healthy food environments, compared with international best practices. Study findings could strengthen civil society advocacies for government accountability to create a healthier food environment.


Treatment of 153 Japanese patients with Q-switched alexandrite laser.
We have recently used Q-switched alexandrite laser for the treatment of various kinds of pigmented skin lesions. We retrospectively compared therapeutic outcomes of 153 Japanese patients who consulted our department. This approach was not very efficient for nevus spilus/café-au-lait spots, which seemed laser-resistant, especially when the pigmentation had appeared after 1 year of age, was treated after 5 years of age, was located on the face, was oval with a smooth border, and the patient was male. This approach was equally effective for senile lentigo, nevus of Ota, and Mongolian spots, but less effective for acquired bilateral nevus of Ota-like macules. Some patients with sacral Mongolian spots or those with light-colored, senile lentigo developed severe post-inflammatory hyperpigmentation after treatment. As a whole, good therapeutic outcome was achieved after multiple treatment sessions. However, the use of other lasers or other treatment modalities should be considered to treat nevus spilus/café-au-lait spots.


Ensemble learning with active example selection for imbalanced biomedical data classification.
In biomedical data, the imbalanced data problem occurs frequently and causes poor prediction performance for minority classes. It is because the trained classifiers are mostly derived from the majority class. In this paper, we describe an ensemble learning method combined with active example selection to resolve the imbalanced data problem. Our method consists of three key components: 1) an active example selection algorithm to choose informative examples for training the classifier, 2) an ensemble learning method to combine variations of classifiers derived by active example selection, and 3) an incremental learning scheme to speed up the iterative training procedure for active example selection. We evaluate the method on six real-world imbalanced data sets in biomedical domains, showing that the proposed method outperforms both the random under sampling and the ensemble with under sampling methods. Compared to other approaches to solving the imbalanced data problem, our method excels by 0.03-0.15 points in AUC measure.


A threat in the air. How stereotypes shape intellectual identity and performance.
A general theory of domain identification is used to describe achievement barriers still faced by women in advanced quantitative areas and by African Americans in school. The theory assumes that sustained school success requires identification with school and its subdomains; that societal pressures on these groups (e.g., economic disadvantage, gender roles) can frustrate this identification; and that in school domains where these groups are negatively stereotyped, those who have become domain identified face the further barrier of stereotype threat, the threat that others' judgments or their own actions will negatively stereotype them in the domain. Research shows that this threat dramatically depresses the standardized test performance of women and African Americans who are in the academic vanguard of their groups (offering a new interpretation of group differences in standardized test performance), that it causes disidentification with school, and that practices that reduce this threat can reduce these negative effects.


Classification and prediction of survival in patients with the leukemic phase of cutaneous T cell lymphoma.
We have used cDNA arrays to investigate gene expression patterns in peripheral blood mononuclear cells from patients with leukemic forms of cutaneous T cell lymphoma, primarily Sezary syndrome (SS). When expression data for patients with high blood tumor burden (Sezary cells >60% of the lymphocytes) and healthy controls are compared by Student's t test, at P < 0.01, we find 385 genes to be differentially expressed. Highly overexpressed genes include Th2 cells-specific transcription factors Gata-3 and Jun B, as well as integrin beta1, proteoglycan 2, the RhoB oncogene, and dual specificity phosphatase 1. Highly underexpressed genes include CD26, Stat-4, and the IL-1 receptors. Message for plastin-T, not normally expressed in lymphoid tissue, is detected only in patient samples and may provide a new marker for diagnosis. Using penalized discriminant analysis, we have identified a panel of eight genes that can distinguish SS in patients with as few as 5% circulating tumor cells. This suggests that, even in early disease, Sezary cells produce chemokines and cytokines that induce an expression profile in the peripheral blood distinctive to SS. Finally, we show that using 10 genes, we can identify a class of patients who will succumb within six months of sampling regardless of their tumor burden.


Breast cancer resistance protein (ABCG2) and drug disposition: intestinal expression, polymorphisms and sulfasalazine as an in vivo probe.
Breast cancer resistance protein (BCRP) is an efflux transporter expressed in tissues that act as barriers to drug entry. Given that single nucleotide polymorphisms (SNPs) in the ABCG2 gene encoding BCRP are common, the possibility exists that these genetic variants may be a determinant of interindividual variability in drug response. The objective of this study is to confirm the human BCRP-mediated transport of sulfasalazine in vitro, evaluate interindividual variation in BCRP expression in human intestine and to determine the role of ABCG2 SNPs to drug disposition in healthy patients using sulfasalazine as a novel in vivo probe. To evaluate these objectives, pinch biopsies were obtained from 18 patients undergoing esophagogastro-duodenoscopy or colonoscopy for determination of BCRP expression in relation to genotype. Wild-type and variant BCRP were expressed in a heterologous expression system to evaluate the effect of SNPs on cell-surface trafficking. A total of 17 healthy individuals participated in a clinical investigation to determine the effect of BCRP SNPs on sulfasalazine pharmacokinetics. In vitro, the cell surface protein expression of the common BCRP 421 C>A variant was reduced in comparison with the wild-type control. Intestinal biopsy samples revealed that BCRP protein and mRNA expression did not significantly differ between patients with 34GG/421CC versus patients with 34GG/421CA genotypes. Remarkably, in subjects with 34GG/421CA genotype, sulfasalazine area under the concentration-time curve was 2.4-fold greater compared with 34GG/421CC subjects (P<0.05). This study links commonly occurring SNPs in BCRP with significantly increased oral sulfasalazine plasma exposure in humans. Accordingly, sulfasalazine may prove to have utility as in vivo probe for assessing the clinical impact of BCRP for the disposition and efficacy of drugs.


Regulation of early endocytic vesicle motility and fission in a reconstituted system.
We previously established conditions to reconstitute kinesin-dependent early endocytic vesicle motility and fission on microtubules in vitro. The present study examined the question whether motility and fission are regulated in this system. Screening for proteins by immunofluorescence microscopy revealed that the small G protein, Rab4, was associated with 80% of hepatocyte-derived early endocytic vesicles that contain the ligand asialoorosomucoid (ASOR). By contrast, other markers for early endocytic vesicles including clathrin, Rab5 and EEA1 were present in the preparation but did not colocalize with the ASOR vesicles. Guanine nucleotides exchanged into the Rab4 present on the vesicles as shown by solubilization of Rab4 by Rab-GDI; solubilization was inhibited by incubation with GTP-gamma-S and promoted by GDP. Pre-incubation of vesicles with GDP increased the number of vesicles moving on microtubules and markedly increased vesicle fission. This increase in motility from GDP was shown to be towards the minus end of microtubules, possibly through activation of the minus-end-directed kinesin, KIFC2. Pre-incubation of vesicles with GTP-gamma-S, by contrast, repressed motility. Addition of exogenous GST-Rab4- GTP-gamma-S led to a further repression of motility and fission. Repression was not seen with addition of GST-Rab4-GDP. Treatment of vesicles with Rab4 antibody also repressed motility, and repression was not seen when vesicles were pre-incubated with GDP. Based on these results we hypothesize that endogenous Rab4-GTP suppresses motility of ASOR-containing vesicles in hepatocytes and that conversion of Rab4-GTP to Rab4-GDP serves as a molecular switch that activates minus-end kinesin-based motility, facilitating early endosome fission and consequent receptor-ligand segregation.


Dendritic cells can present antigen in vivo in a tolerogenic or immunogenic fashion.
Dendritic cells (DC) are unmatched among APCs in their ability to bind, process, and present Ag. Presentation by such potent APCs, if always immunogenic and never tolerogenic, might stimulate pathogenic autoimmune responses. To determine whether Ag presentation by DC can induce tolerance, mice were injected with a rat IgG2b anti-splenic DC mAb, 33D1, and challenged 13 to 28 days later with a stimulatory rat IgG2b mAb. Injection of mice with 1 ng/100 micrograms of 33D1 rarely induced an anti-rat IgG2b Ab response and, in most mice, induced rat IgG2b-specific T cell and B cell tolerance. Tolerant mice had decreased ability to secrete Ab and make both type 1 and type 2 cytokine mRNA and protein in response to immunization with rat IgG2b. 33D1 was 100- to 1000-fold more potent as a tolerogen than an isotype-matched control rat IgG2b mAb. Injecting mice with aggregated 33D1, 33D1 plus anti-IgD mAb, or 33D1 plus IL-1 induced an IgG1 anti-rat IgG2b Ab response rather than tolerance. IL-1 injected 3 days after 33D1 still induced an Ab response rather than tolerance. Not all anti-DC mAbs are tolerogenic. Injection of a DC-specific hamster anti-CD11c mAb (N418) stimulates an IgG anti-hamster response, and injection of 33D1 plus N418 stimulates both anti-hamster and anti-rat IgG2b responses. These observations indicate that DCs can present Ag in either a tolerogenic or stimulatory manner and suggest that inflammatory stimuli can convert an otherwise tolerogenic signal to a stimulatory signal.


PNPLA3 genetic variation in alcoholic steatosis and liver disease progression.
Alcoholic liver disease (ALD) accounts for the majority of chronic liver diseases in Western countries, and alcoholic cirrhosis is among the premier causes of liver failure, hepatocellular carcinoma (HCC) and liver-related mortality causes. Studies in different genders and ethnic groups, as well as in twins provide strong evidence for a significant contribution of host genetic factors to liver disease development in drinkers. The intense quest for genetic modifiers of alcohol-induced fibrosis progression have identified and repeatedly confirmed a genetic polymorphism in the gene coding for patatin-like phospholipase domain-containing 3 (PNPLA3; adiponutrin; rs738409 C/G, M148I) as a risk factor for alcoholic cirrhosis and its related complication, HCC, in different populations. Although carriership of one or both mutated PNPLA3 alleles does not explain the entire liver phenotypic variability in drinkers, it clearly represents one of the strongest single genetic modulators in a complex trait such as ALD. As more genetic data supporting its important role aggregates, novel insight as to PNPLA3's function and that of its genetic variation in liver injury is unveiled pointing to an important novel pathway in alcohol-mediated hepatic lipid turnover with strong implications on inflammation, extra cellular matrix remodelling, and hepatocarcinogenesis. Future study shall decipher whether the gathered knowledge can be translated into therapeutic benefits of patients.


Mitral commissural prolapse.
Mitral commissural prolapse or flail, either isolated or combined with more extensive degenerative valve disease, imposes several challenges both on its diagnosis and management while being a risk factor for valve reoperation after mitral valve repair. Accurate identification of the prolapsing segment is often not feasible with transthoracic 2D echocardiography, with transesophageal 3D imaging then required for correct diagnosis and surgical planning. Various surgical techniques employed alone or in combination have yielded good results in the repair of commissural prolapse. Herein, we analyze the specific characteristics of commissural disease focusing our attention on 2D and 3D echocardiographic findings and we briefly comment on techniques employed for surgical correction of the disease.


Influence of hydrazine sulfate on abnormal carbohydrate metabolism in cancer patients with weight loss.
Thirty-eight patients with advanced cancer and weight loss were tested in a prospectively randomized, double-blind, placebo-controlled trial to evaluate the influence of hydrazine sulfate on carbohydrate metabolism in cancer cachexia. All patients had an initial 3-day inpatient metabolic evaluation including: standard 5-hr p.o. glucose tolerance test, hormone studies, and total glucose production by infusion of [6-3H]glucose. After 30 days of treatment with capsules containing either placebo or hydrazine sulfate in a 60-mg, 3 times/day dosage, inpatient evaluation was repeated. A total of 62 metabolic inpatient evaluations were performed. The pretreatment characteristics of age, sex, prior therapy experience, nutritional parameters and tumor types were comparable in placebo and hydrazine treatment groups. On initial evaluation, abnormal glucose tolerance and increased glucose production were frequently seen. Serial assessment of glucose tolerance showed no improvement after 30 days of placebo treatment. However, the glucose tolerance was significantly improved in patients receiving 30 days of hydrazine sulfate [2-hr glucose; initial 169 +/- 24 (S.E.) mg/dl versus final 128 +/- 12 mg/dl; p less than 0.05]. In addition, the rate of total glucose production was significantly decreased after 30 days of hydrazine sulfate compared to placebo treatment (2.46 mg/kg/min versus 3.07 mg/kg/min, respectively; p less than 0.05). Toxic effects of hydrazine sulfate were minimal. Our results suggest that hydrazine sulfate can influence the abnormal carbohydrate metabolism associated with weight loss in patients with cancer.


Effect of global system for mobile communication (gsm)-like radiofrequency fields on vascular permeability in mouse brain.
The effect of global system for mobile communication (GSM) radiofrequency fields on vascular permeability in the brain was studied using a purpose-designed exposure system at 898.4 MHz. Mice (n= 30) were given a single far field, whole body exposure for 60 minutes at a specific absorption rate of 4 W/kg. Control mice were also sham-exposed (n = 10) or permitted free movement in a cage (n = 10) to exclude any stress-related effects. Vascular permeability changes were detected using albumin immunohistochemistry and the efficacy of this vascular tracer was confirmed with a positive control group exposed to a clostridial toxin known to increase vascular permeability in the brain. No significant difference in albumin extravasation was detected between any of the groups at the light microscope level using the albumin marker.


Factors affecting public prejudice and social distance on mental illness: analysis of contextual effect by multi-level analysis.
While there have been many quantitative studies on the public's attitude towards mental illnesses, it is hard to find quantitative study which focused on the contextual effect on the public's attitude. The purpose of this study was to identify factors that affect the public's beliefs and attitudes including contextual effects. We analyzed survey on the public's beliefs and attitudes towards mental illness in Korea with multi-level analysis. We analyzed the public's beliefs and attitudes in terms of prejudice as an intermediate outcome and social distance as a final outcome. Then, we focused on the associations of factors, which were individual and regional socio-economic factors, familiarity, and knowledge based on the comparison of the intermediate and final outcomes. Prejudice was not explained by regional variables but was only correlated with individual factors. Prejudice increased with age and decreased by high education level. However, social distance controlling for prejudice increased in females, in people with a high education level, and in regions with a high education level and a high proportion of the old. Therefore, social distance without controlling for prejudice increased in females, in the elderly, in highly educated people, and in regions with a high education and aged community. The result of the multi-level analysis for the regional variables suggests that social distance for mental illness are not only determined by individual factors but also influenced by the surroundings so that it could be tackled sufficiently with appropriate considering of the relevant regional context with individual characteristics.


Sexual and reproductive health status and related knowledge among female migrant workers in Guangzhou, China: a cross-sectional survey.
The objective of this study was to investigate the current sexual and reproductive health (SRH) status including SRH-related knowledge and associated factors, self-reported symptoms of reproductive tract infection (RTI), medical assistance seeking behavior, sexual experience and contraceptive use, reproductive information approach and reproductive service utilization among female migrant workers in Huangpu district, Guangzhou city, China. A cross-sectional study was conducted in 2008 in eight factories, which were selected randomly from 32 eligible factories in the Huangpu district in Guangzhou. Descriptive statistics were used to describe the SRH status of migrant workers. Factors associated with the level of SRH knowledge were determined by a logistic regression model. Of 1346 female migrant workers, 831(61.7%) were unmarried and 515 (38.3%) were married. 27.2% of the unmarried respondents and 40.2% of the married respondents had suffered self-reported RTI symptoms. Among unmarried respondents, the median knowledge score was 5 points, compared to 8 points for the married. For unmarried migrant workers, factors associated with the knowledge level were age, education level, access to SRH information and service, sexual experiences and RTI symptoms. For married migrant workers, factors associated with the knowledge level were age, education level, access to SRH services and RTI symptoms. A high prevalence of self-reported RTI symptoms and a low knowledge level were found among young female migrant workers. Unmarried migrant workers are more vulnerable to SRH problems. Those findings demand more specific interventions targeting female migrants and in particular the unmarried.


ATP-sensitive K+ channels in human isolated pancreatic B-cells.
Glucose-stimulated insulin release from rodent pancreatic B-cells is thought to be initiated by the closing of ATP-sensitive K+ channels in the plasma membrane as a consequence of glucose metabolism. We have identified an ATP-sensitive K+ channel in membrane patches excised from human B-cells which is similar to that found in rodent B-cells in conductance, kinetics, ATP sensitivity and its inhibition by sulphonylureas. In man, the ATP-sensitive K+ channel may also have a central role in glucose-stimulated insulin secretion and may be (linked to) the receptor for the hypoglycemic sulphonylureas.


Barbiturates for acute traumatic brain injury.
Raised intracranial pressure (ICP) is an important complication of severe brain injury, and is associated with high mortality. Barbiturates are believed to reduce ICP by suppressing cerebral metabolism, thus reducing cerebral metabolic demands and cerebral blood volume. However, barbiturates also reduce blood pressure and may, therefore, adversely effect cerebral perfusion pressure. To assess the effects of barbiturates in reducing mortality, disability and raised ICP in people with acute traumatic brain injury. To quantify any side effects resulting from the use of barbiturates. The following electronic databases were searched on 26 September 2012: CENTRAL (The Cochrane Library), MEDLINE (Ovid SP), PubMed, EMBASE (Ovid SP), PsycINFO (Ovid SP), PsycEXTRA (Ovid SP), ISI Web of Science: Science Citation Index and Conference Proceedings Citation Index-Science. Searching was not restricted by date, language or publication status. We also searched the reference lists of the included trials and review articles. We contacted researchers for information on ongoing studies. Randomised controlled trials of one or more of the barbiturate class of drugs, where study participants had clinically diagnosed acute traumatic brain injury of any severity. Two review authors screened the search results, extracted data and assessed the risk of bias in the trials. Data from seven trials involving 341 people are included in this review.For barbiturates versus no barbiturate, the pooled risk ratio (RR) of death from three trials was 1.09 (95% confidence interval (CI) 0.81 to 1.47). Death or disability, measured using the Glasgow Outcome Scale was assessed in two trials, the RR with barbiturates was 1.15 (95% CI 0.81 to 1.64). Two trials examined the effect of barbiturate therapy on ICP. In one, a smaller proportion of patients in the barbiturate group had uncontrolled ICP (68% versus 83%); the RR for uncontrolled ICP was 0.81 (95% CI 0.62 to 1.06). In the other, mean ICP was also lower in the barbiturate group. Barbiturate therapy results in an increased occurrence of hypotension (RR 1.80; 95% CI 1.19 to 2.70). For every four patients treated, one developed clinically significant hypotension. Mean body temperature was significantly lower in the barbiturate group.In one study of pentobarbital versus mannitol there was no difference in death between the two study groups (RR 1.21; 95% CI 0.75 to 1.94). Pentobarbital was less effective than mannitol for control of raised ICP (RR 1.75; 95% CI 1.05 to 2.92).In one study the RR of death with pentobarbital versus thiopental was 1.78 (95% CI 1.03 to 3.08) in favour of thiopental. Fewer people had uncontrollable ICP with thiopental (RR 1.64; 95% CI 1.03 to 2.60). There was no significant difference in the effects of pentobarbital versus thiopental for death or disability, measured using the Glasgow Outcome Scale (RR 1.31; 95% CI 0.88 to 1.94), or hypotension (RR 0.95; 95% CI 0.81 to 1.12). There is no evidence that barbiturate therapy in patients with acute severe head injury improves outcome. Barbiturate therapy results in a fall in blood pressure in one in four patients. This hypotensive effect will offset any ICP lowering effect on cerebral perfusion pressure.


Heterogeneous genomic molecular clocks in primates.
Using data from primates, we show that molecular clocks in sites that have been part of a CpG dinucleotide in recent past (CpG sites) and non-CpG sites are of markedly different nature, reflecting differences in their molecular origins. Notably, single nucleotide substitutions at non-CpG sites show clear generation-time dependency, indicating that most of these substitutions occur by errors during DNA replication. On the other hand, substitutions at CpG sites occur relatively constantly over time, as expected from their primary origin due to methylation. Therefore, molecular clocks are heterogeneous even within a genome. Furthermore, we propose that varying frequencies of CpG dinucleotides in different genomic regions may have contributed significantly to conflicting earlier results on rate constancy of mammalian molecular clock. Our conclusion that different regions of genomes follow different molecular clocks should be considered when inferring divergence times using molecular data and in phylogenetic analysis.


Basal promoter of the rat connexin 32 gene: identification and characterization of an essential element and its DNA-binding protein.
The connexin 32 (Cx32) gene, a member of a multigene family, is expressed preferentially in the liver. The basal promoter complex of the rat Cx32 gene was previously localized to a 146-bp region (map positions [mp] -179 to -34) immediately upstream of the first exon. To investigate the biochemical factors contributing to the basal promoter activity, nuclear protein-DNA complexes within this region (mp -177 to -106) were investigated by using a DNA mobility shift assay. Three DNA-protein binding activities, termed Cx32-B1, Cx32-B2, and Cx32-B3, were identified with nuclear protein extracts from hepatoma cell lines, HuH7 and FAO-1. However, only Cx32-B2 binding activity was detected in nuclear protein extract from normal rat liver tissue. This activity was significantly more abundant in rat liver tissue than in hepatoma cell lines and tissues from various other organs. By using methylation interference footprinting, the Cx32-B2 complex was localized to the region between mp -152 and -127 and a DNA probe containing this region bound to a 60-kDa protein in rat liver nuclear extracts. Mutation of two nucleotides in the Cx32-B2 binding site abrogated the formation of the Cx32-B2 protein-DNA complex and significantly reduced the transcriptional activity of the Cx32 promoter. These results indicate that the Cx32-B2 complex is an essential component of the rat Cx32 basal promoter and is likely a major factor in the preferential expression of this gene in the liver.


Mesenchymal stromal cells are present in the heart and promote growth of adult stem cells in vitro.
For many years the human heart has been considered a terminally differentiated organ with no regenerative potential after injury. Recent studies, however, have cast doubt on this long-standing dogma. The objective of this study was to investigate the presence of and characterize mesenchymal stromal cells (MSC) in the adult mouse heart. The impact of MSC on growth and differentiation of adult cardiac stem cells (CSC) was also analyzed. A combination of lineage-negative/c-kit-negative (Lin(-)/c-kit(-)) immunoselection with a plastic-adhesion technique was used to isolate cardiac-derived MSC. The differentiation capacity and expression of surface markers were analyzed. To investigate the impact of MSC on growth and differentiation of adult CSC, Green Fluorescent Protein (GFP(+)) adult CSC were co-cultured with GFP(-) cardiac-derived MSC. MSC were present in the adult mouse heart and they met the criteria established to define mouse MSC. They expressed surface markers and were able to differentiate, in a controlled manner, into multiple lineages. In addition, cardiac-derived MSC promoted the survival and expansion of adult CSC in vitro. MSC can be isolated from the mouse heart and they promote growth and differentiation of adult CSC. The findings from this study could have a significant beneficial impact on future heart failure treatment. Co-culture and co-implantation of cardiac-derived MSC with adult CSC could provide extensive cardiac regeneration and maintenance of the CSC population after implanted into the heart.


Caste fate conflict in swarm-founding social hymenoptera: an inclusive fitness analysis.
A caste system in which females develop into morphologically distinct queens or workers has evolved independently in ants, wasps and bees. Although such reproductive division of labour may benefit the colony it is also a source of conflict because individual immature females can benefit from developing into a queen in order to gain greater direct reproduction. Here we present a formal inclusive fitness analysis of caste fate conflict appropriate for swarm-founding social Hymenoptera. Three major conclusions are reached: (1) when caste is self-determined, many females should selfishly choose to become queens and the resulting depletion of the workforce can substantially reduce colony productivity; (2) greater relatedness among colony members reduces this excess queen production; (3) if workers can prevent excess queen production at low cost by controlled feeding, a transition to nutritional caste determination should occur. These predictions generalize results derived earlier using an allele-frequency model [Behav. Ecol. Sociobiol. (2001) 50: 467] and are supported by observed levels of queen production in various taxa, especially stingless bees, where caste can be either individually or nutritionally controlled.


Uridine nucleotide selectivity of three phospholipase C-activating P2 receptors: identification of a UDP-selective, a UTP-selective, and an ATP- and UTP-specific receptor.
Observation that the G protein-coupled P2U receptor (P2Y2 receptor) is activated by UTP as well as ATP provided the first indication that a class of uridine nucleotide-responsive receptors might exist. This hypothesis was confirmed by our identification of a uridine nucleotide-specific receptor on C6-2B rat glioma cells and by the recent cloning of two uridine nucleotide-responsive receptors, the P2Y6 receptor [J. Biol. Chem. 270:26152-26158 (1995)] and the P2Y4 receptor [J. Biol. Chem. 270:30849-30852 (1995) and J. Biol. Chem. 270:30845-30848 (1995)]. The relative nucleotide selectivities of these uridine nucleotide-activated receptors have not been established. Therefore, we cloned and expressed the P2Y6 and P2Y4 receptors in 1321N1 human astrocytoma cells and compared their relative selectivities for UDP, UTP, and other uridine and adenine nucleotides with that of the P2Y2 receptor expressed in the same cells. These comparisons were made by measuring inositol phosphate accumulation under conditions in which the initial purity and stability of agonists were rigidly ensured and quantitatively assessed. The data indicate that the P2Y2 receptor is activated with similar potencies by ATP and UTP but not by ADP or UDP; the P2Y6 receptor is activated most potently by UDP but weakly by UTP, ATP, and ADP; and the P2Y4 receptor is activated most potently by UTP, less potently by ATP, and not at all by nucleotide diphosphates. Furthermore, the P2Y6 receptor, which displays a uridine nucleotide selectivity essentially identical to that of the uridine nucleotide-specific receptor in C6-2B cells, was shown to be natively expressed in C6-2B cells and to account for the uridine nucleotide responses originally identified in these cells. These results define the uridine nucleotide selectivity of three phospholipase C-linked receptors: a receptor that is selectively activated by UDP (P2Y6 receptor), selectively activated by UTP (P2Y4 receptor), and activated by UTP and ATP but not by diphosphate nucleotides (P2Y2 receptor).


Young people, alcohol, and designer drinks: quantitative and qualitative study.
To examine the appeal of "designer drinks" to young people. Qualitative and quantitative research comprising group discussions and questionnaire led interviews with young people accompanied by a self completion questionnaire. Argyll and Clyde Health Board area, west Scotland. Eight groups aged 12-17 years; 824 aged 12-17 recruited by multistage cluster probability sample from the community health index. Young people were familiar with designer drinks, especially MD 20/20 and leading brands of strong white cider. Attitudes towards these drinks varied quite distinctly with age, clearly reflecting their attitudes towards and motivations for drinking in general. The brand imagery of designer drinks-in contrast with that of more mainstream drinks-matched many 14 and 15 year olds' perceptions and expectations of drinking. Popularity of designer drinks peaked between the ages of 13 and 16 while more conventional drinks showed a consistent increase in popularity with age. Consumption of designer drinks tended to be in less controlled circumstances and was associated with heavier alcohol intake and greater drunkenness. Designer drinks are a cause for concern. They appeal to young people, often more so than conventional drinks, and are particularly attractive to 14-16 year olds. Consumption of designer drinks is also associated with drinking in less controlled environments, heavier drinking, and greater drunkenness. There is a need for policy debate to assess the desirability of these drinks and the extent to which further controls on their marketing are required.


Health-related quality of life and postoperative length of stay for patients with colorectal cancer.
Length of stay (LOS) after surgery is a major determinant of resource utilization for colorectal cancer (CRC). The purpose of this study was to examine the association between pretreatment health-related quality of life (HRQL) scores and postoperative hospital LOS in a cohort of patients undergoing surgery for CRC. Seventy patients with biopsy-proven CRC were enrolled in an IRB-approved, prospective study. Information was collected concerning standard perioperative variables. Prior to surgery, all patients also completed the CRC-specific module of the Functional Assessment of Cancer Therapy (FACT-C). Perioperative variables and FACT-C scores were compared with LOS in both univariate and multivariate analysis. LOS for those patients scoring in the lowest quartile on FACT-C was compared with LOS for patients scoring in the remaining quartiles. Median length of stay for the entire group was 6 (range 3-25) days. In univariate analysis, surgical complications (10.6 vs 6.6 days; P = 0.001) and with poorer FACT-C individual scale scores for Physical Well-Being (9.1 vs 7.3 days; P = 0.04), Functional Well-Being (9.6 vs 7.1 days; P = 0.006), and Colorectal Cancer Concerns (9.5 vs 7.1 days; P = 0.01) were all significantly associated with increased length of stay. In multivariate analysis, surgical morbidity (OR = 5.6; 95% CI 1.5-21.4), age >72 (OR = 6.0; 95% CI 1.6-23.5), and low FACT-C total score (OR = 4.2; 95% CI 1.1-15.6) were independently associated with increased LOS. Pretreatment HRQL scores as measured by FACT-C may be of benefit in the prediction of LOS. Such information may be an important and currently neglected means of risk-adjusting populations undergoing surgery for colorectal cancer for this outcome.


Model for adhesion clutch explains biphasic relationship between actin flow and traction at the cell leading edge.
Cell motility relies on the continuous reorganization of a dynamic actin-myosin-adhesion network at the leading edge of the cell, in order to generate protrusion at the leading edge and traction between the cell and its external environment. We analyze experimentally measured spatial distributions of actin flow, traction force, myosin density, and adhesion density in control and pharmacologically perturbed epithelial cells in order to develop a mechanical model of the actin-adhesion-myosin self-organization at the leading edge. A model in which the F-actin network is treated as a viscous gel, and adhesion clutch engagement is strengthened by myosin but weakened by actin flow, can explain the measured molecular distributions and correctly predict the spatial distributions of the actin flow and traction stress. We test the model by comparing its predictions with measurements of the actin flow and traction stress in cells with fast and slow actin polymerization rates. The model predicts how the location of the lamellipodium-lamellum boundary depends on the actin viscosity and adhesion strength. The model further predicts that the location of the lamellipodium-lamellum boundary is not very sensitive to the level of myosin contraction.


Crosses of NOD mice with the related NON strain. A polygenic model for IDDM.
Chromosome locations of non-major histocompatibility complex (MHC) genes contributing to insulin-dependent diabetes mellitus (IDDM) in mice have been determined by outcrossing NOD mice to other inbred strains congenic for the NOD MHC haplotype (H2g7). At least nine non-MHC IDDM susceptibility genes (Idd) were previously identified at first backcross (BC1) after outcross of NOD to C57BL/10.H2g7 congenic mice (B10.H2g7). We investigated whether the same set of Idd loci segregated with IDDM susceptibility after outcross of NOD to NON.H2g7 congenic mice. Since the outcrosses to NON.H2g7 and B10.H2g7 were performed in the same vivarium, direct comparisons were made of the chromosomal locations and relative strengths of Idd alleles in diabetic progeny from the two different outcrosses. In comparison with the NOD x B10.H2g7 outcross, the NOD x NON.H2g7 outcross produced significantly higher IDDM frequencies in F1, F2, and BC1 generations. The high F2 diabetes frequency allowed evaluation of the effects of homozygous expression of both the susceptibility and the resistance allele at Idd loci. This analysis demonstrated that no single non-MHC Idd locus was essential for the onset of diabetes in this cross. After outcross to NON.H2g7, Idd4 (chromosome [Chr] 11), Idd5 (Chr 1), and Idd8 (Chr 14) did not segregate with IDDM in either the BC1 or the F2 generation. Diabetogenic NOD-derived alleles at Idd2 (Chr 9), Idd3 (Chr 3), and Idd10 (Chr 3) were segregating in the BC1. An NON-derived allele contributing to susceptibility on Chr 7 (Idd7) was also detected. Dominant traits, detectable only in the F2 cross, were encoded by Chr 4 (Idd9) and two newly mapped loci on Chr 13 (Idd14) and 5 (Idd15). A third dominant trait was encoded by Chr 6 (possibly Idd6), but here, in contrast to Idd9, Idd14, and Idd15, the NON allele was diabetogenic. Stepwise logistic regression analysis of the BC1 and F2 data confirmed that the ability to identify certainty of the non-MHC Idd loci was contingent on the extent of homozygosity for NOD background genes. This study shows that the diabetogenic phenotype can be achieved through the actions of variable combinations of MHC-unlinked genes and a diabetogenic MHC haplotype.


Modulation of Allostery with Multiple Mechanisms by Hotspot Mutations in TetR.
Modulating allosteric coupling offers unique opportunities for biomedical applications. Such efforts can benefit from efficient prediction and evaluation of allostery hotspot residues that dictate the degree of co-operativity between distant sites. We demonstrate that effects of allostery hotspot mutations can be evaluated qualitatively and semi-quantitatively by molecular dynamics simulations in a bacterial tetracycline repressor (TetR). The simulations recapitulate the effects of these mutations on abolishing the induction function of TetR and provide a rationale for the different degrees of rescuability observed to restore allosteric coupling of the hotspot mutations. We demonstrate that the same non-inducible phenotype could be the result of perturbations in distinct structural and energetic properties of TetR. Our work underscore the value of explicitly computing the functional free energy landscapes to effectively evaluate and rank hotspot mutations despite the prevalence of compensatory interactions, and therefore provide quantitative guidance to allostery modulation for therapeutic and engineering applications.


Realizing human rights-based approaches for action on the social determinants of health.
Health inequities are clear evidence of violations of the right to health. Yet despite this common ground, action on the social determinants of health aiming to reduce health inequities is sometimes disconnected from implementation of human rights-based approaches. This is explained in part by differing histories, disciplines, and epistemologies. The capacity of human rights instruments to alter policies on social determinants can seem limited. An absolutist focus on individuals and processes can seem at odds with the attention to differences in population health outcomes central to the concern for health equity. However, developments in rights-based approaches have seen the terrain of human rights increasingly address social determinants. Human rights provide a firm legal basis for tackling the inequities in power and resources that the Commission on Social Determinants of Health identifies as fundamental to achieving health equity. Indicators and benchmarks developed for rights-based approaches to health systems can be developed further within health sectors and translated to other sectors and disciplines. The discourse and evidence base of social determinants can also contribute to implementing rights-based approaches, as its resultant policy momentum can provide essential levers to realize the right to health. Therefore, there is no clear-cut delineation between the human rights and health equity movements, and both may constructively work together to realize their goals. Such constructive collaboration will not prove straightforward; it will, instead, require profound engagement and innovations in both theory and practice. Yet this effort represents an important opportunity for those who seek social justice in health.


HINTS to diagnose stroke in the acute vestibular syndrome: three-step bedside oculomotor examination more sensitive than early MRI diffusion-weighted imaging.
Acute vestibular syndrome (AVS) is often due to vestibular neuritis but can result from vertebrobasilar strokes. Misdiagnosis of posterior fossa infarcts in emergency care settings is frequent. Bedside oculomotor findings may reliably identify stroke in AVS, but prospective studies have been lacking. The authors conducted a prospective, cross-sectional study at an academic hospital. Consecutive patients with AVS (vertigo, nystagmus, nausea/vomiting, head-motion intolerance, unsteady gait) with >or=1 stroke risk factor underwent structured examination, including horizontal head impulse test of vestibulo-ocular reflex function, observation of nystagmus in different gaze positions, and prism cross-cover test of ocular alignment. All underwent neuroimaging and admission (generally <72 hours after symptom onset). Strokes were diagnosed by MRI or CT. Peripheral lesions were diagnosed by normal MRI and clinical follow-up. One hundred one high-risk patients with AVS included 25 peripheral and 76 central lesions (69 ischemic strokes, 4 hemorrhages, 3 other). The presence of normal horizontal head impulse test, direction-changing nystagmus in eccentric gaze, or skew deviation (vertical ocular misalignment) was 100% sensitive and 96% specific for stroke. Skew was present in 17% and associated with brainstem lesions (4% peripheral, 4% pure cerebellar, 30% brainstem involvement; chi(2), P=0.003). Skew correctly predicted lateral pontine stroke in 2 of 3 cases in which an abnormal horizontal head impulse test erroneously suggested peripheral localization. Initial MRI diffusion-weighted imaging was falsely negative in 12% (all <48 hours after symptom onset). Skew predicts brainstem involvement in AVS and can identify stroke when an abnormal horizontal head impulse test falsely suggests a peripheral lesion. A 3-step bedside oculomotor examination (HINTS: Head-Impulse-Nystagmus-Test-of-Skew) appears more sensitive for stroke than early MRI in AVS.


Valuations of genetic test information for treatable conditions: the case of colorectal cancer screening.
The value of the information that genetic testing services provide can be questioned for insurance-based health systems. The results of genetic tests oftentimes may not lead to well-defined clinical interventions; however, Lynch syndrome, a genetic mutation for which carriers are at an increased risk for colorectal cancer, can be identified through genetic testing, and meaningful health interventions are available via increased colonoscopic surveillance. Valuations of test information for such conditions ought to account for the full impact of interventions and contingent outcomes. To conduct a discrete-choice experiment to elicit individuals' preferences for genetic test information. A Web-enabled discrete-choice experiment survey was administered to a representative sample of US residents aged 50 years and older. In addition to specifying expenditures on colonoscopies, respondents were asked to make a series of nine selections between two hypothetical genetic tests or a no-test option under the premise that a relative had Lynch syndrome. The hypothetical genetic tests were defined by the probability of developing colorectal cancer, the probability of a false-negative test result, privacy of the result, and out-of-pocket cost. A model specification identifying necessary interactions was derived from assumptions of risk behavior and the decision context and was estimated using random-parameters logit. A total of 650 respondents were contacted, and 385 completed the survey. The monetary equivalent of test information was approximately $1800. Expenditures on colonoscopies to reduce mortality risks affected valuations. Respondents with lower income or who reported being employed significantly valued genetic tests more. Genetic testing may confer benefits through the impact of subsequent interventions on private individuals.


A home-visiting intervention targeting determinants of infant mental health: the study protocol for the CAPEDP randomized controlled trial in France.
Several studies suggest that the number of risk factors rather than their nature is key to mental health disorders in childhood. The objective of this multicentre randomized controlled parallel trial (PROBE methodology) is to assess the impact in a multi-risk French urban sample of a home-visiting program targeting child mental health and its major determinants. This paper describes the protocol of this study. In the study, pregnant women were eligible if they were: living in the intervention area; able to speak French, less than 26 years old; having their first child; less than 27 weeks of amenorrhea; and if at least one of the following criteria were true: less than twelve years of education, intending to bring up their child without the presence of the child's father, and 3) low income. Participants were randomized into either the intervention or the control group. All had access to usual care in mother-child centres and community mental health services free of charge in every neighbourhood. Psychologists conducted all home visits, which were planned on a weekly basis from the 7th month of pregnancy and progressively decreasing in frequency until the child's second birthday. Principle outcome measures included child mental health at 24 months and two major mediating variables for infant mental health: postnatal maternal depression and the quality of the caring environment. A total of 440 families were recruited, of which a subsample of 120 families received specific attachment and caregiver behaviour assessment. Assessment was conducted by an independent assessment team during home visits and, for the attachment study, in a specifically created Attachment Assessment laboratory. The CAPEDP study is the first large-scale randomised, controlled infant mental health promotion programme to take place in France. A major specificity of the program was that all home visits were conducted by specifically trained, supervised psychologists rather than nurses. Significant challenges included designing a mental health promotion programme targeting vulnerable families within one of the most generous but little assessed health and social care systems in the Western World. Current Clinical trial number is NCT00392847.


GenMAPP 2: new features and resources for pathway analysis.
Microarray technologies have evolved rapidly, enabling biologists to quantify genome-wide levels of gene expression, alternative splicing, and sequence variations for a variety of species. Analyzing and displaying these data present a significant challenge. Pathway-based approaches for analyzing microarray data have proven useful for presenting data and for generating testable hypotheses. To address the growing needs of the microarray community we have released version 2 of Gene Map Annotator and Pathway Profiler (GenMAPP), a new GenMAPP database schema, and integrated resources for pathway analysis. We have redesigned the GenMAPP database to support multiple gene annotations and species as well as custom species database creation for a potentially unlimited number of species. We have expanded our pathway resources by utilizing homology information to translate pathway content between species and extending existing pathways with data derived from conserved protein interactions and coexpression. We have implemented a new mode of data visualization to support analysis of complex data, including time-course, single nucleotide polymorphism (SNP), and splicing. GenMAPP version 2 also offers innovative ways to display and share data by incorporating HTML export of analyses for entire sets of pathways as organized web pages. GenMAPP version 2 provides a means to rapidly interrogate complex experimental data for pathway-level changes in a diverse range of organisms.


Evaporated inhomogeneous thin films.
A theoretical and experimental investigation of inhomogeneous CeO(2)-MgF(2) and ZnS-Na(3)AIF(6) anti-reflection coatings for the visible spectrai region has been made. By a proper choice of the refractive index profile, the reflectance may be considerably reduced. In the infrared region, Ge-MgF(2) films on Ge substrates are shown to give a low reflectance with a small wavelength dependence.


The Use of Circulating Tumor DNA for Prognosis of Gastrointestinal Cancers.
Gastrointestinal cancers, including oesophageal, gastric and colorectal cancers (CRC) have high rates of disease recurrence despite curative resection. There are a number of recent studies that have investigated the use of circulating tumor DNA (ctDNA) for prognostic value in these cancers. We reviewed studies that had been published prior to March 2018 that assessed the prognostic values of ctDNA in patients with oesophageal and gastric cancers, gastrointestinal stromal tumors (GIST) and CRC. We identified 63 eligible clinical studies that focussed on recurrence and survival. Studies assessed investigated various ctDNA biomarkers in patients with different stages of cancer undergoing surgical resection, chemotherapy and no treatment. For oesophageal squamous cell carcinoma and oesophageal adenocarcinoma, methylation of certain genes such as APC and DAPK have been highlighted as promising biomarkers for prognostication, but these studies are limited and more comprehensive research is needed. Studies focusing on gastric cancer patients showed that methylation of ctDNA in SOX17 and APC were independently associated with poor survival. Two studies demonstrated an association between ctDNA and recurrence and survival in GIST patients, but more studies are needed for this type of gastrointestinal cancer. A large proportion of the literature was on CRC which identified both somatic mutations and DNA methylation biomarkers to determine prognosis. ctDNA biomarkers that identified somatic mutations were more effective if they were personalized based on mutations found in the primary tumor tissue, but ctDNA methylation studies identified various biomarkers that predicted increased risk of recurrence, poor disease free survival and overall survival. While the use of non-invasive ctDNA biomarkers for prognosis is promising, larger studies are needed to validate the clinical utility for optimizing treatment and surveillance strategies to reduce mortality from gastrointestinal cancers.


Real-time droplet DNA amplification with a new tablet platform.
We present a novel droplet-based tablet platform for temporal polymerase chain reaction (PCR) in microliter droplets. The simple design of the device does not require extensive processing or external equipment, which allows for greater ease of use and integration as a point-of-care diagnostic. We demonstrate its functionality to perform both PCR and reverse-transcription PCR for λ phage DNA and H3 influenza RNA with ramp rates and cycle times consistent with traditional PCR thermal cyclers. We additionally investigate the effect of performing PCR in small volumes on the reaction performance by specifically examining adsorption of reagents at the oil/water interface. We determined that adsorption of Taq polymerase at the biphasic interface reduces yield and impairs reaction performance at standard concentrations. Thus, microdroplet PCR reactions require additional polymerase to achieve sufficient amplification and we project for applications utilizing nanodroplets or picodroplets like digital applications, even greater concentrations of polymerase are required to achieve desired results. Following the adsorption investigation, we evaluated the sensitivity of λ phage PCR on our platform to be less than 2.0 copies/μL with an efficiency of 104.4% and similar sensitivity for reverse-transcription PCR for influenza H3 RNA.


Targeting the mitochondrial respiratory chain of Cryptococcus through antifungal chemosensitization: a model for control of non-fermentative pathogens.
Enhanced control of species of Cryptococcus, non-fermentative yeast pathogens, was achieved by chemosensitization through co-application of certain compounds with a conventional antimicrobial drug. The species of Cryptococcus tested showed higher sensitivity to mitochondrial respiratory chain (MRC) inhibition compared to species of Candida. This higher sensitivity results from the inability of Cryptococcus to generate cellular energy through fermentation. To heighten disruption of cellular MRC, octyl gallate (OG) or 2,3-dihydroxybenzaldehyde (2,3-DHBA), phenolic compounds inhibiting mitochondrial functions, were selected as chemosensitizers to pyraclostrobin (PCS; an inhibitor of complex III of MRC). The cryptococci were more susceptible to the chemosensitization (i.e., PCS + OG or 2,3-DHBA) than the Candida with all Cryptococcus strains tested being sensitive to this chemosensitization. Alternatively, only few of the Candida strains showed sensitivity. OG possessed higher chemosensitizing potency than 2,3-DHBA, where the concentration of OG required with the drug to achieve chemosensitizing synergism was much lower than that required of 2,3-DHBA. Bioassays with gene deletion mutants of the model yeast Saccharomyces cerevisiae showed that OG or 2,3-DHBA affect different cellular targets. These assays revealed mitochondrial superoxide dismutase or glutathione homeostasis plays a relatively greater role in fungal tolerance to 2,3-DHBA or OG, respectively. These findings show that application of chemosensitizing compounds that augment MRC debilitation is a promising strategy to antifungal control against yeast pathogens.


Determinants of occupational multisite musculoskeletal disorders: a cross sectional study among 254 patients.
The aim was to describe the profile of workers with occupational multi-site musculoskeletal disorders (MSMSD) and study the relationship between these lesions and socio-professional factors. This is a cross-sectional study involving 254 subjects with occupational musculoskeletal disorders (MSD), identified in the Department of Occupational Medicine at the University Hospital of Mahdia, in Tunisia, over a period of 10 years from 2005 to 2014. The study population was subdivided into two groups; mono-site MSD and multi-site (≥2 sites) groups. Data collection was based on a questionnaire prepared beforehand and covered the description of sociodemographic and professional characteristics. To study psychosocial constraints at work, we have used the Karasek questionnaire. MS-MSD was correlated to the number of dependent children (p=0.02), job/place of work (p=0.00), qualification (p=0.02), taking a rest period (p=0.03), decision latitude (p=0.00), mental demands (p=0.002), social support (p=0.00) and job stress (p=0.04). After binary logistic regression, MS-MSD depended significantly on the number of dependent children (p=0.013; OR=0,33; IC=0,17-0,83), working spouse (p=0.05; OR=0.35; IC=0.12-0.99), job/place of work (p=0.00; OR=4.16; IC=1.95-8.88), qualification (p=0.008; OR=0.28; IC=0.11-0.72), taking a break during work (p=0.04; OR=3.10; IC=1.04-9.22) and social support (p=0.00; OR=7,1; IC=1,9-25,3). When individual risk factors are fixed, the prevention of MS-MSD must target modifiable levers, related to the professional environment of the employees.


A clue on bee glue: New insight into the sources and factors driving resin intake in honeybees (Apis mellifera).
Honeybees (Apis mellifera) are threatened by numerous pathogens and parasites. To prevent infections they apply cooperative behavioral defenses, such as allo-grooming and hygiene, or they use antimicrobial plant resin. Resin is a chemically complex and highly variable mixture of many bioactive compounds. Bees collect the sticky material from different plant species and use it for nest construction and protection. Despite its importance for colony health, comparatively little is known about the precise origins and variability in resin spectra collected by honeybees. To identify the botanical resin sources of A. mellifera in Western Europe we chemically compared resin loads of individual foragers and tree resins. We further examined the resin intake of 25 colonies from five different apiaries to assess the effect of location on variation in the spectra of collected resin. Across all colonies and apiaries, seven distinct resin types were categorized according to their color and chemical composition. Matches between bee-collected resin and tree resin indicated that bees used poplar (Populus balsamifera, P. x canadensis), birch (Betula alba), horse chestnut (Aesculus hippocastanum) and coniferous trees (either Picea abies or Pinus sylvestris) as resin sources. Our data reveal that honeybees collect a comparatively broad and variable spectrum of resin sources, thus assuring protection against a variety of antagonists sensitive to different resins and/or compounds. We further unravel distinct preferences for specific resins and resin chemotypes, indicating that honeybees selectively search for bioactive resin compounds.