Nurse coaching to explore and modify patient attitudinal barriers interfering with effective cancer pain management.
To describe a complex coaching intervention to help patients with cancer pain explore beliefs and attitudinal barriers interfering with pain management. Patients were coached to explore beliefs about pain, communications about pain management, and the use of analgesics and nonpharmacologic interventions. Published journal articles, abstracts, and psychology textbooks. Personal beliefs, related attitudinal barriers, and associated behaviors impede patient adherence to and success with pain management treatments. Interventions targeting beliefs help patients overcome attitudinal barriers, improve treatment adherence, and obtain better pain relief. Coaching patients to explore beliefs reduces ineffective behaviors and improves pain treatment adherence. A coaching intervention incorporating assessment of patient beliefs promotes self-management, self-efficacy, and adherence to pain management treatment plans. Advanced practice nurses should consider incorporating this intervention into their communications with patients experiencing cancer pain.


A large-scale study on research code quality and execution.
This article presents a study on the quality and execution of research code from publicly-available replication datasets at the Harvard Dataverse repository. Research code is typically created by a group of scientists and published together with academic papers to facilitate research transparency and reproducibility. For this study, we define ten questions to address aspects impacting research reproducibility and reuse. First, we retrieve and analyze more than 2000 replication datasets with over 9000 unique R files published from 2010 to 2020. Second, we execute the code in a clean runtime environment to assess its ease of reuse. Common coding errors were identified, and some of them were solved with automatic code cleaning to aid code execution. We find that 74% of R files failed to complete without error in the initial execution, while 56% failed when code cleaning was applied, showing that many errors can be prevented with good coding practices. We also analyze the replication datasets from journals' collections and discuss the impact of the journal policy strictness on the code re-execution rate. Finally, based on our results, we propose a set of recommendations for code dissemination aimed at researchers, journals, and repositories.


Applications of the marchbanks transcranial-cerebral sonography technique in neurootology: preliminary report.
Transcranial-cerebral sonography (TCCS) is a noninvasive technique that allows clinicians to detect nanoliter (billionths of a liter) displacements of the tympanic membrane. This technique was developed to assess cerebrospinal fluid (CSF) pressure in cases of shunted hydrocephalus; it takes advantage of the CSF connection to the inner ear through the cochlear aqueduct. The movements of the tympanic membrane that are observed in TCCS are those evoked by the acoustic stapedius reflex and those spontaneous movements generated by intracranial arterial, venous, and respiratory pulses transmitted through the inner ear to the stapes and thence to the tympanic membrane. Analysis of the amplitude and direction of these displacements has enabled neurosurgeons and neurologists to estimate CSF pressures accurately in patients evaluated by TCCS. TCCS allows for applications in neurootology, particularly in those patients who present with symptoms of pulsating tinnitus, dizziness and imbalance, or hearing loss. This preliminary report describes the test and its application in a series of patients whose diagnoses included pulsating tinnitus, idiopathic intracranial hypertension, Ménière's disease, perilymphatic fistula, perilymphatic hypertension, arterial stenosis, and Arnold-Chiari syndrome. We conclude that TCCS is a valuable addition to the armamentarium of neurootologists.


Anion binding by p-aminoazobenzene-derived aromatic amides: spectroscopic and electrochemical studies.
The synthesis and complexing properties of p-aminoazobenzene-derived mono-, bis-, and trisamides were described. Ligands 3 and 4 bind anions, including fluorides, chlorides, bromides, acetates, benzoates, dihydrogen phosphates, hydrogen sulfates, and p-toluenesulfonates, in chloroform forming 1 : 1 complexes. The highest value of stability constant was evaluated for the 4-F- complex (log K = 5.63 ± 0.21). On the basis of 1H NMR, and FTIR spectroscopy, the possible nature of the ligand-anion interactions was proposed. The E ⇄ Z isomerization process of tripodal amide 4 in chloroform was studied. The effect of anions on Z to E thermal back isomerization was investigated.


Ethanolic extract of Moringa oleifera Lam. leaves protect the pre-pubertal spermatogonial cells from cyclophosphamide-induced damage.
Moringa oleifera Lam. is widely cultivated in Asian and African countries for its medicinal and dietary significance. The leaves are highly nutritious and are known to possess various biological activities. Pre-pubertal Swiss albino male mice were injected with single dose of cyclophosphamide (CP, 200mg/kg body weight) or ethanolic extract of Moringa oleifera leaves (MOE, 100mg/kg body weight) intraperitoneally. In combination group, MOE was administered 24h prior to CP injection. CP induced a significant decrease in testicular weight (p<0.01) and depletion of germ cells (p<0.001) and higher level of DNA damage (p<0.001) compared to control. The expression of P53, Bax, Cytochrome C (Cyt C) was increased while there was a decrease in the expression of Bcl2, c-Kit and Oct4. Administration of MOE 24h prior to CP treatment ameliorated the depletion (p<0.001), DNA damage (p<0.001) and apoptosis (p<0.01) of germ cells induced by CP. The mitigating effect of MOE appears to be mediated by up-regulating the expression of c-Kit and Oct4 transcripts in P53-independent manner. MOE protects the spermatogonial cells from CP-induced damage by modulating the apoptotic response elicited by CP and therefore can be considered as an efficient method of male fertility preservation.


Reducing black/white disparity: changes in injury mortality in the 15-24 year age group, United States, 1999-2005.
In 1999, the injury death rate for black males aged 15-24 in the USA was 80% greater than for white males: 148.5 vs 82.5/100,000, a difference of 66/100,000. Injury-specific changes between 1999 and 2005 in death rates for the 15-24 age group and in racial disparity were analysed using data from CDC's WISQARS. The gap between black and white all-injury death rates in males was reduced by 24%, to a difference of 50/100,000, largely because of greater decreases in the rates for motor vehicle crashes and firearm suicide in young black men than young white men, and large increases in suicide by suffocation and unintentional poisoning in the latter. Among females, despite a reduction in the black/white gap in firearm homicide rates, the gap between the races in total injury rates changed from a small black excess to a higher rate in young white women, which was due primarily to greater increases in these white women than black women in unintentional poisoning and suicide by suffocation, and greater decreases in black women than white women in firearm suicide.


Limited polymorphisms in k13 gene in Plasmodium falciparum isolates from Dakar, Senegal in 2012-2013.
The emergence of Plasmodium falciparum resistance to artemisinin and its derivatives, manifested as delayed parasite clearance following the treatment, has developed in Southeast Asia. The spread of resistance to artemisinin from Asia to Africa may be catastrophic for malaria control and elimination worldwide. Recently, mutations in the propeller domain of the Kelch 13 (k13) gene (PF3D71343700) were associated with in vitro resistance to artemisinin and with delayed clearance after artemisinin treatment in southern Asia. The aim of the study was to characterize the genetic variability of k13 and to evaluate the molecular resistance to artemisinin for the first time in Senegal. Plasmodium falciparum isolates were collected from 138 malaria patients in Dakar and its districts during the rainy season of October 2012 to January 2013 at the Hôpital Principal de Dakar. The k13 gene was amplified using nested PCR and sequenced. A very limited variability within the k13 gene in Senegalese P. falciparum isolates was identified. No polymorphism was detected in the six k13-propeller blades. Only two mutations, T149S (6.3%) and K189T (42.2%), and one (N) or two (NN) asparagine insertion at the codon 142 (4.7 and 6.3%, respectively) were detected in the Plasmodium/Apicomplexa-specific domain. None of the polymorphisms associated with artemisinin resistance in Southeast Asia was detected in the 138 P. falciparum from Dakar. The present data do not suggest widespread artemisinin resistance in Dakar in 2012-2013. Notably, the C580Y, R539T or Y493H substitutions that were associated with in vitro resistance or delayed parasite clearance in Southeast Asia were not observed in Dakar, nor were any of the polymorphisms observed in parasites from Southeast Asia, nor the M476I mutation that was selected in vitro with artemisinin pressure in a African parasite line.


Genetic mutations in adipose triglyceride lipase and myocardial up-regulation of peroxisome proliferated activated receptor-γ in patients with triglyceride deposit cardiomyovasculopathy.
Adipose triglyceride lipase (ATGL, also known as PNPLA2) is an essential molecule for hydrolysis of intracellular triglyceride (TG). Genetic ATGL deficiency is a rare multi-systemic neutral lipid storage disease. Information regarding its clinical profile and pathophysiology, particularly for cardiac involvement, is still very limited. A previous middle-aged ATGL-deficient patient in our institute (Case 1) with severe heart failure required cardiac transplantation (CTx) and exhibited a novel phenotype, "Triglyceride deposit cardiomyovasculopathy (TGCV)". Here, we tried to elucidate molecular mechanism underlying TGCV. The subjects were two cases with TGCV, including our second case who was a 33-year-old male patient (Case 2) with congestive heart failure requiring CTx. Case 2 was homozygous for a point mutation in the 5' splice donor site of intron 5 in the ATGL, which results in at least two types of mRNAs due to splicing defects. The myocardium of both patients (Cases 1 and 2) showed up-regulation of peroxisome proliferated activated receptors (PPARs), key transcription factors for metabolism of long chain fatty acids (LCFAs), which was in contrast to these molecules' lower expression in ATGL-targeted mice. We investigated the intracellular metabolism of LCFAs under human ATGL-deficient conditions using patients' passaged skin fibroblasts as a model. ATGL-deficient cells showed higher uptake and abnormal intracellular transport of LCFA, resulting in massive TG accumulation. We used these findings from cardiac specimens and cell-biological experiments to construct a hypothetical model to clarify the pathophysiology of the human disorder. In patients with TGCV, even when hydrolysis of intracellular TG is defective, the marked up-regulation of PPARγ and related genes may lead to increased uptake of LCFAs, the substrates for TG synthesis. This potentially vicious cycle of LCFAs could explain the massive accumulation of TG and severe clinical course for this rare disease.


Implementing pelvic floor muscle training in women's childbearing years: A critical interpretive synthesis of individual, professional, and service issues.
Antenatal pelvic floor muscle training (PFMT) may be effective for the prevention and treatment of urinary and fecal incontinence both in pregnancy and postnatally, but it is not routinely implemented in practice despite guideline recommendations. This review synthesizes evidence that exposes challenges, opportunities, and concerns regarding the implementation of PFMT during the childbearing years, from the perspective of individuals, healthcare professionals (HCPs), and organizations. Critical interpretive synthesis of systematically identified primary quantitative or qualitative studies or research syntheses of women's and HCPs attitudes, beliefs, or experiences of implementing PFMT. Fifty sources were included. These focused on experiences of postnatal urinary incontinence (UI) and perspectives of individual postnatal women, with limited evidence exploring the views of antenatal women and HCP or wider organizational and environmental issues. The concept of agency (people's ability to effect change through their interaction with other people, processes, and systems) provides an over-arching explanation of how PFMT can be implemented during childbearing years. This requires both individual and collective action of women, HCPs, maternity services and organizations, funders and policymakers. Numerous factors constrain women's and HCPs capacity to implement PFMT. It is unrealistic to expect women and HCPs to implement PFMT without reforming policy and service delivery. The implementation of PFMT during pregnancy, as recommended by antenatal care and UI management guidelines, requires policymakers, organizations, HCPs, and women to value the prevention of incontinence throughout women's lives by using low-risk, low-cost, and proven strategies as part of women's reproductive health.


Validation of a new outcome measure for orthopaedic trauma inpatients.
Functional performance is the most relevant outcome for trauma patients. Currently, no functional outcome measure for trauma inpatients exists. To report on reliability and validity testing of a new functional outcome measure for orthopaedic trauma inpatients. Descriptive study conducted to test selected psychometric properties of the new Functional Scale for Trauma Inpatients at Rashid Hospital in Dubai, United Arab Emirates. A total of 100 eligible male and female orthopaedic trauma inpatients aged 17 years and older were consecutively sampled. Physiotherapists administered the new scale to eligible patients. Descriptive statistics were used to describe the sample. Internal consistency was assessed by calculating Cronbach's α statistics. Sensitivity to change was assessed using one-way analysis of variance. Eighty-eight per cent of the 100 eligible subjects were male (mean age 34.75 ± 14.46 years). Sixty-three per cent of the patients had trauma to the lower extremity. The Cronbach's α-values were good (range 0.76-0.97). There was a significant difference in the mean scores over three administrations of 'bed', 'out of bed' and 'activities of daily living (ADL)' activity items of Functional Scale for Trauma Inpatients (P = 0.0000). Floor effects were noted at discharge for 'bed activities' and at discharge for 'ADL activities'. The only ceiling effect was noted at admission for 'out of bed activities'. The psychometric tests indicated that the tool is reliable, has sound internal consistency and has the ability to detect changes in functional performance of trauma inpatients. The scale fills an important need for physiotherapists working in trauma inpatient wards. Psychometric properties of the new scale should be tested in other countries.


Infrared radiance and solar glint at the ocean-sky horizon.
An analytic model is developed for the mean and clutter infrared radiance emitted from the ocean surface near the horizon and in the presence of solar glint. The model is based on the identification of a characteristic facet dimension over which the ocean surface is essentially flat. Fluctuations in the facet orientation generated by the water wave motion are modeled by a parameterized wave height power spectral density that provides the two orthogonal wave slope variances. The mean and root-meansquare facet radiances are calculated with Gaussian probability-density functions for the wave slopes. One can determine the number of facets within the field of view of a single detector by estimating the exposed ocean area and dividing by the facet area. This estimation takes into account shadowing effects of the swell wave, the swell wavelength, and the transverse detector field of view. The number of exposed facets together with the central-limit theorem permits computation of the radiance clutter as a function of look-down angle below the horizon. Vertical radiance profiles, parameterized by the azimuthal offset from the solar position, are calculated over a sensor look-down angle range of ±50 mrad about the horizon. The results of this analysis are compared with infrared radiance measurements of the ocean surface near the horizon and in the presence of solar glint. Agreement between the measured and calculated values of the mean and clutter radiances is good.


Plasma membrane-porating domain in poliovirus 2B protein. A short peptide mimics viroporin activity.
Picornavirus 2B, a non-structural protein required for effective viral replication, has been implicated in cell membrane permeabilization during the late phases of infection. Here, we have approached the molecular mechanism of this process by assessing the pore-forming activity of an overlapping peptide library that spanned the complete 2B sequence. At non-cytopathic concentrations, only the P3 peptide, spanning 2B residues 35-55, effectively assembled hydrophilic pores that allowed diffusion of low molecular mass solutes across the cell plasma membrane (IC(50) approximately 4x10(-7) M) and boundary liposome bilayers (starting at peptide to lipid molar ratios>1:10(4)). Circular dichroism data were consistent with its capacity to fold as a helix in a membrane-like environment. Furthermore, addition of this peptide to a sealed plasma-membrane model, consisting of retinal rod outer segments patch-clamped in a whole-cell configuration, induced ion channel activity within seconds at concentrations as low as 10(-8) M. Thus, we have established a "one-helix" 2B version that possesses the intrinsic pore-forming activity required to directly and effectively permeabilize the cell plasma membrane. We conclude that 2B viroporin can be classified as a genuine pore-forming toxin of viral origin, which is produced intracellularly at certain times post infection.


Relationship between dry eye symptoms and pain sensitivity.
Dry eye disease (DED) is common, but little is known about factors contributing to symptoms of dry eye, given the poor correlation between these symptoms and objective signs at the ocular surface. To explore whether pain sensitivity plays a role in patients' experience of DED symptoms. A population-based cross-sectional study of 1635 female twin volunteers, aged 20 to 83 years, from the TwinsUK adult registry. Dry eye disease was diagnosed if participants had at least 1 of the following: (1) a diagnosis of DED by a clinician, (2) the prescription of artificial tears, and/or (3) symptoms of dry eyes for at least 3 months. A subset of 689 women completed the Ocular Surface Disease Index (OSDI) questionnaire. Quantitative sensory testing using heat stimulus on the forearm was used to assess pain sensitivity (heat pain threshold [HPT]) and pain tolerance (heat pain suprathreshold [HPST]). Of the 1622 participants included, 438 (27.0%) were categorized as having DED. Women with DED showed a significantly lower HPT (P = .03) and HPST (P = .003)--and hence had higher pain sensitivity--than those without DED. A strong significant association between the presence of pain symptoms on the OSDI and the HPT and HPST was found (P = .008 for the HPT and P = .003 for the HPST). In addition, participants with an HPT below the median had DED pain symptoms almost twice as often as those with an HPT above the median (31.2% vs 20.5%; odds ratio, 1.76; 95% CI, 1.15-2.71; P = .01). High pain sensitivity and low pain tolerance are associated with symptoms of DED, adding to previous associations of the severity of tear insufficiency, cell damage, and psychological factors. Management of DED symptoms is complex, and physicians need to consider the holistic picture, rather than simply treating ocular signs.


Reliability and validity of ultrasound to estimate muscles: A comparison between different transducers and parameters.
We aimed to investigate the test-retest reliability and validity of ultrasound for two commonly used types of transducer, using different methods for the estimation of muscle size and echo intensity (EI). Fourteen healthy adults were included in this study. Ultrasound images of the rectus femoris size (thickness in cm and cross-sectional area [CSA] in cm2), obtained at the mid-thigh, were validated against MRI. Both a linear and a curved array transducer were used to assess rectus femoris size and EI (values 0-255, higher scores indicating increased intramuscular fat and interstitial fibrous tissue). To assess test-retest reliability of ultrasound, participants were tested twice, with a one-week interval. Validity and reliability were evaluated using paired sample t-tests, intraclass correlation coefficient (ICC), and Bland-Altman plots. No significant differences between the repeated evaluations of rectus femoris thickness, CSA and EI were found. Reliability for thickness and CSA evaluations was excellent for both transducers (ICC = 0.87-0.97) and moderate for EI (ICC = 0.42-0.44). Mean difference between MRI and ultrasound for CSA (curved = 0.59 cm2, p = 0.086; linear = 2.1 cm2, p = 0.002) and thickness (curved = 0.31 cm, p = 0.01; linear = 0.21 cm, p = 0.043) were small but significant, except for CSA using a curved transducer. Agreement between ultrasound and MRI ranged from moderate for thickness (ICC = 0.45) to excellent for CSA (ICC = 0.92). Our study demonstrates that the test-retest reliability and validity of muscle size estimation by ultrasound for both curved and linear array transducers seems to be adequate. Future studies should focus on the longitudinal evaluation of muscle size and EI by ultrasound.


Probable role of endogenous endotoxins in hepatocytolysis during murine hepatitis caused by frog virus 3.
Three new observations bear out the role of endogenous endotoxins in the pathogenesis of murine hepatitis caused by frog virus 3. First, the LD50 of endotoxin is 20 times lower in mice pretreated for 2.5 hr with a sublethal dose of frog virus 3 than in untreated mice. Animals inoculated with one sublethal dose of lipopolysaccharide 2.5 hr after injection of one sublethal dose of virus die, all having developed extensive hepatocellular necrosis. This hypersensitivity varies according to the intensity of virus-induced destruction of Kupffer cells, which are the intrahepatic target of the virus. Second, mortality is significantly lower and the interval between infection and death longer in axenic mice, which are largely protected from portal endotoxemia. Third, the impairment of some biologic activities of endotoxin (through treatment with polymyxin B or indomethacin, for example) protects mice against hepatic damage and death. Likewise, mice rendered tolerant to endotoxins, and C3H/HeJ mice, which are genetically resistant to endotoxins, survive challenge with frog virus 3 and are refractory with regard to hepatocytolysis . These findings suggest that, in hepatitis caused by frog virus 3, endogenous endotoxins are responsible for extensive hepatocytolysis since virus-induced damage to the hepatic reticuloendothelial system prevents their detoxification.


Bioinformatic characterisation of the effector repertoire of the strawberry pathogen Phytophthora cactorum.
The oomycete pathogen Phytophthora cactorum causes crown rot, a major disease of cultivated strawberry. We report the draft genome of P. cactorum isolate 10300, isolated from symptomatic Fragaria x ananassa tissue. Our analysis revealed that there are a large number of genes encoding putative secreted effectors in the genome, including nearly 200 RxLR domain containing effectors, 77 Crinklers (CRN) grouped into 38 families, and numerous apoplastic effectors, such as phytotoxins (PcF proteins) and necrosis inducing proteins. As in other Phytophthora species, the genomic environment of many RxLR and CRN genes differed from core eukaryotic genes, a hallmark of the two-speed genome. We found genes homologous to known Phytophthora infestans avirulence genes including Avr1, Avr3b, Avr4, Avrblb1 and AvrSmira2 indicating effector sequence conservation between Phytophthora species of clade 1a and clade 1c. The reported P. cactorum genome sequence and associated annotations represent a comprehensive resource for avirulence gene discovery in other Phytophthora species from clade 1 and, will facilitate effector informed breeding strategies in other crops.


External causes of death among persons with developmental disability: the effect of residential placement.
The authors analyzed death rates from external causes (accidents, injuries, homicides, etc.) for persons with developmental disability in California. There were 520 such deaths during the 1981-1995 study period, based on 733,705 person-years of exposure; this represents all persons who received any services from the state. Compared with the general California population, persons with developmental disability were at lower risk of homicide, suicide, and poisonings (standardized mortality ratios, 0.31-0.68), but higher risk of pedestrian accidents, falls, fires, and, especially, drowning (standardized mortality ratio=6.22). A major focus of the study was comparisons between different residential settings. Persons in semi-independent living had significantly higher risk than did those in their family home or group homes, with homicides rates being three times higher and pedestrian accidents rates being doubled, while persons in institutions had much lower risks with respect to most causes. Of the 28 deaths due to drug and medication overdoses, 79 percent occurred in supported living or small-group homes. Avoidable deaths could be reduced by making direct care staff more aware of the risks and better trained in acute care, along with improved monitoring of special incidents.


Healthcare-seeking preferences of patients with sexually transmitted infection attending a tertiary care center in South Kerala.
Sexually transmitted infections (STIs) are a major public health problem in developing countries. These diseases are associated with increased risk of transmission of human immunodeficiency virus as well as adverse outcomes on pregnancy and reproductive health. Sexual behavior and healthcare-seeking behavior are identified as the true risk factors of STIs. Hospital-based cross-sectional study design was adopted. Eighty-five STI patients were studied regarding the inappropriate treatment-seeking behavior, the nature of the first point of contact with the health care, the appropriateness of treatment and the concerns of the patient regarding the services rendered by government health-care facilities. Among the 85 patients studied, 55.3% were males and 44.7% were females. Inappropriate treatment-seeking behavior was seen in 29.8% of males and 36.8% of females. About 59.6% of males and 81.6% of females sought appropriate treatment from modern medicine practitioners before attending our institution. Only 7.1% of males and 3.2% of females received appropriate treatment. The government sector was the choice of treatment for 46.4% males and 93.5% females and this difference was statistically significant (P = 0.00081). Lack of free medicines, issues of confidentiality, and privacy were the major service-related issues in the public sector. Appropriate treatment at the first point of contact with the health system is an important measure to prevent further transmission and development of complications. Health providers from both private and public sector should be given frequent periodic training regarding syndromic management of STIs and the training should stress on the need for risk reduction and condom promotion messages along with medical management. Program planners should take necessary steps to ensure adequate and continuous supply of free drugs and tackle issues of confidentiality and privacy.


Role of nonworking muscle on blood metabolites and ions with intense intermittent exercise.
The exchange of ions between blood and inactive muscle was studied in healthy males (n = 11) who performed four 30-s bouts of maximum isokinetic leg exercise, with 4 min of rest between bouts; recovery was followed for 90 min. Blood was sampled from the brachial artery and antecubital vein from inactive arm, and biopsies were taken from the nonworking deltoid muscles. During exercise, whole blood and plasma arteriovenous (a-v) differences across the arm showed increased net uptake of lactate (Lac-), K+, Na+, Cl-, glycerol, and O2. Arm glucose uptake also increased during exercise and the last 30 min of recovery, but there was no change in muscle glycogen content. Deltoid intracellular Lac- concentration [( Lac-]) increased 2.5-fold to 8.2 +/- 1.4 meq/l during exercise and 25 min of recovery. Despite evidence of uptake from blood and plasma a-v differences, deltoid intracellular K+, Na+, and Cl- concentrations did not change. During recovery, arm a-v differences showed no Lac- release, but Cl- and Na+ were released to venous blood for at least 90 min of recovery. During repeated exercise, the elevated Lac- uptake (a-v [Lac-] is 5-8 meq/l) and a HCO3- release (a-v HCO3- concentration is -8-11 meq/l) by the arm was suggestive of a possible anion exchange (Lac-/HCO3-). It is concluded that during heavy exercise non-working muscles take up and oxidize significant amounts of Lac-. Other inactive tissues also play a role in the regulation of ion and metabolite concentrations of blood.


Altered intracellular calcium homeostasis and endoplasmic reticulum redox state in Saccharomyces cerevisiae cells lacking Grx6 glutaredoxin.
Glutaredoxin 6 (Grx6) of Saccharomyces cerevisiae is an integral thiol oxidoreductase protein of the endoplasmic reticulum/Golgi vesicles. Its absence alters the redox equilibrium of the reticulum lumen toward a more oxidized state, thus compensating the defects in protein folding/secretion and cell growth caused by low levels of the oxidase Ero1. In addition, null mutants in GRX6 display a more intense unfolded protein response than wild-type cells upon treatment with inducers of this pathway. These observations support a role of Grx6 in regulating the glutathionylation of thiols of endoplasmic reticulum/Golgi target proteins and consequently the equilibrium between reduced and oxidized glutathione in the lumen of these compartments. A specific function influenced by Grx6 activity is the homeostasis of intracellular calcium. Grx6-deficient mutants have reduced levels of calcium in the ER lumen, whereas accumulation occurs at the cytosol from extracellular sources. This results in permanent activation of the calcineurin-dependent pathway in these cells. Some but not all the phenotypes of the mutant are coincident with those of mutants deficient in intracellular calcium transporters, such as the Golgi Pmr1 protein. The results presented in this study provide evidence for redox regulation of calcium homeostasis in yeast cells.


Characterization of human papillomavirus (HPV) 16 E6 seropositive individuals without HPV-associated malignancies after 10 years of follow-up in the UK Biobank.
Antibodies against the HPV16 oncoprotein E6 are promising biomarkers for HPV16-driven oropharyngeal cancer (HPV16-OPC) due to their high sensitivity and specificity, and prospective manifestation. In previous studies, 0•7% of controls without HPV-associated malignancies were HPV16 E6 seropositive of which only a minority is expected to develop HPV16-driven cancer. We aimed to characterise HPV16 E6 antibodies in individuals without HPV-associated malignancies. We analysed serum antibodies against HPV16 E6, E7, L1 and HPV18 L1 in a random sample (n = 9,695) of the prospective UK Biobank cohort (UKB). Excluding individuals with potentially HPV-associated malignancies (n = 192), we assessed risk factors for seropositivity by logistic regression. In individuals without potentially HPV-associated malignancies (n = 9,503), the HPV16 E6 seroprevalence was 0•8%. Seropositivity against HPV16 E6 and all other HPV antigens was strongly associated with sexual behaviour. The seroprevalence of HPV16 E6, L1 and HPV18 L1 increased with the number of lifetime sex partners (ptrend<0•005), and all HPV antibodies were associated with same-sex intercourse (ORE6 3•1, 95%CI 1•4-6•9; reference category: no same-sex intercourse). HPV16 E6 and L1 seropositivity were associated with young age (≤17 years) at sexual debut (ORE6 2•0, 95%CI 1•1-3•7) compared with individuals reporting sexual debut at age ≥20 years. This is the first study characterising HPV16 E6 antibodies in the general UK population. Their strong association with sexual behaviour, and overlapping risk factor profiles with other HPV antibodies support their relevance for HPV16-OPC disease prediction. However, additional risk stratification will be required to identify individuals at highest risk to develop HPV16-OPC.


Interaction between smoking and functional polymorphism in the TGFB1 gene is associated with ischaemic heart disease and myocardial infarction in patients with rheumatoid arthritis: a cross-sectional study.
Transforming growth factor-beta1 (TGF-beta1) is a pleiotropic cytokine that plays important roles in immunity and inflammation. Some studies have suggested that polymorphism in the TGFB1 gene is associated with heart disease in the general population. The purpose of the present study was to determine whether common single-nucleotide polymorphisms (SNP) in the TGFB1 gene are associated with ischaemic heart disease (IHD) and/or myocardial infarction (MI) in patients with rheumatoid arthritis (RA), and to investigate the influence of smoking on any association. PCR-based assays were used to determine the genotypes of TGFB1 SNPs including TGFB1-509 C/T (rs1800469, in the promoter region), +868 T/C (rs1800470, in exon 1) and +913 G/C (rs1800471, in exon 1) in 414 subjects with established RA. Genotyping for the +868 SNP was also carried out on a second study population of RA patients (n = 259) with early disease. Serum levels of TGF-beta1 were measured using a commercial ELISA kit. Smoking history and IHD/MI status were obtained on each patient. Associations with IHD/MI were assessed using contingency tables and logistic regression analyses. The heterozygous genotype of TGFB+868 was associated with an increased risk of IHD (OR 2.14, 95% CI 1.30 - 3.55) and MI (OR 2.42, 95% CI 1.30-4.50), compared to the homozygous genotypes combined. Smoking was an independent risk for IHD and MI, and evidence of interaction between smoking and TGFB+868 was found. Multivariate analyses indicated that the strongest associations with IHD and MI were due to the combined effect of the TGFB1+868 TC genotype and smoking (OR 2.75, 95% CI 1.59-4.75; and OR 2.58 95% CI 1.33-4.99, respectively), independent of other cardiovascular risk factors. The association of the +868 TC genotype and evidence of +868 TC-smoking interaction with IHD were replicated in a second population of RA patients with early disease. Serum TGF-beta1 levels were not associated with TGFB1 genetic variations, smoking or IHD/MI status. Interaction between smoking and polymorphism in the TGFB1 gene may influence the risk of IHD and MI in patients with RA.


Determination of itraconazole and hydroxyitraconazole in human serum and plasma by micellar electrokinetic chromatography.
The electrokinetic separation of the hydrophobic antimycotic drug itraconazole (ITC) and its major metabolite, hydroxyitraconazole (HITC), by a binary aqueous-organic solvent medium containing sodium dodecylsulfate, by microemulsion electrokinetic chromatography (MEEKC) and by micellar electrokinetic chromatography (MEKC) was studied. The results suggest that the first approach is difficult to apply and that there is no substantial difference between separations performed using MEEKC and MEKC modified with n-butanol. The simpler MEKC method is more than adequate and was thus employed for the analysis of ITC and HITC in human serum and plasma. Separation was achieved in plain fused-silica capillaries having a low-pH buffer (pH 2.2) with sodium dodecyl sulfate micelles and reversed polarity. The addition of 2-propanol and n-butanol enhanced analyte solubility and altered the selectivity of the separation by influencing the magnitude of the electrophoretic component in the separation mechanism. Under optimised conditions and using head-column field-amplified sample stacking, an internal standard, ITC and two forms of HITC could be separated in under 9 min, with detection limits less than 0.01 microg/mL. Analysis of samples from patients currently prescribed ITC revealed a different HITC peak area ratio to that of the standards, suggesting a stereoselective component of ITC metabolisation. Comparison of MEKC data with those of a HPLC method employed on a routine basis showed excellent agreement, indicating the potential of this approach for therapeutic drug monitoring of ITC.


A link between protein translation and body weight.
Nutrient overload induces obesity, a primary risk factor for type 2 diabetes. Ribosomal biogenesis and protein synthesis, which are controlled by the mammalian target of rapamycin (mTOR), are primary energy-consuming processes in cells. mTOR phosphorylates and inactivates members of the eukaryotic translation initiation factor 4E-binding (eIF4E-binding) protein (4E-BP) family, which are translational repressors of 5' cap-dependent protein synthesis. In this issue of the JCI, Le Bacquer et al. report that simultaneous deletion of both 4E-BP1 and 4E-BP2 in mice results in insulin resistance, decreased energy expenditure, and increased adipogenesis (see the related article beginning on page 387). These findings link protein synthesis, insulin sensitivity, and body weight.


Endoscopic morphologic features of ulcerative colitis-associated dysplasia classified according to the SCENIC consensus statement.
Recent advances in endoscopic equipment and diagnostic techniques have made possible the detection of early dysplasia in the inflamed mucosa of ulcerative colitis (UC). The SCENIC consensus statement recommends the use of unified terminology for the morphology of dysplasia. In this study, we investigated the endoscopic features of high-grade dysplasia (HGD) in a clinical setting. We retrospectively identified 62 patients with UC who were diagnosed with colitis-associated cancer or HGD between 1997 and 2015. A total of 39 lesions of HGD detected by targeted biopsy sampling in 31 patients were reviewed, and the endoscopic morphology was classified according to the SCENIC guidelines. In total, 0 (0%), 6 (15.4%), 19 (48.7%), 12 (30.8%), and 2 (5.1%) lesions with HGD were classified as pedunculated, sessile, superficial elevated, flat, and depressed, respectively. Nearly 80% of the lesions were located in the rectum or sigmoid colon. All flat and depressed lesions were red in color. Typically, sessile/superficial elevated lesions accompanied a flat area (Is+IIb/IIa+IIb). Ulceration was observed in 2 depressed lesions (5.1%). Although the borders were indistinct in 21 lesions (53.8%) without the use of magnifying colonoscopy, all lesions could be distinguished from the surrounding mucosa using magnifying endoscopy. This is the first study to classify the morphologic features of HGD using the SCENIC guidelines in a clinical setting. Based on our findings, endoscopists should recognize that HGD is frequently associated with a flat/superficial elevated area and red discoloration and should inspect particularly carefully in the rectum and sigmoid colon. The findings of chromoendoscopy and magnifying colonoscopy may also be useful in distinguishing lesions from the surrounding mucosa.


[Evolution and virulence of primate lentiviruses].
While the AIDS epidemic caused by human immunodeficiency viruses (HIV) has resulted in the death of over 20 million people worldwide, simian immunodeficiency virus (SIV) infection, found in numerous African primate species, does not induce disease symptoms. The factors accounting for this difference between humans and natural host of SIV remain poorly understood. The entangled nature of the host/virus relationship could be the answer, rather than independent virus or host factors. Such a relationship is as a consequence of host/virus adaptation which has evolved over long periods in naturally infected primate species.


Management of the mother-infant dyad with suspected or confirmed SARS-CoV-2 infection in a highly epidemic context.
In the context of SARS-CoV-2 pandemic, the hospital management of mother-infant pairs poses to obstetricians and neonatologists previously unmet challenges. In Lombardy, Northern Italy, 59 maternity wards networked to organise the medical assistance of mothers and neonates with suspected or confirmed SARS-CoV-2 infection. Six "COVID-19 maternity centres" were identified, the architecture and activity of obstetric and neonatal wards of each centre was reorganised, and common assistance protocols for the management of suspected and proven cases were formulated. Here, we present the key features of this reorganization effort, and our current management of the mother-infant dyad before and after birth, including our approach to rooming-in practice, breastfeeding and neonatal follow-up, based on the currently available scientific evidence. Considered the rapid diffusion of COVID-19 all over the world, we believe that preparedness is fundamental to assist mother-infant dyads, minimising the risk of propagation of the infection through maternity and neonatal wards.


Genome-wide microRNA screening in Nile tilapia reveals pervasive isomiRs' transcription, sex-biased arm switching and increasing complexity of expression throughout development.
MicroRNAs (miRNAs) are key regulators of gene expression in multicellular organisms. The elucidation of miRNA function and evolution depends on the identification and characterization of miRNA repertoire of strategic organisms, as the fast-evolving cichlid fishes. Using RNA-seq and comparative genomics we carried out an in-depth report of miRNAs in Nile tilapia (Oreochromis niloticus), an emergent model organism to investigate evo-devo mechanisms. Five hundred known miRNAs and almost one hundred putative novel vertebrate miRNAs have been identified, many of which seem to be teleost-specific, cichlid-specific or tilapia-specific. Abundant miRNA isoforms (isomiRs) were identified with modifications in both 5p and 3p miRNA transcripts. Changes in arm usage (arm switching) of nine miRNAs were detected in early development, adult stage and even between male and female samples. We found an increasing complexity of miRNA expression during ontogenetic development, revealing a remarkable synchronism between the rate of new miRNAs recruitment and morphological changes. Overall, our results enlarge vertebrate miRNA collection and reveal a notable differential ratio of miRNA arms and isoforms influenced by sex and developmental life stage, providing a better picture of the evolutionary and spatiotemporal dynamics of miRNAs.


Validation of input-noise model for simulations of supercontinuum generation and rogue waves.
A new model of pump noise in supercontinuum and rogue wave generation is presented. Simulations are compared with experiments and show that the new model provides significantly better agreement than the currently ubiquitously used one-photon-per-mode model. The new model also allows for a study of the influence of the pump spectral line width on the spectral broadening mechanisms. Specifically, it is found that for four-wave mixing (FWM) a narrow spectral line width ( similar 0.1 nm) initially leads to a build-up of FWM from quantum noise, whereas a broad spectral line width (succeeds, similar 1 nm) initially leads to a gradual broadening of the pump spectrum. Since the new model provides better agreement with experiments and is still simple to implement, it is particularly important that it is used for future studies of the statistical properties of nonlinear spectral broadening, such as the formation of rogue waves.


Endovascular aneurysm repair versus open repair in patients with abdominal aortic aneurysm (EVAR trial 1): randomised controlled trial.
Although endovascular aneurysm repair (EVAR) has a lower 30-day operative mortality than open repair, the long-term results of EVAR are uncertain. We instigated EVAR trial 1 to compare these two treatments in terms of mortality, durability, health-related quality of life (HRQL), and costs for patients with large abdominal aortic aneurysm (AAA). We did a randomised controlled trial of 1082 patients aged 60 years or older who had aneurysms of at least 5.5 cm in diameter and who had been referred to one of 34 hospitals proficient in the EVAR technique. We assigned patients who were anatomically suitable for EVAR and fit for an open repair to EVAR (n=543) or open repair (n=539). Our primary endpoint was all-cause mortality, with secondary endpoints of aneurysm related mortality, HRQL, postoperative complications, and hospital costs. Analyses were by intention to treat. 94% (1017 of 1082) of patients complied with their allocated treatment and 209 died by the end of follow-up on Dec 31, 2004 (53 of aneurysm-related causes). 4 years after randomisation, all-cause mortality was similar in the two groups (about 28%; hazard ratio 0.90, 95% CI 0.69-1.18, p=0.46), although there was a persistent reduction in aneurysm-related deaths in the EVAR group (4%vs 7%; 0.55, 0.31-0.96, p=0.04). The proportion of patients with postoperative complications within 4 years of randomisation was 41% in the EVAR group and 9% in the open repair group (4.9, 3.5-6.8, p<0.0001). After 12 months there was negligible difference in HRQL between the two groups. The mean hospital costs per patient up to 4 years were UK pound sterling 13,257 for the EVAR group versus pound sterling 9946 for the open repair group (mean difference pound sterling 3311, SE 690). Compared with open repair, EVAR offers no advantage with respect to all-cause mortality and HRQL, is more expensive, and leads to a greater number of complications and reinterventions. However, it does result in a 3% better aneurysm-related survival. The continuing need for interventions mandates ongoing surveillance and longer follow-up of EVAR for detailed cost-effectiveness assessment.


The presence of lymphovascular and perineural infiltration after neoadjuvant therapy and oesophagectomy identifies patients at high risk for recurrence.
In patients treated for oesophageal cancer the importance of lymphovascular and perineural invasion (PNI) after neoadjuvant therapy has yet to be established. The aim of this study was to assess the incidence and prognostic significance of these factors in a consecutive series of patients with cancer of the oesophagus or gastro-oesophageal junction (GOJ) who underwent neoadjuvant therapy followed by oesophagectomy. Clinical and pathology results from patients with potentially curable adenocarcinoma, or squamous cell carcinoma of the oesophagus or GOJ were reviewed. Patients were treated with neoadjuvant chemotherapy or chemoradiation followed by transthoracic oesophagectomy and two-field lymphadenectomy. The presence of venous invasion (VI), lymph vessel invasion (LI) and perineural invasion (PNI) were correlated with clinical outcomes. A total of 396 patients underwent oesophagectomy after neoadjuvant therapy for oesophageal cancer. Venous invasion was identified in 150 (38%) of patients, LI in 203 (51%) patients and PNI in 204 (52%) patients. In all, 123 (31%) patients had no evidence of either VI, LI or PNI. A total of 96 (24%) had a combination of two factors and 94 (24%) had all three factors. The presence of VI, LI and PNI was significantly related to tumour stage (P=0.001). Median overall survival was 170.8 months when all three factors were absent, 44.0 months when one factor was present, 27.1 months when two factors were present and 16.0 months when all were present. Multivariate analyses revealed VI, LI and PNI or a combination of these factors were independent predictors of prognosis. In oesophageal cancer patients treated with neoadjuvant therapy followed by oesophagectomy the presence of VI, LI and PNI has an important prognostic impact and may identify patients at high risk of recurrence who would benefit from adjuvant therapies.


Detection of serum neutralizing antibodies to Simbu sero-group viruses in cattle in Tanzania.
Orthobunyaviruses belonging to the Simbu sero-group occur worldwide, including the newly recognized Schmallenberg virus (SBV) in Europe. These viruses cause congenital malformations and reproductive losses in ruminants. Information on the presence of these viruses in Africa is scarce and the origin of SBV is unknown. The aim of this study was to investigate the presence of antibodies against SBV and closely related viruses in cattle in Tanzania, and their possible association with reproductive disorders. In a cross-sectional study, serum from 659 cattle from 202 herds collected in 2012/2013 were analyzed using a commercial kit for SBV ELISA, and 61 % were positive. Univariable logistic regression revealed significant association between ELISA seropositivity and reproductive disorders (OR = 1.9). Sera from the same area collected in 2008/2009, before the SBV epidemic in Europe, were also tested and 71 (54.6 %) of 130 were positive. To interpret the ELISA results, SBV virus neutralization test (VNT) was performed on 110 sera collected in 2012/2013, of which 51 % were positive. Of 71 sera from 2008/2009, 21 % were positive. To investigate potential cross reactivity with related viruses, 45 sera from 2012/2013 that were positive in SBV ELISA were analyzed in VNTs for Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda, Simbu and Tinaroo viruses. All 45 sera were positive for one or more of these viruses. Twenty-nine sera (64.4 %) were positive for SBV, and one had the highest titer for this virus. This is the first indication that Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda and Tinaroo viruses circulate and cause negative effect on reproductive performance in cattle in Tanzania. SBV or a closely related virus was present before the European epidemic. However, potential cross reactivity complicates the interpretation of serological studies in areas where several related viruses may circulate. Virus isolation and molecular characterization in cattle and/or vectors is recommended to further identify the viruses circulating in this region. However, isolation in cattle is difficult due to short viremic period of 2 to 6 days, and isolation in vectors does not necessarily reflect the situation in cattle.


Student public commitment in a school-based diabetes prevention project: impact on physical health and health behavior.
As concern about youth obesity continues to mount, there is increasing consideration of widespread policy changes to support improved nutritional and enhanced physical activity offerings in schools. A critical element in the success of such programs may be to involve students as spokespeople for the program. Making such a public commitment to healthy lifestyle program targets (improved nutrition and enhanced physical activity) may potentiate healthy behavior changes among such students and provide a model for their peers. This paper examines whether student's "public commitment"--voluntary participation as a peer communicator or in student-generated media opportunities--in a school-based intervention to prevent diabetes and reduce obesity predicted improved study outcomes including reduced obesity and improved health behaviors. Secondary analysis of data from a 3-year randomized controlled trial conducted in 42 middle schools examining the impact of a multi-component school-based program on body mass index (BMI) and student health behaviors. A total of 4603 students were assessed at the beginning of sixth grade and the end of eighth grade. Process evaluation data were collected throughout the course of the intervention. All analyses were adjusted for students' baseline values. For this paper, the students in the schools randomized to receive the intervention were further divided into two groups: those who participated in public commitment activities and those who did not. Students from comparable schools randomized to the assessment condition constituted the control group. We found a lower percentage of obesity (greater than or equal to the 95th percentile for BMI) at the end of the study among the group participating in public commitment activities compared to the control group (21.5% vs. 26.6%, p = 0.02). The difference in obesity rates at the end of the study was even greater among the subgroup of students who were overweight or obese at baseline; 44.6% for the "public commitment" group, versus 53.2% for the control group (p = 0.01). There was no difference in obesity rates between the group not participating in public commitment activities and the control group (26.4% vs. 26.6%). Participating in public commitment activities during the HEALTHY study may have potentiated the changes promoted by the behavioral, nutrition, and physical activity intervention components. ClinicalTrials.gov number, NCT00458029.


Preclinical evaluation of the imipridone family, analogs of clinical stage anti-cancer small molecule ONC201, reveals potent anti-cancer effects of ONC212.
Anti-cancer small molecule ONC201 upregulates the integrated stress response (ISR) and acts as a dual inactivator of Akt/ERK, leading to TRAIL gene activation. ONC201 is under investigation in multiple clinical trials to treat patients with cancer. Given the unique imipridone core chemical structure of ONC201, we synthesized a series of analogs to identify additional compounds with distinct therapeutic properties. Several imipridones with a broad range of in vitro potencies were identified in an exploration of chemical derivatives. Based on in vitro potency in human cancer cell lines and lack of toxicity to normal human fibroblasts, imipridones ONC206 and ONC212 were prioritized for further study. Both analogs inhibited colony formation, and induced apoptosis and downstream signaling that involves the integrated stress response and Akt/ERK, similar to ONC201. Compared to ONC201, ONC206 demonstrated improved inhibition of cell migration while ONC212 exhibited rapid kinetics of activity. ONC212 was further tested in >1000 human cancer cell lines in vitro and evaluated for safety and anti-tumor efficacy in vivo. ONC212 exhibited broad-spectrum efficacy at nanomolar concentrations across solid tumors and hematological malignancies. Skin cancer emerged as a tumor type with improved efficacy relative to ONC201. Orally administered ONC212 displayed potent anti-tumor effects in vivo, a broad therapeutic window and a favorable PK profile. ONC212 was efficacious in vivo in BRAF V600E melanoma models that are less sensitive to ONC201. Based on these findings, ONC212 warrants further development as a drug candidate. It is clear that therapeutic utility extends beyond ONC201 to include additional imipridones.


The evolution of Ebola virus: Insights from the 2013-2016 epidemic.
The 2013-2016 epidemic of Ebola virus disease in West Africa was of unprecedented magnitude and changed our perspective on this lethal but sporadically emerging virus. This outbreak also marked the beginning of large-scale real-time molecular epidemiology. Here, we show how evolutionary analyses of Ebola virus genome sequences provided key insights into virus origins, evolution and spread during the epidemic. We provide basic scientists, epidemiologists, medical practitioners and other outbreak responders with an enhanced understanding of the utility and limitations of pathogen genomic sequencing. This will be crucially important in our attempts to track and control future infectious disease outbreaks.


Nephrectomy in autosomal dominant polycystic kidney disease: a patient with exceptionally large, still functioning kidneys.
Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disease. It is characterized by progressive cyst formation in both kidneys, often leading to end-stage kidney disease. Indications for surgical removal of an ADPKD kidney include intractable pain, hematuria, infection, or exceptional enlargement and small abdominal cavity hampering implantation of a donor kidney. We report the case of an extraordinarily large ADPKD kidney weighing 8.7 kg (19.3 lb) with a maximal length of 48 cm (19 inch), and with cysts filled with both clear and bloody fluid.


Perceptions of unfairness in allocations between multiple recipients.
This paper introduces a new model to explain perceptions of unfairness in resource allocations between multiple recipients. The model yields several novel predictions, all confirmed in a series of new empirical tests. For instance, while much prior research focuses on the differences between the judge's share and others' shares, we argue that people also care about differences between others' shares. In particular, the presence of a single loser increases perceptions of unfairness. We also study individual variation in sensitivity to the single-loser dimension. Most centrally, we offer empirical support for the existence - indeed the prevalence - of ostraphobics, individuals with an acute sensitivity to being "ostracized" as a sole loser. We show that ostraphobics perceive unfairness more strongly than other types, are higher in need to belong and fear of negative evaluation, and are more prone to a heretofore unrecognized type of preference reversal with respect to fairness.


Measuring caregiver outcomes in palliative care: a construct validation study of two instruments for use in economic evaluations.
Providing care to patients nearing the end of life can place a considerable burden on caregivers. Hence, policy decisions on interventions in palliative care should be guided by information on this burden. This study investigates construct validation of two preference-based caregiver outcome instruments suitable for economic evaluations: the Carer Experience Scale (CES) and the Care-related Quality of Life (CarerQol) instrument. Moreover, this study reports caregiver experiences in end-of-life care. Data were collected with written questionnaires among caregivers of patients receiving palliative care services in the Southern metropolitan area of Adelaide, South Australia (n = 97). The effect of caregiving on caregivers was measured with the CES, CarerQol, Process Utility (PU) and Caregiver Strain Index (CSI). Convergent, discriminative and clinical validity were studied. As hypothesized, higher negative effect of caregiving measured on the CES was associated with higher negative effect on the CarerQol. Both the CES and CarerQol were associated in the expected positive direction with less strain from caregiving (CSI), more positive care experiences and more PU from caring. Caregivers' and care recipients' health status and duration of caregiving were negatively associated with caring experiences. Our findings suggest that the CES and CarerQol validly assess the effect of caregiving on caregivers in end-of-life care. Economic evaluations in end-of-life care should attempt to incorporate such instruments to provide a more holistic assessment of the true impact of interventions, especially where family and friends are heavily involved in caregiving.


Indole production promotes Escherichia coli mixed-culture growth with Pseudomonas aeruginosa by inhibiting quorum signaling.
Indole production by Escherichia coli, discovered in the early 20th century, has been used as a diagnostic marker for distinguishing E. coli from other enteric bacteria. By using transcriptional profiling and competition studies with defined mutants, we show that cyclic AMP (cAMP)-regulated indole formation is a major factor that enables E. coli growth in mixed biofilm and planktonic populations with Pseudomonas aeruginosa. Mutants deficient in cAMP production (cyaA) or the cAMP receptor gene (crp), as well as indole production (tnaA), were not competitive in coculture with P. aeruginosa but could be restored to wild-type competitiveness by supplementation with a physiologically relevant indole concentration. E. coli sdiA mutants, which lacked the receptor for both indole and N-acyl-homoserine lactones (AHLs), showed no change in competitive fitness, suggesting that indole acted directly on P. aeruginosa. An E. coli tnaA mutant strain regained wild-type competiveness if grown with P. aeruginosa AHL synthase (rhlI and rhlI lasI) mutants. In contrast to the wild type, P. aeruginosa AHL synthase mutants were unable to degrade indole. Indole produced during mixed-culture growth inhibited pyocyanin production and other AHL-regulated virulence factors in P. aeruginosa. Mixed-culture growth with P. aeruginosa stimulated indole formation in E. coli cpdA, which is unable to regulate cAMP levels, suggesting the potential for mixed-culture gene activation via cAMP. These findings illustrate how indole, an early described feature of E. coli central metabolism, can play a significant role in mixed-culture survival by inhibiting quorum-regulated competition factors in P. aeruginosa.


Use of novel sensors combining local positioning and acceleration to measure feeding behavior differences associated with lameness in dairy cattle.
Time constraints for dairy farmers are an important factor contributing to the under-detection of lameness, resulting in delayed or missed treatment of lame cows within many commercial dairy herds. Hence, a need exists for flexible and affordable cow-based sensor systems capable of monitoring behaviors such as time spent feeding, which may be affected by the onset of lameness. In this study a novel neck-mounted mobile sensor system that combines local positioning and activity (acceleration) was tested and validated on a commercial UK dairy farm. Position and activity data were collected over 5 consecutive days for 19 high-yield dairy cows (10 lame, 9 nonlame) that formed a subset of a larger (120 cow) management group housed in a freestall barn. A decision tree algorithm that included sensor-recorded position and accelerometer data was developed to classify a cow as doing 1 of 3 categories of behavior: (1) feeding, (2) not feeding, and (3) out of pen for milking. For each classified behavior the mean number of bouts, the mean bout duration, and the mean total duration across all bouts was determined on a daily basis, and also separately for the time periods in between milking (morning = 0630-1300 h; afternoon = 1430-2100 h; night = 2230-0500 h). A comparative analysis of the classified cow behaviors was undertaken using a Welch t-test with Benjamini-Hochberg post-hoc correction under the null hypothesis of no differences in the number or duration of behavioral bouts between the 2 test groups of lame and nonlame cows. Analysis showed that mean total daily feeding duration was significantly lower for lame cows compared with non-lame cows. Behavior was also affected by time of day with significantly lower mean total duration of feeding and higher total duration of nonfeeding in the afternoons for lame cows compared with nonlame cows. The results demonstrate how sensors that measure both position and acceleration are capable of detecting differences in feeding behavior that may be associated with lameness. Such behavioral differences could be used in the development of predictive algorithms for the prompt detection of lameness as part of a commercially viable automated behavioral monitoring system.


Epithelial cell organization suppresses Myc function by attenuating Myc expression.
c-Myc is an oncogene transcription factor that causes cancer in many settings, including solid tumors that arise in the context of organized tissue structures. Given that disruption of tissue architecture frequently occurs in cancer, there is considerable interest in how cell organization impacts oncogene function. A previous report found that organization of mammary epithelial cells into defined 3-dimensional structures renders them insensitive to the effects of retrovirus-mediated overexpression of Myc, leading to the notion that organization tempers the sensitivity of individual cells to Myc activity. In this article, we report that epithelial cell organization does not profoundly alter Myc activity but, instead, suppresses Myc by modulating its expression. We show that the morphogenesis of mammary epithelial cells into organized acinar structures in vitro is accompanied by widespread changes in gene expression patterns, including a substantial decrease in the expression of Myc. Concomitant with the decrease in endogenous Myc expression, we observe a decrease in transcription from retroviral vectors during morphogenesis and find that Myc transgene expression in acini is much lower than in unorganized cells. This decrease in Myc transgene activity is responsible for the apparent recalcitrance of organized cells to ectopic Myc, as adenovirus-mediated expression of Myc in organized structures potently induces apoptosis. These observations reveal that organization does not alter the inherent response of epithelial cells to Myc and suggest that other tumor suppression mechanisms, apart from structure, antagonize Myc in the development of solid tumors.


VLA-4 integrin cross-linking on human monocytic THP-1 cells induces tissue factor expression by a mechanism involving mitogen-activated protein kinase.
Adhesion molecules such as VLA-4 are important not only for monocyte adhesion to extracellular matrix proteins, but also for subsequent cell activation. Monocyte adherence to fibronectin or engagement of VLA-4 has been demonstrated to stimulate production of potent inflammatory mediators such as tumor necrosis factor-alpha, interleukin-1, and the procoagulant tissue factor protein. However, the intracellular signaling cascades leading to gene expression have not been elucidated. Using the human monocytic THP-1 cell line, VLA-4 cross-linking by monoclonal antibodies directed against its alpha4 and beta1 subunits produced a time-dependent increase in tyrosine phosphorylation of a broad range of cellular proteins. Using Western blot analysis directed against the phosphorylated form of the extracellular signal-related kinase (ERK) mitogen-activated protein (MAP) kinase proteins, as well as immunoprecipitation and in vitro kinase assays, we found that VLA-4 cross-linking increased ERK1/ERK2 tyrosine phosphorylation and activity. In conjunction, integrin cross-linking also increased NF-kappaB nuclear translocation and 4-h expression of tissue factor. Inhibition of tyrosine kinase activity with genistein (10 microg/ml) as well as selective MAP kinase inhibition with the MEK-1 inhibitor PD98059 abolished the VLA-4-dependent ERK tyrosine phosphorylation, inhibited NF kappaB nuclear binding, and abrogated tissue factor expression induced by both VLA-4 cross-linking and adhesion to fibronectin in THP-1 cells and human peripheral blood monocytes. These studies point to the involvement of the MAP kinase pathway in the activation of monocytic cells during transmigration to inflammatory sites.


Bench-to-bedside review: Citrate for continuous renal replacement therapy, from science to practice.
To prevent clotting in the extracorporeal circuit during continuous renal replacement therapy (CRRT) anticoagulation is required. Heparin is still the most commonly used anticoagulant. However, heparins increase the risk of bleeding, especially in critically ill patients. Evidence has accumulated that regional anticoagulation of the CRRT circuit with citrate is feasible and safe. Compared to heparin, citrate anticoagulation reduces the risk of bleeding and requirement for blood products, not only in patients with coagulopathy, but also in those without. Metabolic complications are largely prevented by the use of a strict protocol, comprehensive training and integrated citrate software. Recent studies indicate that citrate can even be used in patients with significant liver disease provided that monitoring is intensified and the dose is carefully adjusted. Since the citric acid cycle is oxygen dependent, patients at greatest risk of accumulation seem to be those with persistent lactic acidosis due to poor tissue perfusion. The use of citrate may also be associated with less inflammation due to hypocalcemia-induced suppression of intracellular signaling at the membrane and avoidance of heparin, which may have proinflammatory properties. Whether these beneficial effects increase patient survival needs to be confirmed. However, other benefits are the reason that citrate should become the first choice anticoagulant for CRRT provided that its safe use can be guaranteed.


Wolbachia Do Not Induce Reactive Oxygen Species-Dependent Immune Pathway Activation in Aedes albopictus.
Aedes albopictus is a major vector of dengue (DENV) and chikungunya (CHIKV) viruses, causing millions of infections annually. It naturally carries, at high frequency, the intracellular inherited bacterial endosymbiont Wolbachia strains wAlbA and wAlbB; transinfection with the higher-density Wolbachia strain wMel from Drosophila melanogaster led to transmission blocking of both arboviruses. The hypothesis that reactive oxygen species (ROS)-induced immune activation plays a role in arbovirus inhibition in this species was examined. In contrast to previous observations in Ae. aegypti, elevation of ROS levels was not observed in either cell lines or mosquito lines carrying the wild-type Wolbachia or higher-density Drosophila Wolbachia strains. There was also no upregulation of genes controlling innate immune pathways or with antioxidant/ROS-producing functions. These data suggest that ROS-mediated immune activation is not an important component of the viral transmission-blocking phenotype in this species.


Vcsa1 acts as a marker of erectile function recovery after gene therapeutic and pharmacological interventions.
We identified molecular markers of erectile function, particularly those responding to erectile dysfunction treatment. Sprague-Dawley retired breeder rats were intracorporeally injected with pVAX-hSlo, pSMAA-hSlo or the control plasmid pVAX. One week later the intracorporeal pressure-to-blood pressure ratio and gene expression were determined by microarray analysis and quantitative reverse transcriptase-polymerase chain reaction. Rat corporeal cells were transfected in vitro with pVAX-hSlo, pSMAA-hSlo or pVAX and the change in gene expression was determined. We also determined whether Vcsa1 expression was changed after pharmacotherapy using tadalafil. Animals treated with vectors expressing hSlo had significantly improved erectile function compared to that in controls, accompanied by changed expression of a subset of genes. Vcsa1 was one of the genes that was most changed in expression (the third of approximately 31,000 with greater than 10-fold up-regulation). Changes in gene expression were different than those observed in corporeal cells transfected in vitro, distinguishing gene expression changes that were a direct effect of hSlo over expression. When tadalafil was administered in retired breeder rats, the Vcsa1 transcript increased 4-fold in corporeal tissue compared to that in untreated controls. Our study identifies a set of genes that are changed in response to improved erectile function, rather than as a direct effect of treatment. We noted Vcsa1 may act as marker of the restoration of erectile function after gene transfer and pharmacotherapy.


Structural Basis for the Regulation of Biofilm Formation and Iron Uptake in A. baumannii by the Blue-Light-Using Photoreceptor, BlsA.
The opportunistic human pathogen, A. baumannii, senses and responds to light using the blue light sensing A (BlsA) photoreceptor protein. BlsA is a blue-light-using flavin adenine dinucleotide (BLUF) protein that is known to regulate a wide variety of cellular functions through interactions with different binding partners. Using immunoprecipitation of tagged BlsA in A. baumannii lysates, we observed a number of proteins that interact with BlsA, including several transcription factors. In addition to a known binding partner, the iron uptake regulator Fur, we identified the biofilm response regulator BfmR as a putative BlsA-binding partner. Using microscale thermophoresis, we determined that both BfmR and Fur bind to BlsA with nanomolar binding constants. To better understand how BlsA interacts with and regulates these transcription factors, we solved the X-ray crystal structures of BlsA in both a ground (dark) state and a photoactivated light state. Comparison of the light- and dark-state structures revealed that, upon photoactivation, the two α-helices comprising the variable domain of BlsA undergo a distinct conformational change. The flavin-binding site, however, remains largely unchanged from dark to light. These structures, along with docking studies of BlsA and Fur, reveal key mechanistic details about how BlsA propagates the photoactivation signal between protein domains and on to its binding partner. Taken together, our structural and biophysical data provide important insights into how BlsA controls signal transduction in A. baumannii and provides a likely mechanism for blue-light-dependent modulation of biofilm formation and iron uptake.


In vivo femtosecond endosurgery: an intestinal epithelial regeneration-after-injury model.
Regeneration of the intestinal epithelium after injury or during pathogenesis is a dynamic cellular process critical for host immunity. However, current epithelial injury models provide poor spatial control, complicating the study of precise cellular responses. Here we developed endoscopic femtosecond-laser surgery capable of generating acute tissue injury. A side-view probe provides a convenient access to the distal colon in the mouse in vivo and allows real-time intraoperative monitoring as well as pre- and post-surgery examinations via multiphoton imaging. The photo-induced damage showed a nonlinear dependence on laser intensity. At an optical power of 200 mW (2.5 nJ per pulse), scanning the beam focus over 300x300 µm(2) area in the colonic mucosa generated substantial vascular damages within 30 s. We confirmed the localized tissue damage and the physiologic regeneration of the disrupted epithelium by in situ barrier function assays, validating the animal model for epithelial regeneration following injury. The femtosecond endosurgery technique is applicable to various experimental models based on laser-induced perturbations.


Cocaine affinity decreased by mutations of aromatic residue phenylalanine 105 in the transmembrane domain 2 of dopamine transporter.
Dopamine transporter (DAT) is a major target of cocaine, one of the most abused drugs. Major efforts have been focused on defining residues in DAT involved in cocaine binding. We have isolated the Drosophila melanogaster DAT (dDAT) cDNA, which is 10-fold less sensitive to cocaine than the mammalian DATs. Replacing transmembrane domain 2 (TM2) of mouse DAT (mDAT) with dDAT sequence reduced cocaine sensitivity. The reciprocal construct exhibited increased cocaine sensitivity. Switching residue 105 in TM2, a phenylalanine conserved in all mammalian DATs, to methionine, the corresponding residue in dDAT, resulted in a functional transporter with cocaine sensitivity 4-fold lower. Replacing F105 with alanine, leucine, isoleucine, serine, threonine, asparagine, or glutamine resulted in transporters with low transport activity. In contrast, changing F105 to the other aromatic residues tyrosine or tryptophan retained more than 75% transport activity and high cocaine sensitivity. Most significantly, the reciprocal construct, switching the methionine in dDAT at the corresponding residue to phenylalanine, increased cocaine sensitivity 3-fold. Finally, the mDAT mutant with a cysteine at this position had normal transport activity but exhibited cocaine sensitivity that was 15-fold lower. These results suggest that F105 in mDAT contributes to high-affinity cocaine binding. The functional cocaine-insensitive mutants provide tools for the study of the mechanism of cocaine addiction.


In vitro antimicrobial activity of propolis samples from different geographical origins against certain oral pathogens.
Propolis is an agent having antimicrobial properties, however, its composition can vary depending on the area where it is collected. In the present study, the antimicrobial activity of five propolis samples, collected from four different regions in Turkey and from Brazil, against nine anaerobic strains was evaluated. Ethanol extracts of propolis (EEP) were prepared from propolis samples and we determined minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEP on the growth of test microorganisms by using agar dilution method. All strains were susceptible and MIC values ranged from 4 to 512 microg/ml for propolis activity. Propolis from Kazan-Ankara showed most effective MIC values to the studied microorganisms. MBC values of Kazan-Ankara EEP samples were ranged from 8 to 512 microg/ml. Death was observed within 4 h of incubation for Peptostreptococcus anaerobius and micros and Lactobacillus acidophilus and Actinomyces naeslundii, while 8 h for Prevotella oralis and Prevotella melaninogenica and Porphyromonas gingivalis, 12 h for Fusobacterium nucleatum, 16 h for Veillonella parvula. It was shown that propolis samples were more effective against Gram positive anaerobic bacteria than Gram negative ones. The organic chemical compositions of EEPs were determined by high-resolution gas chromatography coupled to mass spectrometry (GC-MS). The main compounds of EEPs were flavonoids such as pinobanksin, quercetin, naringenin, galangine, chrysin and aromatic acids such as cafeic acid. Because of increased antimicrobial resistance, propolis may be kept in mind in the treatment of oral cavity diseases.


Staphylococcal Bap Proteins Build Amyloid Scaffold Biofilm Matrices in Response to Environmental Signals.
Biofilms are communities of bacteria that grow encased in an extracellular matrix that often contains proteins. The spatial organization and the molecular interactions between matrix scaffold proteins remain in most cases largely unknown. Here, we report that Bap protein of Staphylococcus aureus self-assembles into functional amyloid aggregates to build the biofilm matrix in response to environmental conditions. Specifically, Bap is processed and fragments containing at least the N-terminus of the protein become aggregation-prone and self-assemble into amyloid-like structures under acidic pHs and low concentrations of calcium. The molten globule-like state of Bap fragments is stabilized upon binding of the cation, hindering its self-assembly into amyloid fibers. These findings define a dual function for Bap, first as a sensor and then as a scaffold protein to promote biofilm development under specific environmental conditions. Since the pH-driven multicellular behavior mediated by Bap occurs in coagulase-negative staphylococci and many other bacteria exploit Bap-like proteins to build a biofilm matrix, the mechanism of amyloid-like aggregation described here may be widespread among pathogenic bacteria.


Spectroscopic studies of protein-heme interactions accompanying the allosteric transition in methemoglobins.
Resonance Raman, optical absorption, and circular dichroism spectroscopic techniques have been used to examine the effect of the addition of inositol hexaphosphate (IHP) to a series of carp and human methemoglobin derivatives. Markers of spin equilibrium in the high-frequency region (1450-1650 cm-1) of the resonance Raman spectrum yield high/low-spin ratios consistent with direct magnetic susceptibility measurements. Changes in the low-frequency region (100-600 cm-1) of the resonance Raman spectrum appear to correlate with the quaternary structure transition. Changes in the ultraviolet absorption spectra and the circular dichroism spectra also appear to be related to the quaternary structure change. By using the resonance Raman spin markers, we find that those derivatives of carp methemoglobin which are in spin equilibrium have a larger ratio of high-spin to low-spin populations than the corresponding derivatives of human methemoglobin. Upon the addition of IHP to the methemoglobins the spin equilibrium is shifted toward a larger high-spin population. This change in equilibrium is larger for the carp protein than for the human protein. We obtain an IHP-induced change in the free energy difference between the high-spin and low-spin states of 300 cal/mol for those human methemoglobins in which a quaternary structure change occurs and 600 cal/mol for carp methemoglobins. Our data are consistent with a quaternary structure change induced by IHP in all the carp methemoglobins studied (F-, H2O, SCN-, NO2-, N3-, and CN-) and in the F-, H2O, and SCN- derivatives of the human protein but not in the NO2-, N3-, and CN- derivatives. The Fe-CN stretching mode has been identified by isotopic substitution and found to be unchanged in frequency in carp CN- metHb when the quaternary structure is changed. On the basis of our results we conclude that the protein forces at the heme due to the addition of IHP do not significantly affect the position of the iron atom with respect to the heme plane. Rather, the changes in spin equilibrium may be caused by protein-induced changes in the orientation of the proximal histidine or tertiary structure changes in the heme pocket which affect the porphyrin macrocycle. Either of these changes, or a combination thereof, leads to changes in the iron d orbital energies and concomitant changes in the spin equilibrium.


The effects of temperature priming on cooperation in the iterated prisoner's dilemma.
Based on initial research findings by Williams and Bargh (2008) and Kang, Williams, Clark, Gray and Bargh (2011) on the interaction between interpersonal and physical warmth, theoretical models such as cognitive scaffolding and the importance of evaluations of interpersonal warmth in trust-based decisions, this experiment investigated the effect of temperature priming on 30 pairs of British university students with hot and cold objects on frequency of cooperation in a game of iterated Prisoner's Dilemma. Participants were found to cooperate significantly more frequently when primed with hot objects than with cold objects, supporting the assertion that physical warmth sensation positively affects interpersonal trust evaluation. No support was found for the prediction that male-male pairs would cooperate less than female-female pairs. The implications of these findings to evolutionary and developmental theories of interpersonal warmth are discussed.


Myc enforces overexpression of EZH2 in early prostatic neoplasia via transcriptional and post-transcriptional mechanisms.
EZH2 is part of the PRC2 polycomb repressive complex that is overexpressed in multiple cancer types and has been implicated in prostate cancer initiation and progression. Here, we identify EZH2 as a target of the MYC oncogene in prostate cancer and show that MYC coordinately regulates EZH2 through transcriptional and post-transcriptional means. Although prior studies in prostate cancer have revealed a number of possible mechanisms of EZH2 upregulation, these changes cannot account for the overexpression EZH2 in many primary prostate cancers, nor in most cases of high grade PIN. We report that upregulation of Myc in the mouse prostate results in overexpression of EZH2 mRNA and protein which coincides with reductions in miR-26a and miR-26b, known regulators of EZH2 in some non-prostate cell types, albeit not in others. Further, in human prostate cancer cells, Myc negatively regulates miR-26a and miR-26b via direct binding to their parental Pol II gene promoters, and forced overexpression of miR-26a and miR-26b in prostate cancer cells results in decreased EZH2 levels and suppressed proliferation. In human clinical samples, miR-26a and miR-26b are downregulated in most primary prostate cancers. As a separate mechanism of EZH2 mRNA upregulation, we find that Myc binds directly to and activates the transcription of the EZH2 promoter. These results link two major pathways in prostate cancer by providing two additional and complementary Myc-regulated mechanisms by which EZH2 upregulation occurs and is enforced during prostatic carcinogenesis. Further, the results implicate EZH2-driven mechanisms by which Myc may stimulate prostate tumor initiation and disease progression.


Work force policy perspectives: registered nurses.
If the decline in full-time labor force participation by registered nurses in hospitals is to be reversed, the issue of equal pay for comparable work must be addressed. Under pressure for cost containment, policies tend to focus on labor force economics rather than on limitations of services. While the two are interrelated, wage policies must be considered independently. This article describes the network which determines how nurse salaries are set: the relationship between the private sector, the general schedule and the Veteran's Administration. The effects of this system are documented, using testimony from a case in the tenth circuit, as well as comparisons with other reference groups: policemen, teachers, laborers, and VA career fields. The evidence suggests that there is a need for policy intervention. Prime areas for action are the comparability practices by governments, particularly in the areas of classification standards and pay setting. Hospital personnel practices which continue past effects of occupational segregation also should be changed.


The association between weight stability and parenteral nutrition characteristics and survival in patients with colorectal cancer.
Knowledge about the impact of metabolic disturbances and parenteral nutrition (PN) characteristics on the survival of cancer patients receiving PN is limited. We aimed to assess the association between clinical and PN characteristics and survival in colorectal-cancer patients receiving PN support. Our study included 572 consecutive colorectal-cancer patients who had received PN support between 2008 and 2013. Patient characteristics, body mass index, weight, medical/surgical history, indication for PN, PN data and survival were recorded. Associations between clinical and PN characteristics and survival were analysed with important confounding factors. The final cohort included 437 evaluable patients, with a mean age of 57 years. Eighty-one percent of the study population had advanced stage of colorectal cancer. Unstable weight (weight change ≥2.5%) prior to PN initiation [hazard ratio (HR) = 1.41, P = 0.023] was adversely associated with survival after adjusting for multiple factors including cancer stage. Bowel obstruction (HR = 1.75, P = 0.017) as a PN indication was associated with worse survival when compared with without bowel obstruction. Higher PN amino acid by ideal body weight (g•kg-1) (HR = 0.59, P = 0.029) was associated with longer survival, whereas a higher percentage of non-PN intravenous calories (HR = 1.04, P = 0.011) was associated with shorter survival independently of confounding factors. Body mass index and weight stability can be useful nutritional indices for survival prediction in cancer patients receiving PN. PN planning should take into account of non-PN calories to achieve optimal energy support and balance. Future research is needed to define optimal PN amino-acid requirement and energy balance.


Triple combination chemotherapy with cisplatin, docetaxel, and irinotecan for advanced non-small cell lung cancer: a phase I/II trial.
To determine the recommended dose and evaluate the response rate and toxicity of triplet chemotherapy using cisplatin, docetaxel, and irinotecan for non-small cell lung cancer (NSCLC) patients with stage IIIB or IV. A total of 65 patients (33 men and 32 women) with advanced NSCLC, a good performance status, and 65 years of age or younger were included in these phase I/II studies. The median age was 52 years. Most patients had performance status 1 (49/65) and stage IV disease (49/65). Adenocarcinoma was the most common tumor histology (55 patients). Cisplatin and docetaxel were given on day 1 and irinotecan on day 2; the cycles were repeated every 3 weeks. In the phase I study, the maximum tolerated doses of combination cisplatin/docetaxel/irinotecan were, respectively, 80/60/60 (mg/m) and the recommended doses for the phase II study were determined to be 60/60/60 (mg/m), respectively. The dose-limiting toxicities were neutropenia, neutropenic fever, and diarrhea. In the phase II study, 157 cycles of chemotherapy were delivered to 49 patients (median three cycles per patient). The objective response rate was 57.1% (95% confidence interval: 43.1%-71.1%). The median survival time and the actual 2-, 3- and estimated 5-year survival rates were 17 months, 33%, 25%, and 18%, respectively. Grade 3/4 toxicities consisted of neutropenia (92%), neutropenic fever (45%), nausea/vomiting (27%), diarrhea (35%), and hepatic toxicity (2%); there were no cases of treatment-related death. This triplet chemotherapy has shown a promising activity against advanced NSCLC according to admission-based treatment with adequate supportive care. The principal toxicity was neutropenic fever, but supportive care should be explored to reduce this incidence.


In vitro study of the influence of dentin desensitizing and sealing on the shear bond strength of two universal resin cements.
To test whether or not dentin desensitizing or sealing methods influence the bond strength of universal resin cements. The bond strength of two universal resin cements (RelyXUnicem, Self-adhesive Prototype Cement) to human dentin was assessed after different dentin surface treatments (test). A conventional resin cement (Panavia 21) served as control. The influence of the following pretreatment methods was tested and compared to freshly ground dentin: dentin desensitizing by means of glutaraldehyde-containing primers (Gluma Desensitizer, Syntac Primer/Adhesive), or dentin sealing by means of bonding agents (Heliobond, ClearfilSE Bond). Additionally, the influence of provisional cement (Freegenol) was analyzed after its application and subsequent mechanical removal. Bond strength after thermocycling was measured in a shear test (n = 10). Data were analyzed with one-way ANOVA, followed by a post-hoc Tukey test (p < 0.05). Both test cements exhibited significantly lower bond strength to freshly ground dentin than the control cement. The desensitization methods did not affect the bond strength of the test cements. Furthermore, the sealing methods significantly increased the bond strength of both test cements as compared to freshly ground dentin. The application and subsequent removal of the provisional cement only had a minor influence on the bond strength of the test cements. The contamination with provisional cement, however, significantly affected the bond strength of the control cement compared to the values achieved on freshly ground dentin. Desensitization or sealing of dentin have a beneficial effect on the bond strength of universal resin cements. The application of a provisional cement prior to adhesive cementation exhibits no significant influence on the bond strength of the universal resin cements.


National trends and disparities in the incidence of hepatocellular carcinoma, 1998-2003.
Previous studies indicate that the incidence of hepatocellular carcinoma in the United States is increasing. These reports, however, have contained limited information on population groups other than whites and blacks. We assessed recent incidence rates and trends for hepatocellular carcinoma by using newly available national data from cancer registries participating in the Centers for Disease Control and Prevention's National Program of Cancer Registries and the National Cancer Institute's Surveillance, Epidemiology, and End Results Program. Data from registries in 38 states and the District of Columbia met our criteria; these data covered 83% of the U.S. population. We computed age-adjusted incidence rates and annual percentages of change from 1998 through 2003. The registries that we used reported 48,048 cases of hepatocellular carcinoma (3.4 cases per 100,000 population per year) for the study period. Whites accounted for three-fourths of cases. The incidence rate for blacks was 1.7 times higher than that for whites, and the rate for Asians/Pacific Islanders was 4 times higher than that for whites. Hispanics had 2.5 times the risk of non-Hispanics. Among Asian/Pacific Islander subgroups, rates were highest for people of Vietnamese and Korean origin. For all races/ethnicities combined, the annual percentages of change were 4.8% for males and 4.3% for females (P < .05). The annual percentage of change was highest for people aged 45-59 years (9.0%, P < .05). The annual percentage of change for Asians/Pacific Islanders was statistically unchanged. We document rising incidence rates of hepatocellular carcinoma in the United States during a time when the overall incidence of cancer has stabilized. Efforts to collect representative etiologic data on new hepatocellular carcinoma cases are needed to enable better characterization of trends and to guide the planning and evaluation of prevention programs.


Range-dependent flexibility in the acoustic field of view of echolocating porpoises (Phocoena phocoena).
Toothed whales use sonar to detect, locate, and track prey. They adjust emitted sound intensity, auditory sensitivity and click rate to target range, and terminate prey pursuits with high-repetition-rate, low-intensity buzzes. However, their narrow acoustic field of view (FOV) is considered stable throughout target approach, which could facilitate prey escape at close-range. Here, we show that, like some bats, harbour porpoises can broaden their biosonar beam during the terminal phase of attack but, unlike bats, maintain the ability to change beamwidth within this phase. Based on video, MRI, and acoustic-tag recordings, we propose this flexibility is modulated by the melon and implemented to accommodate dynamic spatial relationships with prey and acoustic complexity of surroundings. Despite independent evolution and different means of sound generation and transmission, whales and bats adaptively change their FOV, suggesting that beamwidth flexibility has been an important driver in the evolution of echolocation for prey tracking.


Trends in the association between smoking history and general/central obesity in Catalonia, Spain (1992-2003).
To examine trends in the relationship between smoking history and both general and central fatness in adults from a Mediterranean setting. Materials and methods: The ENCAT 1992-1993 and 2002-2003 surveys were used; samples consisted of 482 men, 589 women from 1992-1993, and 515 men, 613 women from 2002-2003, aged 25-60 years. Measured anthropometry and self-reported data on smoking habits, diet, lifestyle and SES were collected. General fatness was defined as WHO's BMI overweight and obesity, and central fatness was defined as WHO's Increased-Risk-for-metabolic-complications Waist Circumference (IR WC) and Substantially-Increased-Risk WC (SIR WC). Simple logistic regression was used to estimate multivariate-adjusted associations between general/central fatness and smoking history. By 2002-2003, most associations observed in 1992-1993 had been strongly attenuated: only male current-heavy-smoking remained associated with IR/SIR WC (three-fold) and female current-moderate-smokers were 0.57 times less likely to have an IR/SIR WC (p < 0.10). Although causality cannot be established, results suggest a positive association between heavy smoking and central fatness among men, but no association between former smoking and general/central fatness; findings strengthen arguments for promoting smoking cessation to reduce smoking -and obesity- associated morbidity and mortality.


Mycobacteriophage L5 infection of Mycobacterium bovis BCG: implications for phage genetics in the slow-growing mycobacteria.
Mycobacteriophage L5 is a well-characterized temperate phage that forms stable lysogens in Mycobacterium smegmatis. The host range of L5 is, however, unclear because previous reports suggested that it does not infect slow-growing mycobacteria such as Mycobacterium tuberculosis and bacille Calmette-Guerin (BCG). Moreover, luciferase reporter phage derivatives of L5 failed to produce light from BCG, suggesting that infection is blocked at or before the stage of DNA injection. In this study, we demonstrate that L5 infection of slow growing mycobacteria specifically requires a high concentration of Ca2+, conditions that differs from those required for infection of M. smegmatis by L5 and for infection of BCG by the closely related phage D29. In addition, we show that there are specific genetic determinants of L5 that confer the ability to infect slow growing mycobacteria, without altering infection of M. smegmatis. These observations extend the use of phage L5 for the diagnosis and analysis of tuberculosis and other mycobacterial diseases.