Cross-validation of murine UV signal transduction pathways in human skin.
Acute UVB irradiation of mouse skin results in activation of phospatidyinositol-3 (PI-3) kinase and mitogen-activated protein kinase (MAPK) pathways leading to altered protein phosphorylation and downstream transcription of genes. We determined whether activation of these pathways also occurs in human skin exposed to 4x minimal erythemic dose of UVB in 23 volunteers. Biopsies were taken prior to, at 30 min, 1 and 24 h post-UVB. In agreement with mouse studies, the earliest UV-induced changes in epidermis were seen in phospho-CREB (two- and five-fold at 30 min and 1 h) and in phospho-MAPKAPK-2 (three-fold at both 30 min and 1 h). At 1 h, phospho-c-JUN and phospho-p38 were increased five- and two-fold, respectively. Moreover, phospho-c-JUN and phospho-p38 were further increased at 24 h (12- and six-fold, respectively). Phospho-GSK-3beta was similarly increased at all time points. Increases in phospho-p53 (12-fold), COX-2 (four-fold), c-FOS (14-fold) and apoptosis were not seen until 24 h. Our data suggest that UVB acts through MAPK p38 and PI-3 kinase with phosphorylation of MAPKAPK-2, CREB, c-JUN, p38, GSK-3beta and p53 leading to marked increases in c-FOS, COX-2 and apoptosis. Validation of murine models in human skin will aid in development of effective skin cancer chemoprevention and prevention strategies.


Apolipoprotein E polymorphism in Omani dyslipidemic patients with and without coronary artery disease.
Apolipoprotein E (APOE) polymorphism is a predictor of interindividual variability in plasma levels of lipids and lipoproteins and a predictor of risk of coronary artery disease (CAD). We studied the relationship between APOE polymorphism and lipid profiles and risk of CAD in Omani dyslipidemic patients. This retrospective study included 244 dyslipidemic patients, of whom 67 had CAD. Fasting blood glucose, lipids, and plasma lipoprotein levels were measured using standard methods, and APOE genotypes were detected by PCR-RFLP. The dyslipidemic patients had the following APOE allele frequencies: APOE*2, 0.030; APOE*3, 0.894; and APOE*4, 0.076. APOE allele frequencies between patients with and without CAD showed no significant differences. Compared to APOE*3/*3 homozygotes, APOE*4 allele patients had higher mean levels of low-density lipoprotein (LDL) cholesterol (p = 0.014), apoB (p = 0.031), lower mean levels of apoA1 (p = 0.043), and a trend of higher mean level of total cholesterol (p = 0.084). Thirty-one percent of patients with CAD had the APOE*4 allele compared to 26% with the APOE*3 allele, but this difference was not significant. Compared with APOE*3/*3 homozygotes, patients with the APOE*4 allele had 1.3 times higher risk for CAD after ignoring dyslipidemia, but this risk was modified after adjusting for dyslipidemia. In conclusion, among dyslipidemic patients, carriers of APOE*4 compared to homozygous carriers of APOE*3 had significantly higher levels of LDL cholesterol and apoB, but no relationship with CAD was found.


Explaining variation in adult Anopheles indoor resting abundance: the relative effects of larval habitat proximity and insecticide-treated bed net use.
Spatial determinants of malaria risk within communities are associated with heterogeneity of exposure to vector mosquitoes. The abundance of adult malaria vectors inside people's houses, where most transmission takes place, should be associated with several factors: proximity of houses to larval habitats, structural characteristics of houses, indoor use of vector control tools containing insecticides, and human behavioural and environmental factors in and near houses. While most previous studies have assessed the association of larval habitat proximity in landscapes with relatively low densities of larval habitats, in this study these relationships were analysed in a region of rural, lowland western Kenya with high larval habitat density. 525 houses were sampled for indoor-resting mosquitoes across an 8 by 8 km study area using the pyrethrum spray catch method. A predictive model of larval habitat location in this landscape, previously verified, provided derivations of indices of larval habitat proximity to houses. Using geostatistical regression models, the association of larval habitat proximity, long-lasting insecticidal nets (LLIN) use, house structural characteristics (wall type, roof type), and peridomestic variables (cooking in the house, cattle near the house, number of people sleeping in the house) with mosquito abundance in houses was quantified. Vector abundance was low (mean, 1.1 adult Anopheles per house). Proximity of larval habitats was a strong predictor of Anopheles abundance. Houses without an LLIN had more female Anopheles gambiae s.s., Anopheles arabiensis and Anopheles funestus than houses where some people used an LLIN (rate ratios, 95% CI 0.87, 0.85-0.89; 0.84, 0.82-0.86; 0.38, 0.37-0.40) and houses where everyone used an LLIN (RR, 95% CI 0.49, 0.48-0.50; 0.39, 0.39-0.40; 0.60, 0.58-0.61). Cooking in the house also reduced Anopheles abundance across all species. The number of people sleeping in the house, presence of cattle near the house, and house structure modulated Anopheles abundance, but the effect varied with Anopheles species and sex. Variation in the abundance of indoor-resting Anopheles in rural houses of western Kenya varies with clearly identifiable factors. Results suggest that LLIN use continues to function in reducing vector abundance, and that larval source management in this region could lead to further reductions in malaria risk by reducing the amount of an obligatory resource for mosquitoes near people's homes.


The impact of sample non-normality on ANOVA and alternative methods.
In this journal, Zimmerman (2004, 2011) has discussed preliminary tests that researchers often use to choose an appropriate method for comparing locations when the assumption of normality is doubtful. The conceptual problem with this approach is that such a two-stage process makes both the power and the significance of the entire procedure uncertain, as type I and type II errors are possible at both stages. A type I error at the first stage, for example, will obviously increase the probability of a type II error at the second stage. Based on the idea of Schmider et al. (2010), which proposes that simulated sets of sample data be ranked with respect to their degree of normality, this paper investigates the relationship between population non-normality and sample non-normality with respect to the performance of the ANOVA, Brown-Forsythe test, Welch test, and Kruskal-Wallis test when used with different distributions, sample sizes, and effect sizes. The overall conclusion is that the Kruskal-Wallis test is considerably less sensitive to the degree of sample normality when populations are distinctly non-normal and should therefore be the primary tool used to compare locations when it is known that populations are not at least approximately normal.


Single-cell dissection reveals the role of aggrephagy patterns in tumor microenvironment components aiding predicting prognosis and immunotherapy on lung adenocarcinoma.
Lung adenocarcinoma (LUAD) is one of the leading malignant cancers. Aggrephagy plays a critical role in key genetic events for various cancers; yet, how aggrephagy functions within the tumor microenvironment (TME) in LUAD remains to be elucidated. In this study, by sequential non-negative matrix factorization (NMF) algorithm, pseudotime analysis, cell-cell interaction analysis, and SCENIC analysis, we have shown that aggrephagy genes demonstrated various patterns among different cell types in LUAD TME. LUAD and Immunotherapy cohorts from public repository were used to determine the prognosis and immune response of aggrephagy TME subtypes. The aggrephagy-deprived prognostic score (ADPS) was quantified based on machine learning algorithms. The cancer-associated fibroblasts (CAFs), tumor-associated macrophages (TAMs), and CD8+ T cells have various aggrephagy patterns, which enhance the intensity of intercellular communication and transcription factor activation. Furthermore, based on the signatures of the newly defined aggrephagy cell subtypes and expression profiles of large cohorts in LUAD patients, we determine that DYNC1I2+CAF-C1, DYNLL1+CAF-C2, PARK7+CAF-C3, VIM+Mac-C1, PARK7+Mac-C2, VIM+CD8+T_cells-C1, UBA52+CD8+T_cells-C2, TUBA4A+CD8+T_ cells-C3, and TUBA1A+CD8+T_cells-C4 are crucial prognostic factors for LUAD patients. The developed ADPS could predict survival outcomes and immunotherapeutic response across ten cohorts (n = 1838), and patients with low ADPS owned a better prognosis, lower genomic alterations, and are more sensitive to immunotherapy. Meanwhile, based on PRISM, CTRP, and CMAP databases, PLK inhibitor BI-2536, may be a potential agent for patients with high ADPS. Taken together, our novel and systematic single-cell analysis has revealed the unique role of aggrephagy in remodeling the TME of LUAD. As a newly demonstrated biomarker, the ADPS facilitates the clinical management and individualized treatment of LUAD.


Prognostic and diagnostic significance of beta-catenin nuclear immunostaining in colorectal cancer.
In the present study, we investigated the prognostic and diagnostic significance of beta-catenin nuclear immunostaining in 60 specimens of normal colorectal tissue; 180 specimens of colorectal polyps, adenomas, and carcinomas; and 40 specimens from patients with the simultaneous occurrence of polyps, adenomas, and carcinomas. Additional specimens from 59 patients with colorectal carcinoma and 14 patients with adenoma who subsequently developed carcinoma were examined for possible survival study. Immunohistochemical staining showed that the occurrence of nuclear beta-catenin correlated with the sequential stages in colorectal carcinogenesis, in which positive staining was observed in 0% of normal tissues, 8% of polyps, 92% of adenomas, and 100% of carcinomas. High immunohistochemical scores in colorectal carcinoma were significantly associated with lymph node metastasis and poor survival. Adenomas associated with synchronous or metachronous carcinomas showed significantly higher levels of nuclear beta-catenin compared with adenomas without associated carcinomas. Nuclear translocation of beta-catenin was rare or absent in other types of cytokeratin 20 positive adenocarcinomas examined (99 cases). Thus, it was positive in only 7% of colonic mucinous adenocarcinomas, 3% of pancreatic adenocarcinomas, 8% of ovarian mucinous cystadenocarcinomas, and 0% of gastric adenocarcinomas. However, 100% of primary and metastatic colorectal adenocarcinomas were positive for nuclear staining for beta-catenin. Thus, nuclear staining for beta-catenin may serve as an additional parameter to help distinguish colorectal adenocarcinomas from adenocarcinomas of other tissue sites. Collectively, the present large-scale study has clearly addressed the clinical significance of beta-catenin nuclear translocation with respect to tumor progression, survival, and differential diagnosis.


Occurrence of Meloidogyne fallax in North America, and Molecular Characterization of M. fallax and M. minor from U.S. Golf Course Greens.
Several species of root-knot nematodes (Meloidogyne spp.) are known to have significant presence on turfgrass in golf course greens, particularly in the western United States. Nematodes isolated from a golf course in King County, WA were identified as Meloidogyne minor based on analysis of the large ribosomal subunit (LSU 28S D2-D3 expansion segment), the internal transcribed spacers 1 and 2 (ITS rDNA), the intergenic spacer region 2 (IGS2), and the nuclear protein-coding gene Hsp90. Sequence-characterized amplified region (SCAR) primers that were originally designed to be specific for M. fallax were found to cross-react with M. minor. A population from California was determined to be M. fallax based on juvenile tail morphology and analysis of the ribosomal markers and Hsp90, comprising the first report of this species in North America. Using trees based on Hsp90 genomic alignments, the phylogenetic relationships of these populations and known root-knot nematode species were congruent with previous trees based on ribosomal genes. Resolution of M. fallax and M. chitwoodi using Hsp90 was equivalent to species separation obtained with 28S or 18S rDNA alignments. The strengths and weaknesses of ribosomal and Hsp90 markers, and the use of SCAR polymerase chain reaction as diagnostic tools are discussed.


Vasopressin mediates the pressor effects of endothelin in the subfornical organ of the rat.
The potent vasoconstrictor peptide endothelin may affect central cardiovascular function in areas with incomplete blood-brain barrier such as the subfornical organ (SFO). In these studies, we determine whether microinjection of endothelin-1 (ET-1) into the SFO increases blood pressure (BP) in a dose-related manner and investigate the potential involvement of sympathetic and vasopressinergic mechanisms. In urethane-anesthetized Sprague-Dawley rats, BP was recorded intra-arterially, and ET-1 (0.125-6.0 pmol/60 nl) was microinjected stereotaxically into the SFO. Whereas vehicle (60 nl) did not change mean BP or HR, ET-1 evoked a dose-related pressor and bradycardic effect. The maximal changes were noted at the 1-pmol dose. No significant hemodynamic effects were observed with ET-1 microinjection in areas immediately surrounding the SFO or into the SFO of rats pretreated with a specific endothelin antagonist. In animals instrumented for recording of renal sympathetic nerve activity (RSNA), the administration of ET-1 (1 pmol/60 nl) evoked pressor (14 +/- 5 mm Hg) and bradycardic (-41 +/- 12 bpm) effects with a decrease in RSNA (16% +/- 3%). The effects on HR and RSNA seem to be mediated by baroreflex changes because in sino-aortic denervated rats, ET-1 pressor effects occur without inhibition of HR or RSNA. We documented the involvement of vasopressin in ET-1 actions by using vasopressin antagonists that inhibited the effects evoked by ET-1 administration. In addition, increases in vasopressin plasma levels were demonstrated at the time of the maximal effect of this peptide. These results indicate that ET-1 acting in the SFO increases BP by a vasopressinergic mechanism.


Myc is dispensable for cardiomyocyte development but rescues Mycn-deficient hearts through functional replacement and cell competition.
Myc is considered an essential transcription factor for heart development, but cardiac defects have only been studied in global Myc loss-of-function models. Here, we eliminated Myc by recombining a Myc floxed allele with the Nkx2.5Cre driver. We observed no anatomical, cellular or functional alterations in either fetuses or adult cardiac Myc-deficient mice. We re-examined Myc expression during development and found no expression in developing cardiomyocytes. In contrast, we confirmed that Mycn is essential for cardiomyocyte proliferation and cardiogenesis. Mosaic Myc overexpression in a Mycn-deficient background shows that Myc can replace Mycn function, recovering heart development. We further show that this recovery involves the elimination of Mycn-deficient cells by cell competition. Our results indicate that Myc is dispensable in cardiomyocytes both during cardiogenesis and for adult heart homeostasis, and that Mycn is exclusively responsible for cardiomyocyte proliferation during heart development. Nonetheless, our results show that Myc can functionally replace Mycn We also show that cardiomyocytes compete according to their combined Myc and Mycn levels and that cell competition eliminates flawed cardiomyocytes, suggesting its relevance as a quality control mechanism in cardiac development.


Association of plasma leptin concentrations with adiposity measurements in rural Chinese adolescents.
There is evidence that leptin is involved in the etiology of obesity-related cardiovascular disease in adults. This raises the question of whether leptin levels in adolescence are indicative of adiposity-related cardiovascular risk. This study investigated gender-specific patterns of plasma leptin during adolescence, assessed which adiposity measurements are most strongly associated with plasma leptin, and estimated to what degree leptin-adiposity associations are influenced by genetic factors. Plasma leptin concentrations were determined using a sandwich immunoassay. Associations between plasma leptin and several adiposity measurements were examined using generalized estimating equations. Genetic contribution to leptin-adiposity association was estimated by Cholesky decomposition models using twin design. Plasma leptin levels were higher in females than males at all Tanner stages. In females, body mass index, waist circumference, fat mass index (FMI), and percentage body fat (%BF) had similar associations with leptin levels. In males, %BF and FMI were more strongly associated with leptin levels than body mass index and waist circumference. In both males and females, percentage trunk fat had the weakest association with leptin among the five adiposity measures. Shared genetic factors account for more than 80% of the phenotypic correlation between %BF and leptin. We found gender differences in leptin levels and leptin-adiposity associations. In both genders, leptin levels were strongly associated with %BF and FMI, particularly in males. Shared genetic factors contributed largely to the phenotypic correlation between leptin and %BF. Our findings underscored the importance of further investigation of leptin as a biomarker of adiposity in adolescents.


Impact of coffee consumption on the gut microbiota: a human volunteer study.
The impact of a moderate consumption of an instant coffee on the general composition of the human intestinal bacterial population was assessed in this study. Sixteen (16) healthy adult volunteers consumed a daily dose of 3 cups of coffee during 3 weeks. Faecal samples were collected before and after the consumption of coffee, and the impact of the ingestion of the product on the intestinal bacteria as well as the quantification of specific bacterial groups was assessed using nucleic acid-based methods. Although faecal profiles of the dominant microbiota were not significantly affected after the consumption of the coffee (Dice's similarity index=92%, n=16), the population of Bifidobacterium spp. increased after the 3-week test period (P=0.02). Moreover, in some subjects, there was a specific increase in the metabolic activity of Bifidobacterium spp. Our results show that the consumption of the coffee preparation resulting from water co-extraction of green and roasted coffee beans produce an increase in the metabolic activity and/or numbers of the Bifidobacterium spp. population, a bacterial group of reputed beneficial effects, without major impact on the dominant microbiota.


Efavirenz dose reduction is safe in patients with high plasma concentrations and may prevent efavirenz discontinuations.
To establish whether efavirenz dose reduction in patients with high plasma concentrations prevents toxicity-induced efavirenz discontinuations. HIV-infected patients with a high efavirenz plasma concentration (>or=4.0 mg/L) while using efavirenz 600 mg once daily as part of their highly active antiretroviral therapy regimen were selected from the AIDS Therapy Evaluation in The Netherlands cohort study. These patients were classified into 2 groups. The reduced-dose group contained all patients who underwent dose reduction after the high plasma concentration measurement; the standard-dose group consisted of patients who had no dose reduction. Kaplan-Meier and Cox proportional hazards analysis were used to assess the impact of dose reduction on toxicity-induced efavirenz discontinuations. One hundred eighty patients with high plasma efavirenz levels were included, 47 of them subsequently had their efavirenz dose reduced from 600 mg to 400 mg once-daily, which resulted in a 41% decrease in the median efavirenz plasma concentration. At week 48, the Kaplan-Meier estimated cumulative incidence of toxicity-induced efavirenz discontinuations was 11.5% in patients who continued the standard dose versus 2.3% in patients who had a dose reduction; P = 0.066 (log-rank test). Dose reduction was not associated with loss of virological suppression. Dose reduction may prevent toxicity-induced discontinuations in patients with high efavirenz plasma concentrations, whereas not compromising virological efficacy.


[Extramedullary diseases as presenting features of aleukemic acute monocytic leukemia].
We present a case of acute monocytic leukemia presenting with extramedullary disease clinically resembling lymphoma. A 36 year-old man presented with arthralgia and was found to have skin eruption, nasopharyngeal mass, hepatosplenomegaly and superficial lymphadenopathy. The biopsies of those lesions as well as bone marrow revealed infiltration of CD45 positive large atypical cells and were interpreted as malignant lymphoma. At that time blood film showed leuko-erythroblastic picture, but no atypical cells were noted. He was treated with CHOP followed by VEPA chemotherapy and achieved partial remission. Four months later headache and double vision occurred with the same atypical cell in the cerebrospinal fluid. The cells were identified as monoblasts by cytochemistry and immunophenotype. The diagnosis of aleukemic acute monocytic leukemia was made and its CNS involvement was successfully treated with chemotherapy with DHAP and intrathecal MTX. However, two months later, blasts finally appeared in the peripheral blood and the patient died of multiorgan failure eight months after presentation. The blasts were positive for T cell markers (CD2, 4, 8) and NK marker (NKH-1) as well as monocytic markers. This finding, together with those by other investigators, may indicate the association between these immunophenotypes and extramedullary manifestations of acute monocytic leukemia.


Music listening in families and peer groups: benefits for young people's social cohesion and emotional well-being across four cultures.
Families are central to the social and emotional development of youth, and most families engage in musical activities together, such as listening to music or talking about their favorite songs. However, empirical evidence of the positive effects of musical family rituals on social cohesion and emotional well-being is scarce. Furthermore, the role of culture in the shaping of musical family rituals and their psychological benefits has been neglected entirely. This paper investigates musical rituals in families and in peer groups (as an important secondary socialization context) in two traditional/collectivistic and two secular/individualistic cultures, and across two developmental stages (adolescence vs. young adulthood). Based on cross-sectional data from 760 young people in Kenya, the Philippines, New Zealand, and Germany, our study revealed that across cultures music listening in families and in peer groups contributes to family and peer cohesion, respectively. Furthermore, the direct contribution of music in peer groups on well-being appears across cultural contexts, whereas musical family rituals affect emotional well-being in more traditional/collectivistic contexts. Developmental analyses show that musical family rituals are consistently and strongly related to family cohesion across developmental stages, whereas musical rituals in peer groups appear more dependent on the developmental stage (in interaction with culture). Contributing to developmental as well as cross-cultural psychology, this research elucidated musical rituals and their positive effects on the emotional and social development of young people across cultures. The implications for future research and family interventions are discussed.


Premixed calcium silicate cement for endodontic applications: injectability, setting time and radiopacity.
Calcium silicate-based materials (also called MTA) are increasingly being used in endodontic applications. However, the handling properties of MTA are not optimal when it comes to injectability and cohesion. Premixing the cements using glycerol avoids these issues. However, there is a lack of data on the effect of common cement variables on important properties of premixed cements for endodontic applications. In this study, the effects of liquid-to-powder ratio, amount of radiopacifier and amount of calcium sulfate (added to control the setting time) were screened using a statistical model. In the second part of the study, the liquid-to-powder ratio was optimized for cements containing three different amounts of radiopacifier. Finally, the effect of using glycerol rather than water was evaluated in terms of radiopacity. The setting time was found to increase with the amount of radiopacifier when the liquid-to-powder ratio was fixed. This was likely due to the higher density of the radiopacifier in comparison to the calcium silicate, which gave a higher liquid-to-powder ratio in terms of volume. Using glycerol rather than water to mix the cements led to a decrease in radiopacity of the cement. In conclusion, we were able to produce premixed calcium silicate cements with acceptable properties for use in endodontic applications.


Identifying appropriate subjects for abuse liability studies using prestudy pharmacological testing.
Drug abuse liability testing usually involves a subject population of individuals who are current or former drug abusers. To determine whether choosing subjects from a wider and more diverse population of nondrug abusers results in subjective response patterns that are pharmacologically associated with drug abuse liability, a series of studies were conducted using the prescription opiate hydromorphone and the barbiturate secobarbital to prescreen subjects before their acceptance into large multidrug, multidose abuse liability studies. The results of these prescreening studies show that there were some subjects who were not able to report consistently and reliably relevant drug effects, even though they received pharmacologically active doses as measured by objective indexes such as pupil diameter for the opiate study and psychomotor impairment for the barbiturate study. To test the consistency of these results, several of the subjects from the hydromorphone screening were retested with hydrocodone. The results demonstrate that subjects who were unable to report positive subjective effects of hydromorphone were also unable to respond appropriately to hydrocodone, and subjects who responded appropriately to hydromorphone in the prescreening study consistently responded to another opiate. Using this prescreening methodology has allowed sensitive and reliable data on the abuse liability of benzodiazepine agonists and partial agonists, barbiturates, carbamate compounds, amphetamines, selective serotonin reuptake inhibitors and prescription opiate compounds to be generated using a nondrug abusing population. In conclusion, due to the relative heterogeneity of a population of nondrug abusers compared with a drug abusing population, it is necessary to prescreen subjects for their ability to detect and report subjective drug effects, and to distinguish the effects of an active drug from those of placebo. Selecting subjects from this population has several advantages over selecting subjects from the drug abusing population that outweigh any inconvenience of this simple prescreening methodology.


What women say about antenatal care in St Petersburg, Russian Federation.
A capacity building program for professionals in maternal and child health care in St Petersburg, Russian Federation, commenced in 1994. The program was designed to provide 12 workshops in evidence-based antenatal and intrapartum care, as well as appropriate preparation of parents for pregnancy, birth and parenthood, for health care professionals in the city. As part of this program women's observations on the quality of their maternity care services during their transition to parenthood were explored. These reports reflect maternity care practices in the former Soviet Union.


Electrophysiological characterization of the flounder type II Na+/Pi cotransporter (NaPi-5) expressed in Xenopus laevis oocytes.
The two electrode voltage clamp technique was used to investigate the steady-state and presteady-state kinetic properties of the type II Na+/Pi cotransporter NaPi-5, cloned from the kidney of winter flounder (Pseudopleuronectes americanus) and expressed in Xenopus laevis oocytes. Steady-state Pi-induced currents had a voltage-independent apparent K(m) for Pi of 0.03 mM and a Hill coefficient of 1.0 at neutral pH, when superfusing with 96 mM Na+. The apparent K(m) for Na+ at 1 mM Pi was strongly voltage dependent (increasing from 32 mM at -70 mV to 77 mM at -30 mV) and the Hill coefficient was between 1 and 2, indicating cooperative binding of more than one Na+ ion. The maximum steady-state current was pH dependent, diminishing by 50% or more for a change from pH 7.8 to pH 6.3. Voltage jumps elicited presteady-state relaxations in the presence of 96 mM Na+ which were suppressed at saturating Pi (1 mM). Relaxations were absent in non-injected oocytes. Charge was balanced for equal positive and negative steps, saturated at extremes of potential and reversed at the holding potential. Fitting the charge transfer to a Boltzmann relationship typically gave a midpoint voltage (V0.5) close to zero and an apparent valency of approximately 0.6. The maximum steady-state transport rate correlated linearly with the maximum Pi-suppressed charge movement, indicating that the relaxations were NaPi-5-specific. The apparent transporter turnover was estimated as 35 sec-1. The voltage dependence of the relaxations was Pi-independent, whereas changes in Na+ shifted V0.5 to -60 mV at 25 mM Na+. Protons suppressed relaxations but contributed to no detectable charge movement in zero external Na+. The voltage dependent presteady-state behavior of NaPi-5 could be described by a 3 state model in which the partial reactions involving reorientation of the unloaded carrier and binding of Na+ contribute to transmembrane charge movement.


Altered prostaglandin E2 regulation of cytokine production in atopic dermatitis.
The monocyte-derived inflammatory mediator, prostaglandin E2 (PGE2), can reduce IFN-gamma production, and this in turn may relate to IL-4 up-regulation of IgE synthesis and impaired delayed hypersensitivity in atopy. These abnormalities may relate to the cyclic nucleotide dysregulation in atopic dermatitis (AD), where monocyte cyclic AMP-phosphodiesterase (PDE) activity is increased and the consequent reduction in cAMP levels allows increased inflammatory responsiveness. In this study, we assessed the relationship between PGE2 and IFN-gamma production along with abnormal PDE activity in AD monocytes. Blood mononuclear leukocytes (MNL) from normal and AD donors were cultured for 24 hours, and supernatants were assayed for PGE2 and IFN-gamma by RIA. Spontaneous PGE2, but not leukotriene C4 release, was significantly increased in AD MNL (p < 0.05), although IFN-gamma levels were reduced (p < 0.05). In contrast, purified AD T cells, after removal of PGE2-producing monocytes, produced levels of IFN-gamma significantly higher than in normal T cell cultures. Inhibition of PGE2 synthesis by indomethacin caused increased IFN-gamma production by MNL cultures. We noted a strong negative correlation (r = 0.77) between PDE activity and IFN-gamma production in MNL cultures. We speculate that abnormal cyclic nucleotide metabolism caused by increased PDE activity may allow elevated levels of PGE2 production by AD monocytes. This study demonstrates a regulatory interaction between monocytes and T cells in AD and suggests that PGE2 may be an extracellular messenger between these cells to modulate IFN-gamma production.


Alpha-adrenergic receptor blockade with prazosin. Consideration of hypertension, heart failure, and potential new applications.
Prazosin, an orally active alpha-1 selective adrenergic antagonist, has been of value in treating patients with hypertension and congestive heart failure. In contrast to non-subtype-selective alpha-adrenergic antagonists and direct-acting vasodilators, prazosin's hypotensive action is accompanied by little or no increase in heart, rate, plasma renin, or plasma norepinephrine. Prazosin is a versatile drug that may be used alone or in combination to treat mild, moderate, or severe hypertension. The antihypertensive effect is sustained, and may increase during long-term therapy. The major side effect, postural hypotension after the first drug administration, is related to drug dose and intravascular volume depletion. Other side effects are mild and seldom limit therapy. In patients with congestive heart failure, prazosin results in balanced venous and arterial dilation, similar to that produced by nitroprusside. Attenuation of some or all of prazosin's initial hemodynamic effects has been seen during multiple short-term administrations. However, chronic studies have shown sustained symptomatic and hemodynamic improvement during long-term administration; initial hemodynamic attenuation may be transient or partial, and does not preclude long-term effectiveness, particularly during exercise. Preliminary studies indicate that prazosin may also be effective in treating patients with peripheral vasospasm due to Raynaud's phenomenon or ergotamine overdose.


Hypochlorite-mediated fragmentation of hyaluronan, chondroitin sulfates, and related N-acetyl glycosamines: evidence for chloramide intermediates, free radical transfer reactions, and site-specific fragmentation.
Myeloperoxidase released from activated phagocytes reacts with H(2)O(2) in the presence of chloride ions to give hypochlorous acid. This oxidant has been implicated in the fragmentation of glycosaminoglycans, such as hyaluronan and chondroitin sulfates. In this study it is shown that reaction of HOCl with glycosaminoglycans and model compounds yields chloramides derived from the N-acetyl function of the glycosamine rings. The results of EPR spin trapping and product studies are consistent with the formation of amidyl radicals from these chloramides via both metal ion-dependent and -independent processes. In the case of glycosaminoglycan-derived amidyl radicals, evidence has been obtained in studies with model glycosides that these radicals undergo rapid intramolecular abstraction reactions to give carbon-centered radicals at C-2 on the N-acetyl glycosamine rings (via a 1,2-hydrogen atom shift) and at C-4 on the neighboring uronic acid residues (via 1,5-hydrogen atom shifts). The C-4 carbon-centered radicals, and analogous species derived from model glycosides, undergo pH-independent beta-scission reactions that result in glycosidic bond cleavage. With N-acetyl glucosamine C-1 alkyl glycosides, product formation via this mechanism is near quantitative with respect to chloramide loss. Analogous reactions with the glycosaminoglycans result in selective fragmentation at disaccharide intervals, as evidenced by the formation of "ladders" on gels; this selectivity is less marked under atmospheric oxygen concentrations than under anoxic conditions, due to competing peroxyl radical reactions. As the extracellular matrix plays a key role in mediating cell adhesion, growth, activation, and signaling, such HOCl-mediated glycosaminoglycan fragmentation may play a key role in disease progression and resolution, with the resulting fragments modulating the magnitude and quality of the immune response in inflammatory conditions.


Noise, cost and speed-accuracy trade-offs: decision-making in a decentralized system.
Many natural and artificial decision-making systems face decision problems where there is an inherent compromise between two or more objectives. One such common compromise is between the speed and accuracy of a decision. The ability to exploit the characteristics of a decision problem in order to vary between the extremes of making maximally rapid, or maximally accurate decisions, is a useful property of such systems. Colonies of the ant Temnothorax albipennis (formerly Leptothorax albipennis) are a paradigmatic decentralized decision-making system, and have been shown flexibly to compromise accuracy for speed when making decisions during house-hunting. During emigration, a colony must typically evaluate and choose between several possible alternative new nest sites of differing quality. In this paper, we examine this speed-accuracy trade-off through modelling, and conclude that noise and time-cost of assessing alternative choices are likely to be significant for T. albipennis. Noise and cost of such assessments are likely to mean that T. albipennis' decision-making mechanism is Pareto-optimal in one crucial regard; increasing the willingness of individuals to change their decisions cannot improve collective accuracy overall without impairing speed. We propose that a decentralized control algorithm based on this emigration behaviour may be derived for applications in engineering domains and specify the characteristics of the problems to which it should be suited, based on our new results.


Multinational study of pneumococcal serotypes causing acute otitis media in children.
Streptococcus pneumoniae is a major cause of acute otitis media (AOM) in young children. More than 90 immunologically distinct pneumococcal serotypes have been identified, but limited information is available regarding their relative importance in AOM. We analyzed nine existing datasets comprising pneumococcal isolates from middle ear fluid samples collected from 1994 through 2000 from 3,232 children with AOM from Finland, France, Greece, Israel, several East European countries, the US and Argentina. We examined the distribution of pneumococcal serotypes in relation to several demographic and epidemiologic variables, including gender, age, antibiotic resistance and source of culture material. The major serotypes identified included 19F and 23F, each comprising 13 to 25% of pneumococcal middle ear fluid isolates in most datasets; 14 and 6B, comprising 6 to 18%; whereas 6A, 19A and 9V each comprised 5 to 10%. Despite differences in location, study design and antibiotic susceptibility, each major serotype was prominent in most age groups of each dataset. Serotypes represented in the 7-valent pneumococcal conjugate vaccine (PCV-7, 4, 6B, 9V, 14, 18C, 19F, 23F) accounted for 60 to 70% of all pneumococcal isolates in the 6- to 59-month age range, but only 40 to 50% of isolates in children <6 or >/=60 months old. Serotype 3 and, in certain datasets, serotypes 1 and 5, were more important in the <6- and >/=60-month age groups. In each age group vaccine-related serotypes (mainly 6A and 19A) comprised an additional 10 to 15% of all pneumococcal isolates. Four serotypes (23F, 19F, 14 and 6B) accounted for 83% of all penicillin-resistant observations. This analysis of several geographically diverse datasets indicates that a limited number of serotypes, largely represented in PCV-7, accounted for the majority of episodes of pneumococcal AOM in children between 6 and 59 months of age. Certain serotypes appeared to be relatively more significant in children <6 months or >59 months of age.


Molecular models of the open and closed states of the whole human CFTR protein.
Cystic fibrosis transmembrane conductance regulator (CFTR), involved in cystic fibrosis (CF), is a chloride channel belonging to the ATP-binding cassette (ABC) superfamily. Using the experimental structure of Sav1866 as template, we previously modeled the human CFTR structure, including membrane-spanning domains (MSD) and nucleotide-binding domains (NBD), in an outward-facing conformation (open channel state). Here, we constructed a model of the CFTR inward-facing conformation (closed channel) on the basis of the recent corrected structures of MsbA and compared the structural features of those two states of the channel. Interestingly, the MSD:NBD coupling interfaces including F508 (DeltaF508 being the most common CF mutation) are mainly left unchanged. This prediction, completed by the modeling of the regulatory R domain, is supported by experimental data and provides a molecular basis for a better understanding of the functioning of CFTR, especially of the structural features that make CFTR the unique channel among the ABC transporters.


Attitudes on end-of-life care and advance care planning in the lesbian and gay community.
Gay men and lesbians have special interests in documenting their preferences regarding advance care planning and end-of-life care. A 64-item survey instrument was developed to ascertain the preferences of this community regarding approaches to end-of-life care, viewpoints on physician-assisted suicide (PAS) and euthanasia, and practices regarding advance care planning. The survey was completed by 575 participants recruited through community-based health care and social service organizations serving the lesbian and gay community, primarily in the New York metropolitan area. Respondents represent a diverse group of women (36%) and men (63%) from various age, racial/ethnic, and religious/spiritual backgrounds; 10% were human immunodeficiency virus (HIV)-positive. Respondents' perspectives on end-of-life care are generally consistent with findings from other attitudinal studies of U.S. adults: a majority supported legalization of PAS and preferred a palliative approach to end-of-life care. However, the gay community sample revealed even stronger support for assisted suicide and palliative care. Although respondents completed advance directives at a higher rate than adults generally, the legal importance for gay men and lesbians to execute directives should encourage health care providers and community organizations to assume a larger educational role on advance care planning. Results confirm other reports on the need to address provider communication skills. It is speculated that the HIV epidemic was a major influence behind these results because of the overwhelming personal impact of the epidemic on most gay men and lesbians during the past two decades.


Tunability of the CO adsorption energy on a Ni/Cu surface: Site change and coverage effects.
The adsorption energy of carbon monoxide on Ni ad-islands and ultra-thin films grown on the Cu(110) surface can be finely tuned via a complex interplay among diffusion, site change mechanisms, and coverage effects. The observed features of CO desorption can be explained in terms of migration of CO molecules from Cu to Ni islands, competition between bridge and on-top adsorption sites, and repulsive lateral adsorbate-adsorbate interactions. While the CO adsorption energy on clean Cu(110) is of the order of 0.5 eV, Ni-alloying allows for its controlled, continuous tunability in the 0.98-1.15 eV range with Ni coverage. Since CO is a fundamental reactant and intermediate in many heterogeneous catalytic (electro)-conversion reactions, insight into these aspects with atomic level detail provides useful information to potentially drive applicative developments. The tunability range of the CO adsorption energy that we measure is compatible with the already observed tuning of conversion rates by Ni doping of Cu single crystal catalysts for methanol synthesis from a CO2, CO, and H2 stream under ambient pressure conditions.


Developing the NASA food system for long-duration missions.
Even though significant development has transformed the space food system over the last 5 decades to attain more appealing dietary fare for low-orbit space crews, the advances do not meet the need for crews that might travel to Mars and beyond. It is estimated that a food system for a long-duration mission must maintain organoleptic acceptability, nutritional efficacy, and safety for a 3- to 5-y period to be viable. In addition, the current mass and subsequent waste of the food system must decrease significantly to accord with the allowable volume and payload limits of the proposed future space vehicles. Failure to provide the appropriate food or to optimize resource utilization introduces the risk that an inadequate food system will hamper mission success and/or threaten crew performance. Investigators for the National Aeronautics and Space Administration (NASA) Advanced Food Technology (AFT) consider identified concerns and work to mitigate the risks to ensure that any new food system is adequate for the mission. Yet, even with carefully planned research, some technological gaps remain. NASA needs research advances to develop food that is nutrient-dense and long-lasting at ambient conditions, partial gravity cooking processes, methods to deliver prescribed nutrients over time, and food packaging that meets the mass, barrier, and processing requirements of NASA. This article provides a brief review of research in each area, details the past AFT research efforts, and describes the remaining gaps that present barriers to achieving a food system for long exploration missions.


Colon Cancer Family Registry: an international resource for studies of the genetic epidemiology of colon cancer.
Family studies have served as a cornerstone of genetic research on colorectal cancer. The Colorectal Cancer Family Registry (Colon CFR) is an international consortium of six centers in North America and Australia formed as a resource to support studies on the etiology, prevention, and clinical management of colorectal cancer. Differences in design and sampling schemes ensures a resource that covers the continuum of disease risk. Two separate recruitment strategies identified colorectal cancer cases: population-based (incident case probands identified by cancer registries; all six centers) and clinic-based (families with multiple cases of colorectal cancer presenting at cancer family clinics; three centers). At this time, the Colon CFR is in year 10 with the second phase of enrollment nearly complete. In phase I recruitment (1998-2002), population-based sampling ranged from all incident cases of colorectal cancer to a subsample based on age at diagnosis and/or family cancer history. During phase II (2002-2007), population-based recruitment targeted cases diagnosed before the age of 50 years are more likely attributable to genetic factors. Standardized protocols were used to collect information regarding family cancer history and colorectal cancer risk factors, and biospecimens were obtained to assess microsatellite instability (MSI) status, expression of mismatch repair proteins, and other molecular and genetic processes. Of the 8,369 case probands enrolled to date, 2,602 reported having one or more colorectal cancer-affected relatives and 799 met the Amsterdam I criteria for Lynch syndrome. A large number of affected (1,324) and unaffected (19,816) relatives were enrolled, as were population-based (4,108) and spouse (983) controls. To date, 91% of case probands provided blood (or, for a few, buccal cell) samples and 75% provided tumor tissue. For a selected sample of high-risk subjects, lymphocytes have been immortalized. Nearly 600 case probands had more than two affected colorectal cancer relatives, and 800 meeting the Amsterdam I criteria and 128, the Amsterdam II criteria. MSI testing for 10 markers was attempted on all obtained tumors. Of the 4,011 tumors collected in phase I that were successfully tested, 16% were MSI-high, 12% were MSI-low, and 72% were microsatellite stable. Tumor tissues from clinic-based cases were twice as likely as population-based cases to be MSI-high (34% versus 17%). Seventeen percent of phase I proband tumors and 24% of phase II proband tumors had some loss of mismatch repair protein, with the prevalence depending on sampling. Active follow-up to update personal and family histories, new neoplasms, and deaths in probands and relatives is nearly complete. The Colon CFR supports an evolving research program that is broad and interdisciplinary. The greater scientific community has access to this large and well-characterized resource for studies of colorectal cancer.


Cesarean birth and maternal morbidity among Black women and White women after implementation of a blended payment policy.
To test whether Minnesota's blended payment policy had differential effects on cesarean use and maternal morbidity among black women and white women in Minnesota, as compared to six control states. Claims data from births to Medicaid fee-for-service beneficiaries, 2006-2012, in Minnesota (policy state) and six control states (Wisconsin, Iowa, Illinois, Oregon, Idaho, and Montana). The key study intervention was Minnesota's blended payment policy, which established one single payment rate for uncomplicated vaginal and cesarean births in 2009. The primary outcome was cesarean birth, and secondary outcomes were maternal morbidity (composite), postpartum hemorrhage, and chorioamnionitis. Policy effects were assessed using race-stratified comparative interrupted time series analysis. Following policy implementation, cesarean use decreased among both black and white women in Minnesota compared to control states; this decline was larger among black women (-2.88 percent 3-year cumulative decline, from a prepolicy cesarean rate of 22.2 percent) than among white women (-1.32 percent, P = .0013). Postpartum hemorrhage increased, with larger increases among black women (1.20 percent 3-year cumulative increase), compared with white women (0.48 percent, P < .001) in Minnesota compared with control states. Policy-related declines in cesarean use after Minnesota's blended payment policy were larger in black women. Increases in postpartum hemorrhage signal potential unintended consequences of policy-related cesarean reduction.


How cohabitation, marriage, separation, and divorce influence BMI: A prospective panel study.
This study examines how changes in cohabitation or marital status affect Body Mass Index (BMI) over time in a large representative sample. Participants were 20,950 individuals (50% female; 19 to 100 years), representative of the German population, who provided 81,926 observations over 16 years. Face-to-face interviews were used to obtain demographic data, including cohabitation and marital status, height, body weight, and weight-relevant behaviors (exercise, healthy eating, and smoking). Control variables included age, notable changes in status (life events such as having children or change in employment status), perceived stress, and subjective health. Cohabitation led to significant weight gain in men and women-after four years or longer, about twice the gain associated with marriage (controlling for weight-related behaviors, age, children, employment, stress, and health). BMI after separation was largely comparable to BMI before starting cohabitation; women lost some weight in the first year, men gained some weight after four or more years of separation. Divorce generally predicted weight gain. Changes in exercise, healthy eating, and smoking did not attenuate the effect of changes in relationship status on BMI. This is among the first longitudinal studies to directly compare the effects of key changes in relationship status on BMI. The findings extend and qualify previous results by showing that the benefits of marriage or cohabitation do not necessarily include a healthier BMI. They also suggest that relationship transitions-particularly moving in with a partner and divorce-may be important time windows for weight gain prevention. (PsycINFO Database Record


Change of blood pressure and urine flow rate during cardiopulmonary bypass and its relation to postoperative renal function.
The relationship between the perfusion flow, the mean arterial pressure (MAP) and the urine flow rate during cardiopulmonary bypass (CPB) and the effect of oliguria developed during CPB on the postoperative renal dysfunction were studied prospectively in 69 open heart surgery patients. The MAP, the perfusion flow and the urine flow rate were monitored every five minutes during the first 45 minutes after the commencement of CPB and after the removal of the aortic cross clamp (AX). The serum creatinine (Cr), creatinine clearance (CCr) and blood urea nitrogen were measured before operation, as well as on the first, second and third postoperative days. The dosage of catecholamines and diuretics used and the duration of intubation and hospitalization in the intensive care unit were also recorded. The urine flow rate correlated with MAP much better than the perfusion flow during CPB (r = 0.4768, p < 0.0001). The urine flow rate and MAP decreased significantly after the initiation of CPB and after the release of the AX; however, oliguria developed only during the first 30 minutes of CPB. There were no differences in postoperative Cr, postoperative CCr, doses of catecholamines or diuretics, and the duration of intubation between patients with or without development of oliguria during CPB. Parameters measured during CPB could not predict CCr during the first three postoperative days. We conclude that it is MAP rather than perfusion flow which correlates with the urine flow rate during CPB. Periods of oliguria during CPB did not correlate with or help in the prediction of the development of postoperative renal dysfunction.


Whole-genome amplification using Φ29 DNA polymerase.
The cornerstones of any genetic analysis study are the quality and quantity of the DNA samples. DNA is a precious limited resource, and in human disease studies the accessibility of sample DNA is often governed by the isolation method and the human source. Additionally, forensic analysis and archaeological research are generally infeasible without intact sample DNA. Therefore, mechanisms to preserve or enhance the quantity of the DNA stock are crucial to the success of these studies. Historically, to preserve and maintain DNA stocks, costly and labor-intensive Epstein-Barr-virus-transformed cell lines were produced. The creation of cell lines can be valuable for a number of reasons in addition to creating a renewable resource of DNA, but the cost and effort to create them, as well as the requirement of intact cells to begin with, limit the utility of this approach. More recently, whole-genome amplification (WGA), utilizing the unique property of the enzyme Φ29 DNA polymerase, has been used to generate robust high-fidelity copies of the genome. As described in this protocol, WGA using Φ29 DNA polymerase allows unbiased representation of the genome via multiple-strand displacement, followed by rolling-circle amplification on random primers.


Graduated driver licensing (GDL) in the United States in 2016: A literature review and commentary.
This is the sixth in a series of reviews of research on graduated driver licensing (GDL) published in the Journal of Safety Research, the present review covering the period mid-2012 through 2016. In the two decades since GDL programs began to be introduced on a widespread basis in the United States, a vast amount of research has been published. The current review discusses recent research and the present state of knowledge on the following topics: characteristics of the novice driver population; effects of GDL on crashes for ages 16-19; the learner and intermediate periods; night and passenger restrictions; cellphone laws; GDL for older novices; enforcement of GDL rules; and programs attempting to influence GDL compliance and safe driving practices in general. GDL stands out as a successful policy for reducing teen driver crashes and is worth building on to extend its benefits. Strengthening existing GDL programs has the most potential for producing further crash reductions.


Experimental models of epilepsy in young animals.
Seizures occur more frequently early in life. Some of these early seizures may eventually become epilepsy. Others are reactive seizures due to excessive environmental stimuli that, in any other age group, might not have elicited a similar response. To understand the developmental aspects of seizures and epilepsy in humans, it is important to study these processes in animals of equivalent ages. In this paper, we describe several animal models of developmental seizures, including their electroclinical manifestations and their validity in respect to human epileptic syndromes. There are several factors that may account for the increased seizure susceptibility of the immature brain, including the delayed development of effective systems or synaptic networks that are involved in the suppression of seizures. A better insight of the basic pathophysiology of seizures as a function of age in animal models will lead to the development of new therapeutic approaches for the treatment of age-specific epileptic disorders in humans.


Effect of variable carbohydrate intake on exercise performance in female endurance cyclists.
This study measured the effect of variable carbohydrate intake on time to exhaustion, variations in heart rate (HR), respiratory exchange ratio (RER), and rating of perceived exertion (RPE) in female endurance cyclists during an exercise trial. Subjects were 11 eumenorrheic women with maximal oxygen consumption (VO2max) 60.1 +/- 5.1 ml/kg who habitually cycled at least 100 miles per week. In a crossover design, each woman was randomly assigned to a eucaloric diet providing 8, 5, or 3 g of CHO/kg of body weight. Subjects cycled at least 100 miles while adhering to the diet for 6 days. The exercise trial was performed on the 7th day, consisting of a 60 min cycle at 70% VO2max, followed by an increase in intensity to 90% VO2max until the intensity could no longer be maintained. Results indicated no difference in mean time to exhaustion, heart rate, or RPE. RER increased over time-elapsed (F = 40.4, p < .001) and across diets (F = 6.1, p = .015). Female endurance cyclists did not experience a difference in time to exhaustion, HR, or RPE with different levels of CHO intake during an endurance trial. RER varied with diet at submaximal intensities. Further research is needed to determine the optimal level of CHO intake for this population.


A randomized controlled trial of filgrastim during remission induction and consolidation chemotherapy for adults with acute lymphoblastic leukemia: CALGB study 9111.
Recombinant human granulocyte colony-stimulating factor (G-CSF; filgrastim) shortens the time to neutrophil recovery after intensive chemotherapy, but its role in the treatment of adults with acute lymphoblastic leukemia (ALL) is uncertain. We randomly assigned 198 adults with untreated ALL (median age, 35 years; range, 16 to 83) to receive either placebo or G-CSF (5 microgram/kg/d) subcutaneously, beginning 4 days after starting intensive remission induction chemotherapy and continuing until the neutrophil count was >/=1, 000/microL for 2 days. The study assignment was unblinded as individual patients achieved a complete remission (CR). Patients initially assigned to G-CSF then continued to receive G-CSF through 2 monthly courses of consolidation therapy. Patients assigned to placebo received no further study drug. The median time to recover neutrophils >/=1,000/microL during the remission induction course was 16 days (interquartile range [IQR], 15 to 18 days) for the patients assigned to receive G-CSF and 22 days (IQR, 19 to 29 days) for the patients assigned to placebo (P < .001). Patients in the G-CSF group had significantly shorter durations of neutropenia (<1, 000/microL) and thrombocytopenia (<50,000/microL) and fewer days in the hospital (median, 22 days v 28 days; P = .02) compared with patients receiving placebo. The patients assigned to receive G-CSF had a higher CR rate and fewer deaths during remission induction than did those receiving placebo (P = .04 by the chi-square test for trend). During Courses IIA and IIB of consolidation treatment, patients in the G-CSF group had significantly more rapid recovery of neutrophils >/=1,000/microL than did the control group by approximately 6 to 9 days. However, the patients in the G-CSF group did not complete the planned first 3 months of chemotherapy any more rapidly than did the patients in the placebo group. Overall toxicity was not lessened by the use of G-CSF. After a median follow-up of 4. 7 years, there were no significant differences in either the disease-free survival (P = .53) or the overall survival (P = .25) for the patients assigned to G-CSF (medians, 2.3 years and 2.4 years, respectively) compared with those assigned to placebo (medians, 1.7 and 1.8 years, respectively). Adults who received intensive chemotherapy for ALL benefited from G-CSF treatment, but its use did not markedly affect the ultimate outcome.


Comparison of a human portable blood glucose meter, veterinary portable blood glucose meter, and automated chemistry analyzer for measurement of blood glucose concentrations in dogs.
To compare blood glucose concentrations measured with 2 portable blood glucose meters (PBGMs) validated for use in dogs (PBGM-D) and humans (PBGM-H) and an automated chemistry analyzer. Validation study. 92 samples of fresh whole blood and plasma from 83 dogs with various diseases. Each PBGM was used to measure whole blood glucose concentration, and the automated analyzer was used to measure plasma glucose concentration. Passing-Bablok linear regression and Bland-Altman plots were used to determine correlations and bias between the PBGMs and the automated analyzer. Calculated acceptability limits based on combined inherent instrument imprecision were used with Bland-Altman plots to determine agreement. Clinical relevance was assessed via error grid analysis. Although correlation between results of both PBGMs and the standard analyzer was > 0.90, disagreement was greater than could be explained by instrument imprecision alone. Mean difference between PBGM-H and chemistry-analyzer values was -15.8 mg/dL. Mean difference between PBGM-D and chemistry-analyzer values was 2.4 mg/dL. Linear regression analysis revealed proportional bias of PBGM-H (greater disagreement at higher glucose concentrations); no proportional bias was detected for PBGM-D. No constant bias was detected for either PBGM. Error grid analysis revealed all measurements from both PBGMs were within zones without an anticipated effect on clinical outcome. Neither PBGM had exact agreement with the automated analyzer; however, the disagreement detected did not have serious clinical consequences. Our findings stressed the importance of using the same device for monitoring trends in dogs and using instrument-specific reference ranges.


Role of US LI-RADS in the LI-RADS Algorithm.
The US Liver Imaging Reporting and Data System (LI-RADS) was released in 2017 and is the newest of the four American College of Radiology (ACR) LI-RADS algorithms. US LI-RADS provides standardized terminology, technical recommendations, and a reporting framework for US examinations performed for screening or surveillance in patients at risk for developing hepatocellular carcinoma (HCC). The appropriate patient population for screening and surveillance includes individuals who are at risk for developing HCC but do not have known or suspected cancer. This includes patients with cirrhosis from any cause and subsets of patients with chronic hepatitis B virus infection in the absence of cirrhosis. In an HCC screening or surveillance study, US LI-RADS recommends assigning two scores that apply to the entire study: the US category, which determines follow-up, and a visualization score, which communicates the expected level of sensitivity of the examination but does not affect management. Three US categories are possible: US-1 negative, a study with no evidence of HCC; US-2 subthreshold, a study in which an observation less than 10 mm is depicted that is not definitely benign; and US-3 positive, a study in which an observation greater than or equal to 10 mm or a new thrombus in vein is identified, for which diagnostic contrast material-enhanced imaging is recommended. Three visualization scores are possible: A (no or minimal limitations), B (moderate limitations), and C (severe limitations). ©RSNA, 2019.


The role of temperature and dispersal in moss-microarthropod community assembly after a catastrophic event.
There is a clear crisis in the maintenance of biodiversity. It has been generated by a multitude of factors, notably habitat loss, now compounded by the effects of climate change. Predicted changes in climate include increased severity and frequency of extreme climatic events. To manage landscapes, an understanding of the processes that allow recovery from these extreme events is required. Understanding these landscape-scale processes of community assembly and disassembly is hindered by the large scales at which they operate. Model systems provide a means of studying landscape scale processes at tractable scales. Here, we assess the combined effects of temperature and habitat-patch isolation on assembly of naturally diverse moss microarthropod communities after a high-temperature event. We show that community assembly depends on temperature and on degree of habitat isolation. Heated communities were heavily dominated in abundance by two species, one of them relatively large. The resulting size-structure is unlike that seen in the field. Community composition in habitat fragments appears also to have been influenced by the source pool of recolonizing fauna. Our results highlight the value of dispersal in disturbed landscapes and the potential for habitat connectivity to buffer communities from the effects of climate change.


A Tool for Interactive Data Visualization: Application to Over 10,000 Brain Imaging and Phantom MRI Data Sets.
In this paper we propose a web-based approach for quick visualization of big data from brain magnetic resonance imaging (MRI) scans using a combination of an automated image capture and processing system, nonlinear embedding, and interactive data visualization tools. We draw upon thousands of MRI scans captured via the COllaborative Imaging and Neuroinformatics Suite (COINS). We then interface the output of several analysis pipelines based on structural and functional data to a t-distributed stochastic neighbor embedding (t-SNE) algorithm which reduces the number of dimensions for each scan in the input data set to two dimensions while preserving the local structure of data sets. Finally, we interactively display the output of this approach via a web-page, based on data driven documents (D3) JavaScript library. Two distinct approaches were used to visualize the data. In the first approach, we computed multiple quality control (QC) values from pre-processed data, which were used as inputs to the t-SNE algorithm. This approach helps in assessing the quality of each data set relative to others. In the second case, computed variables of interest (e.g., brain volume or voxel values from segmented gray matter images) were used as inputs to the t-SNE algorithm. This approach helps in identifying interesting patterns in the data sets. We demonstrate these approaches using multiple examples from over 10,000 data sets including (1) quality control measures calculated from phantom data over time, (2) quality control data from human functional MRI data across various studies, scanners, sites, (3) volumetric and density measures from human structural MRI data across various studies, scanners and sites. Results from (1) and (2) show the potential of our approach to combine t-SNE data reduction with interactive color coding of variables of interest to quickly identify visually unique clusters of data (i.e., data sets with poor QC, clustering of data by site) quickly. Results from (3) demonstrate interesting patterns of gray matter and volume, and evaluate how they map onto variables including scanners, age, and gender. In sum, the proposed approach allows researchers to rapidly identify and extract meaningful information from big data sets. Such tools are becoming increasingly important as datasets grow larger.


All-conjugated triblock polyelectrolytes.
All-conjugated triblock polyfluorenes with well-defined molecular weights and low polydispersities are synthesized via chain-growth Suzuki-Miyaura polymerization. Ionization of pendant alkylbromide chains by pyridine affords amphiphilic triblock polyelectrolytes with neutral/charged/neutral or charged/neutral/charged segments. The immiscible blocks lead to aggregation in polar and nonpolar solvents, and to complex surface morphologies depending on the polarity of the substrate. These triblock polyelectrolytes can also be used as interfacial layers in polymer light-emitting diodes to facilitate electron injection from aluminum.


Enhanced expression and secretion of an epithelial membrane antigen (MA5) in a human mucinous breast tumor line (BT549).
The mouse monoclonal antibody MA5, generated versus a membrane-enriched extract of breast cancer metastatic to liver, detects one or two high molecular weight species (greater than 200 kD) in breast tumor membranes, human milk fat globule membranes, and various breast tumor cell lines. From comparative studies of five breast carcinoma lines (BT20, BT549, MCF-7, T47D, and ZR75-1), as well as an epithelial line established from milk (HBL-100), we report the stimulation of expression of MA5-reactive antigen in a mucinous breast tumor cell line (BT549) through the use of a culture medium supplemented with charcoal-absorbed fetal calf serum, insulin, and hydrocortisone. Large amounts of aggregated MA5-reactive antigen are secreted into the culture medium and can be recovered from the media for further purification by centrifugation. These findings suggest that BT549 cells, grown in the special nutritive medium, may be useful in providing an ample source of epithelial membrane antigen (also termed polymorphic epithelial mucin) for standardization of clinical assay protocols, as well as provide a model system for studies of the regulation of expression for this class of antigens in breast carcinoma.


Biodiversity of the Great Barrier Reef-how adequately is it protected?
The Great Barrier Reef (GBR) is the world's most iconic coral reef ecosystem, recognised internationally as a World Heritage Area of outstanding significance. Safeguarding the biodiversity of this universally important reef is a core legislative objective; however, ongoing cumulative impacts including widespread coral bleaching and other detrimental impacts have heightened conservation concerns for the future of the GBR. Here we review the literature to report on processes threatening species on the GBR, the status of marine biodiversity, and evaluate the extent of species-level monitoring and reporting. We assess how many species are listed as threatened at a global scale and explore whether these same species are protected under national threatened species legislation. We conclude this review by providing future directions for protecting potentially endangered elements of biodiversity within the GBR. Most of the threats identified to be harming the diversity of marine life on the GBR over the last two-three decades remain to be effectively addressed and many are worsening. The inherent resilience of this globally significant coral reef ecosystem has been seriously compromised and various elements of the biological diversity for which it is renowned may be at risk of silent extinction. We show at least 136 of the 12,000+ animal species known to occur on the GBR (approximately 20% of the 700 species assessed by the IUCN) occur in elevated categories of threat (Critically Endangered, Endangered or Vulnerable) at a global scale. Despite the wider background level of threat for these 136 species, only 23 of them are listed as threatened under regional or national legislation. To adequately protect the biodiversity values of the GBR, it may be necessary to conduct further targeted species-level monitoring and reporting to complement ecosystem management approaches. Conducting a vigorous value of information analysis would provide the opportunity to evaluate what new and targeted information is necessary to support dynamic management and to safeguard both species and the ecosystem as a whole. Such an analysis would help decision-makers determine if further comprehensive biodiversity surveys are needed, especially for those species recognised to be facing elevated background levels of threat. If further monitoring is undertaken, it will be important to ensure it aligns with and informs the GBRMPA Outlook five-year reporting schedule. The potential also exists to incorporate new environmental DNA technologies into routine monitoring to deliver high-resolution species data and identify indicator species that are cursors of specific disturbances. Unless more targeted action is taken to safeguard biodiversity, we may fail to pass onto future generations many of the values that comprise what is universally regarded as the world's most iconic coral reef ecosystem.


Substantial observer variability in the differentiation between primary intracerebral hemorrhage and hemorrhagic transformation of infarction on CT brain imaging.
CT remains the most commonly used imaging technique in acute stroke but is often delayed after minor stroke. Interobserver reliability in distinguishing hemorrhagic transformation of infarction from intracerebral hemorrhage may depend on delays to CT but has not been reported previously despite the clinical importance of this distinction. Initial CT scans with intraparenchymal hematoma from the first 1000 patients with stroke in the Oxford Vascular Study were independently categorized as intracerebral hemorrhage or hemorrhagic transformation of infarction by 5 neuroradiologists, both blinded and unblinded to clinical history. Thirty scans were reviewed twice. Agreement was quantified by the kappa statistic. Seventy-eight scans showed intraparenchymal hematoma. Blinded pairwise interrater agreements for a diagnosis of intracerebral hemorrhage ranged from kappa=0.15 to 0.48 with poor overall agreement (kappa=0.35; 95% CI, 0.15 to 0.54) even after unblinding (kappa=0.41; 0.21 to 0.60). Blinded intrarater agreements ranged from kappa=0.21 to 0.92. Lack of consensus after unblinding was greatest in patients scanned >or=24 hours after stroke onset (67% versus 25%, P=0.001) and in minor stroke (National Institutes of Health Stroke Scale <or=5: 56% versus 29%, P=0.04) with disagreement in 75% of patients scanned >or=24 hours after minor stroke and in 48% of all 30-day stroke survivors in whom reliable diagnosis would be expected to influence long-term management. Reliability of diagnosis of intraparenchymal hematoma on CT brain scan in minor stroke is poor, particularly if scanning is delayed. Immediate brain imaging is justified in patients with minor stroke.


Impact of repeat ablation of ventricular tachycardia in patients with structural heart disease.
Recurrences of ventricular tachycardia (VT) after initial catheter ablation is a significant clinical problem. In this study, we report the efficacy and risks of repeat VT ablation in patients with structural heart disease (SHD) in a tertiary single centre over a 7-year period. Two hundred ten consecutive patients referred for repeat VT ablation after previous ablation in our institution were included in the analysis (53% ischaemic cardiomyopathy, 91% males, median age 65 years, mean left ventricular ejection fraction 35%). After performing repeat ablation, the clinical VTs were acutely eliminated in 82% of the patients, but 46% of the cohort presented with VT recurrence during the 25-month follow-up. Repeat ablation led to a 73% reduction of shock burden in the first year and 61% reduction until the end of follow-up. Similarly, VT burden was reduced 55% in the first year and 36% until the end of the study. Fifty-two patients (25%) reached the combined endpoint of ventricular assist device implantation, heart transplantation, or death. Advanced New York Heart Association functional class, anteroseptal substrate, and periprocedural complication after repeat ablation were associated with worse prognosis independently of the type of cardiomyopathy. While complete freedom from VT after repeat ablation in SHD was difficult to achieve, ablation led to a significant reduction in VT and shock burden. Besides advanced heart failure characteristics, anteroseptal substrate and periprocedural complications predicted a worse outcome.


Histomorphometric and transcriptome evaluation of early healing bone treated with a novel human particulate dentin powder.
Human particulate dentin (HPD) shows potential as an alternative bone grafting material. However, the mechanism of bone healing at the molecular level after grafting with HPD is unclear. This study assessed the histological and global gene expression of bone tissues grafted with HPD. The HPD was prepared to 250-500 µm in size. X-ray diffraction (XRD) and energy dispersive x-ray spectroscopy (EDX) were performed to confirm the crystal structure, organic compound residues, and surface morphology, respectively. Bony defects were created on the heads of 24 New Zealand White rabbits. Sterilized HPD was used as the grafting material. The quality and quantity of new bone formation was evaluated using micro-CT and histologic analyses during the 8 week experimental periods. For microarray assay, bone tissue and blood samples were taken at 3, 5 and 7 d post-implantation. The results of XRD and EDX showed that HPD exhibited physical and chemical properties similar to natural hydroxyapatite. New bone formation was observed after HPD implantation compared to the controls, as shown on hematoxylin and eosin staining and micro-CT. The bone volume of HPD treated animals was higher than that of the control group at all observation times. Microarray analysis showed that vascular development coupled with immune and inflammatory related genes were expressed in the early healing stage. The gene coding for the IL-1 antagonist, IL1RN, was expressed to inhibit the inflammatory response, and at the same time, the CCL2 gene was activated to 2.3 times the normal level. BMP2, RUNX2, COL1A, and OPN expression were also up-regulated. CCL2 predominated in osteoblastogenesis of the HPD-treated bony defect in the early stage of healing. HPD accelerated bone regeneration and augmentation. These results suggested that HPD provided potential as a bone graft resource during the bone healing process.


A Survey of Tele-Critical Care State and Needs in 2019 and 2020 Conducted among the Members of the Society of Critical Care Medicine.
The study's objective was to assess facilitators and barriers of Tele-Critical Care (TCC) perceived by SCCM members. By utilizing a survey distributed to SCCM members, a cross-sectional study was developed to analyze survey results from December 2019 and July 2020. SCCM members responded to the survey (n = 15,502) with a 1.9% response rate for the first distribution and a 2.54% response rate for the second survey (n = 9985). Participants (n = 286 and n = 254) were almost equally distributed between non-users, providers, users, and potential users of TCC services. The care delivery models for TCC were similar across most participants. Some consumers of TCC services preferred algorithmic coverage and scheduled rounds, while reactive and on-demand models were less utilized. The surveys revealed that outcome-driven measures were the principal form of TCC performance evaluation. A 1:100 (provider: patients) ratio was reported to be optimal. Factors related to costs, perceived lack of need for services, and workflow challenges were described by those who terminated TCC services. Barriers to implementation revolved around lack of reimbursement and adequate training. Interpersonal communication was identified as an essential TCC provider skill. The second survey introduced after the onset pandemic demonstrated more frequent use of advanced practice providers and focus on performance measures. Priorities for effective TCC deployment include communication, knowledge, optimal operationalization, and outcomes measurement at the organizational level. The potential effect of COVID-19 during the early stages of the pandemic on survey responses was limited and focused on the need to demonstrate TCC value.


Validity of conducting clinical dysphagia assessments for patients with normal to mild cognitive impairment via telerehabilitation.
To assess the validity of conducting clinical dysphagia assessments via telerehabilitation, 40 individuals with dysphagia from various etiologies were assessed simultaneously by a face-to-face speech-language pathologist (FTF-SLP) and a telerehabilitation SLP (T-SLP) via an Internet-based videoconferencing telerehabilitation system. Dysphagia status was assessed using a Clinical Swallowing Examination (CSE) protocol, delivered via a specialized telerehabilitation videoconferencing system and involving the use of an assistant at the patient's end of the consultation to facilitate the assessment. Levels of agreement between the FTF-SLP and T-SLP revealed that the majority of parameters reached set levels of clinically acceptable levels of agreement. Specifically, agreement between the T-SLP and FTF-SLP ratings for the oral, oromotor, and laryngeal function tasks revealed levels of exact agreement ranging from 75 to 100% (kappa = 0.36-1.0), while the parameters relating to food and fluid trials ranged in exact agreement from 79 to 100% (kappa = 0.61-1.0). Across the parameters related to aspiration risk and clinical management, exact agreement ranged between 79 and 100% (kappa = 0.49-1.0). The data show that a CSE conducted via telerehabilitation can provide valid and reliable outcomes comparable to clinical decisions made in the FTF environment.


Evaluation of recombinant human bone morphogenetic protein-2 in oral applications including the use of endosseous implants: 3-year results of a pilot study in humans.
This study evaluated patients who had been treated with recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded in an absorbable collagen sponge (ACS) in human extraction sites or in sites that required alveolar ridge augmentation. An earlier report on the same patients revealed that after 4 months, implantation of rhBMP-2/ACS was safe, as determined by clinical, radiographic, systemic, and immunological analyses. In this longer-term follow-up, eligible patients were restored with endosseous dental implants in the area treated with rhBMP-2/ACS and bone biopsy samples were taken for histological analysis of the treated human bone tissue. The primary objective was to monitor the long-term safety of patients treated with rhBMP-2/ACS. Another objective was to evaluate the dental implants placed in the sites treated with rhBMP-2. Patient safety was evaluated by clinical examinations, periapical radiographs, and occurrence of adverse experiences. Dental implants were evaluated by radiographic and clinical examination. All 12 patients have been followed for 3 years. Two years following surgical implantation of rhBMP-2/ACS, no serious or unexpected adverse experiences occurred. The adverse experiences that did occur were mostly benign and compatible with the dental implant surgeries performed in these patients. No adverse experiences were deemed as related to the rhBMP-2/ACS. Furthermore, no safety concerns in the local area of rhBMP-2/ACS placement were noted, based on oral wound examinations. In the 10 patients (6 extraction socket patients and 4 augmentation patients) who received endosseous implants, all implants were clinically stable at all assessments and all 10 patients have been functionally restored. Histological evaluation of the human bone core biopsies revealed normal bone tissue formation identical to the surrounding native bone. Three-year follow-up clinical examinations revealed that all implants had stable marginal bone levels and healthy peri-implant tissues. These 3-year results demonstrate that rhBMP-2/ACS can be used safely in human patients. Human bone biopsies reveal normal bone formation in areas treated with rhBMP-2/ACS. Endosseous implants placed in these areas were all stable with no radiographic or clinical complications. The results from this study suggest that rhBMP-2/ACS (0.43 mg/ml) can be safely used in tooth extraction sites and in local ridge augmentation procedures and that endosseous dental implants placed in bony areas treated with rhBMP-2/ACS are stable and can be functionally restored without complication.


Single-Molecule Analysis of Lipid-Protein Interactions in Crude Cell Lysates.
Recognition of signaling phospholipids by proteins is a critical requirement for the targeting and initiation of many signaling cascades. Most biophysical methods for measuring protein interactions with signaling phospholipids use purified proteins, which do not take into account the effect of post-translational modifications and other cellular components on these interactions. To potentially circumvent these problems, we have developed a single-molecule fluorescence approach to analyzing lipid-protein interactions in crude cell extracts. As a proof of principle for this assay, we show that a variety of lipid-binding domains (LBDs) can be recruited from cell lysates specifically onto their target phospholipids. With single-molecule analysis in real-time, our assay allows direct determination of binding kinetics for transient lipid-protein interactions and has revealed unique assembly properties and multiple binding modes of different LBDs. Whereas single-copy LBDs display transient interaction with lipid vesicles, tandem-repeat LBDs, often used as lipid biosensors, tend to form stable interactions that accumulate over time. We have extended the assay to study a cellular protein, Akt, and discovered marked differences in the lipid binding properties of the full-length protein compared to its PH domain. Importantly, we have found that phosphorylation of Akt at T308 and S473 does not affect the lipid binding behaviors of Akt, contrary to the long-standing model of Akt regulation. Overall, this work establishes the single-molecule lipid pulldown assay as a simple and highly sensitive approach to interrogating lipid-protein interactions in a setting that at least partly mimics the cellular environment.


A Liquid Chromatography/Mass Spectrometry Method for Screening Disulfide Tethering Fragments.
We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a percent bound value for each compound and identifies false positives by calculating significance of detected masses (signal significance). An example pipeline is available through Biovia's ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1 to 2 days.


TGF-β/Smad3 inhibit vascular smooth muscle cell apoptosis through an autocrine signaling mechanism involving VEGF-A.
We have previously shown that in the presence of elevated Smad3, transforming growth factor-β (TGF-β) transforms from an inhibitor to a stimulant of vascular smooth muscle cell (SMC) proliferation and intimal hyperplasia (IH). Here we identify a novel mechanism through which TGF-β/Smad3 also exacerbates IH by inhibiting SMC apoptosis. We found that TGF-β treatment led to inhibition of apoptosis in rat SMCs following viral expression of Smad3. Conditioned media from these cells when applied to naive SMCs recapitulated this effect, suggesting an autocrine pathway through a secreted factor. Gene array of TGF-β/Smad3-treated cells revealed enhanced expression of vascular endothelial growth factor (VEGF), a known inhibitor of endothelial cell apoptosis. We then evaluated whether VEGF is the secreted mediator responsible for TGF-β/Smad3 inhibition of SMC apoptosis. In TGF-β/Smad3-treated cells, VEGF mRNA and protein as well as VEGF secretion were increased. Moreover, recombinant VEGF-A inhibited SMC apoptosis and a VEGF-A-neutralizing antibody reversed the inhibitory effect of conditioned media on SMC apoptosis. Stimulation of SMCs with TGF-β led to the formation of a complex of Smad3 and hypoxia-inducible factor-1α (HIF-1α) that in turn activated the VEGF-A promoter and transcription. In rat carotid arteries following arterial injury, Smad3 and VEGF-A expression were upregulated. Moreover, Smad3 gene transfer further enhanced VEGF expression as well as inhibited SMC apoptosis. Finally, blocking either the VEGF receptor or Smad3 signaling in injured carotid arteries abrogated the inhibitory effect of Smad3 on vascular SMC apoptosis. Taken together, our study reveals that following angioplasty, elevation of both TGF-β and Smad3 leads to SMC secretion of VEGF-A that functions as an autocrine inhibitor of SMC apoptosis. This novel pathway provides further insights into the role of TGF-β in the development of IH.


Single-center evaluation of the single-dose pharmacokinetics of the angiotensin II receptor antagonist azilsartan medoxomil in renal impairment.
Azilsartan medoxomil (AZL-M) is a potent angiotensin II receptor blocker that decreases blood pressure in a dose-dependent manner. It is a pro-drug and not detected in blood after oral administration because of rapid hydrolysis to the active moiety, azilsartan (AZL). AZL undergoes further metabolism to the major metabolite M-II and minor metabolites. The objective of this study was to determine the effect of renal impairment on the pharmacokinetics of AZL and its major metabolite. This was a single-center, open-label, phase I parallel-group study which examined the single-dose (40-mg) pharmacokinetics of AZL and M-II in 24 subjects with mild, moderate, or severe renal impairment or end-stage renal disease requiring hemodialysis (n = 6 per group), respectively, and healthy matched subjects (n = 24). Renal impairment/disease did not cause clinically meaningful increases in exposure to AZL. M-II exposure was higher in all renally impaired subjects and highest in those with severe impairment (approx fivefold higher vs. control). M-II is pharmacologically inactive; increased exposure was not considered important in dose selection for AZL-M in subjects with renal impairment. Hemodialysis did not significantly remove AZL or M-II. Renal impairment had no clinically meaningful effect on the plasma protein binding of AZL or M-II. Single doses of AZL-M 40 mg were well tolerated in all subject groups. Based on the pharmacokinetic and tolerability findings, no dose adjustment of AZL-M is required for subjects with any degree of renal impairment, including end-stage renal disease.


Measurement of protein using bicinchoninic acid.
Bicinchoninic acid, sodium salt, is a stable, water-soluble compound capable of forming an intense purple complex with cuprous ion (Cu1+) in an alkaline environment. This reagent forms the basis of an analytical method capable of monitoring cuprous ion produced in the reaction of protein with alkaline Cu2+ (biuret reaction). The color produced from this reaction is stable and increases in a proportional fashion over a broad range of increasing protein concentrations. When compared to the method of Lowry et al., the results reported here demonstrate a greater tolerance of the bicinchoninate reagent toward such commonly encountered interferences as nonionic detergents and simple buffer salts. The stability of the reagent and resulting chromophore also allows for a simplified, one-step analysis and an enhanced flexibility in protocol selection. This new method maintains the high sensitivity and low protein-to-protein variation associated with the Lowry technique.


Assessing the power of tag SNPs in the mapping of quantitative trait loci (QTL) with extremal and random samples.
Recent studies have indicated that the human genome could be divided into regions with low haplotype diversity interspersed with regions of high haplotype diversity. In regions of low haplotype diversity, a small fraction of SNPs (tag SNPs) are sufficient to account for most of the haplotype diversity of the human genome. These tag SNPs can be extremely useful for testing the association of a marker locus with a qualitative or quantitative trait locus in that it may not be necessary to genotype all the SNPs. When tag SNPs are used to reduce the genotyping effort in association studies, it is important to know how much power is lost. It is also important to know how much power is gained when tag SNPs instead of the same number of randomly chosen SNPs are used. We design a simulation study to tackle these problems for a variety of quantitative association tests using either case-parent samples or unrelated population samples. First, the samples are generated based on the quantitative trait model with the assumption of either an extremal sampling scheme or a random sampling scheme. Second, a small number of samples are selected to determine the haplotype blocks and the tag SNPs. Third, the statistical power of the tests is evaluated using four kinds of data: (1) all the SNPs and the corresponding haplotypes, (2) the tag SNPs and the corresponding haplotypes, (3) the same number of evenly spaced SNPs with minor allele frequency greater than a threshold and the corresponding haplotypes, (4) the same number of randomly chosen SNPs and their corresponding haplotypes. Our results suggest that in most situations genotyping efforts can be significantly reduced by using tag SNPs for mapping the QTL in association studies without much loss of power, which is consistent with previous studies on association mapping of qualitative traits. For all situations considered, two-locus haplotype analysis using tag SNPs are more powerful than those using the same number of randomly selected SNPs, but the degree of such power differences depends upon the sampling scheme and the population history.


"Reactivity to stimuli" is a temperamental factor contributing to canine aggression.
Canine aggression is one of the most frequent problems in veterinary behavioral medicine, which in severe cases may result in relinquishment or euthanasia. As it is important to reveal underlying factors of aggression for both treatment and prevention, we recently developed a questionnaire on aggression and temperamental traits and found that "reactivity to stimuli" was associated with aggression toward owners, children, strangers, and other dogs of the Shiba Inu breed. In order to examine whether these associations were consistent in other breeds, we asked the owners of insured dogs of Anicom Insurance Inc. to complete our questionnaire. The top 17 contracted breeds were included. The questionnaire consisted of dogs' general information, four items related to aggression toward owners, children, strangers, and other dogs, and 20 other behavioral items. Aggression-related and behavioral items were rated on a five-point frequency scale. Valid responses (n = 5610) from owners of dogs aged 1 through 10 years were collected. Factor analyses on 18 behavioral items (response rate over 95%) extracted five largely consistent factors in 14 breeds: "sociability with humans," "fear of sounds," "chase proneness," "reactivity to stimuli," and "avoidance of aversive events." By stepwise multiple regression analyses, using the Schwartz's Bayesian information criterion (BIC) method with aggression points as objective variables and general information and temperamental factor points as explanatory variables, "reactivity to stimuli," i.e., physical reactivity to sudden movement or sound at home, was shown to be significantly associated with owner-directed aggression in 13 breeds, child-directed aggression in eight breeds, stranger-directed aggression in nine breeds, and dog-directed aggression in five breeds. These results suggest that "reactivity to stimuli" is simultaneously involved in several types of aggression. Therefore, it would be worth taking "reactivity to stimuli" into account in the treatment and prevention of canine aggression.


Expression of prolyl-hydroxylases PHD-1, 2 and 3 and of the asparagine hydroxylase FIH in non-small cell lung cancer relates to an activated HIF pathway.
The oxygen sensitive prolyl-hydroxylase domain enzymes (PHDs) and the asparagines hydroxylase (FIH, factor inhibiting HIF) regulate the transcriptional activity of HIFs. We assessed the expression of these enzymes in a series of 73 non-small cell lung carcinomas (NSCLC). A direct association of PHDs with FIH and of the PHDs/FIH with HIFs expression was noted. Thirty three of 73 cases had high HIF/PHD expression, predicting intense HIF activity; 19/73 cases had low HIF and high PHD expression mimicking the normal bronchial pattern; and 18/73 cases had low HIF/PHD (inactive HIF pathway). High lactate dehydrogenase LDH5 expression was noted in cases with high HIF/PHD phenotype. The value of such a classification in defining different metabolic phenotypes of NSCLC deserves further evaluation.