[Treatment of senile macular degeneration with Ginkgo biloba extract. A preliminary double-blind drug vs. placebo study].
Senile macular degeneration is a frequent cause of blindness for which there is no satisfactory medical treatment. A double-blind trial comparing Ginkgo biloba extract with a placebo was conducted in 10 out-patients at the Hôpital Foch. Drug effectiveness was assessed on the results of fundoscopy and of measurements of visual acuity and visual field. In spite of the small population sample, a statistically significant improvement in long distance visual acuity was observed after treatment with Ginkgo biloba extract. The assumed pathogenesis of senile macular degeneration is discussed with emphasis on free oxygenated radicals.


A comprehensive NMR methodology to assess the composition of biobased and biodegradable polymers in contact with food.
A nuclear magnetic resonance (NMR) methodology was assessed regarding the identification and quantification of additives in three types of polylactide (PLA) intended as food contact materials. Additives were identified using the LNE/NMR database which clusters NMR datasets on more than 130 substances authorized by European Regulation No. 10/2011. Of the 12 additives spiked in the three types of PLA pellets, 10 were rapidly identified by the database and correlated with spectral comparison. The levels of the 12 additives were estimated using quantitative NMR combined with graphical computation. A comparison with chromatographic methods tended to prove the sensitivity of NMR by demonstrating an analytical difference of less than 15%. Our results therefore demonstrated the efficiency of the proposed NMR methodology for rapid assessment of the composition of PLA.


Enzymatic oxidation of cholesterol aggregates in supercritical carbon dioxide.
Fundamental studies of enzyme-solvent interactions can be conducted with supercritical fluids because small changes in pressure or temperature may bring about great changes in the properties of a single solvent near its critical point. Cholesterol oxidase is active in supercritical carbon dioxide and supercritical carbon dioxide-cosolvent mixtures. Variations in solvent power caused by pressure changes or by the addition of dopants affected the rate of enzymatic oxidation of cholesterol by altering the structure of cholesterol aggregates.


Factors affecting skim milk progesterone assay results.
Five studies were performed to determine factors affecting progesterone concentration in skim milk. Results of the first study indicated that progesterone concentration was higher in skim milk of samples kept 16 hours in an ice bath (0 C) than of those left at room temperature (21 C). In the second study, this temperature effect was found to be reversible, with skim milk progesterone concentration increasing when whole milk samples were cooled prior to centrifugation. In the third study, [3H]-labeled progesterone was used to determine the relationship between fat content of foremilk (the first milk obtained from the teats), midmilk (milk obtained midway through milking), and strippings (milk obtained immediately after milking machines have been removed) samples and temperature (4 C and 21 C) on the percentage of progesterone in the skim milk fraction. The relationship between percentage of butterfat and percentage of progesterone in skim milk was linear when the log of these variables was used for calculations. In the fourth study, assayable progesterone in the skim milk fraction of foremilk, midmilk, and strippings was affected by temperature. In the fifth study, a multiple-regression procedure was used to determine the amount of variation in percentage of radioactive progesterone in the skim milk fraction. Independent variables (whole milk butterfat and temperature of incubation [1, 3, 13, 22, 37, and 50 C]) and the natural log of each variable, were entered into a stepwise multiple-regression analysis.(ABSTRACT TRUNCATED AT 250 WORDS)


Organizational sensemaking about risk controls: the case of offshore hydrocarbons production.
In the same way that individuals' risk perceptions can influence how they behave toward risks, how organizational members make sense of risk controls is an important influence on how they apply and maintain such controls. In this article, we describe an analysis of sensemaking about the control of risk in offshore hydrocarbons production, an industry that continues to produce disasters of societal significance. A field study of 80 interviews was conducted in five offshore oil and gas companies and the agency that regulates them. The interviews were analyzed using qualitative template analysis. This provided a categorization of the many ways of acting through which informants made sense of the risk control task, and indicated that the organizations placed substantially different emphases on different ways of acting. Nevertheless, this sensemaking fell into two broad classes: that which tended to limit or be pessimistic about organizational controls, and that which tended to extend or be optimistic about organizational controls. All the participating organizations collectively placed a balanced emphasis on these two classes. We argue that this balanced sensemaking is an adaptation rather than a deliberate choice, but that it is an important element of controlling risk in its own right.


Draft Genome Sequence of the Acidophilic, Halotolerant, and Iron/Sulfur-Oxidizing Acidihalobacter prosperus DSM 14174 (Strain V6).
The principal genomic features of Acidihalobacter prosperus DSM 14174 (strain V6) are presented here. This is a mesophilic, halotolerant, and iron/sulfur-oxidizing acidophile that was isolated from seawater at Vulcano, Italy. It has potential for use in biomining applications in regions where high salinity exists in the source water and ores.


Healing effects of Aegle marmelos (L.) Correa fruit extract on experimental colitis.
Graded doses of 50% ethanolic extract of dried fruit pulp of Aegle marmelos (AME) (100, 200 and 400 mg/kg) daily for 14 days in acetic acid (AA)-induced colitis in rats showed 200 mg/kg of AME as an optimal effective dose against AA-induced colonic damage score and weight. This dose (200 mg/kg; po) was further studied in AA-induced colitis for its effects on various physical (mucous/blood in stool, food and water intake and body weight changes), histology, antibacterial activity and biochemical parameters like free radicals (nitric oxide and lipid peroxidation), antioxidants (superoxide dismutase, catalase and reduced glutathione) and myeloperoxidase (acute-inflammatory marker) activities in rat colonic tissue. AME decreased colonic mucosal damage and inflammation (macroscopic and microscopic), mucous/bloody diarrhea, fecal frequency and increased body weight affected in AA-induced colitis. AME showed significant antibacterial activity and enhanced the antioxidants but decreased free radicals and myeloperoxidase activities thereby decreasing tissue damage and inflammation and thus, affording ulcer healing. The above effects of A. marmelos authenticated its use in indigenous system of Medicine.


Characterization of rare lens epithelium-derived growth factor/p75 genetic variants identified in HIV-1 long-term nonprogressors.
Lens epithelium-derived growth factor (LEDGF)/p75 is a cellular binding partner of HIV-1 integrase and a crucial cofactor for HIV-1 replication. Here, we study two LEDGF/p75 exonic variants I436S and T473I, identified in HIV-1 long-term nonprogressors, together with Q472L. In-vitro binding assays, cell culture complementation, and functional rescue. Binding affinities of wild-type, I436S, T473I, and Q472L LEDGF/p75 for HIV-1 integrase were comparable. All LEDGF/p75 variants bound equally well to LEDGF/p75 interacting partners JPO2 and PogZ. In addition, HIV-1 replication was evaluated in human somatic LEDGF/p75-knockout cells and LEDGF/p75-knockdown cells complemented with either wild-type LEDGF/p75 or the respective LEDGF/p75 variants. All variants rescued HIV-1 replication to wild-type levels, whereas LEDGF/p75 D366N, defective for interaction with HIV-1 integrase, did not. Although identified in a cohort of long-term nonprogressors, our study did not indicate that the I436S or T473I mutation in LEDGF/p75 affects the interaction with HIV-1 integrase.


Essential Fatty Acid Deficiency in 2015: The Impact of Novel Intravenous Lipid Emulsions.
The fatty acids, linoleic acid (18:2ω-6) and α-linolenic acid (18:3ω-3), are essential to the human diet. When these essential fatty acids are not provided in sufficient quantities, essential fatty acid deficiency (EFAD) develops. This can be suggested clinically by abnormal liver function tests or biochemically by an elevated Mead acid and reduced linoleic acid and arachidonic acid level, which is manifested as an elevated triene/tetraene ratio of Mead acid/arachidonic acid. Clinical features of EFAD may present later. With the introduction of novel intravenous (IV) lipid emulsions in North America, the proportion of fatty acids provided, particularly the essential fatty acids, varies substantially. We describe a case series of 3 complicated obese patients who were administered parenteral nutrition (PN), primarily using ClinOleic 20%, an olive oil-based lipid emulsion with reduced amounts of the essential fatty acids, linoleic and α-linolenic, compared with more conventional soybean oil emulsions throughout their hospital admission. Essential fatty acid profiles were obtained for each of these patients to investigate EFAD as a potential cause of abnormal liver enzymes. Although the profiles revealed reduced linoleic acid and elevated Mead acid levels, this was not indicative of the development of essential fatty acid deficiency, as reflected in the more definitive measure of triene/tetraene ratio. Instead, although the serum fatty acid panel reflected the markedly lower but still adequate dietary linoleic acid content and greatly increased oleic acid content in the parenteral lipid emulsion, the triene/tetraene ratio remained well below the level, indicating EFAD in each of these patients. The availability and use of new IV lipid emulsions in PN should encourage the clinician to review lipid metabolism based on the quantity of fatty acids provided in specific parenteral lipid emulsions and the expected impact of these lipid emulsions (with quite different fatty acid composition) on measured fatty acid profiles.


A20 Attenuates FFAs-induced Lipid Accumulation in Nonalcoholic Steatohepatitis.
A20 is a ubiquitin-editing enzyme that attenuates the activity of proximal signaling complexes at pro-inflammatory receptors. It has been well documented that A20 protein plays an important role in response to liver injury and hepatocytes apoptosis in pro-inflammatory pathways. However, there was little evidence showing that A20 protein was involving in fatty-acid homeostasis except the up-regulation of two fatty acid metabolism regulatory genes at mRNA level (PPARa and CPT1a) by adenovirus-mediated A20 protein overexpression. In this study we found that: 1) the expression level of A20 protein was significantly higher in the steatotic liver from MCD-fed mice than the controls; 2) Overexpression of A20 protein suppressed FFAs-stimulated triglyceride deposition in HepG2 cells while under expression of A20 protein increased FFAs-stimulated triglyceride deposition; 3) Overexpression of A20 protein in HepG2 cells upregulated genes that promote β-oxidation and decreased the mRNA levels of key lipogenic genes such as fatty acid synthase (FAS), indicating A20 function as anti-steatotic factor by the activation of mitochondrial β-oxidation and attenuation of de novo lipogenesis; 4) Nonalcoholic steatohepatitis (NASH) patients showed significantly higher A20 expression level in liver compared with control individuals. Our results demonstrated that A20 protein plays an important role in fatty-acid homeostasis in human as well as animals. In addition, our data suggested that the pathological function of A20 protein in hepatocyte from lipotoxicity to NASH is by the alleviation of triglyceride accumulation in hepatocytes. Elevated expression of A20 protein could be a potential therapeutic strategy for preventing the progression of nonalcoholic steatohepatitis.


Pharmacokinetics of oral terbinafine in adult horses.
The primary study objective was to compare the pharmacokinetics of p.o. terbinafine alone to p.o. terbinafine administered with p.o. cimetidine in healthy adult horses. The second objective was to assess the pharmacokinetics of terbinafine when administered per rectum in two different suspensions at 30 mg/kg to adult horses. Six healthy adult horses were included in this crossover study. Plasma terbinafine concentrations were quantified with liquid chromatography and mass spectrometry. The half-life (geometric mean) was 8.38 and 10.76 h, for p.o. alone and p.o. with cimetidine, respectively. The mean maximum plasma concentrations were 0.291 μg/mL at 1.54 h and 0.418 μg/mL at 1.28 h for p.o. alone and p.o. with cimetidine, respectively. Terbinafine with cimetidine had an average CMAX 44% higher and the relative F was 153% compared p.o. terbinafine alone, but was not statistically different (P > 0.05). Terbinafine was infrequently detected when administered per rectum in two different suspensions (water or olive oil). Minor adverse effects included oral irritation, fever, and colic. All resolved spontaneously. More pharmacokinetic studies are indicated assessing drug-drug interactions and using multiple dosing intervals to improve our knowledge of effective oral dosing, the potential for drug accumulation, and systemic adverse effect of terbinafine in horses.


Risk factors for abnormal anal cytology in young heterosexual women.
Although anal cancers are up to four times more common in women than men, little is known about the natural history of anal human papillomavirus (HPV) infections and HPV-related anal lesions in women. This study reports on the prevalence of and risks for anal cytological abnormalities over a 1-year period in a cohort of young women participating in a study of the natural history of cervical HPV infection. In addition to their regularly scheduled sexual behavior interviews and cervical testing, consenting women received anal HPV DNA and cytological testing. Anal cytology smears were obtained from 410 women whose mean age was 22.5 +/- 2.5 years at the onset of the study. Sixteen women (3.9%) were found to have abnormal anal cytology: 4 women had low-grade squamous intraepithelial lesions (SILs) or condyloma; and 12 women had atypical cells of undetermined significance. Factors found to be significantly associated with abnormal anal cytology were a history of anal sex [odds ratio (OR), 6.90; 95% confidence interval (CI), 1.7-47.2], a history of cervical SILs (OR, 4.13; 95% CI, 1.3-14.9), and a current anal HPV infection (OR, 12.28; 95% CI, 3.9-43.5). The strong association between anal intercourse and the development of HPV-induced SILs supports the role of sexual transmission of HPV in anal SILs. Young women who had engaged in anal intercourse or had a history of cervical SILs were found to be at highest risk.


Adolescents sex-typed friendship experiences: does having a sister versus a brother matter?
This study examines the connections between having a sister versus a brother and coming from a same-sex versus an opposite-sex sibling dyad and the degree of sex-typing in adolescents' friendship experiences, including the qualities of their friendships (i.e., intimacy, control) and their friends' personal attributes (i.e., sex-typed leisure interests, expressive and instrumental personality qualities). Participants were 159 firstborn-secondborn adolescent sibling pairs (M = 14.94 years and M = 12.43 years, respectively) and a close friend of each sibling (N = 636, including siblings and friends). Data were collected during home visits with siblings and telephone interviews with friends of siblings. The results suggested that sisters may learn control tactics from their brothers that they apply in their friendships; boys, however, were less likely to model the emotional intimacy that characterized their sisters' experiences with friends. In addition, coming from an opposite-sex sibling dyad was linked to sex-typing in friends' personal attributes, particularly their masculine leisure interests and instrumental personality qualities. Sisters and brothers may provide unique opportunities to learn about sex-typed relationship experiences in early adolescence, a time when gender segregation in the peer context is pervasive.


Pentraxin-chromatin interactions: serum amyloid P component specifically displaces H1-type histones and solubilizes native long chromatin.
Pure serum amyloid P component (SAP) and native long chromatin, mixed together at wt/wt ratios between 1:1 and 1:2 in the presence of physiological concentrations of NaCl and calcium, both remained in solution, whereas each alone precipitates rapidly under these conditions. This solubilization accompanies the binding of SAP to chromatin and the displacement of H1-type histones, which are essential for condensation and higher order folding of chromatin. Such binding of SAP to chromatin is remarkable since displacement of H1 and H5 by salt alone requires approximately 0.5 M NaCl. SAP also bound to nucleosome core particles forming soluble complexes with an apparent stoichiometry of 1:2, a result that is compatible with attachment of SAP at the nucleosome dyad, the site of H1 in intact chromatin. SAP thus undergoes a specific, avid interaction with chromatin that promotes its solubilization and may thereby contribute to the physiological handling of chromatin released from cells in vivo. In contrast, C-reactive protein (CRP) did not bind significantly to either chromatin or to core particles at physiological ionic strength. Incubation of chromatin with either normal serum, or acute phase human serum containing raised levels of CRP, did not induce complement activation regardless of the presence of added SAP or CRP, nor was any cleavage of DNA observed.


Two heads are better than one: Australian tobacco control experts' and mental health change champions' consensus on addressing the problem of high smoking rates among people with mental illness.
Objective The aims of the present study were to explore the beliefs of Australian experts in tobacco control and change champions working in mental health and tobacco cessation, and to identify measures for addressing the problem of high smoking rates for people with mental illness. Methods Qualitative interviews were undertaken to explore participants' views, and the Delphi technique was used to achieve consensus on ways in which the problem would be best addressed. Results This consensus centred on the need for leadership within the mental health system. The problem was reconceptualised from being solely the responsibility of the mental health sector into an issue that requires the combined resources of a partnership and shared leadership between government and non-government services, public health leaders, policy makers and people with mental illness and their families. Conclusions Collaboration would raise the priority of the issue, reduce the debilitating effect of stigma and discrimination within the mental health sector and would place smoking reduction firmly on the political and public agenda. A recovery-orientated focus would increase the skill base and be inclusive of workers, families and carers of people with mental illness who face smoking issues on a daily basis. Reconceptualising this as an issue that would benefit from cooperation and partnerships would disrupt the notion that the problem is solely the responsibility of the mental health sector. What is known about the topic? Rates of smoking have remained high for people with mental illness despite population-wide public health strategies successfully reducing smoking rates in the general population. For people with mental illness, the benefits of quitting smoking for both their mental and physical health are overshadowed by concerns about the complexity of their needs. There is a lack of knowledge about how smoking cessation support can be improved to increase success rates in smokers with mental illness. What does this paper add? The present study is the first to bring a cross-sector lens of public health and mental health 'experts' together to discuss the reasons for the high rates of smoking among people with mental illness and to obtain their shared agreement on solutions. This Australian-specific study analyses participants' responses to the problem representation and reveals what the issue is considered to be, where action should occur and how the problem should be resolved. What are the implications for practitioners? For the Australian context, there is a need for leadership and a consistent smoke-free message about the benefits of not smoking. Staff working in mental health require training in providing brief interventions, motivational interviewing and pharmacological support. Joining together as a partnership of government and non-government services, including public health leaders and policy makers, and involving people with mental illness and their families, would benefit all concerned.


Persistent Disease Activity in Patients With Long-Standing Glomerular Disease.
Glomerular diseases are characterized by variable disease activity over many years. We aimed to analyze the relationship between clinical disease activity and duration of glomerular disease. Disease activity in adults with chronic minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, and IgA nephropathy (IgAN; first diagnostic biopsy >5 years before enrollment; Of Longstanding Disease [OLD] cohort, n = 256) followed at Columbia University Medical Center (CUMC), was compared with disease activity of an internal and external cohort of patients with first diagnostic biopsy <5 years before enrollment drawn from the Cure Glomerulonephropathy Network (CureGN cohort, n = 1182; CUMC-CureGN cohort, n = 362). Disease activity was defined by (i) Kidney Disease: Improving Global Outcomes-recommended threshold criteria for initiation of immunosuppression in primary glomerulonephropathy (GN) and (ii) CureGN's Disease Activity Working Group definitions for activity. No significant differences were detected among the 3 cohorts in terms of age, sex, serum creatinine, and urinary protein-to-creatinine ratio. For each GN subtype, disease activity in the OLD cohort was comparable with disease activity in the entire CureGN and the CUMC-CureGN cohort. When limiting our comparisons to disease activity in incident CUMC-CureGN patients (first diagnostic biopsy within 6 months of enrollment), OLD patients demonstrated similar activity rates as incident patients. Disease activity did not differ among patients with shorter versus longer duration of disease. Such survivor patients, with long-term but persistent disease, are potentially highly informative for understanding the clinical course and pathogenesis of GN and may help identify factors mediating more chronic subtypes of disease.


Age, sex, material deprivation and respiratory mortality.
The aim of this study was to examine the effects of age, sex and social deprivation on mortality rates for respiratory infection. An ecological study was undertaken, using official public health mortality data and population census data for the West Midlands health region, UK. Postcodes at the time of death were used to assign Townsend deprivation scores and the resulting deprivation quintile. Poisson regression analysis was used to estimate the association between respiratory mortality, deprivation quintile, age and sex. In most age groups there was a statistically significant trend of increasing mortality with increasing deprivation. The relative risk for the most deprived was highest in the 45-64 year age-group (RR=4.4, 95% CI 4.0, 4.8). However, the absolute risks were greater in those aged 75-84 years (RR=1.3, 95% CI 1.3, 1.4) where the annual death rate was 669 per 100,000. Consistently higher mortality rates were seen in males. These results suggest that the risk of mortality from respiratory infection varies by sex and generally increases with increasing age and deprivation quintile. The identified association between deprivation and mortality from respiratory infections is consistent with the effect of deprivation on many other diseases. Addressing the social determinants of ill health may help to reduce the high burden of respiratory mortality in the UK. However, individual level studies and examination of other areas are needed to explain the mechanisms by which deprivation increases the risk of mortality from respiratory infection, and thereby identify target groups for effective interventions.


R164C mutation in FOXQ1 H3 domain affects formation of the hair medulla.
A number of single gene mutations in laboratory mice produce hair follicle defects resulting in deformed hair shafts. The radiation-induced (SB/LeJ-Foxq1(sa)) satin mutant mice have a satin-like sheen to their hair and dilute colouration. This sheen is due to failure of the hair shafts to develop normal medullas, while the pigment dilution is due to the unrelated beige (lysosomal trafficking regulator, Lyst(bg)) mutation. A new allelic mutation, Foxq1(sa-J), arose spontaneously on the albino (tyrosinase, Tyr(c)) MRL/MpJ-Fas(lpr) background. The Foxq1(sa-J) allele has a C to T transition at position 490. By contrast, the Foxq1(sa) mutant allele was confirmed to be a 67 base pair deletion followed by two base changes (GA to AT). Morphologic changes were similar to those seen in Hoxc13 transgenic and targeted mutant mice. This new allelic mutation provides yet another tool to investigate formation of the interior structures of hair shafts.


The integrated continence system: a manual therapy approach to the treatment of stress urinary incontinence.
Stress urinary incontinence (SUI) constitutes a large-scale public health concern. The integrated continence system (ICS) developed by the authors is an evidence-based model that demonstrates how urinary incontinence is maintained through the interaction of three structural systems (intrinsic urethral closure, urethral support and lumbopelvic stability) and three modifiable factors (motor control, musculoskeletal and behavioural). The purpose of the ICS is first, to demonstrate the important role that manual physiotherapists can play in the treatment of SUI and second, to guide clinical practice decisions in order to improve clinical outcomes among women with SUI.


Minerals and Sarcopenia; The Role of Calcium, Iron, Magnesium, Phosphorus, Potassium, Selenium, Sodium, and Zinc on Muscle Mass, Muscle Strength, and Physical Performance in Older Adults: A Systematic Review.
Minerals may contribute to prevent and treat sarcopenia, the age-related loss of muscle mass, muscle strength, and physical performance. So far, there is no comprehensive review on the impact of minerals on sarcopenia outcomes. The aim of this systematic review is to evaluate the role of calcium, iron, magnesium, phosphorus, potassium, selenium, sodium, and zinc on muscle mass, muscle strength, and physical performance in older adults. A systematic search was conducted between March 2016 and July 2016, in the PubMed database using predefined search terms. Articles on the role of dietary mineral intake or mineral serum concentrations on muscle mass, muscle strength, physical performance, and/or the prevalence of sarcopenia in healthy or frail older adults (average age ≥ 65 years) were selected. Only original research publications were included. The search and data extraction were conducted in duplicate by 2 independent researchers. The Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement was followed in constructing this systematic review. The Effective Public Health Practice Project (EPHPP) Quality Assessment Tool for Quantitative Studies was used to evaluate the quality of the selected articles. From the 3346 articles found, a total of 10 studies met the inclusion criteria. Observational studies showed that serum selenium (n = 1) and calcium intake (n = 1) were significantly associated with muscle mass, and magnesium (n = 1), selenium (n = 1), iron (n = 1), and zinc (n = 1) intake were significantly and positively associated with physical performance in older adults. Furthermore, magnesium (n = 2), selenium (n = 2), calcium (n = 2), and phosphorus (n = 1) intake were associated with the prevalence of sarcopenia. Magnesium supplementation improved physical performance based on one randomized controlled trial. No studies on the role of sodium or potassium on muscle mass, muscle strength, or physical performance were found. Minerals may be important nutrients to prevent and/or treat sarcopenia. Particularly, magnesium, selenium, and calcium seem to be most promising. Most of the included studies, however, were observational studies. Therefore, more randomized controlled trials are needed to elucidate the potential benefits of mineral intake to prevent and/or treat sarcopenia and support healthy aging.


Caffeine urinary metabolite ratios as markers of enzyme activity: a theoretical assessment.
At least six urinary metabolite ratios of caffeine have been proposed as probes for in vivo CYP1A2 activity and three for in vivo NAT2 activity. Claims for the frequency distribution of the activity of CYP1A2 based on these empirical ratios have varied from log-normal to trimodal. We have examined the validity of these nine ratios by developing computer simulations using values reported in the literature for the kinetic parameters of caffeine and its metabolites. The results show that the sensitivity of the ratios to confounding variables is, in some cases, greater than their sensitivity to the activity of the enzyme that they are intended to mark. The six CYP1A2 ratios did not exhibit the same pattern of dependency on confounding variables which, in turn, resulted in different shapes of population distributions for each ratio as enzyme activity was varied systematically. Although the dependency of the three NAT2 ratios on confounding variable was less marked, they also showed different patterns of dependency. The outcomes of the simulations were consistent with much of the experimental data on caffeine metabolite ratios. To support the findings from the simulations, simplified equations for each metabolite ratio were derived which emphasize the dominant determinants. With some of the CYP1A2 ratios urine flow was significant to the point where its variance and heterogeneity between populations could lead to spurious detection of polymorphism in CYP1A2 function. Also, if the variability of a dominant confounding factor was high and sensitivity of the ratio to intrinsic CYP1A2 activity was low, any polymorphism in the latter would be obscured. When a specific time interval was defined for urine collection, this time was shown to be a critical factor in the ability to discriminate bimodality in some of the ratios, when a marked polymorphism in enzyme activity was assumed. Those ratios which have shown no evidence for bimodality in CYP1A2 function in experimental studies are inherently more discriminant of such heterogeneity compared to those ratios which have been claimed to detect polymorphism of CYP1A2 from experimental data. While recommending a 'best buy' from amongst the caffeine urinary metabolite ratios, we favour plasma/saliva indices (caffeine half-life or paraxanthine/caffeine ratio in a spot sample).


Fifteen-year Outcomes Following Conservative Management Among Men Aged 65 Years or Older with Localized Prostate Cancer.
To understand the threat posed by localized prostate cancer and the potential impact of surgery or radiation, patients and healthcare providers require information on long-term outcomes following conservative management. To describe 15-yr survival outcomes and cancer therapy utilization among men 65 years and older managed conservatively for newly diagnosed localized prostate cancer. This is a population-based cohort study with participants living in predefined geographic areas covered by the Surveillance, Epidemiology, and End Results program. The study includes 31 137 Medicare patients aged ≥65 yr diagnosed with localized prostate cancer in 1992-2009 who initially received conservative management (no surgery, radiotherapy, cryotherapy, or androgen deprivation therapy [ADT]). All patients were followed until death or December 31, 2009 (for prostate cancer-specific mortality [PCSM]) and December 31, 2011 (for overall mortality). Competing-risk analyses were used to examine PCSM, overall mortality, and utilization of cancer therapies. The 15-yr risk of PCSM for men aged 65-74 yr diagnosed with screening-detected prostate cancer was 5.7% (95% confidence interval [CI] 3.7-8.0%) for T1c Gleason 5-7 and 22% (95% CI 16-35%) for Gleason 8-10 disease. After 15 yr of follow-up, 24% (95% CI 21-27%) of men aged 65-74 yr with screening-detected Gleason 5-7 cancer received ADT. The corresponding result for men with Gleason 8-10 cancer was 38% (95% CI 32-44%). The major study limitations are the lack of data for men aged <65 yr and detailed clinical information associated with secondary cancer therapy. The 15-yr outcomes following conservative management of newly diagnosed Gleason 5-7 prostate cancer among men aged ≥65 yr are excellent. Men with Gleason 8-10 disease managed conservatively face a significant risk of PCSM. We examined the long-term survival outcomes for a large group of patients diagnosed with localized prostate cancer who did not have surgery, radiotherapy, cryotherapy, or androgen deprivation therapy in the first 6 mo after cancer diagnosis. We found that the 15-yr disease-specific survival is excellent for men diagnosed with Gleason 5-7 disease. The data support conservative management as a reasonable choice for elderly patients with low-grade localized prostate cancer.


Vaccine safety issues at the turn of the 21st century.
Global gains in vaccination coverage during the early 21st century have been threatened by the emergence of antivaccination groups that have questioned the effectiveness of vaccines to generate public distrust of vaccines and immunisation programmes. This manuscript summarises six key topics that have been at the centre of global discussions on vaccine safety during the early 21st century: thiomersal in multi-dose non-live vaccines, aluminium adjuvants used with several non-live vaccines, autism and auto-immune conditions as possible consequences of vaccination, a risk of immune overload with increasing numbers of vaccinations, and detrimental non-specific effects (NSEs) of vaccination. For each topic, we describe the hypothesis behind the public concern, the evidence reviewed by the WHO's Global Advisory Committee for Vaccine Safety (GACVS) during 1999-2019, and any significant new data that has emerged since GACVS conclusions were made. Although the scientific evidence on these issues overwhelmingly supports the safety of vaccines, communication messages to caregivers and providers need to condense and convey scientific information in an appropriate way to address concerns contributing to vaccine distrust. In addition, there is need for further studies specifically designed to address both positive and negative NSE of vaccination. The role of GACVS will be increasingly important in evaluating the evidence and engaging the global community in promoting and assuring the safety of vaccines in the decades to come as we move into an era in which we use new vaccination platforms, antigens and formulations.


Osmotic balance in the eggs of the turtle Chelodina rugosa during developmental arrest under water.
The tropical Australian turtle Chelodina rugosa normally lays its hard-shelled eggs in mud, under shallow freshwater, during the monsoon season. The eggs undergo developmental arrest until the water recedes and oxygen is able to diffuse into the embryo. This period of arrest can exceed 12 wk without embryonic mortality. To understand how the eggs avoid osmotic absorption of water leading to shell rupture and embryonic death, this study investigates the solute concentrations and volumes of the albumen and yolk compartments during submergence in distilled water. The albumen loses considerable sodium through the shell, particularly during the first week, and its osmotic concentration drops from 234 mmol/kg at laying to about 23 mmol/kg. Meanwhile, water from the albumen slowly moves through the vitelline membrane into the yolk compartment, which enlarges at a constant rate until it approaches the inside of the shell at about 22 wk. Osmotic uptake dilutes yolk solutes, decreasing the osmotic concentration from 281 mmol/kg at laying to 132 mmol/kg at 157 d. Loss of embryonic viability is associated with contact of the vitelline membrane with the inside of the shell. The principal adaptation of this species for protracted developmental arrest under water is a vitelline membrane of such low permeability to water that the expansion of the yolk compartment occurs about 10 times more slowly than in other chelonians.


5-hydroxymethylfurfural-embedded poly (vinyl alcohol)/sodium alginate hybrid hydrogels accelerate wound healing.
Treating large acute or chronic wounds remains a challenging task because of a lack of effective methods to accelerate wound healing. Though growth factor-based wound products are found to be effective, they are costly and are potentially associated with increased cancer mortality. Effective, safe, and affordable strategies to tackle large or chronic wounds need to further be designed and developed. Here, we designed a new antioxidant-embedded hydrogel system for speeding up the wound healing process. We prepared multifunctional poly (vinyl alcohol)/sodium alginate (PVA/SA) hydrogels with the incorporation of 5-hydroxymethylfurfural (5-HMF) and silver nanoparticles (Ag-NPs), and investigated their physiochemical and biological properties in vitro and in vivo. PVA, SA, 5-HMF, and Ag-NPs were employed for good mechanical properties, good biocompatibility, anti-inflammation, and anti-bacterial activity, respectively. 5-HMF is commonly found in many food products (e.g. honey, coffee, and black garlic) and is known as an antioxidant. In our in vitro study, 5-HMF was found to effectively facilitate the proliferation and migration of human skin fibroblasts (HSF), and collagen production. Besides, 5-HMF-embedded PVA/SA hybrid hydrogels supported controlled release and good cell compatibility, and more importantly accelerated wound healing in vivo through ameliorated inflammation, enhanced angiogenesis/vascularization, increased collagen production, and promoted re-epithelialization.


Enhanced dynamic nuclear polarization via swept microwave frequency combs.
Dynamic nuclear polarization (DNP) has enabled enormous gains in magnetic resonance signals and led to vastly accelerated NMR/MRI imaging and spectroscopy. Unlike conventional cw-techniques, DNP methods that exploit the full electron spectrum are appealing since they allow direct participation of all electrons in the hyperpolarization process. Such methods typically entail sweeps of microwave radiation over the broad electron linewidth to excite DNP but are often inefficient because the sweeps, constrained by adiabaticity requirements, are slow. In this paper, we develop a technique to overcome the DNP bottlenecks set by the slow sweeps, using a swept microwave frequency comb that increases the effective number of polarization transfer events while respecting adiabaticity constraints. This allows a multiplicative gain in DNP enhancement, scaling with the number of comb frequencies and limited only by the hyperfine-mediated electron linewidth. We demonstrate the technique for the optical hyperpolarization of 13C nuclei in powdered microdiamonds at low fields, increasing the DNP enhancement from 30 to 100 measured with respect to the thermal signal at 7T. For low concentrations of broad linewidth electron radicals [e.g., TEMPO ((2,2,6,6-tetramethylpiperidin-1-yl)oxyl)], these multiplicative gains could exceed an order of magnitude.


Eating disorder risk, exercise dependence, and body weight dissatisfaction among female nutrition and exercise science university majors.
Past research has examined eating disorder risk among college students majoring in Nutrition and has suggested an increased risk, while other studies contradict these results. Exercise Science majors, however, have yet to be fully examined regarding their risk for eating disorders and exercise dependence. Based on pressures to fit the image associated with careers related to these two disciplines, research is warranted to examine the potential risk for both eating disorder and exercise dependence. The purpose of this study is to compare eating disorder risk, exercise dependence, and body weight dissatisfaction (BWD) between Nutrition and Exercise Science majors, compared to students outside of these career pathways. Participants (n = 89) were divided into three groups based on major; Nutrition majors (NUTR; n = 31), Exercise Science majors (EXSC; n = 30), and other majors (CON; n = 28). Participants were given the EAT-26 questionnaire and the Exercise Dependence Scale. BWD was calculated as the discrepancy between actual BMI and ideal BMI. The majority of participants expressed a desire to weigh less (83%) and EXSC had significantly (p = .03) greater BWD than NUTR. However, there were no significant differences in eating disorder risk or exercise dependence among majors. This study suggested there was no significant difference in eating disorder risk or exercise dependence between the three groups (NUTR, EXSC, and CON).


Changes in blood pressure during the normal menstrual cycle.
1. Changes in blood pressure during the normal menstrual cycle are not well documented, and previous studies have given conflicting results. 2. Thirty normotensive women and ten mildly hypertensive women measured their blood pressure at home each morning for 6 weeks, under standardized conditions, using a UA-751 semi-automatic sphygmomanometer. All had normal menstrual cycles and subjects entered the study at different phases of the cycle. 3. Blood pressure was higher at the onset of menstruation than at most other phases of the cycle (systolic blood pressure, P less than 0.05; diastolic blood pressure, P less than 0.001). Adjusted diastolic blood pressure was higher in the follicular than in the luteal phase (mean difference 1.23 mmHg, P less than 0.001). Similarly, blood pressure was lower during days 17-26 than during the remainder of the cycle (adjusted mean difference in systolic blood pressure -0.65 mmHg, P = 0.07; adjusted mean difference in diastolic blood pressure -1.19 mmHg, P less than 0.001). 4. Similar patterns were seen in normotensive and hypertensive subjects, and changes in plasma 17 beta-oestradiol and progesterone concentrations were also similar in the two groups.


Ypt/rab gtpases: regulators of protein trafficking.
Ypt/Rab guanosine triphosphatases (GTPases) have emerged in the last decade as key regulators of protein transport in all eukaryotic cells. They seem to be involved in all aspects of vesicle trafficking: vesicle formation, motility, and docking, and membrane remodeling and fusion. The functions of Ypt/Rabs are themselves controlled by upstream regulators that stimulate both their nucleotide cycling and their cycling between membranes. Ypt/Rabs transmit signals to downstream effectors in a guanosine triphosphate (GTP)-dependent manner. The identity of upstream regulators and downstream effectors is known for a number of Ypt/Rabs, and models for their mechanisms of action are emerging. In at least two cases, Ypt/Rab upstream regulators and downstream effectors are found together in a single complex. In agreement with the idea that Ypt/Rabs function in all aspects of vesicular transport, their diverse effectors have recently been shown to function in all identified aspects of vesicle transport. Activators and effectors for individual Ypt/Rabs share no similarity, but are conserved between yeast and mammalian cells. Finally, cross talk demonstrated among the various Ypt/Rabs, and between Ypt/Rabs and other signaling factors, suggests possible coordination among secretory steps, as well as between protein transport and other cellular processes.


SU-E-T-390: Monitorization of the Dynamic Multi-Leaf Collimator Performance Using Log Files - a Quality Assurance Tool for Intensity Modulated Radiotherapy.
Commission baseline values of the multi-leaf collimator (MLC) using Varian's dynalog file viewer software (DFV) and establish tolerance levels to use as a periodic quality assurance tool in intensity modulated radiotherapy (IMRT). A sliding window test was created to have all leafs moving at a constant high speed of 2.5cm/s. This test was performed for 100 days at gantry angles of 0°, 90° and 270° during the morning quality checks in an accelerator performing RapidArc treatments with a Millennium 120 MLC. The DFV was used to collect the percentage of counts in bins 1 (0.05mm) to 8 (deviations in steps of 0.5mm) and the root mean square error (RMS). These were analyzed statistically and a baseline value and tolerance levels were established. Results show that the greater percentage of counts is found at bin 4 with an average of 97%. Bins 1 to 3 have less than 1 % and bin 5 presents an average of 1.7%. As for bin 6, the average was below 0.1%. Bins 7 and 8 presented no counts. When comparing the results of the different gantry angles, significant differences are only found in bin 4 and 5, with a better performance for the 0° gantry. As for the RMS error, no difference was found between different carriages or gantry angles and the average value was 0.129. A tolerance level was established for bins that correspond to greater deviations - bins 5 to 8, with an upper limit equivalent to twice the standard deviation of each bin. With the introduction of IMRT, the MLC quality assurance became more extensive and thorough. However a quantitative analysis is often difficult to interpret and time consuming. The purposed method allows the monitorization of the stability and performance of the MLC in a quantitative and easy way.


Nano-analyses of wear particles from metal-on-metal and non-metal-on-metal dual modular neck hip arthroplasty.
Increased failure rates due to metallic wear particle-associated adverse local tissue reactions (ALTR) is a significant clinical problem in resurfacing and total hip arthroplasty. Retrieved periprosthetic tissue of 53 cases with corrosion/conventional metallic wear particles from 285 revision operations for ALTR was selected for nano-analyses. Three major classes of hip implants associated with ALTR, metal-on-metal hip resurfacing arthroplasty (MoM HRA) and large head total hip replacement (MoM LHTHA) and non-metal-on-metal dual modular neck total hip replacement (Non-MoM DMNTHA) were included. The size, shape, distribution, element composition, and crystal structure of the metal particles were analyzed by conventional histological examination and electron microscopy with analytic tools of 2D X-ray energy dispersive spectrometry and X-ray diffraction. Distinct differences in size, shape, and element composition of the metallic particles were detected in each implant class which correlate with the histological features of severity of ALTR and variability in implant performance.


Need for and use of contraception by women before and during COVID-19 in four sub-Saharan African geographies: results from population-based national or regional cohort surveys.
Although hindrances to the sexual and reproductive health of women are expected because of COVID-19, the actual effect of the pandemic on contraceptive use and unintended pregnancy risk in women, particularly in sub-Saharan Africa, remains largely unknown. We aimed to examine population-level changes in the need for and use of contraception by women during the COVID-19 pandemic, determine if these changes differed by sociodemographic characteristics, and compare observed changes during the COVID-19 pandemic with trends in the 2 preceding years. In this study, we used four rounds of Performance Monitoring for Action (PMA) population-based survey data collected in four geographies: two at the country level (Burkina Faso and Kenya) and two at the subnational level (Kinshasa, Democratic Republic of the Congo and Lagos, Nigeria). These geographies were selected for this study as they completed surveys immediately before the onset of COVID-19 and implemented a follow-up specific to COVID-19. The first round comprised the baseline PMA panel survey implemented between November, 2019, and February, 2020 (referred to as baseline). The second round comprised telephone-based follow-up surveys between May 28 and July 20, 2020 (referred to as COVID-19 follow-up). The third and fourth rounds comprised two previous cross-sectional survey rounds implemented in the same geographies between 2017 and 2019. Our analyses were restricted to 7245 women in union (married or living with a partner, as if married) who were interviewed at baseline and COVID-19 follow-up. The proportion of women in need of contraception significantly increased in Lagos only, by 5·81 percentage points (from 74·5% to 80·3%). Contraceptive use among women in need increased significantly in the two rural geographies, with a 17·37 percentage point increase in rural Burkina Faso (30·7% to 48·1%) and a 7·35 percentage point increase in rural Kenya (71·6% to 78·9%). These overall trends mask several distinct patterns by sociodemographic group. Specifically, there was an increase in the need for contraception among nulliparous women across all geographies investigated. Our findings do not support the anticipated deleterious effect of COVID-19 on access to and use of contraceptive services by women in the earliest stages of the pandemic. Although these results are largely encouraging, we warn that these trends might not be sustainable throughout prolonged economic hardship and service disruptions. Bill & Melinda Gates Foundation. For the French translation of the abstract see Supplementary Materials section.


The political economy of the welfare state in developed capitalist countries.
The article analyzes the social, economic, and political changes taking place in developed capitalist countries that are affecting their welfare states, in particular the changes in the family, in people's life cycles, in economic and social structures, and, most importantly, in the political contexts. The author shows how these changes take place and how the ways in which various countries respond to them depend mainly on the correlation of forces (of which class forces continue to be of great importance) and their expression in the political space. The dominant theoretical frame (promoted by international agencies and many governments) assumes that all governments, regardless of their political coloration, are forced to follow the same policies because of the need to be competitive in the globalized economy, where international markets (whether financial or commercial) determine what governments can and must do. Questioning this economic determinism, the article recovers the importance of politics, putting politics at the center of the explanation of what is happening in the welfare states of the developed capitalist countries, including the neoliberal aggression against them.


NFAT is well placed to direct both enhancer looping and domain-wide models of enhancer function.
Nuclear factor of activated T cells (NFAT) plays a central role in activating gene expression at the level of chromatin structure. A study now reveals that NFAT may also help to organize chromatin domains and enable enhancer-promoter communication. In activated T cells, inducible intrachromosomal looping occurs between the tumor necrosis factor-alpha (TNF-alpha) gene promoter and two NFAT-dependent enhancers located at -9 kb and +3 kb. This topology places the TNF-alpha gene and the adjacent lymphotoxin (LT) genes in separate loops, thereby allowing independent regulation of the TNF-alpha gene within a multigene locus. These findings build on other studies that indicate that NFAT is intimately associated with activities that disrupt nucleosomes within enhancers and mobilize nucleosomes across extensive chromatin domains linking enhancers and promoters. Taken together, these studies highlight NFAT as a factor that creates a chromatin environment that is permissive for both the recruitment and the clustering of factors that control transcription at promoters and enhancers.


Interphotoreceptor retinoid-binding protein: biochemistry and molecular biology.
Interphotoreceptor retinoid-binding protein (IRBP) is a recent and still somewhat enigmatic newcomer in the family of photoreceptor-specific proteins involved in vision. It has been isolated, characterized, cloned and sequenced from several species in less than a decade. Its extracellular localization and the increased amount of bound all-trans retinol following illumination are consistent with the proposed function of IRBP in extracellular transport or buffering of retinol and retinal isomers in the IPM. Its capacity to bind a great many other hydrophobic ligands, however, may indicate a multi-purpose function. Many questions concerning biosynthesis and processing, secretion, tertiary structure and ligand binding remain to be answered. Perhaps most interesting, from a molecular biology point of view, will be the regulation of transcription and tissue-specific expression of this photoreceptor protein, and possible involvement of its gene in human retinal degenerations. Constructs consisting of IRBP regulatory sequences and foreign antigens incorporated into the germ line of transgenic mice will help in the characterization of promoters and enhancer elements, while suppression of IRBP gene expression and its consequences may give us a more precise answer about IRBP function.


Optimal activation of an endogenous gene by HOX11 requires the NH2-terminal 50 amino acids.
The HOX11 homeobox gene was first identified through studies of the t(7;10) and t(10;14) chromosomal translocations of acute T-cell leukemia. In addition, analysis of Hox11-/- mice has demonstrated a critical role for this gene in murine spleen development. A possible mode of in vivo function for the HOX11 protein in these two situations is regulation of target genes following DNA binding via the homeodomain, but little is known about how HOX11 regulates transcription in vivo. By performing transcriptional studies in yeast and mammalian one-hybrid systems, a modular transcriptional transactivation region at the NH2 terminus of HOX11 has been functionally dissected from other parts of the protein. This NH2-terminal region includes the previously identified short conserved Hep motif, which itself activates transcription in one-hybrid assays. The importance of the NH2-terminal region for the function of HOX11 in vivo was assayed by activating a HOX11-dependent gene in NIH 3T3 cells. Activation of this gene was found to be dependent upon an intact homeodomain in HOX11, but maximal activation was obtained only when the NH2-terminal 50 amino acids of HOX11 was present, showing that this region of HOX11 is important for in vivo transcriptional control of a chromosomal target gene.


Generalization of the Ronchi, sinusoidal, and triangular rulings for Gaussian-laser-beam-diameter measurements.
Transmittance (or reflectance) characteristics of generalized gratings are described for Gaussian-laserbeam-diameter measurements. The generalized gratings provide for accurate measurement of both small and large Gaussian beam diameters and are superior to the Ronchi, the triangular, and the sinusoidal rulings. Also the proposed generalized gratings account for all width variations of the opaque, transparent, and transitional regions within a selected ruling.


Critical role of soluble amyloid-β for early hippocampal hyperactivity in a mouse model of Alzheimer's disease.
Alzheimer's disease (AD) is characterized by a progressive dysfunction of central neurons. Recent experimental evidence indicates that in the cortex, in addition to the silencing of a fraction of neurons, other neurons are hyperactive in amyloid-β (Aβ) plaque-enriched regions. However, it has remained unknown what comes first, neuronal silencing or hyperactivity, and what mechanisms might underlie the primary neuronal dysfunction. Here we examine the activity patterns of hippocampal CA1 neurons in a mouse model of AD in vivo using two-photon Ca(2+) imaging. We found that neuronal activity in the plaque-bearing CA1 region of older mice is profoundly altered. There was a marked increase in the fractions of both silent and hyperactive neurons, as previously also found in the cortex. Remarkably, in the hippocampus of young mice, we observed a selective increase in hyperactive neurons already before the formation of plaques, suggesting that soluble species of Aβ may underlie this impairment. Indeed, we found that acute treatment with the γ-secretase inhibitor LY-411575 reduces soluble Aβ levels and rescues the neuronal dysfunction. Furthermore, we demonstrate that direct application of soluble Aβ can induce neuronal hyperactivity in wild-type mice. Thus, our study identifies hippocampal hyperactivity as a very early functional impairment in AD transgenic mice and provides direct evidence that soluble Aβ is crucial for hippocampal hyperactivity.


Adrenal weight and catecholamine content in renal and calciferol-induced hypertension in rabbits.
1. Hypertension was produced in rabbits by unilateral renal encapsulation combined with contralateral nephrectomy or by the administration of calciferol and calcium lactate. 2. Weighed post mortem, the adrenal glands from the two hypertensive groups were significantly heavier than those from normotensive animals of similar weight. There was a direct correlation between the adrenal gland weight and the mean arterial blood pressure measured at the time of the terminal experiment. 3. The catecholamine content of the adrenal glands from the two groups of hypertensive animals was similar to that of the normotensive animals. 4. There was histological evidence of adrenal cortical hypertrophy in the glands of the hypertensive groups of animals.


Electrostatic interactions play a minor role in the binding of ExoS to 14-3-3 proteins.
14-3-3 proteins belong to a family of conserved molecules expressed in all eukaryotic cells that play an important role in a multitude of signalling pathways. 14-3-3 proteins bind either to phosphoserine/phosphothreonine residues or to sequence-specific non-phosphorylated motifs in more than 200 interaction partners [Pozuelo Rubio, Geraghty, Wong, Wood, Campbell, Morrice and Mackintosh (2004) Biochem. J. 379, 395-408]. These interactions result in cell-cycle regulation, apoptosis, stress responses, cell metabolism and malignant transformation. One example of a phosphorylation-independent interaction is the binding of 14-3-3 to ExoS (exoenzyme S), a bacterial ADP-ribosyltransferase toxin of Pseudomonas aeruginosa. In the present study, we have utilized additional biochemical and infection analyses to define further the structural basis of the interaction between ExoS and 14-3-3. An ExoS leucine-substitution mutant dramatically reduced the interaction potential with 14-3-3 suggesting that Leu422, Leu423, Leu426 and Leu428 of ExoS are important for its interaction with 14-3-3, its enzymatic activity and cytotoxicity. However, ExoS substitution mutants of residues that interact with 14-3-3 through an electrostatic interaction, such as Ser416, His418, Asp424 and Asp427, showed no reduction in their interaction potential with 14-3-3. These ExoS substitution mutants were also as aggressive as wild-type ExoS at inducing cell death and to modify endogenous ExoS target within the cell. In conclusion, electrostatic interaction between ExoS and 14-3-3 via polar residues (Ser416, His418, Asp424 and Asp427) appears to be of secondary importance. Thus the interaction between the 'roof' of the groove of 14-3-3 and ExoS relies more on hydrophobic interaction forces, which probably contributes to induce cell death after ExoS infection and activation.


Intrarenal Transplantation of Hypoxic Preconditioned Mesenchymal Stem Cells Improves Glomerulonephritis through Anti-Oxidation, Anti-ER Stress, Anti-Inflammation, Anti-Apoptosis, and Anti-Autophagy.
To confer further therapeutic potential and prevent some adverse effects by the mesenchymal stem cells (MSCs) transplantation, we explored the effects of locally intrarenal arterial administration of hypoxic preconditioned MSCs in the anti-Thy1.1 induced rat glomerulonephritis. Proteinuria, histochemical staining, and western blotting were used to explore the therapeutic effects and mechanisms. Locally intrarenal arterial MSCs transplantation successfully implanted the fluorescent or CD44 labeled MSCs in the nephritic glomeruli, ameliorated proteinuria, and glomerulosclerosis in nephritic rats. Hypoxic preconditioning significantly upregulated hypoxic inducible factor-1α/VEGF (HIF-1α/VEGF) in the MSCs and was more efficient than normoxic MSCs in reducing the degree of urinary protein, glomerulosclerosis, fibrosis, macrophage/monocyte infiltration, GRP78 mediated endoplasmic reticulum stress, Beclin-1/LC3-II mediated autophagy, and Bax/Bcl-2/caspase 3 mediated apoptosis. Hypoxic MSCs could further promote intranuclear nuclear factor (erythroid-derived 2, Nrf2) and reduce nuclear factor kappa B expression in nephritic kidneys. As compared to normoxic MSCs, hypoxic MSCs transplantation significantly upregulated the renal expression of anti-oxidative response elements/enzymes including glutamate-cysteine ligase catalytic subunit, glutamate-cysteine ligase modifier subunit, glutathione peroxidase, catalase, Mn, and Cu/Zn superoxide dismutase. In summary, intrarenal hypoxic preconditioning MSCs transplantation was more effective to activate hypoxic inducible factor-1α/VEGF/Nrf2 (HIF-1α/VEGF/Nrf2) signaling, preserve anti-oxidant proteins and anti-oxidative responsive element proteins, and subsequently reduce glomerular apoptosis, autophagy, and inflammation.


Exploring the local elastic properties of bilayer membranes using molecular dynamics simulations.
Membrane mechanical elastic properties regulate a variety of cellular processes involving local membrane deformation, such as ion channel function and vesicle fusion. In this work, we used molecular dynamics simulations to estimate the local elastic properties of a membrane. For this, we calculated the energy needed to extract a DOPE lipid molecule, modified with a linker chain, from a POPC bilayer membrane using the umbrella sampling technique. Although the extraction energy entails several contributions related not only to elastic deformation but also to solvation, careful analysis of the potential of mean force (PMF) allowed us to dissect the elastic contribution. With this information, we calculated an effective linear spring constant of 44 ± 4 kJ·nm(-2)·mol(-1) for the DOPC membrane, in agreement with experimental estimates. The membrane deformation profile was determined independently during the stretching process in molecular detail, allowing us to fit this profile to a previously proposed continuum elastic model. Through this approach, we calculated an effective membrane spring constant of 42 kJ·nm(-2)·mol(-1), which is in good agreement with the PMF calculation. Furthermore, the solvation energy we derived from the data is shown to match the solvation energy estimated from critical micelle formation constants. This methodology can be used to determine how changes in lipid composition or the presence of membrane modifiers can affect the elastic properties of a membrane at a local level.


Horizontal gene transfer is a significant driver of gene innovation in dinoflagellates.
The dinoflagellates are an evolutionarily and ecologically important group of microbial eukaryotes. Previous work suggests that horizontal gene transfer (HGT) is an important source of gene innovation in these organisms. However, dinoflagellate genomes are notoriously large and complex, making genomic investigation of this phenomenon impractical with currently available sequencing technology. Fortunately, de novo transcriptome sequencing and assembly provides an alternative approach for investigating HGT. We sequenced the transcriptome of the dinoflagellate Alexandrium tamarense Group IV to investigate how HGT has contributed to gene innovation in this group. Our comprehensive A. tamarense Group IV gene set was compared with those of 16 other eukaryotic genomes. Ancestral gene content reconstruction of ortholog groups shows that A. tamarense Group IV has the largest number of gene families gained (314-1,563 depending on inference method) relative to all other organisms in the analysis (0-782). Phylogenomic analysis indicates that genes horizontally acquired from bacteria are a significant proportion of this gene influx, as are genes transferred from other eukaryotes either through HGT or endosymbiosis. The dinoflagellates also display curious cases of gene loss associated with mitochondrial metabolism including the entire Complex I of oxidative phosphorylation. Some of these missing genes have been functionally replaced by bacterial and eukaryotic xenologs. The transcriptome of A. tamarense Group IV lends strong support to a growing body of evidence that dinoflagellate genomes are extraordinarily impacted by HGT.


Development and trends of biosurfactant analysis and purification using rhamnolipids as an example.
During the last few decades, increasing interest in biological surfactants led to an intensification of research for the cost-efficient production of biosurfactants compared with traditional petrochemical surface-active components. The quest for alternative production strains also is associated with new demands on biosurfactant analysis. The present paper gives an overview of existing analytical methods, based on the example of rhamnolipids. The methods reviewed range from simple colorimetric testing to sophisticated chromatographic separation coupled with detection systems like mass spectrometry, by means of which detailed structural information is obtained. High-performance liquid chromatography (HPLC) coupled with mass spectrometry currently presents the most precise method for rhamnolipid identification and quantification. Suitable approaches to accelerate rhamnolipid quantification for better control of biosurfactant production are HPLC analysis directly from culture broth by adding an internal standard or Fourier transform infrared attenuated total reflectance spectroscopy measurements of culture broth as a possible quasi-online quantification method in the future. The search for alternative rhamnolipid-producing strains makes a structure analysis and constant adaptation of the existing quantification methods necessary. Therefore, simple colorimetric tests based on whole rhamnolipid content can be useful for strain and medium screening. Furthermore, rhamnolipid purification from a fermentation broth will be considered depending on the following application.


Placement of removable metal biliary stent in post-orthotopic liver transplantation anastomotic stricture.
Postoperative biliary strictures are the most common cause of benign biliary stricture in Western countries, secondary to either operative injury or bile duct anastomotic stricture following orthotopic liver transplantation (OLT). Surgery or endoscopic interventions are the mainstay of treatment for benign biliary strictures. We aim to report the outcome of 2 patients with refractory anastomotic biliary stricture post-OLT, who had successful temporary placement of a prototype removable covered self-expandable metal stent (RCSEMS). These 2 patients (both men, aged 44 and 53 years) were given temporary placement of a prototype RCSEMS (8.5 Fr gauge delivery system, 8 mm x 40 mm stent dimensions) in the common bile duct across the biliary stricture. There was no morbidity associated with stent placement and removal in these 2 cases. Clinical parameters improved after the RCSEMS placement. Long-term biliary patency was achieved in both the patients. No further biliary intervention was required within 14 and 18 mo follow-up after stent removal.


Bioaugmentation with engineered endophytic bacteria improves contaminant fate in phytoremediation.
Phytoremediation of volatile organic contaminants often proves not ideal because plants and their rhizosphere microbes only partially degrade these compounds. Consequently, plants undergo evapotranspiration that contaminates the ambient air and, thus, undermines the merits of phytoremediation. Under laboratory conditions, endophytic bacteria equipped with the appropriate degradation pathways can improve in planta degradation of volatile organic contaminants. However, several obstacles must be overcome before engineered endophytes will be successful in field-scale phytoremediation projects. Here we report the first in situ inoculation of poplar trees, growing on a TCE-contaminated site, with the TCE-degrading strain Pseudomonas putida W619-TCE. In situ bioaugmentation with strain W619-TCE reduced TCE evapotranspiration by 90% under field conditions. This encouraging result was achieved after the establishment and enrichment of P. putida W619-TCE as a poplar root endophyte and by further horizontal gene transfer of TCE metabolic activity to members of the poplar's endogenous endophytic population. Since P. putida W619-TCE was engineered via horizontal gene transfer, its deliberate release is not restricted under European genetically modified organisms (GMO) regulations.


Metabolic tumor volume is an independent prognostic factor in patients treated definitively for non-small-cell lung cancer.
Fluorine-18 flurodeoxyglucose positron emission tomography (FDG-PET) imaging has rapidly become the standard of care for staging patients with lung cancer. We evaluated the prognostic value of metabolic tumor volume (MTV), a measure of tumor burden on FDG-PET imaging, in patients with non-small-cell lung cancer (NSCLC) treated definitively. A retrospective review identified 61 patients with NSCLC who underwent FDG-PET imaging for pretreatment staging. Metabolically active tumor regions were segmented on the PET scans semiautomatically to calculate the total body MTV. We determined the relationship of overall survival (OS) and progression-free survival (PFS) with MTV in the entire cohort, and in the subgroup treated definitively. The estimated median PFS and OS for the entire cohort were 11.1 months and 18.9 months. Higher MTV was significantly associated with worse OS (P = 0.00075) and PFS (P = 0.00077). For definitively treated patients, when MTV was analyzed as a binary value above or below the median value, 2-year PFS was 60% versus 39.7% (median PFS 34.9 vs. 11.9 months) and 2-year OS was 79.7% versus 33.3% (median OS 41.9 vs. 18.9 months), respectively (log-rank P = 0.12 for PFS and P = 0.066 for OS). When MTV was analyzed as a continuous variable, multivariate Cox proportional hazards analysis demonstrated a trend to worse PFS (hazard ratio [HR] = 1.31; P = 0.12) and significantly worse OS (HR = 1.53; P = 0.018) with increasing MTV after controlling for known prognostic variables. Tumor burden as assessed by MTV yields prognostic information on survival beyond that of established prognostic factors in patients with NSCLC treated definitively.


Treating anxiety disorders in children with high functioning autism spectrum disorders: a controlled trial.
A family-based, cognitive behavioural treatment for anxiety in 47 children with comorbid anxiety disorders and High Functioning Autism Spectrum Disorder (HFA) was evaluated. Treatment involved 12 weekly group sessions and was compared with a waiting list condition. Changes between pre- and post-treatment were examined using clinical interviews as well as child-, parent- and teacher-report measures. Following treatment, 71.4% of the treated participants no longer fulfilled diagnostic criteria for an anxiety disorder. Comparisons between the two conditions indicated significant reductions in anxiety symptoms as measured by self-report, parent report and teacher report. Discussion focuses on the implications for the use of cognitive behaviour therapy with HFA children, for theory of mind research and for further research on the treatment components.


Linear-scaling symmetry-adapted perturbation theory with scaled dispersion.
We present a linear-scaling symmetry-adapted perturbation theory (SAPT) method that is based on an atomic orbital (AO) formulation of zeroth-order SAPT (SAPT0). The non-dispersive terms are realized with linear-scaling cost using both the continuous fast multipole method (CFMM) and the linear exchange (LinK) approach for integral contractions as well as our efficient Laplace-based coupled-perturbed self-consistent field method (DL-CPSCF) for evaluating response densities. The reformulation of the dispersion term is based on our linear-scaling AO Møller-Plesset second-order perturbation theory (AO-MP2) method, that uses our recently introduced QQR-type screening [S. A. Maurer, D. S. Lambrecht, J. Kussmann, and C. Ochsenfeld, J. Chem. Phys. 138, 014101 (2013)] for preselecting numerically significant energy contributions. Similar to scaled opposite-spin MP2, we neglect the exchange-dispersion term in SAPT and introduce a scaling factor for the dispersion term, which compensates for the error and at the same time accounts for basis set incompleteness effects and intramonomer correlation. We show in extensive benchmark calculations that the new scaled-dispersion (sd-)SAPT0 approach provides reliable results for small and large interacting systems where the results with a small 6-31G** basis are roughly comparable to supermolecular MP2 calculations in a triple-zeta basis. The performance of our method is demonstrated with timings on cellulose fragments, DNA systems, and cutouts of a protein-ligand complex with up to 1100 atoms on a single computer core.


Demand for mental health care and changes in service use patterns in the Netherlands, 1979 to 1995.
Mental health services appear increasingly incapable of satisfying the demand for care, which may cause some segments of the population to be less effectively reached. This study investigated the rates of use of mental health services in the Netherlands from 1979 to 1995 and examined whether particular sociodemographic groups made greater or lesser relative use of these services over time. Data were derived from the Facilities Use Surveys, a series of Dutch cross-sectional population studies that have recorded household characteristics and service use since 1979. More than 28,000 households were included in the analyses. The overall use of mental health services virtually doubled from 1979 to 1995; a particularly steep rise was seen in the first half of the 1980s. Households that had one parent, that had low income, that were dependent on benefits, and that were younger all had greater odds of using both specialized mental health care (for example, prevention programs for mental health problems and psychotherapeutic and social psychiatric treatment offered by psychologists, psychotherapists, or psychiatrists) and social work services (for example, psychosocial counselling and practical support offered by social workers to people with social problems, such as housing, finances, and psychosocial issues). Households with low education were less likely to use specialized mental health care but were more likely to use social work services. Nonreligious households and urban households were more likely to use specialized mental health care and were equally likely to use social work services. Overall, these relative use patterns did not change over time. Despite greater pressures on mental health services and the many changes in service delivery in recent decades, relative patterns of help seeking and referral to mental health services have not varied systematically over time.


Application of kinomic array analysis to screen for altered kinases in atrial fibrillation remodeling.
Dysregulation of protein kinase-mediated signaling is an early event in many diseases, including the most common clinical cardiac arrhythmia, atrial fibrillation (AF). Kinomic profiling represents a promising technique to identify candidate kinases. In this study we used kinomic profiling to identify kinases altered in AF remodeling using atrial tissue from a canine model of AF (atrial tachypacing). Left atrial tissue obtained in a previous canine study was used for kinomic array (containing 1024 kinase pseudosubstrates) analysis. Three groups of dogs were included: nonpaced controls and atrial tachypaced dogs, which were contrasted with geranylgeranylacetone-treated dogs with AF, which are protected from AF promotion, to enhance specificity of detection of putative kinases. While tachypacing changed activity of 50 kinases, 40 of these were prevented by geranylgeranylacetone and involved in differentiation and proliferation (SRC), contraction, metabolism, immunity, development, cell cycle (CDK4), and survival (Akt). Inhibitors of Akt (MK2206) and CDK4 (PD0332991) and overexpression of a dominant-negative CDK4 phosphorylation mutant protected against tachypacing-induced contractile dysfunction in HL-1 cardiomyocytes. Moreover, patients with AF show down- and upregulation of SRC and Akt phosphorylation, respectively, similar to findings of the kinome array. Contrasting kinomic array analyses of controls and treated subjects offer a versatile tool to identify kinases altered in atrial remodeling owing to tachypacing, which include Akt, CDK4, and SRC. Ultimately, pharmacological targeting of altered kinases may offer novel therapeutic possibilities to treat clinical AF.


Epithelial invasion outcompetes hypha development during Candida albicans infection as revealed by an image-based systems biology approach.
Candida albicans is the most common opportunistic fungal pathogen of the human mucosal flora, frequently causing infections. The fungus is responsible for invasive infections in immunocompromised patients that can lead to sepsis. The yeast to hypha transition and invasion of host-tissue represent major determinants in the switch from benign colonizer to invasive pathogen. A comprehensive understanding of the infection process requires analyses at the quantitative level. Utilizing fluorescence microscopy with differential staining, we obtained images of C. albicans undergoing epithelial invasion during a time course of 6 h. An image-based systems biology approach, combining image analysis and mathematical modeling, was applied to quantify the kinetics of hyphae development, hyphal elongation, and epithelial invasion. The automated image analysis facilitates high-throughput screening and provided quantities that allow for the time-resolved characterization of the morphological and invasive state of fungal cells. The interpretation of these data was supported by two mathematical models, a kinetic growth model and a kinetic transition model, that were developed using differential equations. The kinetic growth model describes the increase in hyphal length and revealed that hyphae undergo mass invasion of epithelial cells following primary hypha formation. We also provide evidence that epithelial cells stimulate the production of secondary hyphae by C. albicans. Based on the kinetic transition model, the route of invasion was quantified in the state space of non-invasive and invasive fungal cells depending on their number of hyphae. This analysis revealed that the initiation of hyphae formation represents an ultimate commitment to invasive growth and suggests that in vivo, the yeast to hypha transition must be under exquisitely tight negative regulation to avoid the transition from commensal to pathogen invading the epithelium.


Progress on scaling up integrated services for sexual and reproductive health and HIV.
This paper considers new developments to strengthen sexual and reproductive health and HIV linkages and discusses factors that continue to impede progress. It is based on a previous review undertaken for the United Kingdom Department for International Development in 2006 that examined the constraints and opportunities to scaling up these linkages. We argue that, despite growing evidence that linking sexual and reproductive health and HIV is feasible and beneficial, few countries have achieved significant scale-up of integrated service provision. A lack of common understanding of terminology and clear technical operational guidance, and separate policy, institutional and financing processes continue to represent significant constraints. We draw on experience with tuberculosis and HIV integration to highlight some lessons. The paper concludes that there is little evidence to determine whether funding for health systems is strengthening linkages and we make several recommendations to maximize opportunities represented by recent developments.


External validation of a Cox prognostic model: principles and methods.
A prognostic model should not enter clinical practice unless it has been demonstrated that it performs a useful role. External validation denotes evaluation of model performance in a sample independent of that used to develop the model. Unlike for logistic regression models, external validation of Cox models is sparsely treated in the literature. Successful validation of a model means achieving satisfactory discrimination and calibration (prediction accuracy) in the validation sample. Validating Cox models is not straightforward because event probabilities are estimated relative to an unspecified baseline function. We describe statistical approaches to external validation of a published Cox model according to the level of published information, specifically (1) the prognostic index only, (2) the prognostic index together with Kaplan-Meier curves for risk groups, and (3) the first two plus the baseline survival curve (the estimated survival function at the mean prognostic index across the sample). The most challenging task, requiring level 3 information, is assessing calibration, for which we suggest a method of approximating the baseline survival function. We apply the methods to two comparable datasets in primary breast cancer, treating one as derivation and the other as validation sample. Results are presented for discrimination and calibration. We demonstrate plots of survival probabilities that can assist model evaluation. Our validation methods are applicable to a wide range of prognostic studies and provide researchers with a toolkit for external validation of a published Cox model.


A Single-Cell Transcriptomic Map of the Human and Mouse Pancreas Reveals Inter- and Intra-cell Population Structure.
Although the function of the mammalian pancreas hinges on complex interactions of distinct cell types, gene expression profiles have primarily been described with bulk mixtures. Here we implemented a droplet-based, single-cell RNA-seq method to determine the transcriptomes of over 12,000 individual pancreatic cells from four human donors and two mouse strains. Cells could be divided into 15 clusters that matched previously characterized cell types: all endocrine cell types, including rare epsilon-cells; exocrine cell types; vascular cells; Schwann cells; quiescent and activated stellate cells; and four types of immune cells. We detected subpopulations of ductal cells with distinct expression profiles and validated their existence with immuno-histochemistry stains. Moreover, among human beta- cells, we detected heterogeneity in the regulation of genes relating to functional maturation and levels of ER stress. Finally, we deconvolved bulk gene expression samples using the single-cell data to detect disease-associated differential expression. Our dataset provides a resource for the discovery of novel cell type-specific transcription factors, signaling receptors, and medically relevant genes.


NKG2A, a New Kid on the Immune Checkpoint Block.
One of the major breakthroughs of cancer immunotherapy has come from blocking immune checkpoint molecules on tumor-reactive T cells. Now, two studies examine targeting of a novel immune checkpoint, NKG2A, that can be expressed on both NK cells and on CD8+ T cells, either combined with a tumor-targeting antibody or with a tumor-specific vaccine.


When choice is demotivating: can one desire too much of a good thing?
Current psychological theory and research affirm the positive affective and motivational consequences of having personal choice. These findings have led to the popular notion that the more choice, the better-that the human ability to manage, and the human desire for, choice is unlimited. Findings from 3 experimental studies starkly challenge this implicit assumption that having more choices is necessarily more intrinsically motivating than having fewer. These experiments, which were conducted in both field and laboratory settings, show that people are more likely to purchase gourmet jams or chocolates or to undertake optional class essay assignments when offered a limited array of 6 choices rather than a more extensive array of 24 or 30 choices. Moreover, participants actually reported greater subsequent satisfaction with their selections and wrote better essays when their original set of options had been limited. Implications for future research are discussed.


Whole-Genome Sequencing Reveals the Contribution of Long-Term Carriers in Staphylococcus aureus Outbreak Investigation.
Whole-genome sequencing (WGS) makes it possible to determine the relatedness of bacterial isolates at a high resolution, thereby helping to characterize outbreaks. However, for Staphylococcus aureus, the accumulation of within-host diversity during carriage might limit the interpretation of sequencing data. In this study, we hypothesized the converse, namely, that within-host diversity can in fact be exploited to reveal the involvement of long-term carriers (LTCs) in outbreaks. We analyzed WGS data from 20 historical outbreaks and applied phylogenetic methods to assess genetic relatedness and to estimate the time to most recent common ancestor (TMRCA). The findings were compared with the routine investigation results and epidemiological evidence. Outbreaks with epidemiological evidence for an LTC source had a mean estimated TMRCA (adjusted for outbreak duration) of 243 days (95% highest posterior density interval [HPD], 143 to 343 days) compared with 55 days (95% HPD, 28 to 81 days) for outbreaks lacking epidemiological evidence for an LTC (P = 0.004). A threshold of 156 days predicted LTC involvement with a sensitivity of 0.875 and a specificity of 1. We also found 6/20 outbreaks included isolates with differing antimicrobial susceptibility profiles; however, these had only modestly increased pairwise diversity (mean 17.5 single nucleotide variants [SNVs] [95% confidence interval {CI}, 17.3 to 17.8]) compared with isolates with identical antibiograms (12.7 SNVs [95% CI, 12.5 to 12.8]) (P < 0.0001). Additionally, for 2 outbreaks, WGS identified 1 or more isolates that were genetically distinct despite having the outbreak pulsed-field gel electrophoresis (PFGE) pulsotype. The duration-adjusted TMRCA allowed the involvement of LTCs in outbreaks to be identified and could be used to decide whether screening for long-term carriage (e.g., in health care workers) is warranted. Requiring identical antibiograms to trigger investigation could miss important contributors to outbreaks.


Prevention of colorectal cancer with vitamin D.
The fact that colorectal cancer (CRC) is the second leading cause of cancer mortality in the United States emphasizes the need for more effective preventive and therapeutic modalities. There is growing evidence that vitamin D may reduce the incidence of CRC. Results of epidemiologic, in vitro, in vivo animal and clinical studies suggest that a low serum vitamin D level may be a serious risk factor for CRC and a high serum vitamin D level may reduce the risk of CRC. On a molecular level, vitamin D suppresses CRC development and growth by affecting cell proliferation, differentiation, apoptosis, and angiogenesis. Vitamin D insufficiency and CRC are common in the elderly population. Vitamin D insufficiency is simple to screen for and treatable with vitamin D supplementation. Serum 25-hydroxyvitamin D (calcidiol) is the best measure of vitamin D status and should be checked routinely for individuals with risk factors for CRC. Maintaining serum concentrations of calcidiol above 32 ng/ml (80 nmol/l) in individuals whose serum calcidiol level is low may help prevent CRC as well as osteoporosis, fractures, infections, and cardiovascular disease. Daily calcidiol intake of 1000 International Units can increase serum vitamin D to sufficient levels in most elderly persons and, based on available data, may substantially lower the incidence of CRC with minimal risks.


Activity and safety of brigatinib in ALK-rearranged non-small-cell lung cancer and other malignancies: a single-arm, open-label, phase 1/2 trial.
Anaplastic lymphoma kinase (ALK) gene rearrangements are oncogenic drivers of non-small-cell lung cancer (NSCLC). Brigatinib (AP26113) is an investigational ALK inhibitor with potent preclinical activity against ALK mutants resistant to crizotinib and other ALK inhibitors. We aimed to assess brigatinib in patients with advanced malignancies, particularly ALK-rearranged NSCLC. In this ongoing, single-arm, open-label, phase 1/2 trial, we recruited patients from nine academic hospitals or cancer centres in the USA and Spain. Eligible patients were at least 18 years of age and had advanced malignancies, including ALK-rearranged NSCLC, and disease that was refractory to available therapies or for which no curative treatments existed. In the initial dose-escalation phase 1 stage of the trial, patients received oral brigatinib at total daily doses of 30-300 mg (according to a standard 3 + 3 design). The phase 1 primary endpoint was establishment of the recommended phase 2 dose. In the phase 2 expansion stage, we assessed three oral once-daily regimens: 90 mg, 180 mg, and 180 mg with a 7 day lead-in at 90 mg; one patient received 90 mg twice daily. We enrolled patients in phase 2 into five cohorts: ALK inhibitor-naive ALK-rearranged NSCLC (cohort 1), crizotinib-treated ALK-rearranged NSCLC (cohort 2), EGFRT790M-positive NSCLC and resistance to one previous EGFR tyrosine kinase inhibitor (cohort 3), other cancers with abnormalities in brigatinib targets (cohort 4), and crizotinib-naive or crizotinib-treated ALK-rearranged NSCLC with active, measurable, intracranial CNS metastases (cohort 5). The phase 2 primary endpoint was the proportion of patients with an objective response. Safety and activity of brigatinib were analysed in all patients in both phases of the trial who had received at least one dose of treatment. This trial is registered with ClinicalTrials.gov, number NCT01449461. Between Sept 20, 2011, and July 8, 2014, we enrolled 137 patients (79 [58%] with ALK-rearranged NSCLC), all of whom were treated. Dose-limiting toxicities observed during dose escalation included grade 3 increased alanine aminotransferase (240 mg daily) and grade 4 dyspnoea (300 mg daily). We initially chose a dose of 180 mg once daily as the recommended phase 2 dose; however, we also assessed two additional regimens (90 mg once daily and 180 mg once daily with a 7 day lead-in at 90 mg) in the phase 2 stage. four (100% [95% CI 40-100]) of four patients in cohort 1 had an objective response, 31 (74% [58-86]) of 42 did in cohort 2, none (of one) did in cohort 3, three (17% [4-41]) of 18 did in cohort 4, and five (83% [36-100]) of six did in cohort 5. 51 (72% [60-82]) of 71 patients with ALK-rearranged NSCLC with previous crizotinib treatment had an objective response (44 [62% (50-73)] had a confirmed objective response). All eight crizotinib-naive patients with ALK-rearranged NSCLC had a confirmed objective response (100% [63-100]). Three (50% [95% CI 12-88]) of six patients in cohort 5 had an intracranial response. The most common grade 3-4 treatment-emergent adverse events across all doses were increased lipase concentration (12 [9%] of 137), dyspnoea (eight [6%]), and hypertension (seven [5%]). Serious treatment-emergent adverse events (excluding neoplasm progression) reported in at least 5% of all patients were dyspnoea (ten [7%]), pneumonia (nine [7%]), and hypoxia (seven [5%]). 16 (12%) patients died during treatment or within 31 days of the last dose of brigatinib, including eight patients who died from neoplasm progression. Brigatinib shows promising clinical activity and has an acceptable safety profile in patients with crizotinib-treated and crizotinib-naive ALK-rearranged NSCLC. These results support its further development as a potential new treatment option for patients with advanced ALK-rearranged NSCLC. A randomised phase 2 trial in patients with crizotinib-resistant ALK-rearranged NSCLC is prospectively assessing the safety and efficacy of two regimens assessed in the phase 2 portion of this trial (90 mg once daily and 180 mg once daily with a 7 day lead-in at 90 mg). ARIAD Pharmaceuticals.


Insulin-like growth factor-1 enhances epidermal growth factor receptor activation and renal tubular cell regeneration in postischemic acute renal failure.
Growth factors such as insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), and hepatocyte growth factor have been shown to accelerate the recovery from postischemic acute renal failure (ARF) with a concomitant increase in DNA synthesis. Interactions between growth factors have been demonstrated in a number of in vitro studies. This study examined the effect of exogenous IGF-1 on the DNA synthesis and EGF receptor (EGF-R) activation in postischemic rat kidneys. Thirty minutes after the relief of 30-minute total occlusion of the left renal artery in anesthetized 225 to 300 gm Sprague-Dawley rats, either IGF-1 (75 micrograms/kg) or normal saline solution (NS, 0.2 ml) was given by intravenous bolus, followed by twice daily subcutaneous injections of IGF-1 (50 micrograms/kg) or 0.2 ml NS for 4 days, respectively, in IGF-1-Tx) and NS treated (NS-Tx) groups (n = 8 each). On the day after the completion of treatment, inulin clearance (ml/kg/min) of the postischemic kidneys in the IGF-1-Tx group was significantly higher (p < 0.01) than inulin clearance of kidneys in the NS-Tx group. This was associated with improved kidney morphology. IGF-1 treatment also enhanced the labeling index of 5-bromo-2'-deoxyuridine (percent of stained tubule cells), a marker for active DNA synthesis, in the outer medulla of postischemic kidneys at 1 day and 2 days after the injury. EGF-R tyrosine phosphorylation (which reflects receptor activation) increased in postischemic kidneys in both NS-Tx (n = 5) and IGF-1-Tx (n = 3) groups 1 day after the injury as compared with nonischemic contralateral kidneys. In the IGF-1-Tx group there was also increased iodine 125-labeled EGF binding and EGF-R protein. Our results demonstrate a beneficial effect of IGF-1 on postischemic ARF. Furthermore, they suggest that EGF-R activation is involved in tubular regeneration and that IGF-1 may enhance EGF-R activation by increasing EGF-R expression.


Race, otitis media, and antibiotic selection.
Previous research suggests that physicians may be less likely to diagnose otitis media (OM) and to prescribe broad-spectrum antibiotics for black versus nonblack children. Our objective was to determine whether race is associated with differences in OM diagnosis and antibiotic prescribing nationally. We examined OM visit rates during 2008 to 2010 for children ≤14 years old using the National Ambulatory Medical Care Survey and National Hospital Ambulatory Medical Care Survey. We compared OM visits between black and nonblack children, as percentages of all outpatient visits and visit rates per 1000. We compared antibiotic prescribing by race as the percentage of OM visits receiving narrow-spectrum (eg, amoxicillin) versus broader-spectrum antibiotics. We used multivariable logistic regression to examine whether race was independently associated with antibiotic selection for OM. The percentage of all visits resulting in OM diagnosis was 30% lower in black children compared with others (7% vs 10%, P = .004). However, OM visits per 1000 population were not different between black and nonblack children (253 vs 321, P = .12). When diagnosed with OM during visits in which antibiotics were prescribed, black children were less likely to receive broad-spectrum antibiotics than nonblack children (42% vs 52%, P = .01). In multivariable analysis, black race was negatively associated with broad-spectrum antibiotic prescribing (adjusted odds ratio 0.59; 95% confidence interval, 0.40-0.86). Differences in treatment choice for black children with OM may indicate race-based differences in physician practice patterns and parental preferences for children with OM.


Nose tip refinement using interdomal suture in caucasian nose.
Refinement of the nose tip can be accomplished by a variety of techniques, but currently, the use of sutures in the nasal tip with conservative resection of the alar cartilage is the most frequently recommended approach. To classify the nasal tip and to demonstrate the interdomal suture applied to nasal tip refinement in the Caucasian nose, as well as to provide a simple and practical presentation of the surgical steps. Development of surgical algorithm for nasal tip surgery: 1. Interdomal suture (double binding suture), 2. Interdomal suture with alar cartilage weakening (cross-hatching), 3. Interdomal suture with cephalic removal of the alar cartilage (McIndoe technique) based on the nasal tip type classification. This classification assesses the interdomal distance (angle of domal divergence and intercrural distance), domal arch width, cartilage consistency, and skin type. Interdomal suture is performed through endonasal rhinoplasty by basic technique without delivery (Converse-Diamond technique) under local anesthesia. This classification is simple and facilitates the approach of surgical treatment of the nasal tip through interdomal suture, systematizing and standardizing surgical maneuvers for better refinement of the Caucasian nose.