Care of Healthy as well as Sick Newborns in India: A Narrative Review.
Majority of neonatal deaths occur in developing countries. There is an increase in the proportion of neonatal deaths as part of the under-5 mortality over the past decade. Hence we need to accelerate further to achieve the goal of single digit neonatal mortality rate (NMR) by 2030. The two major arms of NMR reduction include facility-based neonatal care (FBNC) and home-based neonatal care (HBNC). FBNC addresses care at birth, care of the normal newborn, and care of small and sick newborns. HBNC provides continuum of care for newborn and post-natal mothers facilitated by Accredited Social Health Activist (ASHA) workers. One of the main challenges is to maintain good quality of neonatal care. Zero separation, linkage of community & facility and roles of professional bodies are considered way forward to achieve India Newborn Action Plan (INAP) goals. This review summarizes existing programs for newborn health and diseases and provides an over-arching view of the way-forward.


Heat stress-induced effects of photosystem I: an overview of structural and functional responses.
Temperature is one of the main factors controlling the formation, development, and functional performance of the photosynthetic apparatus in all photoautotrophs (green plants, algae, and cyanobacteria) on Earth. The projected climate change scenarios predict increases in air temperature across Earth's biomes ranging from moderate (3-4 °C) to extreme (6-8 °C) by the year 2100 (IPCC in Climate change 2007: The physical science basis: summery for policymakers, IPCC WG1 Fourth Assessment Report 2007; Climate change 2014: Mitigation of Climate Change, IPCC WG3 Fifth Assessment Report 2014). In some areas, especially of the Northern hemisphere, even more extreme warm seasonal temperatures may occur, which possibly will cause significant negative effects on the development, growth, and yield of important agricultural crops. It is well documented that high temperatures can cause direct damages of the photosynthetic apparatus and photosystem II (PSII) is generally considered to be the primary target of heat-induced inactivation of photosynthesis. However, since photosystem I (PSI) is considered to determine the global amount of enthalpy in living systems (Nelson in Biochim Biophys Acta 1807:856-863, 2011; Photosynth Res 116:145-151, 2013), the effects of elevated temperatures on PSI might be of vital importance for regulating the photosynthetic response of all photoautotrophs in the changing environment. In this review, we summarize the experimental data that demonstrate the critical impact of heat-induced alterations on the structure, composition, and functional performance of PSI and their significant implications on photosynthesis under future climate change scenarios.


Prospective cohort study assessing patient satisfaction and diagnostic accuracy in channeled endoscope local anesthetic biopsy in the investigation of head and neck cancer.
There are delays in the head and neck Urgent Suspicion of Cancer (USOC) pathway. Local anesthetic (LA) biopsy with channeled endoscopes in outpatients can reduce time to diagnosis. Questionnaire-based prospective study of LA or general anesthetic (GA) biopsy for investigation of cancer from September 2021 to July 2022. 100% (n = 48) were very satisfied or satisfied with their overall experience. 71% (n = 20) of LA patients reported they would prefer to have a biopsy under LA. LA biopsy was 68% sensitive and 100% specific for cancer diagnosis. 28.6% (8) and 10% (2) of LA and GA patients had palliative MDT outcomes. Median time to diagnosis was 44.5 and 49.0 days for LA and GA biopsy, respectively. LA biopsy had a high satisfaction rate and is sensitive for diagnosing cancer. LA biopsy can expedite treatment pathways. LA biopsy should be considered as an option for first line investigation.


The remarkable UV light invulnerability of thymine GNA dinucleotides.
We herein demonstrate the UV resistance of glycol nucleic acid (GNA) dinucleotides. This resistance sustains the hypothesis of GNA as a nucleic acid prebiotic ancestor on early Earth, a time of intense solar UV light. Such photorobustness, due to the absence of intrastrand base stacking, could offer an opportunity for nanodevice development requiring challenging UV conditions.


Human lung small-cell carcinoma contains bombesin.
The presence of immunoreactive bombesin in a human lung small-cell carcinoma grown in nude mice was established by several criteria: (i) Radioimmunoassay of tissue extracts for bombesin revealed approximately 6.5 pmol/g of tissue; (ii) bombesin was found in 12-14% of the tumor cells by immunohistochemical localization; (iii) gel filtration of small-cell carcinoma extract on Sephadex G-75 and Bio-Gel P-4 gave only a single peak of immunoreactivity, which occurred at the elution volume of bombesin; and (iv) reverse-phase HPLC of acid-solubilized extracts separated the immunoreactive material into three discrete peaks, one of which eluted with a retention time identical to that of synthetic bombesin. The presence of bombesin may represent the ectopic expression of this peptide in small-cell carcinoma, because immunoreactive bombesin was found in human fetal and neonatal lung but apparently not in adult lung tissue [Wharton, J., Polak, J. M., Bloom, S. R., Ghatei, M. A., Solcia, E., Brown, M. R. & Pearse, A. G. E. (1978) Nature (London) 273, 769-770]. The immunoreactive bombesin previously found in mammalian tissues is considerably larger than amphibian bombesin; these data substantiate the presence of a mammalian form of bombesin in a human tumor that may have a structure similar to that of the amphibian peptide.


Propagation in cell culture of the dinoflagellate amyloodinium, an ectoparasite of marine fishes.
Amyloodinium ocellatum, a common dinoflagellate ectoparasite of marine fishes, was successfully propagated on a fish gill cell line. In vitro infections were similar in cytopathology and development to those reported on natural hosts, and large numbers of parasites could be produced. Exposure of parasites in cell culture to an antiprotozoal drug produced a dose-dependent inhibition of infectivity that was much more sensitive than a motility assay previously used to assess the toxic effects of a drug on protozoan ectoparasites. This propagation system may be a useful model for studying the biology and control of protozoan skin parasites of fishes and for quantitatively studying hostparasite interaction at cellular interfaces.


Preventing the development of varices in cirrhosis.
Gastroesophageal varices are a direct consequence of portal hypertension. Nonselective beta-adrenergic blockers decrease portal pressure and are effective in preventing variceal hemorrhage. However, a large multicenter placebo-controlled trial demonstrates that nonselective beta-adrenergic blockers are not effective in preventing the development of varices and are associated with a significant rate of adverse events. This therapy is, therefore, not recommended in compensated cirrhotic patients without varices at large. In this very compensated group of patients with cirrhosis (stage 1, ie, without varices and without ascites or encephalopathy) the predictive value (both for the development of varices and for the development of clinical decompensation) of a baseline hepatic venous pressure gradient greater than 10 mm Hg is confirmed, supporting this threshold level as one that defines a clinically significant portal hypertension. Importantly, reductions in hepatic venous pressure gradient >10% are associated with a significant reduction in the development of varices, a therapeutic goal that could be achieved through the use of beta-blockers or other drugs being developed for the treatment of portal hypertension.


Understanding Trends in Pertussis Incidence: An Agent-Based Model Approach.
We examined the impact of undetected infections, adult immunity, and waning vaccine-acquired immunity on recent age-related trends in pertussis incidence. We developed an agent-based model of pertussis transmission in Dakota County, Minnesota using case data from the Minnesota Department of Health. For outbreaks in 2004, 2008, and 2012, we fit our model to incidence in 3 children's age groups relative to adult incidence. We estimated parameters through model calibration. The duration of vaccine-acquired immunity after completion of the 5-dose vaccination series decreased from 6.6 years in the 2004 model to approximately 3.0 years in the 2008 and 2012 models. Tdap waned after 2.1 years in the 2012 model. A greater percentage of adults were immune in the 2008 model than in the 2004 and 2012 models. On average, only 1 in 10 adult infections was detected, whereas 8 in 10 child infections were detected. The observed trends in relative pertussis incidence in Dakota County can be attributed in part to fluctuations in adult immunity and waning vaccine-acquired immunity. No single factor accounts for current pertussis trends.


Assessment of Adherence to Cardiovascular Medicines in Saudi Population: An Observational Study in Patients Attending a Tertiary Care Hospital.
Drug adherence has been extensively evaluated in many developed countries in the West using different methods of medication adherence measurement; however, there are relatively few reports studying the adherence levels among Saudi patients. Thus, this study will evaluate the adherence to cardiovascular medicines in Saudi patients visiting (PSCC) in Al-Qassim, Saudi Arabia. This cross-sectional observational study relied on self-administered questionnaires. This study used the Morisky, Green, and Levine (MGL) Adherence Scale, also known as the MAQ (Medication Adherence Questionnaire), in PSCC's pharmacy waiting room in Qassim, Saudi Arabia. This study included 993 PSCC pharmacy waiting room patients. The patients were between 11 and 50 years old, and 52.7 percent were male. Most participants (71.2%) were above 50, while 16.3% were 41-50. Non-adherent patients cited traveling or being busy (28.6%), forgetting (18.7%), daily multi-medications (7.1%), being sleepy or sleeping (6.6%), and not repeating the prescription (6.6%). The Medicine Adherence Questionnaire indicated that 62.6 percent of patients fully adhered to their medications, and 21.6 percent usually adhered. Only drug adverse effects affected adherence (p =0.0001). The current study showed that there is a good level of adherence among patients with cardiovascular diseases toward their diseases. The most common reasons for neglecting medications include traveling or being busy, forgetting multiple medications, and being tired or sleeping. Having experience with side effects was the only significant factor affecting adherence to medications.


CRISPR/Cas13a-Powered Electrochemical Microfluidic Biosensor for Nucleic Acid Amplification-Free miRNA Diagnostics.
Noncoding small RNAs, such as microRNAs, are becoming the biomarkers of choice for multiple diseases in clinical diagnostics. A dysregulation of these microRNAs can be associated with many different diseases, such as cancer, dementia, and cardiovascular conditions. The key for effective treatment is an accurate initial diagnosis at an early stage, improving the patient's survival chances. In this work, the first clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a-powered microfluidic, integrated electrochemical biosensor for the on-site detection of microRNAs is introduced. Through this unique combination, the quantification of the potential tumor markers microRNA miR-19b and miR-20a is realized without any nucleic acid amplification. With a readout time of 9 min and an overall process time of less than 4 h, a limit of detection of 10 pm is achieved, using a measuring volume of less than 0.6 µL. Furthermore, the feasibility of the biosensor platform to detect miR-19b in serum samples of children, suffering from brain cancer, is demonstrated. The validation of the obtained results with a standard quantitative real-time polymerase chain reaction method shows the ability of the electrochemical CRISPR-powered system to be a low-cost, easily scalable, and target amplification-free tool for nucleic acid based diagnostics.


Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins.
The aim of this study was to test whether galectin-3 is present in human corneal epithelium and whether lipopolysaccharide (LPS) purified from Pseudomonas aeruginosa ATCC 19660 binds to this animal lectin and/or to another human corneal epithelial protein(s) (HCEP) and to confirm which component of LPS (inner or outer core or lipid A) is important in bacterial binding by using the eye in organ culture. LPS isolated and purified from P. aeruginosa ATCC 19660 and a commercial LPS (serotype 10) differed in polyacrylamide gel analysis but bound similarly to blotted HCEP. Binding was determined to be a receptor-ligand type of interaction by the solid-phase assay, because it was both specific and saturable. Several LPS binding proteins in HCEP were identified by an overlay method. Western blotting with antibody against galectin-3 revealed the presence of this protein in both freshly isolated and cultured transformed human corneal epithelium. Binding inhibition assays showed that antibody specific for the outer core region of LPS and an anti-galectin antibody significantly inhibited bacterial binding in vitro. These data provide further evidence that LPS is an important adhesin of P. aeruginosa, that it binds to protein receptor molecules in HCEP, that one of the LPS binding proteins is galectin-3, and that the outer core portion of the molecule appears to be critical for LPS binding to the eye.


Can cytological features differentiate reactive renal tubular cells from low-grade urothelial carcinoma cells?
To compare the cytomorphological and immunocytochemical features of reactive renal tubular cells and low-grade urothelial carcinoma cells (LG-UCs). We examined 15 cytological parameters in 38 cases with reactive renal tubular cells in renal disease and 20 cases of LG-UCs from bladder cancer that had been diagnosed by histological examination. Voided urine cytological parameters evaluated were as follows: (i) maximum cell numbers of clusters, (ii) cannibalism, (iii) rosette-like arrangement, (iv) hobnail-shaped cells, (v) vacuolated cytoplasm, (vi) intracytoplasmic haemosiderin, (vii) irregular nuclear contours, (viii) chromatin pattern, (ix) prominent nucleoli, (x) cast encasement, (xi) casts, (xii) dysmorphic erythrocytes, (xiii) isomorphic erythrocytes, (xiv) necrosis, and (xv) vimentin reactivity. The above parameters were determined using Mann-Whitney U-test and chi-square test, with differences considered significant at P < 0.05. In reactive renal tubular cells, low to moderate cell numbers of clusters (fewer than 50 cells), rosette-like arrangement, hobnail-shaped cells, vacuolated cytoplasm, intracytoplasmic haemosiderin, euchromatin pattern, prominent nucleoli, dysmorphic erythrocytes and vimentin reactivity were present in significantly higher proportions compared with those in LG-UCs. In LG-UCs, high cell numbers of clusters (50 cells or more), cannibalism, heterochromatin pattern, isomorphic erythrocytes and necrosis were seen in significantly higher proportions. No significant differences were observed in irregular nuclear contours, cast encasement or casts. Based on results of the present study, maximum cell numbers of clusters, cannibalism, rosette-like arrangement, hobnail-shaped cells, vacuolated cytoplasm, intracytoplasmic haemosiderin, chromatin pattern, prominent nucleoli, dysmorphic erythrocytes, isomorphic erythrocytes, necrosis, and vimentin reactivity were capable of distinguishing reactive renal tubular cells from LG-UCs.


Distal 11q monosomy syndrome: a report of two Egyptian sibs with normal parental karyotypes confirmed by molecular cytogenetics.
Jacobsen syndrome is a rare disorder, caused by segmental monosomy for the distal end of the long arm of chromosome 11 with variable phenotypic expressivity. We report on the first male (6 years old) and female (3 years old) sibs with clinical and cytogenetics characterization of Jacobsen syndrome. Their karyotypes showed deletion 11q23.3-qter. Patients presented with growth and psychomotor retardation, facial dysmorphism, eye anomalies, and congenital heart disease (variable degrees of septal defect). Family history revealed a clinically similar brother, who died at 2 months old from cardiac anomalies in the form of single ventricle without being subjected to further investigations. Chromosomal analysis of the parents was normal. Karyotyping for the 2 patients and their parents was confirmed by fluorescence in situ hybridization analysis (FISH) using whole chromosome painting probes for 11 (WCP 11). Relevant investigations for both sibs showed mild thrombocytopenia with normal platelets morphology and striking periventricular demyelination on neuroimaging. Inguinal small testicles as well as focal epileptiform dysfunction were recorded in the male patient only. Abdominal ultrasound, hearing test, and DEXA scan were normal in both patients. Due to of the presence of apparently 3 affected offspring and normal parental karyotypes, an inherited predisposition was highly suspected. The large size of the distal deleted 11q segment in our patients support the recent hypothesis, that Jacobsen syndrome is a chromosomal deletion syndrome with genetic predisposition, due to expansion of p(CCG)n trinucleotide in the folate-sensitive fragile site FRA11B, at breakpoint 11q23.3. In conclusion, identification and further delineation of more similar patients will contribute to understanding the genetic basis of the 11q phenotype.


The chest radiograph and sarcoidosis. Reevaluation of the chest radiograph in assessing activity of sarcoidosis: a preliminary communication.
Classic radiographic staging of sarcoidosis, utilizing stages 0 through III, is a purely descriptive classification and bears no relation to the clinical activity of the disease. We propose a new method of determining sarcoidosis activity based solely on the chest radiograph. This "staging" is based on the presence or absence of air bronchograms, peribronchial cuffing, and subpleural thickening. Adenopathy, reticular, reticulonodular, and nodular opacities are not included in our methodology. Other diagnostic methods currently utilized for staging activity include Gallium scanning, bronchoalveolar lavage, and physiologic studies. In 85 studies performed in 51 patients with biopsy-proven sarcoidosis, this staging method agreed with Gallium scintigraphy results in 68.2%. The system failed in specific and identifiable subgroups of patients. We feel that the chest radiograph compares favorably with other staging methods in detecting the alveolitis phase of sarcoidosis, and is reproducible, noninvasive, and cost efficient.


Parathyroid Hormone Levels in the Prediction of Ischemic Stroke Risk.
Objective. It was examined whether PTH and 25-dihydroxyvitamin D (25(OH)D) levels, together or separately, are indicators of the risk of stroke. Materials and Methods. This prospective study was performed at two centers. In the study, 100 patients diagnosed with acute ischemic stroke and 100 control individuals in the same age range were examined. In addition to neurological examination, cranial imaging, extensive routine blood chemistry, PTH, and 25(OH)D levels were evaluated in all cases. Stroke risk factors were determined. Logistic regression was used for statistical analysis. Results. A total of 60 patients and 79 control individuals were included in the study. Different estimation models were designed in order to examine the relationship between PTH and 25(OH)D levels with stroke. According to modeling results, it was determined that the most effective predictor for risk of stroke was 25(OH)D levels, followed by hypertension and PTH levels, respectively. Conclusion. PTH and 25(OH)D levels together can make important contributions to determination of stroke risk, and further investigations are needed to understand this relationship more fully.


Construction of a prognostic model for lung adenocarcinoma based on heat shock protein-related genes and immune analysis.
Lung adenocarcinoma (LUAD) represents a prevalent form of cancer, with low early diagnosis rates and high mortality rates, posing a global health challenge. Heat shock proteins (HSPs) assume a crucial role within the tumor immune microenvironment (TME) of LUAD. Here, a collection of 97 HSP-related genes (HSPGs) was assembled based on prior literature reports, of which 36 HSPGs were differentially expressed in LUAD. In The Cancer Genome Atlas (TCGA) cohort, we constructed a prognostic model for risk stratification and prognosis prediction by integrating 13 HSPGs. In addition, the prognostic significance and predictive efficacy of the HSP-related riskscore were examined and validated in the Gene Expression Omnibus (GEO) cohort. To facilitate the clinical use of this riskscore, we also established a nomogram scale by verifying its effectiveness through different methods. In light of these outcomes, we concluded a significant correlation between HSPs and TME in LUAD, and the riskscore can be a reliable prognostic indicator. Furthermore, this study evaluated the differences in immunophenoscore, tumor immune dysfunction and exclusion score, and sensitivity to several common chemotherapy drugs among LUAD individuals in different risk groups, which may aid in clinical decision-making for immune therapy and chemotherapy in LUAD individuals.


Assortment of encounters and evolution of cooperativeness.
The method of evolutionary stable strategies (ESS), in its current form, is confronted with a difficulty when it tries to explain how some social behaviors initiate their evolution. We show that this difficulty may be removed by changing the assumption made tacitly in game theory (and in ESS) of randomness of meetings or encounters. In reality, such randomness seems to be rare in nature. Family, population and social structure, customs, and habits impose various types of deviation from randomness. Introducing nonrandomness of meeting in a way formally similar to assortative mating, we show that the bar to initial increase of inherited cooperative or altruistic behaviors can be removed, provided there is sufficient assortment of meetings. Family structure may cause contacts predominantly between certain types of relatives, and one can reconstruct some results of classical kin selection in terms of evolutionary stable strategy with assortative meetings. Neighbor effects and group selection might be similarly treated. Assortment need not be a passive consequence of population and social structure, but it can also be actively pursued. Behaviors favoring the choice of cooperative companions will have the effect of favoring the evolution of cooperativeness. It can be shown that discrimination in the choice of companions, especially if combined with assortment, can favor the development of cooperativeness, making initial increase of cooperative behavior possible even at levels of assortment passively imposed which would not be adequate, per se, to guarantee the increase of cooperativeness. It is possible that, in some cases, cooperativeness and behavior favoring some type of assortment are coselected.


Statistical Modeling of Subjective Sleep Quality.
Effective maintenance/improvement of sleep quality requires knowledge of how sleep quality is connected to quantitative features of sleep and arbitrarily selected habitual lifestyles, which naturally depend on the demographic characteristics of individuals. To fulfill these needs, a regression model of subjective sleep quality was constructed, whereby one might be able to design a practical strategy for achieving comfortable sleep adapted to individual conditions. Based on data obtained from our previous study, fundamental correlation profiles between day-to-day subjective and quantitative features of sleep were estimated. Obtained correlation profiles involving SRSs, quantitative features of sleep, and sleep habits across a week such as bedtime preference (chronotype), discrepancy between chronotype and social time cue (social jetlag), and habitual sleep-wake pattern (HSWP) were characterized specifically for each self-ratings of sleep quality (SRS) category through backward stepwise Linear Mixed Effect (LME) modeling. The LME model represented SRSs with acceptable accuracy, allowing identification of determinant factors for each category of SRS. The SRS is one possible option to clarify sleep status. In this study, we proposed a possible framework including model-based predictors of SRS in which self-awareness of sleep quality could be improved to facilitate healthy sleep practices.


Depression and health-related quality of life among persons with sensory disabilities in a health professional shortage area.
The authors examined depression and health-related quality of life among individuals with self-reported sensory impairments living in a health professional shortage area. Health surveys of residents were conducted in 2006 and 2010. Responses were analyzed by groups of residents reporting vision loss, hearing loss, dual hearing and vision loss, and no sensory loss. In 2006, the total sample size was n = 2,591, and in 2010, it was n = 3,955. The CESD-5 scale (Shrout & Yager, 1989) was included in 2006, and the PHQ-9 (Kroenke, Spitzer, & Williams, 2001) was included in 2010. Rates of depression on the CESD-5 were determined by the recommended cut-off scores and on the PHQ-9 by the recommended algorithm. The Centers for Disease Control (CDC) Healthy Days instrument (Moriarty, Zack, & Kobau, 2003) was used in both surveys to assess health-related quality of life. In both surveys, individuals who reported sensory loss had higher rates of depression and lower health-related quality of life than individuals with no reported sensory loss. Individuals reporting sensory loss had high rates of depression and a compromised quality of life compared to respondents without these impairments. These data imply strategic community-based health care services, including mental health initiatives, may be indicated for individuals with sensory loss living in underserved regions. Implications for rehabilitation psychology research, service, and policy are discussed as innovations in these areas are needed to better understand and address the disparities that may compromise the overall well-being of residents of underserved communities. (PsycINFO Database Record


The association of C-reactive protein and CRP genotype with coronary heart disease: findings from five studies with 4,610 cases amongst 18,637 participants.
It is unclear whether C-reactive protein (CRP) is causally related to coronary heart disease (CHD). Genetic variants that are known to be associated with CRP levels can be used to provide causal inference of the effect of CRP on CHD. Our objective was to examine the association between CRP genetic variant +1444C>T (rs1130864) and CHD risk in the largest study to date of this association. We estimated the association of CRP genetic variant +1444C>T (rs1130864) with CRP levels and with CHD in five studies and then pooled these analyses (N = 18,637 participants amongst whom there were 4,610 cases). CRP was associated with potential confounding factors (socioeconomic position, physical activity, smoking and body mass) whereas genotype (rs1130864) was not associated with these confounders. The pooled odds ratio of CHD per doubling of circulating CRP level after adjustment for age and sex was 1.13 (95%CI: 1.06, 1.21), and after further adjustment for confounding factors it was 1.07 (95%CI: 1.02, 1.13). Genotype (rs1130864) was associated with circulating CRP; the pooled ratio of geometric means of CRP level among individuals with the TT genotype compared to those with the CT/CC genotype was 1.21 (95%CI: 1.15, 1.28) and the pooled ratio of geometric means of CRP level per additional T allele was 1.14 (95%CI: 1.11, 1.18), with no strong evidence in either analyses of between study heterogeneity (I(2) = 0%, p>0.9 for both analyses). There was no association of genotype (rs1130864) with CHD: pooled odds ratio 1.01 (95%CI: 0.88, 1.16) comparing individuals with TT genotype to those with CT/CC genotype and 0.96 (95%CI: 0.90, 1.03) per additional T allele (I(2)<7.5%, p>0.6 for both meta-analyses). An instrumental variables analysis (in which the proportion of CRP levels explained by rs1130864 was related to CHD) suggested that circulating CRP was not associated with CHD: the odds ratio for a doubling of CRP level was 1.04 (95%CI: 0.61, 1.80). We found no association of a genetic variant, which is known to be related to CRP levels, (rs1130864) and having CHD. These findings do not support a causal association between circulating CRP and CHD risk, but very large, extended, genetic association studies would be required to rule this out.


Major vault protein supports glioblastoma survival and migration by upregulating the EGFR/PI3K signalling axis.
Despite their ubiquitous expression and high conservation during evolution, precise cellular functions of vault ribonucleoparticles, mainly built of multiple major vault protein (MVP) copies, are still enigmatic. With regard to cancer, vaults were shown to be upregulated during drug resistance development as well as malignant transformation and progression. Such in a previous study we demonstrated that human astrocytic brain tumours including glioblastoma are generally high in vault levels while MVP expression in normal brain is comparably low. However a direct contribution to the malignant phenotype in general and that of glioblastoma in particular has not been established so far. Thus we address the questions whether MVP itself has a pro-tumorigenic function in glioblastoma. Based on a large tissue collection, we re-confirm strong MVP expression in gliomas as compared to healthy brain. Further, the impact of MVP on human glioblastoma aggressiveness was analysed by using gene transfection, siRNA knock-down and dominant-negative genetic approaches. Our results demonstrate that MVP/vaults significantly support migratory and invasive competence as well as starvation resistance of glioma cells in vitro and in vivo. The enhanced aggressiveness was based on MVP-mediated stabilization of the epidermal growth factor receptor (EGFR)/phosphatidyl-inositol-3-kinase (PI3K) signalling axis. Consequently, MVP overexpression resulted in enhanced growth and brain invasion in human glioblastoma xenograft models. Our study demonstrates, for the first time, that vaults have a tumour-promoting potential by stabilizing EGFR/PI3K-mediated migration and survival pathways in human glioblastoma.


Early Assessment of Colorectal Cancer by Quantifying Circulating Tumor Cells in Peripheral Blood: ECT2 in Diagnosis of Colorectal Cancer.
Circulating tumor cells (CTCs) in peripheral blood is an indication of poor prognosis for patients with different cancer types. However, most of the available technologies for detecting CTCs show low sensitivity and specificity. Therefore, we attempted to find an alternative marker for CTCs of colorectal cancer. We have directly extracted RNA from CTCs contained in 1.5 mL peripheral blood from 90 colorectal cancer patients and 151 healthy donors, and screened these samples for candidate marker genes by nested real-time quantitative polymerase chain reaction (PCR). From genes selected from a public database of microarray analyses, we successfully identified epithelial cell transforming sequence 2 oncogene (ECT2) as a gene that exhibits high differential expression ratios (p < 0.01). ECT2 displays good sensitivity and specificity, with an area under the curve (AUC) value of 0.821. This marker gene also has a high detection rate in patients with serum carcinoembryonic antigen (CEA) concentrations below the diagnostic threshold of 5 ng/mL. The expression of ECT2 can therefore serve as an alternative measurement that can compensate for the inadequacy of the current CEA test in the diagnosis and monitoring of colorectal cancer patients.


Competent jerks, lovable fools, and the formation of social networks.
When looking for help with a task at work, people turn to those best able to do the job. Right? Wrong. New research shows that work partners tend to be chosen not for ability but for likability. Drawing from their study encompassing 10,000 work relationships in five organizations, the authors have classified work partners into four archetypes: the competent jerk, who knows a lot but is unpleasant; the lovable fool, who doesn't know much but is a delight; the lovable star, who's both smart and likable; and the incompetent jerk, who.. .well, that's self-explanatory. Of course, everybody wants to work with the lovable star, and nobody wants to work with the incompetent jerk. More interesting is that people prefer the lovable fool over the competent jerk. That has big implications for every organization, as both of these types often represent missed opportunities. Because they are liked by a disproportionate number of people, lovable fools can bridge gaps between diverse groups that might not otherwise interact. But their networking skills are often developed at the expense of job performance, which can make these employees underappreciated and vulnerable to downsizing. To get the most out of them, managers need to protect them and put them in positions that don't waste their bridge-building talents. As for the competent jerks, too often their expertise goes untapped by people who just can't put up with them. But many can be socialized through coaching or by being made accountable for bad behavior. Others may need to display their competence in more isolated settings. Intriguingly, managers aren't limited to leveraging people that others like and changing those that others loathe. They also can create situations in which people are more apt to like one another, whatever their individual qualities.


North Atlantic summers have warmed more than winters since 1353, and the response of marine zooplankton.
Modeling and measurements show that Atlantic marine temperatures are rising; however, the low temporal resolution of models and restricted spatial resolution of measurements (i) mask regional details critical for determining the rate and extent of climate variability, and (ii) prevent robust determination of climatic impacts on marine ecosystems. To address both issues for the North East Atlantic, a fortnightly resolution marine climate record from 1353-2006 was constructed for shallow inshore waters and compared to changes in marine zooplankton abundance. For the first time summer marine temperatures are shown to have increased nearly twice as much as winter temperatures since 1353. Additional climatic instability began in 1700 characterized by ∼5-65 year climate oscillations that appear to be a recent phenomenon. Enhanced summer-specific warming reduced the abundance of the copepod Calanus finmarchicus, a key food item of cod, and led to significantly lower projected abundances by 2040 than at present. The faster increase of summer marine temperatures has implications for climate projections and affects abundance, and thus biomass, near the base of the marine food web with potentially significant feedback effects for marine food security.


Systematics and geographical distribution of Galba species, a group of cryptic and worldwide freshwater snails.
Cryptic species can present a significant challenge to the application of systematic and biogeographic principles, especially if they are invasive or transmit parasites or pathogens. Detecting cryptic species requires a pluralistic approach in which molecular markers facilitate the detection of coherent taxonomic units that can then be analyzed using various traits (e.g., internal morphology) and crosses. In asexual or self-fertilizing species, the latter criteria are of limited use. We studied a group of cryptic freshwater snails (genus Galba) from the family Lymnaeidae that have invaded almost all continents, reproducing mainly by self-fertilization and transmitting liver flukes to humans and livestock. We aim to clarify the systematics, distribution, and phylogeny of these species with an integrative approach that includes morphology, molecular markers, wide-scale sampling across America, and data retrieved from GenBank (to include Old World samples). Our phylogenetic analysis suggests that the genus Galba originated ca. 22 Myr ago and today comprises six species or species complexes. Four of them show an elongated-shell cryptic phenotype and exhibit wide variation in their genetic diversity, geographic distribution, and invasiveness. The remaining two species have more geographically restricted distributions and exhibit a globose-shell cryptic phenotype, most likely phylogenetically derived from the elongated one. We emphasize that no Galba species should be identified without molecular markers. We also discuss several hypotheses that can explain the origin of cryptic species in Galba, such as convergence and morphological stasis.


Systematic review of guidelines on neonatal hypoglycemia.
In recent years, a series of clinical guidelines on neonatal hypoglycemia have been developed in different countries and regions. This systematic review was aimed at providing evidence for clinical decision-making and providing ideas for future research by comparatively analyzing the contents of various guidelines. A multilateral approach was used, including comprehensive literature searches and online research. The retrieved studies were screened by two independent reviewers according to our inclusion criteria. The two reviewers independently extracted the descriptive data. Four appraisers assessed the guidelines using the AGREE-II instrument. Ten clinical guidelines on neonatal hypoglycemia were included, with a mean score of 45.28%-83.45% in six domains. The guidelines are relatively consistent in their recommendations on clinical symptoms of neonatal hypoglycemia, but different in risk factors, preventive measures, thresholds for clinical management of hypoglycemia, target glucose ranges for its control, and pharmacotherapy. By summarising the recommendations in the guidelines on neonatal hypoglycemia, we found that blood glucose values were not the only observational indicator, and other indicators (e.g., ketone bodies, lactate) related to glucose metabolism should also be considered for a comprehensive assessment. There is still a lack of consensus on thresholds for the clinical management of hypoglycemia and target glucose ranges for its control, and the recommendations on its pharmacotherapy are rather simple and sketchy. In the future, more high-quality studies are required to further improve the early identification of neonatal hypoglycemia and intervention strategies against it.


Actin polymerization and interaction with other proteins in temperature-induced gelation of sea urchin egg extracts.
The gel which forms on warming the extracts of the cytoplasmic proteins of sea urchin eggs has been separated into two fractions, one containing F-actin and the other containing two proteins of 58,000 and 22,000 mol wt. When combined in 0.1 M KCl, even at 0 degrees C, these components will form gel material identical to that formed by warming extracts. This gel is a network of laterally aggregated F-actin filaments which are in register and which display a complex cross-banding pattern generated by the presence of the other two proteins. Low concentrations of calcium block the assembly of these proteins to form this complex structure, which may play some cytoskeletal role in the cytoplasm. This association of F-actin with the other proteins to form a gel is very likely the last step fo the process occurring in warmed extracts. At low temperatures, gelation of extracts is limited by the relative absence of F-actin, as demonstrated by the inability to sediment it at 100,000 g and also by the fact that gelation occurs immediately if exogenous F-actin is added to cold extracts. The transformation of the G-actin present in the extract to the F-form is apparently repressed at low temperatures. This is shown directly by the failure of added G-actin to polymerize at low temperatures in the presence of extract. These observations resemble those which have been reported on preparations from amoeboid cells and may be significant in the involvement of actin and these other proteins in cell division and later developmental processes.


Welfare reform, family support, and child development: perspectives from policy analysis and developmental psychopathology.
This article explores the implications of recent welfare-related policy change for the well-being of children in low-income families, and for research investigating child development processes and outcomes. It provides an overview of current welfare-related policies and explores the implications for developmental researchers. The article also synthesizes early findings from research, highlighting both overall impacts and the more nuanced evidence that while families are transitioning off welfare, only a small number are transitioning out of poverty, and a subgroup of families at risk are not faring well. It then examines, from a theoretical and methodological framework, what developmental psychopathology might bring to the study of welfare-related impacts on children in the context of this complex and changing policy landscape, and what welfare researchers might bring to the field of developmental psychopathology. The article concludes with broad recommendations for both research and policy.


Early authors - R. H. Mole.
Purpose: To reflect on the contributions of R. H. Mole to radiobiology. Robin Mole was a very active radiobiologist when the IJRB was first established and had two papers in the first few issues of the journal in 1959. Conclusions: R. H. Mole made significant contributions to radiobiology and is particularly associated with studies of the effects of low dose exposures on the reproductive health of mammals. Many of his ideas are still debated and remain controversial.


Plasma Clusterin and Lipid Profile: A Link with Aging and Cardiovascular Diseases in a Population with a Consistent Number of Centenarians.
The role of Clusterin in attenuation of inflammation and reverse cholesterol transfer makes this molecule a potential candidate as a marker for cancer, cardiovascular disease, diabetes mellitus, and metabolic syndrome. In elderly subjects cardiovascular diseases represent the primary cause of death and different clinical studies have shown a positive correlation of these diseases with changes in the lipid pattern. This work aimed at evaluating the relationship between circulating clusterin and the biochemical parameters that characterize the lipid profile of a Sardinian population divided into five age groups including centenarians; the high frequency in Sardinia of these long-lived individuals gave us the opportunity to extend the range of the age groups to be analyzed to older ages and to better evaluate the changes in the lipid balance during ageing and its relationship with clusterin concentration in plasma. Our results showed that Clusterin concentration values of the youngest group were more similar with the centenarian's group compared to the other age groups, and a positive correlation arises with LDL. Furthermore given the high prevalence of cardiovascular diseases in the population examined and the association of Clusterin with these pathologies we evaluated Clusterin concentration variation in two groups with or without cardiovascular diseases. In presence of cardiovascular disease, Clusterin is significantly related to the most atherogenic components of lipid profile (total cholesterol and LDL), especially in women, suggesting its potential role in modulating cardiovascular metabolic risk factors.


Structure-activity relationships OF N-methylthiolated beta-lactam antibiotics with C3 substitutions and their selective induction of apoptosis in human cancer cells.
The development of novel anti-cancer drugs that induce apoptosis has long been a focus of drug discovery. Beta-lactam antibiotics have been used for over 60 years to fight bacterial infectious diseases with little or no side effects observed. Recently a new class of N-methylthiolated beta-lactams has been discovered that have potent activity against methicillin resistant Staphylococcus aureas. Most recently, we determined the potential effects of these N-thiolated beta-lactams on tumorigenic cell growth and found that they are apoptosis-inducers in human cancer cell lines. In the current study, we further determined the effects of the substitution of the O-methyl moiety on C3 and stereochemistry of the beta-lactams on the anti-proliferative and apoptosis-inducing abilities. We have found that lactam 18, in which C3 is substituted with an acrylate ester group, is a very effective proliferation inhibitor against human premalignant and malignant breast, leukemic, and simian virus 40-transformed fibroblast cells. Generally speaking, increasing the size of the moiety on C3 decreases its anti-proliferation potency, possibly indicating steric hindrance with the cellular target or decreased permeability through the cell membrane. We also found that the stereochemistry of the beta-lactams plays an important role in their potency. The 3S,4R isomers are more effective than their enantiomers (3R,4S), suggesting that 3S,4R configuration is more favorable for target interaction.


Male germ cell transplantation: promise and problems.
Male germ cell transplantation is a novel technique in which donor male stem germ cells are surgically transferred to the seminiferous tubules of a recipient testis by direct injection or via the rete testis or efferent duct. All germ cells that are destined to become stem spermatogonia are defined as male stem germ cells, including primordial germ cells from the gonadal ridges, and gonocytes and stem spermatogonia from the testis, all of which are transplantable and capable of undergoing normal spermatogenesis. Xenotransplantation of male germ cells from one species into the testis of another species, including human testicular cells in the mouse, has so far proved to be unsuccessful. However, the immunodeficient mouse testis can support rat spermatogenesis and produce apparently normal rat spermatozoa. The underlying mechanisms remain elusive. The present mini-review will focus on the importance of stem spermatogonial transplantation for testicular stem cell biology and discuss the likelihood of immune rejection after transplantation, which may limit the success of all male germ cell transplantation.


Trophic hierarchies illuminated via amino acid isotopic analysis.
Food web ecologists have long sought to characterize the trophic niches of animals using stable isotopic analysis. However, distilling trophic position from isotopic composition has been difficult, largely because of the variability associated with trophic discrimination factors (inter-trophic isotopic fractionation and routing). We circumvented much of this variability using compound-specific isotopic analysis (CSIA). We examined the (15)N signatures of amino acids extracted from organisms reared in pure culture at four discrete trophic levels, across two model communities. We calculated the degree of enrichment at each trophic level and found there was a consistent trophic discrimination factor (~7.6‰). The constancy of the CSIA-derived discrimination factor permitted unprecedented accuracy in the measurement of animal trophic position. Conversely, trophic position estimates generated via bulk-(15)N analysis significantly underestimated trophic position, particularly among higher-order consumers. We then examined the trophic hierarchy of a free-roaming arthropod community, revealing the highest trophic position (5.07) and longest food chain ever reported using CSIA. High accuracy in trophic position estimation brings trophic function into sharper focus, providing greater resolution to the analysis of food webs.


Kinetic mechanism of Clostridium perfringens phospholipase C. Hydrolysis of a thiophosphate analogue of lysophosphatidylcholine.
The hydrolysis of S-[2-(hexadecanoyloxy)ethyl]thiophosphocholine (I), an analogue of lysophosphatidylcholine, by Clostridium perfringens phospholipase C, was followed at pH 7.5, 37 degrees C and I 1.0 (maintained with KCl), in a continuous assay, by monitoring the reduction of 5,5'-dithiobis-(2-nitrobenzoic acid) at 412 nm. Simple saturation kinetics are observed with linear mixed-type slope-intercept effects for the hydrolysis of compound (I) with variable [Ca2+] at fixed concentrations of compound (I) and a simple slope effect as [compound (I)] is varied at fixed concentrations of Ca2+. These data are consistent with a simple ordered rapid-equilibrium mechanism in which Ca2+ binds to the enzyme first followed by substrate. The observed kinetic constants at pH 7.5, 37 degrees C and I 1.0 are K1 = 12.0 mM (Ca2+ dissociation), K2 = 36 microM [compound (I) dissociation] and Vmax. = 552 microM.min-1.mg-1. Alkane diammonium salts inhibit the enzyme by a non-competitive mechanism that involves binding to free enzyme, E.Ca2+ and E.Ca2+.S. The use of the simple micellarized substrate under these conditions allows the determination of kinetic and inhibition constants without complications arising from enzyme-micelle interactions.


Sample Preparation for Probe Electrospray Ionization Mass Spectrometry.
Mass spectrometry (MS) is a powerful tool in analytical chemistry because it provides very accurate information about molecules, such as mass-to-charge ratios (m/z), which are useful to deduce molecular weights and structures. While it is essentially a destructive analytical method, recent advancements in the ambient ionization technique have enabled us to acquire data while leaving tissue in a relatively intact state in terms of integrity. Probe electrospray ionization (PESI) is a so-called direct method because it does not require complex and time-consuming pretreatment of samples. A fine needle serves as a sample picker, as well as an ionization emitter. Based on the very sharp and fine property of the probe tip, destruction of the samples is minimal, allowing us to acquire the real-time molecular information from living things in situ. Herein, we introduce three applications of PESI-MS technique that will be useful for biomedical research and development. One involves the application to solid tissue, which is the basic application of this technique for the medical diagnosis. As this technique requires only 10 mg of the sample, it may be very useful in the routine clinical settings. The second application is for in vitro medical diagnostics where human blood serum is measured. The ability to measure fluid samples is also valuable in various biological experiments where a sufficient volume of sample for conventional analytical techniques cannot be provided. The third application leans toward the direct application of probe needles in living animals, where we can obtain real-time dynamics of metabolites or drugs in specific organs. In each application, we can infer the molecules that have been detected by MS or use artificial intelligence to obtain a medical diagnosis.


Management of severe perioperative bleeding: guidelines from the European Society of Anaesthesiology: First update 2016.
: The management of perioperative bleeding involves multiple assessments and strategies to ensure appropriate patient care. Initially, it is important to identify those patients with an increased risk of perioperative bleeding. Next, strategies should be employed to correct preoperative anaemia and to stabilise macrocirculation and microcirculation to optimise the patient's tolerance to bleeding. Finally, targeted interventions should be used to reduce intraoperative and postoperative bleeding, and so prevent subsequent morbidity and mortality. The objective of these updated guidelines is to provide healthcare professionals with an overview of the most recent evidence to help ensure improved clinical management of patients. For this update, electronic databases were searched without language restrictions from 2011 or 2012 (depending on the search) until 2015. These searches produced 18 334 articles. All articles were assessed and the existing 2013 guidelines were revised to take account of new evidence. This update includes revisions to existing recommendations with respect to the wording, or changes in the grade of recommendation, and also the addition of new recommendations. The final draft guideline was posted on the European Society of Anaesthesiology website for four weeks for review. All comments were collated and the guidelines were amended as appropriate. This publication reflects the output of this work.


Early appearance of activated matrix metalloproteinase-9 and blood-brain barrier disruption in mice after focal cerebral ischemia and reperfusion.
Blood-brain barrier (BBB) disruption is thought to play a critical role in the pathophysiology of ischemia/reperfusion. Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes that can degrade all the components of the extracellular matrix when they are activated. Gelatinase A (MMP-2) and gelatinase B (MMP-9) are able to digest the endothelial basal lamina, which plays a major role in maintaining BBB impermeability. The present study examined the expression and activation of gelatinases before and after transient focal cerebral ischemia (FCI) in mice. Adult male CD1 mice were subjected to 60 min FCI and reperfusion. Zymography was performed from 1 to 23 h after reperfusion using the protein extraction method with detergent extraction and affinity-support purification. MMP-9 expression was also examined by both immunohistochemistry and Western blot analysis, and tissue inhibitors to metalloproteinase-1 was measured by reverse zymography. The BBB opening was evaluated by the Evans blue extravasation method. The 88-kDa activated MMP-9 was absent from the control specimens, while it appeared 3 h after transient ischemia by zymography. At this time point, the BBB permeability alteration was detected in the ischemic brain. Both pro-MMP-9 (96 kDa) and pro-MMP-2 (72 kDa) were seen in the control specimens, and were markedly increased after FCI. A significant induction of MMP-9 was confirmed by both immunohistochemistry and Western blot analysis. The early appearance of activated MMP-9, associated with evidence of BBB permeability alteration, suggests that activation of MMP-9 contributes to the early formation of vasogenic edema after transient FCI.


The relationship between posttraumatic stress disorder, illness cognitions, defence styles, fatigue severity and psychological well-being in chronic fatigue syndrome.
This study investigated, firstly, the rate of posttraumatic stress disorder (PTSD) and the level of psychological well-being amongst people with chronic fatigue syndrome (CFS); and secondly, the extent to which illness cognitions, defence styles and PTSD symptom severity related to fatigue severity and psychological well-being. Seventy-eight participants with a diagnosis of CFS completed the Chalder Fatigue Scale, the General Health Questionnaire-28, the Posttraumatic Stress Diagnostic Scale, the Illness Cognition Questionnaire and the Defence Style Questionnaire. Fifty-nine participants were recruited from the general public to form the non-fatigued control group. CFS participants had significantly higher levels of PTSD symptoms, lower levels of psychological well-being and more traumatic life events compared to the non-fatigued controls. Trauma exposure and PTSD severity both predicted CFS status. However, regression analyses demonstrated no significant relationship between PTSD symptoms and fatigue severity or the degree of psychological well-being. 'Helplessness' predicted both physical and mental fatigue and psychological well-being, whilst the 'mature' defence styles predicted fatigue severity only. The results offer support to previous research showing that the rate of traumatic life events and PTSD are significantly higher amongst the CFS population. The lack of relationship between PTSD symptoms and fatigue severity or psychological well-being indicates that these processes may operate independently of one another, via different appraisal processes. This study focused on fatigue severity, but it may be that the role of pain in CFS is a key element in the previously reported association between PTSD and CFS.


[Concentrations of Acidic Gases, Ammonia and Related Water-Soluble Ions in PM2.5 and Gas-Particle Partitioning in Qingdao].
Fifty-eight samples were collected in Qingdao from November to December 2012 using a PM2.5 sampler. Concentrations of acidic gases, ammonia and their partner ions in PM2.5 were determined, and the correlation between concentrations and the gas-particle partitioning were analyzed. The averaged concentrations of HNO3, HNO2, HCl, SO2 and NH, in atmosphere samples of Qingdao were 1. 36, 1. 64, 1. 46, 27. 58 and 2. 95 µg.m-3, respectively, for the concentrations of NO3-, NO2, Cl-, SO(4)2- and NH: in PM2, were 6. 49, 0. 12, 1. 95, 9. 36 and 6. 04 µg.m-3, respectively. When the measurements made in haze days, compared to those made in clear days, the concentration of these ions in PM2.5 increased more than those of acidic gases and ammonia; In foggy days, the concentrations of gases were lower than those in clear days except HNO2, while the reverse was true for the concentrations of all particulate species in PM2.5. During whole study period, the NH3 level was high and the NH4+ in PM2.s in Qingdao was mainly present as (NH4)2SO4. The analysis results suggested that formation of N03 were mainly determined by HNO3. To further explore the control factors of these species in PM2.5, the thermodynamic equilibrium model of ISORROPIA I was employed. Our sensitivity tests showed that the formation of NH4+ was less sensitive to the change of total NH3 (TNH3), but sensitive to the changes of total H2 SO4(TSO4) and total HNO3 (TNO3). The formation of NO3- and SO(4)2- was sensitive to the changes of TNO3 and TSO4, respectively. Taken together, our findings implied that the reduction of TNO3 and TSO4 rather than TNH3 appears to be more effective in lowering mass concentrations of PM2.5 in Qingdao.


Effects of oligomycin on glucose utilization and calcium transport in African trypanosomes.
Oligomycin (3 microgram ml-1) inhibited glucose utilization in Trypanosoma brucei S42 as shown by measurements of oxygen uptake and pyruvate production. Carbonyl cyanide 3-chlorophenylhydrazone, an uncoupler of oxidative phosphorylation, did not relieve this inhibition, although some relief was afforded by the alternative substrate glycerol. Naturally dyskinetoplastic Trypanosoma evansi MIAG 105 was less sensitive to inhibition by oligomycin although glycerol relief was still observed, relecting the differential sensitivity of the two pathways. With glucose present as the substrate, 45Ca2+ transport was inhibited by oligomycin in T. brucei, but was stimulated in T. evansi. These results are discussed in terms of alternative systems for maintaining cytoplasmic Ca2+ concentrations in normal and dyskinetoplastic strains of trypanosome.


Conditional reprogramming culture conditions facilitate growth of lower-grade glioma models.
The conditional reprogramming cell culture method was developed to facilitate growth of senescence-prone normal and neoplastic epithelial cells, and involves co-culture with irradiated fibroblasts and the addition of a small molecule Rho kinase (ROCK) inhibitor. The aim of this study was to determine whether this approach would facilitate the culture of compact low-grade gliomas. We attempted to culture 4 pilocytic astrocytomas, 2 gangliogliomas, 2 myxopapillary ependymomas, 2 anaplastic gliomas, 2 difficult-to-classify low-grade neuroepithelial tumors, a desmoplastic infantile ganglioglioma, and an anaplastic pleomorphic xanthoastrocytoma using a modified conditional reprogramming cell culture approach. Conditional reprogramming resulted in robust increases in growth for a majority of these tumors, with fibroblast conditioned media and ROCK inhibition both required. Switching cultures to standard serum containing media, or serum-free neurosphere conditions, with or without ROCK inhibition, resulted in decreased proliferation and induction of senescence markers. Rho kinase inhibition and conditioned media both promoted Akt and Erk1/2 activation. Several cultures, including one derived from a NF1-associated pilocytic astrocytoma (JHH-NF1-PA1) and one from a BRAF p.V600E mutant anaplastic pleomorphic xanthoastrocytoma (JHH-PXA1), exhibited growth sufficient for preclinical testing in vitro. In addition, JHH-NF1-PA1 cells survived and migrated in larval zebrafish orthotopic xenografts, while JHH-PXA1 formed orthotopic xenografts in mice histopathologically similar to the tumor from which it was derived. These studies highlight the potential for the conditional reprogramming cell culture method to promote the growth of glial and glioneuronal tumors in vitro, in some cases enabling the establishment of long-term culture and in vivo models.


Spousal caregiving and incident hypertension.
Caring for one's spouse has been associated with poor health, including risk of cardiovascular disease (CVD) onset and mortality. However, few studies have assessed the risk of incident hypertension associated with spousal caregiving. This paper investigates this association in a large, nationally representative sample of American older adults. Married, hypertension-free, Health and Retirement Study (HRS) respondents aged 50+ in 2000, (n = 5,708) were followed up to 8 years (1,708 new self-reported hypertension diagnoses). Current caregiving exposure was defined as assisting a spouse with instrumental or basic activities of daily living (IADLs) 14+ h/week; we define providing ≥14 h/week of care at two consecutive biennial surveys as "long-term caregiving." We used inverse probability weighted discrete-time hazard models with time-updated exposure and covariates to estimate effects of current and long-term caregiving on incident hypertension. We tested for effect modification by race, gender, and recipient memory illness. Sensitivity analyses restricted to respondents whose spouses had care needs. After adjusting for demographic, socioeconomic, and health factors, (including risk behaviors, comorbid conditions, and self-rated health), current caregiving significantly predicted hypertension incidence (risk ratio (RR) = 1.36, 95% confidence interval (CI): 1.01, 1.83). For long-term caregivers, there was significant evidence of risk of hypertension onset associated with caregiving (RR = 2.29, 95% CI: 1.17, 4.49). The risk of hypertension onset associated with both current and long-term caregiving did not vary by race, gender, or recipient memory illness diagnosis. Sensitivity analyses supported the primary findings. Providing IADL care to a spouse significantly predicted hypertension onset in a nationally representative sample of US adults.


Tamoxifen retinopathy.
A 63-year-old female on long-term, high-dose tamoxifen treatment for metastatic breast cancer developed bilateral intraretinal refractile opacities, lesions at the level of the retinal pigment epithelium, and cystoid macular oedema.


Nerve growth in the absence of growth cone filopodia and the effects of a small applied electric field.
Nerve orientation may involve a biasing of the distribution of tension at the growth cone. Chemical and electrical guidance cues cause more filopodia to appear on one side of the growth cone and this may determine turning behaviour. In a small applied electric field, filopodia predominate on the cathodal side of the growth cone and nerves turn towards the cathode. Removing all filopodia by treatment with cytochalasin D did not prevent nerves from continued slow growth and nerves still oriented towards the cathode. It is concluded that nerves can perform some types of orienting behaviour in the complete absence of filopodia.


Interdisciplinary and inter-institutional differences in learning preferences among Malaysian medical and health sciences students.
The learner-centred approach in medical and health sciences education makes the study of learning preferences relevant and important. This study aimed to investigate the interdisciplinary, inter-institutional, gender and racial differences in the preferred learning styles among Malaysian medical and health sciences students in three Malaysian universities, namely SEGi University (SEGi), University of Malaya (UM) and Universiti Tunku Abdul Rahman (UTAR). It also investigated the differences in the preferred learning styles of these students between high achievers and non-high achievers. This cross-sectional study was carried out on medical and health sciences students from three Malaysian universities following the approval of the Research and Ethics Committee, SEGi University. Purposive sampling was used and the preferred learning styles were assessed using the VARK questionnaire. The questionnaire was validated prior to its use. Three disciplines (medicine, pharmacy and dentistry) were chosen based on their entry criteria and some similarities in their course structure. The three participating universities were Malaysian universities with a home-grown undergraduate entry medical program and students from a diverse cultural and socioeconomic background. The data were analysed using the Statistical Package for the Social Sciences (SPSS) software, version 22. VARK subscale scores were expressed as mean+standard deviation. Comparisons of the means were carried out using t-test or ANOVA. A p value of <0.05 was considered as statistically significant, and <0.001 as highly significant. Both statistically significant interdisciplinary and inter-institutional differences in learning preferences were observed. Out of the 337 students, a majority of the participants were unimodal learners (n=263, 78.04%). The most common type of learners was the reading/writing type (n=92, 27.30%) while the kinesthetic subscale (M=6.98, SD=2.85) had the highest mean score. Female students (M=6.86, SD=2.86) scored significantly higher than male students (M=6.08, SD=2.41; t(249), p=0.014) in the auditory subscale, whereas Chinese students (M=5.87, SD=2.65) scored significantly higher than Malay students (M=4.70, SD=2.87; p=0.04) in the visual subscale. However, the mean VARK subscale scores did not differ significantly between high achievers and non-high achievers (p>0.05). This study gives an insight into the learner characteristics of more than one medical school in Malaysia. Such multi-institutional studies are lacking in the published literature and this study gives a better representation of the current situation in the learning preferences among medical students in Malaysia.


Ectopic over expression of kiss1 may compensate for the loss of kiss2.
Kisspeptin (KISS) is a neuropeptide which plays a central role in the regulation of the hypothalamic-pituitary-gonadal axis, and is essential for sexual maturation and fertility in mammals. Unlike mammals, which possess only one KISS gene, two paralogous genes, kiss1 and kiss2, have been identified in zebrafish and other non-mammalian vertebrates. Previous studies suggest that Kiss2, but not Kiss1, is the reproduction relevant form amongst the two. To better understand the role of each of these isoforms in reproduction, a loss of function approach was applied. Two genetic manipulation techniques-clustered regularly interspaced short palindromic repeats (CRISPR) and transcription activator-like effector nucleases (TALEN)-were used to generate kiss1 and kiss2 knockout (KO) zebrafish lines, respectively. Examination of these KO lines showed that reproductive capability was not impaired, confirming earlier observations. Further analysis revealed that KO of kiss2 caused a significant increase in expression levels of kiss1, kiss2r and tac3a, while KO of kiss1 had no effect on the expression of any of the examined genes. In situ hybridization analysis revealed that kiss1 mRNA is expressed only in the habenula in wild type brains, while in kiss2 KO fish, kiss1 mRNA-expressing cells were identified also in the ventral telencephalon, the ventral part of the entopeduncular nucleus, and the dorsal and ventral hypothalamus. Interestingly, these regions are known to express kiss2r, and the ventral hypothalamus normally expresses kiss2. These results suggest that a compensatory mechanism, involving ectopic kiss1 expression, takes place in the kiss2 KO fish, which may substitute for Kiss2 activity.


Striatal 6-[18F]fluorodopa accumulation after combined inhibition of peripheral catechol-O-methyltransferase and monoamine oxidase type B: differing response in relation to presynaptic dopaminergic dysfunction.
The aim was to investigate the effects of inhibition of monoamine oxidase type B (MAO-B) with selegiline alone and the combined inhibition of peripheral catechol-O-methyltransferase (COMT) with entacapone and MAO-B with selegiline on striatal 6-[18F]fluorodopa (FDOPA) accumulation, and whether the effect of entacapone + selegiline on FDOPA uptake differed depending on the severity of the presynaptic dopaminergic dysfunction. Thus, eight healthy controls, eight de novo patients with Parkinson's disease (PD), and 18 levodopa-treated PD patients were investigated with positron emission tomography (PET). Half of the subjects in each population belonged to the selegiline group and half to the entacapone + selegiline group. Both groups were studied twice with PET using FDOPA. After the first (baseline) FDOPA PET investigation, both groups were on 2 weeks of selegiline treatment, 10 mg daily. Thereafter, the second FDOPA PET was performed for all subjects with a premedication administered 60 min before the PET imaging; one group received 10 mg of selegiline, and the other group received a single 400 mg dose of entacapone coadministered with 10 mg of selegiline. Selegiline treatment alone had no significant influence on striatal FDOPA metabolism. The FDOPA accumulation, expressed as striatal-to-occipital ratios and modified decarboxylation coefficients (k3R0), increased significantly after entacapone + selegiline administration in all subject populations. The FDOPA uptake rate constant (Ki) remained virtually unchanged in controls and in de novo patients but decreased significantly in levodopa-treated PD patients after entacapone + selegiline intake. Entacapone + selegiline administration did not influence significantly the unidirectional blood-to-brain clearance for FDOPA (K1D) or the relative dopadecarboxylase activity (k3D). The changes in the studied parameters after entacapone + selegiline administration probably reflect the effects of entacapone, since entacapone alone has caused similar changes in previous PET studies. Response in FDOPA accumulation to entacapone + selegiline was higher in controls and de novo patients compared with levodopa-treated PD patients. The milder response in levodopa-treated patients might reflect the reduced ability of the degenerated dopaminergic neurons to utilize the prolonged FDOPA availability, produced by entacapone.


Peptostreptococcus stomatis sp. nov., isolated from the human oral cavity.
Seven strains of anaerobic Gram-positive cocci isolated from human oral sites were subjected to a comprehensive range of phenotypic and genotypic tests. 16S rRNA gene sequence analysis revealed that the strains constituted a homogeneous group that was distinct from species with validly published names, but related to Peptostreptococcus anaerobius. All oral strains tested belonged to this group, whereas all non-oral strains studied were confirmed as P. anaerobius. A novel species, Peptostreptococcus stomatis sp. nov., is proposed to accommodate these oral strains. P. stomatis is weakly saccharolytic and produces acetic, butyric, isobutyric, isovaleric and isocaproic acids as end products of fermentation. The type strain of P. stomatis is W2278T (=DSM 17678T = CCUG 51858T); the G+C content of the DNA of this strain is 36 mol%.


CD105 positive neovessels are prevalent in early stage carotid lesions, and correlate with the grade in more advanced carotid and coronary plaques.
Previous studies have demonstrated that expression of CD105 is a sensitive marker and indicator of endothelial cell/microvessel activation and proliferation in aggressive solid tumour growth and atherosclerotic plaque lesions. Since intimal neovascularization contributes significantly to subsequent plaque instability, haemorrhage and rupture. We have used immunohistochemical analysis to investigate the expression of CD105-positive vessels in both large (carotid) and medium calibre (coronary and middle cerebral artery, MCAs) diseased vessels in an attempt to identify any correlation with plaque growth, stage and complication/type. Here we show, that carotid arteries expressed intimal neovascularization associated with CD105-positive endothelial cells, concomitant with increased inflammation in early stage lesions, preatheroma (I-III) whilst they were not present in coronary plaques of the same grade. Some of these CD105-positive neovessels were immature, thin walled and without smooth muscle cell coverage making them more prone to haemorrhage and rupture. In high-grade lesions, neovessel proliferation was similar in both arterial types and significantly higher numbers of CD105-positive vasa vasorum were associated with plaque regions in coronary arteries. In contrast, although the MCAs exhibited expanded intimas and established plaques, there were very few CD105 positive neovessels. Our results show that CD105 is a useful marker of angiogenesis within adventitial and intimal vessels and suggest the existence of significant differences in the pathological development of atherosclerosis in separate vascular beds which may have important consequences when considering management and treatment of this disease.


Systemic inflammation and lung function: A longitudinal analysis.
Systemic inflammation is associated with impaired lung function in healthy adults as well as in patients with lung disease. The mechanism for this association is unknown and it is unclear if systemic inflammation leads to impaired lung function or if poor lung function leads to inflammation. We explored the temporal associations between blood C-reactive protein (CRP), fibrinogen, and white blood cells, and lung function in young adults. Spirometry, plethysmography, and diffusion capacity were measured in a population-based cohort at ages 32 and 38 years. High-sensitivity CRP, fibrinogen, and white blood cells were measured at the same ages. Higher levels of CRP and, to a lesser extent, fibrinogen were associated with lower lung volumes in cross-sectional analyses at both ages 32 and 38 years. Higher CRP and fibrinogen at age 32 were associated with higher FEV1 and FEV1/FVC at age 38, but not other measures of lung function. Lower lung volumes (total lung capacity, functional residual capacity, and residual volume) but not airflow obstruction (FEV1/FVC) at age 32 were associated with higher CRP at age 38. Associations between age 32 lung function and fibrinogen at follow-up were weaker, but consistent. There were no longitudinal associations between white blood cells and lung function. We found no evidence that systemic inflammation causes a decline in lung function. However, lower lung volumes were associated with higher CRP and fibrinogen at follow-up indicating that pulmonary restriction may be a risk factor for systemic inflammation. The mechanism for this association remains unclear.


A Bayesian kriging model for estimating residential exposure to air pollution of children living in a high-risk area in Italy.
A core challenge in epidemiological analysis of the impact of exposure to air pollution on health is assessment of the individual exposure for subjects at risk. Geographical information systems (GIS)-based pollution mapping, such as kriging, has become one of the main tools for evaluating individual exposure to ambient pollutants. We applied universal Bayesian kriging to estimate the residential exposure to gaseous air pollutants for children living in a high-risk area (Milazzo- Valle del Mela in Sicily, Italy). Ad hoc air quality monitoring campaigns were carried out: 12 weekly measurements for sulphur dioxide (SO2) and nitrogen dioxide (NO2) were obtained from 21 passive dosimeters located at each school yard of the study area from November 2007 to April 2008. Universal Bayesian kriging was performed to predict individual exposure levels at each residential address for all 6- to 12-years-old children attending primary school at various locations in the study area. Land use, altitude, distance to main roads and population density were included as covariates in the models. A large geographical heterogeneity in air quality was recorded suggesting complex exposure patterns. We obtained a predicted mean level of 25.78 (± 10.61) µg/m(3) of NO2 and 4.10 (± 2.71) µg/m(3) of SO2 at 1,682 children's residential addresses, with a normalised root mean squared error of 28% and 25%, respectively. We conclude that universal Bayesian kriging approach is a useful tool for the assessment of realistic exposure estimates with regard to ambient pollutants at home addresses. Its prediction uncertainty is highly informative and can be used for both designing subsequent campaigns and for improved modelling of epidemiological associations.


Development of white matter fiber covariance networks supports executive function in youth.
During adolescence, the brain undergoes extensive changes in white matter structure that support cognition. Data-driven approaches applied to cortical surface properties have led the field to understand brain development as a spatially and temporally coordinated mechanism that follows hierarchically organized gradients of change. Although white matter development also appears asynchronous, previous studies have relied largely on anatomical tract-based atlases, precluding a direct assessment of how white matter structure is spatially and temporally coordinated. Harnessing advances in diffusion modeling and machine learning, we identified 14 data-driven patterns of covarying white matter structure in a large sample of youth. Fiber covariance networks aligned with known major tracts, while also capturing distinct patterns of spatial covariance across distributed white matter locations. Most networks showed age-related increases in fiber network properties, which were also related to developmental changes in executive function. This study delineates data-driven patterns of white matter development that support cognition.


Increased prevalence of ECG markers for sudden cardiac arrest in refractory epilepsy.
People with epilepsy are at increased risk of sudden cardiac arrest (SCA) due to ECG-confirmed ventricular tachycardia/fibrillation, as seen in a community-based study. We aimed to determine whether ECG-risk markers of SCA are more prevalent in people with epilepsy. In a cross-sectional, retrospective study, we analysed the ECG recordings of 185 people with refractory epilepsy and 178 controls without epilepsy. Data on epilepsy characteristics, cardiac comorbidity, and drug use were collected, and general ECG variables (heart rate (HR), PQ and QRS intervals) assessed. We analysed ECGs for three markers of SCA risk: severe QTc prolongation (male >450 ms, female >470 ms), Brugada ECG pattern, and early repolarisation pattern (ERP). Multivariate regression models were used to analyse differences between groups, and to identify associated clinical and epilepsy-related characteristics. People with epilepsy had higher HR (71 vs 62 bpm, p<0.001) and a longer PQ interval (162.8 vs 152.6 ms, p=0.001). Severe QTc prolongation and ERP were more prevalent in people with epilepsy (QTc prolongation: 5% vs 0%; p=0.002; ERP: 34% vs 13%, p<0.001), while the Brugada ECG pattern was equally frequent in both groups (2% vs 1%, p>0.999). After adjustment for covariates, epilepsy remained associated with ERP (ORadj 2.4, 95% CI 1.1 to 5.5) and severe QTc prolongation (ORadj 9.9, 95% CI 1.1 to 1317.7). ERP and severe QTc prolongation appear to be more prevalent in people with refractory epilepsy. Future studies must determine whether this contributes to increased SCA risk in people with epilepsy.


Phase II study of targeted therapy with temozolomide in acute myeloid leukaemia and high-risk myelodysplastic syndrome patients pre-screened for low O(6) -methylguanine DNA methyltransferase expression.
Resistance to temozolomide is largely mediated by the DNA repair enzyme O(6) -methylguanine DNA methyltransferase (MGMT). We conducted a prospective multicentre study of patients with previously untreated acute myeloid leukaemia (AML) or high-risk myelodysplastic syndrome (MDS) who were not candidates for intensive therapy. Patient selection was based on MGMT expression by Western blot. Patients with MGMT:ACTB (β-actin) ratio <0·2 were eligible to receive temozolomide 200 mg/m(2) /d ×7 d. Patients achieving a complete response (CR) could receive up to 12 monthly cycles of temozolomide ×5/28 d. Of 166 patients screened, 81 (49%) demonstrated low MGMT expression; 45 of these were treated with temozolomide. The overall response rate was 53%; 36% achieved complete clearance of blasts, with 27% achieving a CR/CR with incomplete platelet recovery (CRp). Factors associated with a trend toward a higher response rate included MDS, methylated MGMT promoter and standard cytogenetic risk group. Induction and post-remission cycles were well-tolerated and most patients were treated on an outpatient basis. Patient who achieved CR/CRp had a superior overall survival compared to partial or non-responders. In conclusion, targeted therapy based on pre-selection for low MGMT expression was associated with a higher response rate to temozolomide compared to previous reports of unselected patients.


A power fallacy.
The power fallacy refers to the misconception that what holds on average -across an ensemble of hypothetical experiments- also holds for each case individually. According to the fallacy, high-power experiments always yield more informative data than do low-power experiments. Here we expose the fallacy with concrete examples, demonstrating that a particular outcome from a high-power experiment can be completely uninformative, whereas a particular outcome from a low-power experiment can be highly informative. Although power is useful in planning an experiment, it is less useful-and sometimes even misleading-for making inferences from observed data. To make inferences from data, we recommend the use of likelihood ratios or Bayes factors, which are the extension of likelihood ratios beyond point hypotheses. These methods of inference do not average over hypothetical replications of an experiment, but instead condition on the data that have actually been observed. In this way, likelihood ratios and Bayes factors rationally quantify the evidence that a particular data set provides for or against the null or any other hypothesis.


A modified Jones test: lacrimal scintigram correlation.
To determine the correlation between lacrimal scintigraphy and a modified Jones primary dye drainage test combined with the fluorescein dye disappearance test (DDT) and Jones secondary dye test, in diagnosing the cause of epiphora. A prospective, consecutive clinical trial was conducted. All patients presenting with epiphora were considered, but those with trichiasis, corneal irritation, lower eyelid ectropion, Bell palsy, or previous lacrimal surgery were excluded. A cohort of 39 patients (65 eyes) underwent the fluorescein DDT and the senior author's (R.B.) modified Jones primary dye drainage test (Jones dye test 1 [JDT1]). For this test, an entire fluorescein 2% Minim dose was instilled in the 2 eyes, and the patient was then asked to gently blow each nostril separately in a white tissue; the degree of fluorescein staining was noted. If there was no fluorescein staining of the tissue (negative JDT1), a traditional Jones secondary dye test (Jones dye test 2) was also performed. All patients except for those testing strongly positive on modified testing or with negative DDT and JDT1 and nil drainage from the nose in Jones dye test 2, indicating anatomical obstruction, were referred for lacrimal scintigraphy. In total, 59 of 65 eyes (91%) showed a positive correlation between the diagnosis arrived at after dye testing using the modified primary dye test and diagnosis on lacrimal scintigraphy. The interrater reliability for the raters was found to be κ = 0.770 and 95% confidence interval = 0.594-0.945. The strength of agreement is considered to be "good." The authors found that in the presence of patent lacrimal drainage system dysfunction, modified Jones primary dye test, combined with the fluorescein DDT and Jones secondary dye test, correlated strongly with lacrimal scintigraphy.


When can refuges mediate the genetic effects of fire regimes? A simulation study of the effects of topography and weather on neutral and adaptive genetic diversity in fire-prone landscapes.
Understanding how landscape heterogeneity mediates the effects of fire on biodiversity is increasingly important under global changes in fire regimes. We used a simulation experiment to investigate how fire regimes interact with topography and weather to shape neutral and selection-driven genetic diversity under alternative dispersal scenarios, and to explore the conditions under which microrefuges can maintain genetic diversity of populations exposed to recurrent fire. Spatial heterogeneity in simulated fire frequency occurred in topographically complex landscapes, with fire refuges and fire-prone "hotspots" apparent. Interannual weather variability reduced the effect of topography on fire patterns, with refuges less apparent under high weather variability. Neutral genetic diversity was correlated with long-term fire frequency under spatially heterogeneous fire regimes, being higher in fire refuges than fire-prone areas, except under high dispersal or low fire severity (low mortality). This generated different spatial genetic structures in fire-prone and fire-refuge components of the landscape, despite similar dispersal. In contrast, genetic diversity was only associated with time since the most recent fire in flat landscapes without predictable refuges and hotspots. Genetic effects of selection driven by fire-related conditions depended on selection pressure, migration distance and spatial heterogeneity in fire regimes. Allele frequencies at a locus conferring higher fitness under successional environmental conditions followed a pattern of "temporal adaptation" to contemporary conditions under strong selection pressure and high migration. However, selected allele frequencies were correlated with spatial variation in long-term mean fire frequency (relating to environmental predictability) under weak dispersal, low selection pressure and strong spatial heterogeneity in fire regimes.


[Differences in the mutagenic effectiveness of N-nitrosomethyl- and N-nitrosoethylurea acting on bacterial DNA in vivo].
Chemically induced lethals and reversion frequencies of two mutants of Salmonella typhimurium LT-2 1530 and 1532 were examined. Prototrophic mutants to histidine independence occurred by two different mechanisms and were induced by nitrous acid, hydroxylamine, N-nitrosomethylurea (NMU), N-nitrosoethylurea (NEU) and N, N, nitrosodimethylurea (NDMU). The ability of the mutants to revert was affected by the chemical structure (in the row of N-nitrosoalkylurea - by alkyl group) of the mutagen. NEU is found to induce in bacterial DNA only base pair substitutions; NMU and NDMU - deletions and base pair substitutions. An original model of deletion mechanism with Goldwait endonuclease II participation is proposed.


Maximal growth occurs at a broad range of essential amino acids to total nitrogen ratios in kittens.
Kittens fed diets containing 2.0 and 3.0 times (x) the NRC (1986) essential amino acid (EAA) requirement (EAArq) and 210 to 560 g crude protein (CP)/kg diet had growth rates and plasma amino acid patterns that were not significantly different than kittens fed a control diet (CD) containing 1.5 x EAArq and 350 g CP/kg diet. Growth rates of kittens fed diets containing only EAA (with nontoxic levels of arginine and methionine) and 280 to 460 g CP/kg diet were equivalent to those of kittens fed CD. Kittens fed only EAA and 140 and 210 g CP/kg diet had growth rates that were significantly lower than kittens fed CD. Since the growth rate of kittens fed 1.5 x EAArq and 210 g CP/kg diet in a previous experiment was equivalent to kittens fed CD (Taylor et al., 1997), it is suggested that the requirement for CP is higher (up to 280 g CP/kg diet) when only EAA are fed. The higher crude protein requirement appears to be primarily a consequence of the high obligatory nitrogen loss as urea (especially from arginine) incurred in the conversion of nitrogen from EAA to dispensable amino acids in the liver and secondarily because of a slow rate of catabolism of the EAA. A 3-dimensional plot of weight gains vs. CP levels and EAA to total nitrogen (E:T) ratios of kittens shows a broad range of CP levels and E:T ratios that support optimal growth in the kitten. It is suggested that similar patterns would occur in the chick, rat and other species if adverse effects caused by excesses of specific amino acids are avoided.


Identification of nonpeptide CCR5 receptor agonists by structure-based virtual screening.
A three-dimensional model of the chemokine receptor CCR5 has been built to fulfill structural peculiarities of its alpha-helix bundle and to distinguish known CCR5 antagonists from randomly chosen drug-like decoys. In silico screening of a library of 1.6 million commercially available compounds against the CCR5 model by sequential filters (drug-likeness, 2-D pharmacophore, 3-D docking, scaffold clustering) yielded a hit list of 59 compounds, out of which 10 exhibited a detectable binding affinity to the CCR5 receptor. Unexpectedly, most binders tested in a functional assay were shown to be agonists of the CCR5 receptor. A follow-up database query based on similarity to the most potent binders identified three new CCR5 agonists. Despite a moderate affinity of all nonpeptide ligands for the CCR5 receptor, one of the agonists was shown to promote efficient receptor internalization, which is a process therapeutically favorable for protection against HIV-1 infection.


Active optical depth resolution improvement of the laser tomographic scanner.
Laser scanning tomography can be used to assess retinal nerve fiber layer thickness and optic disc topography of the human eye. A pinhole is located at a plane conjugate to the focal plane of the scanning laser beam. This so-called confocal configuration assures that only the light originating from the illuminated focal plane on the retina passes through the pinhole and is detected by the photomultiplier. Consequently, images with high spatial resolution in all directions are obtained. An active optical system (active mirror) further improves the lateral/depth resolution of the laser tomographic scanner. By partially compensating for the optical aberrations introduced by the cornea and lens, the active optical system allows the illuminating beam to be enlarged to 6 mm, thus improving depth resolution twofold.


Dynamic Chemistry of Disulfide Terminated Oligonucleotides in Duplexes and Double-Crossover Tiles.
Designed nanostructures formed by self-assembly of multiple DNA strands suffer from low stability at elevated temperature and under other denaturing conditions. Here, we propose a method for covalent coupling of DNA strands in such structures by the formation of disulfide bonds; this allows disassembly of the structure under reducing conditions. The dynamic chemistry of disulfides and thiols was applied to crosslink DNA strands with terminal disulfide modifications. The formation of disulfide-linked DNA duplexes consisting of three strands is demonstrated, as well as a more-complex DNA double-crossover tile. All the strands in the fully disulfide-linked structures are covalently and geometrically interlocked, and it is demonstrated that the structures are stable under heating and in the presence of denaturants. Such a reversible system can be exploited in applications where higher DNA stability is needed only temporarily, such as delivery of cargoes to cells by DNA nanostructures.


Cost-effectiveness of screening strategies for Gonorrhea among females in private sector care.
To identify the optimal screening algorithm for gonorrhea infection among females in private sector care, using cost-effectiveness analysis. We compared 6 strategies using decision analysis for urine nucleic acid amplification testing for gonorrhea testing in a theoretical cohort of 10,000 females aged 15-35 years: 1) screen women aged younger than 25 years; 2) screen women aged younger than 30 years; 3) screen women aged younger than 25 years who report any risk (pregnant, drug use, new sexual partner < 30 days); 4) screen women aged younger than 30 years who report any risk; 5) screen women aged younger than 25 years or those who report any risk; and 6) screen women aged younger than 30 years or those who report any risk. Infection prevalence and sensitivity and specificity were by direct observation from a retrospective cohort of females attending the Baltimore City Sexually Transmitted Disease Clinics between 1999 and 2002. The main outcome measures were untreated gonorrhea cases and their sequelae in women, transmission to a male partner, congenital outcomes, and cost to prevent a case. Prevalence of gonorrhea was 3.0%. Not screening would result in 300 untreated cases. Not screening was cost-saving over all screening strategies. Strategy 6 resulted in the fewest cases of untreated infection (82), although Strategy 3 was the most cost-effective of the screening strategies. Univariate sensitivity analysis identified a threshold of 4.75% gonorrhea prevalence, more than which Strategy 3 became cost-saving over not screening. Screening is recommended for females aged younger than 25 years with specific risks in populations with a gonorrhea prevalence of 4.75% or greater. II-2.


Dynamic contrast-enhanced magnetic resonance imaging measurements in renal cell carcinoma: effect of region of interest size and positioning on interobserver and intraobserver variability.
The purpose of this study was to assess the influence of region of interest (ROI) size and positioning on perfusion and permeability parameters as well as on interobserver and intraobserver variability of dynamic contrast-enhanced (DCE-MRI) of primary renal cell carcinoma (RCC) and metastases. Thirty-nine DCE-MRI examinations of 34 patients with primary RCC and 20 examinations of 9 patients with RCC metastases obtained at 1.5 T were evaluated. Pretreatment and posttreatment analysis with antiangiogenic therapy was performed in 4 patients with primary RCCs and 5 patients with metastases. The ROIs of the whole tumor (wROI), the circular edge (cROI), a user-defined arbitrary small region (sROI), and a semiautomated segmented ROI were independently defined by 2 readers on 1 slice on arterial phase DCE-MRI images or on parametric plasma-flow maps. Analysis with a 2-compartment exchange model provided 4 parameters: plasma flow (FP), plasma volume (vp), permeability-surface product (PS), and extravascular-extracellular volume (ve). Interobserver and intraobserver parameter correlations were calculated using the intraclass correlation coefficient, and within-subject variability were considered on the basis of the coefficient of variation. Differences in measurement values of variable ROI size were assessed with paired t test. Mean values of FP and vp with sROIs were significantly higher than those with wROI, cROI, and semiautomated segmented ROI placement in tumor or metastases. Values of ve showed no significant difference between ROI sizes. The highest interobserver and intraobserver correlation with 0.99/0.98 for metastases and 0.97/0.98 for primary RCCs, respectively, was observed for all parameters when defining wROIs on dynamic images. Perfusion parameters of wROI measurements for FP (dynamic, 0.97; parametric maps, 0.96) and vp (0.95/0.89) showed higher interobserver correlation than did permeability parameters ve (0.64/0.6) and PS (0.79/0.5) in primary RCCs. The wROIs showed also the lowest within-subject coefficients of variation for perfusion parameters FP and vp compared with cROI and sROIs in primary RCCs and metastases. The ROI size and positioning do substantially influence quantitative perfusion and permeability parameters in DCE-MRI. The best interobserver and intraobserver correlation can be obtained when defining a whole-tumor ROI. The perfusion parameters are the most reliable, whereas the permeability parameters are more susceptible to interobserver variability. No significant differences between placing ROIs on morphological or parametric images were observed.