Abortion surveillance--United States, 2001.
CDC began abortion surveillance in 1969 to document the number and characteristics of women obtaining legal induced abortions. This report summarizes and describes data voluntarily reported to CDC regarding legal induced abortions obtained in the United States in 2001. For each year since 1969, CDC has compiled abortion data by state or area of occurrence. During 1973-1997, data were received from or estimated for 52 reporting areas in the United States: 50 states, the District of Columbia, and New York City. In 1998 and 1999, CDC compiled abortion data from 48 reporting areas. Alaska, California, New Hampshire, and Oklahoma did not report, and data for these states were not estimated. In 2000 and 2001, Oklahoma again reported these data, increasing the number of reporting areas to 49. A total of 853,485 legal induced abortions were reported to CDC for 2001 from 49 reporting areas, representing a 0.5% decrease from the 857,475 legal induced abortions reported by the same 49 reporting areas for 2000. The abortion ratio, defined as the number of abortions per 1,000 live births, was 246 in 2001, compared with 245 reported for 2000. This represents a 0.4% increase in the abortion ratio. The abortion rate was 16 per 1,000 women aged 15-44 years for 2001, the same as for 2000. For both the 48 and 49 reporting areas, the abortion rate remained relatively constant during 1997-2001. The highest percentages of reported abortions were for women who were unmarried (82%), white (55%) and aged <25 years (52%). Of all abortions for which gestational age was reported, 59% were performed at < or =8 weeks' gestation and 88% at <13 weeks. From 1992 (when detailed data regarding early abortions were first collected) through 2001, steady increases have occurred in the percentage of abortions performed at < or =6 weeks' gestation. A limited number of abortions were obtained at >15 weeks' gestation, including 4.3% at 16-20 weeks and 1.4% at > or =21 weeks. A total of 35 reporting areas submitted data stating that they performed medical (nonsurgical) procedures, making up 2.9% of all reported procedures from the 45 areas with adequate reporting on type of procedure. In 2000 (the most recent year for which data are available), 11 women died as a result of complications from known legal induced abortion. No deaths were associated with known illegal abortion. During 1990-1997, the number of legal induced abortions gradually declined. When the same 48 reporting areas are compared, the number of abortions decreased during 1996-2001. In 2000 and 2001, even with one additional reporting state, the number of abortions declined slightly. In 2000, as in previous years, deaths related to legal induced abortions occurred rarely (less than one death per 100,000 abortions). Abortion surveillance in the United States continues to provide the data necessary for examining trends in numbers and characteristics of women who obtain legal induced abortions and to increase understanding of this pregnancy outcome. Policymakers and program planners use these data to improve the health and well-being of women and infants.


Microscale investigation of thin film surface ageing of bitumen.
This paper investigates the mechanism of bitumen surface ageing, which was validated utilizing the atomic force microscopy and the differential scanning calorimetry. To validate the surface ageing, three different types of bitumen with different natural wax content were conditioned in four different modes: both ultraviolet and air, only ultraviolet, only air and without any exposure, for 15 and 30 days. From the atomic force microscopy investigation after 15 and 30 days of conditioning period, it was found that regardless the bitumen type, the percentage of microstructure on the surface reduced with the degree of exposure and time. Comparing all the four different exposures, it was observed that ultraviolet radiation caused more surface ageing than the oxidation. It was also found that the combined effect was not simply a summation or multiplication of the individual effects. The differential scanning calorimetry investigation showed that the amount of crystalline fractions in bitumen remain constant even after the systematic conditioning. Interestingly, during the cooling cycle, crystallization of wax molecules started earlier for the exposed specimens than the without exposed one. The analysis of the obtained results indicated that the ageing created a thin film upon the exposed surface, which acts as a barrier and creates difficulty for the wax induced microstructures to float up at the surface. From the differential scanning calorimetry analysis, it can be concluded that the ageing product induced impurities in the bitumen matrix, which acts as a promoter in the crystallization process.


A proposed model linking inflammation to obesity, diabetes, and periodontal infections.
Obesity is an important risk factor for diabetes, cardiovascular disease, and periodontal disease. Adipocytes appear to secrete proinflammatory cytokines which may be the molecules linking the pathogenesis of these diseases. We evaluated the relationship between obesity, periodontal disease, and diabetes mellitus insulin resistance as well as the plasma levels of tumor necrosis factor alpha (TNFalpha) and its soluble receptors (sTNFalpha) to assess the relationship of inflammation to obesity, diabetes, and periodontal infections. The relationship between periodontal disease, obesity, and insulin resistance was examined in the Third National Health and Nutrition Examination Survey (NHANES III). In a population of 12,367 non-diabetic subjects, the variable body mass index (BMI) was used as an assessment of obesity and periodontal disease was assessed by mean clinical attachment loss. The plasma levels of TNFalpha and sTNFalpha were assessed in subsets of 1,221 adults from Erie County, New York, who represented the highest and lowest quartile of BMI. These subjects had extensive periodontal and medical evaluations. In the NHANES III portion of the study, BMI was positively related to severity of periodontal attachment loss (P <0.001). Weighted multiple logistic regressions showed that this relationship is likely mediated by insulin resistance, since overweight individuals (with BMI >or=27 kg/m2) with high levels of insulin resistance (IR) exhibited an odds ratio of 1.48 (95% confidence interval 1.13 - 1.93) for severe periodontal disease as compared to overweight subjects with low IR. In the Erie County adult population, the highest levels of TNFalpha and sTNFalpha receptors were found in those individuals in the highest quartile of BMI. A positive correlation of TNFalpha levels with periodontal disease was found only in those in the lowest quartile of BMI. Obesity is a significant predictor of periodontal disease and insulin resistance appears to mediate this relationship. Furthermore, obesity is associated with high plasma levels of TNFalpha and its soluble receptors, which in turn may lead to a hyperinflammatory state increasing the risk for periodontal disease and also accounting in part for insulin resistance. Further studies of the molecular basis of insulin resistance and its relationship to diabetes, periodontal disease, and obesity are necessary to fully test the hypothesis that adipocyte production of proinflammatory cytokines is a pathogenic factor linking obesity to diabetes and periodontal infections.


Sequence heterogeneity within three different regions of the hepatitis G virus genome.
Two sets of primers derived from the 5'-terminal region and the NS5 region of the hepatitis G virus (HGV) genome were used to amplify PCR fragments from serum specimens obtained from different parts of the world. All PCR fragments from the 5'-terminal region (5'-PCR, n = 56) and from the NS5 region (NS5-PCR, n = 85) were sequenced and compared to corresponding published HGV sequences. The range of nucleotide sequence similarity varied from 74 and 78% to 100% for 5'-PCR and NS5-PCR fragments, respectively. Additionally, five overlapping PCR fragments comprising an approximately 2.0-kb structural region of the HGV genome were sequenced from each of five sera obtained from three United States residents. These sequences were compared to 20 published sequences comprising the same region of the HGV genome. Nucleotide and deduced amino acid sequences obtained from different individuals were homologous from 82.9 to 93. 6% and from 90.4 to 99.0%, respectively. Sequences obtained from follow-up specimens were almost identical. Comparative analysis of deduced amino acid sequences of the HGV structural proteins and hepatitis C virus (HCV) structural proteins combined with an analysis of predicted secondary structures and hydrophobic profiles allowed prediction of processing sites within the HGV structural proteins. A phylogenetic sequence analysis performed on the 2.0-kb structural region supports the existence of three previously identified HGV genetic groups. However, phylogenetic analysis performed on only small DNA fragments yielded inconsistent genetic grouping and failed to confirm the existence of genetic groups. Thus, in contrast to HCV where almost any region can be used for genotyping, only large or carefully selected genome fragments can be used to identify consistent HGV genetic groups.


Mechanisms of postprandial hyperlipidaemia--remnants and coronary artery disease.
High plasma concentrations of triglyceride-rich lipoprotein are associated with an increased risk of coronary artery disease (CAD). In the postprandial state, there is a large increase in chylomicron and very low-density lipoprotein (VLDL) concentrations. The accumulation of potentially atherogenic particles is controlled by the balance of their synthesis and clearance. Chylomicrons are rich in triglyceride and secreted by the intestine postprandially. Chylomicrons compete with VLDL for hydrolysis by lipoprotein lipase (LPL). This competition may cause the increase in large plasma concentration of VLDL seen in the postprandial state. Postprandial increases in atherogenic plasma lipoprotein concentrations are accentuated in insulin-resistant states. Insulin resistance is associated with greater flux of free fatty acids, which may in turn lead to enhanced synthesis of VLDL. Alimentary lipidaemia has been shown to effect changes in the coagulation cascade which may provide additional connections between postprandial lipaemia and CAD. Thus, specific postprandial changes in plasma lipids may be indicative of atherogenic risk. Measurement of postprandial plasma triglyceride concentration and the apo B-48 and apo B-100 contents in triglyceride-rich lipoprotein fractions are probably useful indicators of postprandial dyslipidaemia, but prospective studies of how these lipid variables relate to CAD progression are needed.


Diffuse coronary artery disease in diabetic patients: fact or fiction?
To compare angiographically-determined coronary artery disease in diabetic patients with controls, 1,653 patients coming to cardiac catheterization were reviewed retrospectively to find 37 diabetic and 79 control patients matched for sex, age (+/- 3 years), and risk factors (hypertension, hyperlipidemia, and smoking). The severity of coronary artery disease was assessed using an angiographic grading system. The following results were obtained: 16 of 37 diabetic patients (43%) had three-vessel disease compared to 20 of 79 controls (25%). Seventy-six of 111 (68%) diabetic vessels were diseased compared to 110 of 237 control vessels (46%) (P less than 0.005). The total coronary score reflecting total extent of disease for diabetic patients was 371 (mean 10.0 +/- (SEM) compared to 594 for controls (mean 7.5 +/- 0.7, (P less than 0.01). Diabetic patients had a statistically similar number of diffusely diseased vessels as controls (28% vs 22%). There were only three of 76 diabetic vessels (4%) considered inoperable compared to seven of 110 (6%) control vessels. We conclude that diabetic patients with chest pain have more coronary artery disease than nondiabetics, but no more diffuse or inoperable disease.


Degradation of reactive dyes in a photocatalytic circulating-bed biofilm reactor.
Decolorization and mineralization of reactive dyes by intimately coupled TiO₂-photocatalysis and biodegradation (ICPB) on a novel TiO₂-coated biofilm carrier were investigated in a photocatalytic circulating-bed biofilm reactor (PCBBR). Two typical reactive dyes--Reactive Black 5 (RB5) and Reactive Yellow 86 (RY86)--showed similar first-order kinetics when being photocatalytically decolorized at low pH (~4-5) in batch experiments. Photocatalytic decolorization was inhibited at neutral pH in the presence of phosphate or carbonate buffer, presumably due to electrostatic repulsion from negatively charged surface sites on TiO₂, radical scavenging by phosphate or carbonate, or both. Therefore, continuous PCBBR experiments were carried out at a low pH (~4.5) to maintain high photocatalytic efficiency. In the PCBBR, photocatalysis alone with TiO₂-coated carriers could remove target compound RB5 and COD by 97% and 47%, respectively. Addition of biofilm inside macroporous carriers maintained a similar RB5 removal efficiency, but COD removal increased to 65%, which is evidence of ICPB despite the low pH. ICPB was further proven by finding microorganisms inside carriers at the end of the PCBBR experiments. A proposed ICPB pathway for RB5 suggests that a major intermediate, a naphthol derivative, was responsible for most of the residual COD, while most of the nitrogen in the azo-bonds (-N=N-) was oxidized to N₂.


Antisense Oligonucleotide-Based Therapeutic against Menin for Triple-Negative Breast Cancer Treatment.
The tumor suppressor menin has dual functions, acting either as a tumor suppressor or as an oncogene/oncoprotein, depending on the oncological context. Triple-negative breast cancer (TNBC) is characterized by the lack of expression of the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (ERBB2/HER2) and is often a basal-like breast cancer. TNBC is associated with a dismal prognosis and an insufficient response to chemotherapies. Previously, menin was shown to play a proliferative role in ER-positive breast cancer; however, the functions of menin in TNBC remain unknown. Here, we have demonstrated that menin is expressed in various TNBC subtypes with the strongest expression in the TNBC Hs 578T cells. The depletion of menin by an antisense oligonucleotide (ASO) inhibits cell proliferation, enhances apoptosis in Hs 578T cells, highlighting the oncogenic functions of menin in this TNBC model. ASO-based menin silencing also delays the tumor progression of TNBC xenografts. Analysis of the menin interactome suggests that menin could drive TNBC tumorigenesis through the regulation of MLL/KMT2A-driven transcriptional activity, mRNA 3'-end processing and apoptosis. The study provides a rationale behind the use of ASO-based therapy, targeting menin in monotherapy or in combination with chemo or PARP inhibitors for menin-positive TNBC treatments.


Child malaria treatment practices among mothers in Kenya.
A study of 883 mothers with children aged 0-9 years was undertaken in Kilifi district on the Kenyan coast in order to examine child malaria treatment practices. Quantitative and qualitative methods were used to investigate: whether complications of childhood malaria were recognized; decision-making dynamics in treatment-seeking; and the extent and reasons for the use of proprietary treatment. Childhood malaria was perceived as a mild, everyday illness, not preventable but treatable. The link between malaria and mosquitoes was not recognized. Mothers recognized convulsions, anaemia and splenomegaly but did not link them to malaria. Antimalarial drugs were not given or were withdrawn from children suffering from these conditions. Ill children were treated promptly by purchase of over-the-counter drugs at retail outlets. The health education implications of these findings are discussed.


Paths into Sepsis: Trajectories of Presepsis Healthcare Use.
Sepsis is a leading cause of death and disability whose heterogeneity is often cited as a key impediment to translational progress. To test the hypothesis that there are consequential and significant differences in sepsis outcomes that result from differences in a patient's clinical course leading up to sepsis hospitalization. We conducted an observational cohort study of U.S. Health and Retirement Study (HRS) participants in Medicare (1998-2012) and U.S. Department of Veterans Affairs beneficiaries (2009). Using latent profile analysis, we identified patient subtypes based on trajectory of presepsis healthcare facility use. Subtypes were identified in the derivation cohort (1,512 sepsis hospitalizations among earlier HRS participants), then validated them in two additional cohorts (1,992 sepsis hospitalizations among later HRS participants; 32,525 sepsis hospitalizations among U.S. Department of Veterans Affairs beneficiaries). We measured the association between presepsis path and 90-day mortality using chi-square tests and multivariable logistic regression. We identified three subtypes: low use of inpatient healthcare facilities, comprising 84% of the derivation cohort; rising use, 12%; and high use, 4%. The shape and distribution of presepsis trajectories were similar in all three cohorts. In the derivation cohort, 90-day mortality differed by presepsis trajectory as follows: 38% (low use), 63% (rising use), and 48% (high use) (P < 0.001). This association persisted in the validation cohorts (P < 0.001 for each). The rising use class remained an independent predictor of mortality after adjustment for potential confounders, including detailed physiologic data. In national cohorts of patients with sepsis, we have shown that several distinct paths into sepsis exist. These paths, identified by trajectories of presepsis healthcare use, are predictive of 90-day mortality.


Laser-Doppler velocimeter measurements in nonuniform flow: error estimates.
An approximate technique is described for estimating errors in velocity measurements obtained by laser- Doppler velocimeter (LDV) techniques due to flow variation within the finite dimensions of the scattering volume. The analysis is applicable to steady laminar or turbulent velocity profiles of arbitrary form. By suitable adjustments in the evaluation of the equations, the technique is also applicable to both cw and individual realization LDV applications. In the LDV technique, the velocities of many particles are observed, either simultaneously or sequentially depending on the particle concentration. The velocity measured at a point is thus some type of mean velocity that is not necessarily equal to the velocity at the center of the scattering volume. The analysis presents a mathematical model for approximating the above averaging process from which estimates of errors in LDV measurements are obtained due to a nonuniform velocity distribution within the scattering volume. In addition, the analysis is extended to allow estimation of errors obtained when velocity measurements are made at locations sufficiently near the wall that part of the scattering volume is truncated by the wall. Example calculations are presented for an arbitrary second order velocity profile, for laminar parabolic flow, and for turbulent flow in a pipe for both cw and individual realization signals.


Dairy food, calcium and vitamin D intake in pregnancy, and wheeze and eczema in infants.
The present prospective study examined the association between maternal consumption of dairy products, calcium, and vitamin D during pregnancy and the risk of wheeze and eczema in the infants aged 16-24 months. Subjects were 763 Japanese mother-child pairs. Data on maternal intake during pregnancy were assessed with a diet history questionnaire. Symptoms of wheeze and eczema were based on criteria of the International Study of Asthma and Allergies in Childhood. Higher maternal intake of total dairy products, milk, cheese and calcium during pregnancy was significantly related to a decreased risk of infantile wheeze, but not eczema (adjusted ORs (95% CI) between extreme quartiles were 0.45 (0.25-0.79), 0.50 (0.28-0.87), 0.51 (0.31-0.85), and 0.57 (0.32-0.99), respectively). When maternal vitamin D consumption during pregnancy was categorised into two groups using a cut-off point at the 25th percentile, children whose mothers had consumed > or =4.309 microg x day(-1) had a significantly reduced risk of wheeze and eczema (adjusted ORs (95% CI) were 0.64 (0.43-0.97) and 0.63 (0.41-0.98), respectively). Higher consumption of calcium and dairy foods other than yoghurt during pregnancy may reduce the risk of infantile wheeze. Higher vitamin D intake during pregnancy may be protective against childhood wheeze and eczema.


Healing of the aponeurosis during recovery from aponeurotomy: morphological and histological adaptation and related changes in mechanical properties.
Aponeurotomy, which is the transection of an aponeurosis perpendicular to its length, is performed to lengthen spastic and/or short muscles. During recovery, the cut ends of the aponeurosis are reconnected by new connective tissue bridging both ends. The aim of this study is to investigate the histological features of this new connective tissue as well as its mechanical properties after recovery from aponeurotomy. For this purpose, aponeurotomy was performed on the proximal aponeurosis of rat m. gastrocnemius medialis (GM), which was followed by six weeks of recovery. The lengths of aponeurotic tissues were measured as a function of active muscle length. The results are compared to a control group as well as to the acute effects and a sham operated group. Activation of the muscle at increasing lengths after aponeurotomy caused a gap between the cut ends of the aponeurosis. However, after recovery, new connective tissue is formed bridging the aponeurotic ends, consisting of thin collagen fibres, which are densely packed and generally arranged in the direction of the aponeurosis. The number of fibroblasts was three to five times higher than that of aponeurotic tissue of the intact parts as well as that of the acute and sham operated muscles. The strain of the new connective tissue as a function of active muscle length was shown to be about three times higher than that of the aponeurosis. It is concluded that the inserted new aponeurotic tissue is more compliant and that the aponeurosis becomes 10-15% longer than in untreated muscle. As a consequence, the muscle fibres located distally to the new aponeurotic tissue will become shorter than prior to aponeurotomy. This explains a shift of the length-force curve, which favours the restoration of the range of joint motion.


Identifying the deficiencies of current diagnostic criteria for neurofibromatosis 2 using databases of 2777 individuals with molecular testing.
We have evaluated deficiencies in existing diagnostic criteria for neurofibromatosis 2 (NF2). Two large databases of individuals fulfilling NF2 criteria (n = 1361) and those tested for NF2 variants with criteria short of diagnosis (n = 1416) were interrogated. We assessed the proportions meeting each diagnostic criterion with constitutional or mosaic NF2 variants and the positive predictive value (PPV) with regard to definite diagnosis. There was no evidence for usefulness of old criteria "glioma" or "neurofibroma." "Ependymoma" had 100% PPV and high levels of confirmed NF2 diagnosis (67.7%). Those with bilateral vestibular schwannoma (VS) alone aged ≥60 years had the lowest confirmation rate (6.6%) and reduced PPV (80%). Siblings as a first-degree relative, without an affected parent, had 0% PPV. All three individuals with unilateral VS and an affected sibling were proven not to have NF2. The biggest overlap was with LZTR1-associated schwannomatosis. In this category, seven individuals with unilateral VS plus ≥2 nondermal schwannomas reduced PPV to 67%. The present study confirms important deficiencies in NF2 diagnostic criteria. The term "glioma" should be dropped and replaced by "ependymoma." Similarly "neurofibroma" should be removed. Dropping "sibling" from first-degree relatives should be considered and testing of LZTR1 should be recommended for unilateral VS.


Breast reconstruction with the DIEP flap or the muscle-sparing (MS-2) free TRAM flap: is there a difference?
The advantages of breast reconstruction using the deep inferior epigastric perforator (DIEP) flap and the muscle-sparing free transverse rectus abdominis musculocutaneous (TRAM) flap (MS-2) are well recognized. Both techniques optimize abdominal function by maintaining the vascularity, innervation, and continuity of the rectus abdominis muscle. The purpose of this study was to compare these two methods of breast reconstruction and determine whether there is a difference in outcome. The study considered 177 women who have had breast reconstruction using muscle-sparing flaps over a 4-year period. This includes 89 women who had an MS-2 free TRAM flap procedure, of which 65 were unilateral and 24 were bilateral, and 88 women who had a DIEP flap procedure, of which 66 were unilateral and 22 were bilateral. The total number of flaps was 223. Mean follow-up was 23 months (range, 3 to 49 months). For all MS-2 free TRAM flaps (n = 113), outcome included fat necrosis in eight (7.1 percent), venous congestion in three (2.7 percent), and total necrosis in two (1.8 percent). For the women who had an MS-2 free TRAM flap, an abdominal bulge occurred in three women (4.6 percent) after unilateral reconstruction and in five women (21 percent) after bilateral reconstruction. The ability to perform sit-ups was noted in 63 women (97 percent) after unilateral reconstruction and 20 women (83 percent) after bilateral reconstruction. For all DIEP flaps (n = 110), outcome included fat necrosis in seven (6.4 percent), venous congestion in five (4.5 percent), and total necrosis in three (2.7 percent) patients. For the women who had DIEP flap reconstruction, an abdominal bulge occurred in one woman (1.5 percent) after unilateral reconstruction and in one woman (4.5 percent) after bilateral reconstruction. The ability to perform sit-ups was noted in all women after unilateral reconstruction and in 21 women (95 percent) after bilateral reconstruction. These results demonstrate that there are no significant differences in fat necrosis, venous congestion, or flap necrosis after DIEP or MS-2 free TRAM flap reconstruction. The percentage of women who are able to perform sit-ups and the percentage of women who did not develop a postoperative abdominal bulge is increased after DIEP flap reconstruction; however, this difference is not statistically significant.


Characterization of endothelial colony-forming cells from peripheral blood samples of adult horses.
To isolate and characterize endothelial colony-forming cells (ECFCs; a subtype of endothelial progenitor cells) from peripheral blood samples of horses. Jugular venous blood samples from 24 adult horses. Blood samples were cultured in endothelial cell growth medium. Isolated ECFCs were characterized by use of functional assays of fluorescence-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) uptake and vascular tubule formation in vitro. Expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor 2, and von Willebrand factor) and hematopoietic (CD14) cell markers was assessed through indirect immunofluorescence assay and flow cytometry. The number of passages before senescence was determined through serial evaluation of DiI-Ac-LDL uptake, vascular tubule formation, and cell doubling rates. Samples from 3 horses produced colonies at 12 ± 2.5 days with characteristic endothelial single layer cobblestone morphology and substantial outgrowth on expansion. Equine ECFCs formed vascular tubules in vitro and had uptake of DiI-Ac-LDL (74.9 ± 14.7% positive cells). Tubule formation and DiI-Ac-LDL uptake diminished by passage 5. Equine ECFCs tested positive for von Willebrand factor, vascular endothelial growth factor receptor 2, CD34, and CD105 with an immunofluorescence assay and for CD14 and CD105 via flow cytometry. ECFCs can be isolated from peripheral blood of horses and have characteristics similar to those described for other species. These cells may have potential therapeutic use in equine diseases associated with ischemia or delayed vascularization.


Facile methodology of nanoemulsion preparation using oily polymer for the delivery of poorly soluble drugs.
Aqueous solubility of an active pharmaceutical ingredient (API) is a determining factor that has a direct impact on formulation strategies and overall bioavailability. Fabrication of nanoemulsions of poorly soluble drugs is one of the widely utilized approaches to overcome this problem. However, thermodynamic instability and tedious manufacturing processes of nanoemulsions limit their clinical translation. Therefore, this study was focused on circumventing the abovementioned hurdles by utilizing the polymer as an oil phase, instead of conventional oils. The nanoemulsion was prepared via a facile low-energy nanoprecipitation method using renewable poly(δ-decalactone) (PDL), as an oil phase and Pluronic F-68 as surfactant. The prepared nanoemulsions were characterized in terms of size, drug encapsulation efficiency, stability, and toxicity. Five different hydrophobic drugs were utilized to evaluate the drug delivery capability of the PDL nanoemulsion. The prepared nanoemulsions with sizes less than 200 nm were capable to enhance the aqueous solubility of the drugs by 3 to 10 times compared with the well-established Pluronic F-68 micelles. No phase separation or significant changes in size and drug content was observed with PDL nanoemulsions after high-speed centrifugation and 3 months of storage at two different temperatures (20 °C and 50 °C). PDL nanoemulsions were found to be non-heamolytic up to concentrations of 1 mg/mL, and the cell cytotoxicity studies on MDA-MB-231 and MEF cells suggest a concentration and time-dependent toxicity, where the PDL polymer itself induced no cytotoxicity. The results from this study clearly indicate that the PDL polymer has a tremendous potential to be utilized as an oil phase to prepare stable nanoemulsions via a facile methodology, ultimately favouring clinical translations. Graphical abstract TOC graphic.


Collective decision making and social interaction rules in mixed-species flocks of songbirds.
Associations in mixed-species foraging groups are common in animals, yet have rarely been explored in the context of collective behaviour. Despite many investigations into the social and ecological conditions under which individuals should form groups, we still know little about the specific behavioural rules that individuals adopt in these contexts, or whether these can be generalized to heterospecifics. Here, we studied collective behaviour in flocks in a community of five species of woodland passerine birds. We adopted an automated data collection protocol, involving visits by RFID-tagged birds to feeding stations equipped with antennae, over two winters, recording 91 576 feeding events by 1904 individuals. We demonstrated highly synchronized feeding behaviour within patches, with birds moving towards areas of the patch with the largest proportion of the flock. Using a model of collective decision making, we then explored the underlying decision rule birds may be using when foraging in mixed-species flocks. The model tested whether birds used a different decision rule for conspecifics and heterospecifics, and whether the rules used by individuals of different species varied. We found that species differed in their response to the distribution of conspecifics and heterospecifics across foraging patches. However, simulating decisions using the different rules, which reproduced our data well, suggested that the outcome of using different decision rules by each species resulted in qualitatively similar overall patterns of movement. It is possible that the decision rules each species uses may be adjusted to variation in mean species abundance in order for individuals to maintain the same overall flock-level response. This is likely to be important for maintaining coordinated behaviour across species, and to result in quick and adaptive flock responses to food resources that are patchily distributed in space and time.


Partial protection of Simian immunodeficiency virus (SIV)-infected rhesus monkeys against superinfection with a heterologous SIV isolate.
Although there is increasing evidence that individuals already infected with human immunodeficiency virus type 1 (HIV-1) can be infected with a heterologous strain of the virus, the extent of protection against superinfection conferred by the first infection and the biologic consequences of superinfection are not well understood. We explored these questions in the simian immunodeficiency virus (SIV)/rhesus monkey model of HIV-1/AIDS. We infected cohorts of rhesus monkeys with either SIVmac251 or SIVsmE660 and then exposed animals to the reciprocal virus through intrarectal inoculations. Employing a quantitative real-time PCR assay, we determined the replication kinetics of the two strains of virus for 20 weeks. We found that primary infection with a replication-competent virus did not protect against acquisition of infection by a heterologous virus but did confer relative control of the superinfecting virus. In animals that became superinfected, there was a reduction in peak replication and rapid control of the second virus. The relative susceptibility to superinfection was not correlated with CD4(+) T-cell count, CD4(+) memory T-cell subsets, cytokine production by virus-specific CD8(+) or CD4(+) cells, or neutralizing antibodies at the time of exposure to the second virus. Although there were transient increases in viral loads of the primary virus and a modest decline in CD4(+) T-cell counts after superinfection, there was no evidence of disease acceleration. These findings indicate that an immunodeficiency virus infection confers partial protection against a second immunodeficiency virus infection, but this protection may be mediated by mechanisms other than classical adaptive immune responses.


Metabolic engineering of the mixed-acid fermentation pathway of Escherichia coli for anaerobic production of glutamate and itaconate.
Itaconic acid, an unsaturated C5-dicarboxylic acid, is a biobased building block for the polymer industry. The purpose of this study was to establish proof of principle for an anaerobic fermentation process for the production of itaconic acid by modification of the mixed acid fermentation pathway of E. coli. E. coli BW25113 (DE3) and the phosphate acetyltransferase (pta) and lactate dehydrogenase (ldhA) deficient strain E. coli BW25113 (DE3) Δpta-ΔldhA were used to study anaerobic itaconate production in E. coli. Heterologous expression of the gene encoding cis-aconitate decarboxylase (cadA) from A. terreus in E. coli BW25113 (DE3) did not result in itaconate production under anaerobic conditions, but 0.08 mM of itaconate was formed when the genes encoding citrate synthase (gltA) and aconitase (acnA) from Corynebacterium glutamicum were also expressed. The same amount was produced when cadA was expressed in E. coli BW25113 (DE3) Δpta-ΔldhA. The titre increased 8 times to 0.66 mM (1.2 % Cmol) when E. coli BW25113 (DE3) Δpta-ΔldhA also expressed gltA and acnA. In addition, this strain produced 8.5 mM (13 % Cmol) of glutamate. The use of a nitrogen-limited growth medium reduced the accumulation of glutamate by nearly 50 % compared to the normal medium, and also resulted in a more than 3-fold increase of the itaconate titre to 2.9 mM. These results demonstrated that E. coli has potential to produce itaconate and glutamate under anaerobic conditions, closing the redox balance by co-production of succinate or ethanol with H2 and CO2.


Impact of ATM and SLC22A1 Polymorphisms on Therapeutic Response to Metformin in Iranian Diabetic Patients.
Metabolic syndrome and its pathological sequel, type 2 diabetes are considered as important global health problems. Metformin is the most common drug prescribed for patients with this disorder. Consequently, understanding the genetic pathways involved in pharmacokinetics and pharmacodynamics of this drug can have a considerable effect on the personalized treatment of type 2 diabetes. In this study, we evaluated the association between rs11212617 polymorphism of ATM gene and rs628031 of SLC22A1 gene with response to treatment in newly diagnosed type 2 diabetes patients. We genotyped rs11212617 and rs628031 polymorphism by PCR based restriction fragment length polymorphism (RFLP) and assessed the role of this polymorphisms on response to treatment in 140 patients who have been recently diagnosed with type 2 diabetes and were under monotherapy with metformin for 6 months. Response to metformin was defined by HbA1c and fasting blood sugar (FBS) values. Based on such evaluations, patients were divided into two groups: responders (n= 63) and non-responders (n= 77). No significant association was found between these polymorphisms and response to treatment (OR= 0.86, [95% CI 0.52-1.41], P= 0.32) for rs11212617 and (OR= 0.45, [95% CI 0.64-1.76], P= 0.45) for rs 628031. The reported gene variants in ATM and SLC22A1 are not significantly associated with metformin treatment response in type 2 diabetic patients in an Iranian population.


Global implementation of two shared socioeconomic pathways for future sanitation and wastewater flows.
Households are an important source of nutrient loading to surface water. Sewage systems without or with only primary wastewater treatment are major polluters of surface water. Future emission levels will depend on population growth, urbanisation, increases in income and investments in sanitation, sewage systems and wastewater treatment plants. This study presents the results for two possible shared socioeconomic pathways (SSPs). SSP1 is a scenario that includes improvement of wastewater treatment and SSP3 does not include such improvement, with fewer investments and a higher population growth. The main drivers for the nutrient emission model are population growth, income growth and urbanisation. Under the SSP1 scenario, 5.7 billion people will be connected to a sewage system and for SSP3 this is 5 billion. Nitrogen and phosphorus emissions increase by about 70% under both SSP scenarios, with the largest increase in SSP1. South Asia and Africa have the largest emission increases, in the developed countries decrease the nutrient emissions. The higher emission level poses a risk to ecosystem services.


Efficient visual search of videos cast as text retrieval.
We describe an approach to object retrieval which searches for and localizes all the occurrences of an object in a video, given a query image of the object. The object is represented by a set of viewpoint invariant region descriptors so that recognition can proceed successfully despite changes in viewpoint, illumination and partial occlusion. The temporal continuity of the video within a shot is used to track the regions in order to reject those that are unstable. Efficient retrieval is achieved by employing methods from statistical text retrieval, including inverted file systems, and text and document frequency weightings. This requires a visual analogy of a word which is provided here by vector quantizing the region descriptors. The final ranking also depends on the spatial layout of the regions. The result is that retrieval is immediate, returning a ranked list of shots in the manner of Google. We report results for object retrieval on the full length feature films 'Groundhog Day', 'Casablanca' and 'Run Lola Run', including searches from within the movie and specified by external images downloaded from the Internet. We investigate retrieval performance with respect to different quantizations of region descriptors and compare the performance of several ranking measures. Performance is also compared to a baseline method implementing standard frame to frame matching.


Pd-catalyzed Suzuki-Miyaura reactions of aryl halides using bulky biarylmonophosphine ligands.
The following protocol describes the application of a highly active Pd-based catalyst system in the Suzuki-Miyaura cross-coupling reaction of arylboronic acids with aryl chlorides to provide biaryl compounds. The general procedure includes a detailed description of an appropriate reaction setup, two methods for assaying the crude reaction mixture (GC and TLC) and a procedure for the isolation, purification and characterization of the anticipated product. Reagents and catalyst precursors can be manipulated in the air; however, the cross-coupling reactions must be performed in an inert atmosphere. Two Suzuki-Miyaura reactions are included in the text as representative examples of these procedures. Although the reactions can proceed in less than 5 min, the protocols, including workup, generally take 6-30 h to be completed.


Measurement properties of existing clinical assessment methods evaluating scapular positioning and function. A systematic review.
The aims were to compile a schematic overview of clinical scapular assessment methods and critically appraise the methodological quality of the involved studies. A systematic, computer-assisted literature search using Medline, CINAHL, SportDiscus and EMBASE was performed from inception to October 2013. Reference lists in articles were also screened for publications. From 50 articles, 54 method names were identified and categorized into three groups: (1) Static positioning assessment (n = 19); (2) Semi-dynamic (n = 13); and (3) Dynamic functional assessment (n = 22). Fifteen studies were excluded for evaluation due to no/few clinimetric results, leaving 35 studies for evaluation. Graded according to the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN checklist), the methodological quality in the reliability and validity domains was "fair" (57%) to "poor" (43%), with only one study rated as "good". The reliability domain was most often investigated. Few of the assessment methods in the included studies that had "fair" or "good" measurement property ratings demonstrated acceptable results for both reliability and validity. We found a substantially larger number of clinical scapular assessment methods than previously reported. Using the COSMIN checklist the methodological quality of the included measurement properties in the reliability and validity domains were in general "fair" to "poor". None were examined for all three domains: (1) reliability; (2) validity; and (3) responsiveness. Observational evaluation systems and assessment of scapular upward rotation seem suitably evidence-based for clinical use. Future studies should test and improve the clinimetric properties, and especially diagnostic accuracy and responsiveness, to increase utility for clinical practice.


Wireless EEG with individualized channel layout enables efficient motor imagery training.
The study compared two channel-reduction approaches in order to investigate the effects of systematic motor imagery (MI) neurofeedback practice in an everyday environment using a very user-friendly EEG system consisting of individualized caps and highly portable hardware. Sixteen BCI novices were trained over four consecutive days to imagine left and right hand movements while receiving feedback. The most informative bipolar channels for use on the subsequent days were identified on the first day for each individual based on a high-density online MI recording. Online classification accuracy on the first day was 85.1% on average (range: 64.7-97.7%). Offline an individually-selected bipolar channel pair based on common spatial patterns significantly outperformed a pair informed by independent component analysis and a standard 10-20 pair. From day 2 to day 4 online MI accuracy increased significantly (day 2: 69.1%; day 4: 73.3%), which was mostly caused by a reduction in ipsilateral event-related desynchronization of sensorimotor rhythms. The present study demonstrates that systematic MI practice in an everyday environment with a user-friendly EEG system results in MI learning effects. These findings help to bridge the gap between elaborate laboratory studies with healthy participants and efficient home or hospital based MI neurofeedback protocols.


Characterization and clinical significance of right ventricular mechanics in pulmonary hypertension evaluated with cardiovascular magnetic resonance feature tracking.
Prognosis in pulmonary hypertension (PH) is related to right ventricular (RV) function. Quantification of RV mechanics may offer additive value. The objective of our study is to determine the feasibility and clinical and prognostic value of RV strain analysis by cardiovascular magnetic resonance (CMR) based feature tracking (FT) in PH. We retrospectively enrolled 116 patients (age 52.2 ± 12 years, 73.6 % women) referred to CMR for PH evaluation who underwent right heart catheterization within 1 month. Using dedicated FT software, peak global longitudinal and circumferential RV strain and strain rates (GLS, GCS, GLSR, and GCSR, respectively) were quantified from standard cine images. Using multivariate regression analysis, we evaluated the associations of strain with a composite endpoint of death, lung transplantation, or functional class deterioration. RV strain analysis was feasible in 110 (95 %) patients. Patients were classified into: Group A (no PH, normal right ventricular ejection fraction [RVEF]; n = 17), Group B (PH, normal RVEF; n = 26), or Group C (PH, abnormal RVEF; n = 67). All strain and strain rate values were reduced in Group C. Furthermore, GCSR was significantly reduced in Group B (-0.92 [-1.0/-0.7]; p < 0.001) compared to Group A (-1.12 [-1.3/-0.9]; p < 0.001). After adjustment for six clinically meaningful covariates, GLS (hazard ratio 1.06; p = 0.026), GLSR (hazard ratio 2.52; p = 0.04), and GCSR (hazard ratio 4.5; p = 0.01) were independently associated with the composite endpoint. GCSR successfully discriminated patients with and without events (p = 0.01). Quantification of RV strain with CMR-FT is feasible in the majority of patients, correlates with disease severity, and is independently associated with poor outcomes in PH.


Vitamin K2 biosynthetic enzyme, UBIAD1 is essential for embryonic development of mice.
UbiA prenyltransferase domain containing 1 (UBIAD1) is a novel vitamin K2 biosynthetic enzyme screened and identified from the human genome database. UBIAD1 has recently been shown to catalyse the biosynthesis of Coenzyme Q10 (CoQ10) in zebrafish and human cells. To investigate the function of UBIAD1 in vivo, we attempted to generate mice lacking Ubiad1, a homolog of human UBIAD1, by gene targeting. Ubiad1-deficient (Ubiad1(-/-)) mouse embryos failed to survive beyond embryonic day 7.5, exhibiting small-sized body and gastrulation arrest. Ubiad1(-/-) embryonic stem (ES) cells failed to synthesize vitamin K2 but were able to synthesize CoQ9, similar to wild-type ES cells. Ubiad1(+/-) mice developed normally, exhibiting normal growth and fertility. Vitamin K2 tissue levels and synthesis activity were approximately half of those in the wild-type, whereas CoQ9 tissue levels and synthesis activity were similar to those in the wild-type. Similarly, UBIAD1 expression and vitamin K2 synthesis activity of mouse embryonic fibroblasts prepared from Ubiad1(+/-) E15.5 embryos were approximately half of those in the wild-type, whereas CoQ9 levels and synthesis activity were similar to those in the wild-type. Ubiad1(-/-) mouse embryos failed to be rescued, but their embryonic lifespans were extended to term by oral administration of MK-4 or CoQ10 to pregnant Ubiad1(+/-) mice. These results suggest that UBIAD1 is responsible for vitamin K2 synthesis but may not be responsible for CoQ9 synthesis in mice. We propose that UBIAD1 plays a pivotal role in embryonic development by synthesizing vitamin K2, but may have additional functions beyond the biosynthesis of vitamin K2.


Characterisation of clinical manifestations and treatment strategies for invasive beta-haemolytic streptococcal infections in a Swiss tertiary hospital.
Invasive streptococcal infections affect more than half a million patients worldwide every year and have a high lethality. Little is known about the epidemiology and microbiological characteristics of streptococcal infections in Switzerland. This case series study aims to describe the demographics, known risk factors for streptococcal skin and soft tissue infections, clinical presentations, treatment and outcomes of patients admitted to the University Hospital Zurich between 2000 and 2014 with invasive streptococcal infections caused by Streptococcus pyogenes (group A Streptococcus), Streptococcus dysgalactiae ssp. equisimilis or the Streptococcus anginosus group, as well as the microbiological characteristics of the clinical isolates. Data collected retrospectively from patients hospitalised between 2000 and 2014 with invasive streptococcal infections were analysed. M protein gene (emm) typing of the bacterial clinical isolates was carried out according to the Centers for Disease Control and Prevention guidelines. A total of 86 patients with invasive beta-haemolytic streptococcal infections were included in this study, of which 49% presented with necrotising fasciitis (NF). The median age was 44 years and half were female. The most common risk factor was acute skin lesions. C-reactive protein levels were significantly higher in patients with NF, as were acute renal failure and distributive shock. Beta-lactam antibiotics were given to most patients, and intravenous immunoglobulins were given to 18% of patients within the first 24 hours. All patients suffering from NF underwent surgery. The overall case fatality rate was 8.1% at 30 days post admission. All Group A Streptococcus strains were susceptible to penicillin and clindamycin, and we found resistance to tetracycline in 11.9% of strains. The most common emm-type isolated was emm1 (44.4%). Invasive beta-haemolytic streptococcal infections, the most severe presentation of which is NF, remain a serious clinical issue and require rapid diagnosis and treatment. This is the first representative analysis monitoring clinical and microbiological characteristics of patients with a severe invasive beta-haemolytic streptococcal infection and treated in Zurich, Switzerland. In addition to the detailed reporting of various clinical and microbiological characteristics, we show that C-reactive protein levels, acute renal failure and distributive shock were higher in the patients with NF. We also found a low case fatality rate compared to other reports. The detailed clinical data and microbiological characteristics depicted in this study will lead to a better understanding of regional differences in severe invasive streptococcal infections.


Maternal diet during pregnancy: an emerging risk factor for childhood asthma.
Evaluation of: Willers SM, Wijga AH, Brunekreef B et al. Maternal food consumption during pregnancy and the longitudinal development of childhood asthma. Am. J. Respir. Crit. Care Med. 178, 124-131 (2008). It has been hypothesized that the recent marked increase in the prevalence of asthma may, in part, be a consequence of changing diet. There is increasing interest in the possibility that childhood asthma may be influenced by maternal diet during pregnancy and an increasing number of studies have highlighted associations between childhood asthma and maternal intake of certain foods (e.g., fish, fruits and vegetables) and nutrients (e.g., vitamin E, vitamin D, zinc and polyunsaturated fatty acids) during pregnancy. Maternal diet during pregnancy has the potential to influence fetal immune and airway development during a critical period of life with long-term irreversible consequences, such as childhood asthma. Further research, particularly intervention studies, needs to be carried out to establish whether dietary intervention during pregnancy can be used as a healthy, low-cost, public-health measure to reduce the prevalence of childhood asthma.


An overview and proposed framework of social-environmental influences on the physical-activity behavior of women.
The primary purposes of this review were to identify key social-environmental influences on the physical-activity behavior of women and to propose an organizing framework of these influences in the social environment. Computerized searches of MEDLINE, Science Direct, and CINAHL were conducted to identify relevant research. Study Inclusion and Exclusion Criteria. Studies were included if they used a sample of adult women or included gender-specific analysis, identified or measured some aspect of the social environment in relation to physical activity, and were published in English within the last two decades. Characteristics of study design, sample population, and identified social-environmental variables were extracted. Through an iterative review of the data by several researchers, common influences and themes were identified and organized into a social environment framework, which included the following components: social support/social networks, life transitions/ multiple roles, and cultural standards/gender role expectations. Women are exposed to societal messages that indicate physical activity is not a priority and may be inappropriate; they may also lack the social support necessary to adopt and maintain physical activity. This review reveals the complex range of factors in the social environment that influence women, and it provides support for the importance of the social environment on physical-activity behavior in women. A social-environmental framework related to Bronfenbrenner's social ecological model is proposed along with recommended strategies to apply in interventions targeting women.


Phase I pharmacokinetic and pharmacodynamic study of recombinant human endostatin in patients with advanced solid tumors.
Endostatin is the first endogenous angiogenesis inhibitor to enter clinical trials. Laboratory investigations with endostatin have indicated broad antitumor activity coupled with remarkably low toxicity. A phase I trial of recombinant human endostatin was designed to evaluate toxicity and explore biologic effectiveness in patients with refractory solid tumors. Endostatin was administered as a 1-hour intravenous infusion given daily for a 28-day cycle. A starting dose of 30 mg/m2 was explored with subsequent dose escalations of 60, 100, 150, 225, and 300 mg/m2. Assessment of serum pharmacokinetics was performed on all 21 patients. Western blot assay and mass spectroscopy were employed to evaluate endostatin metabolism. Circulating levels of endogenous proangiogenic growth factors were examined. Tumor and tumor blood supply were imaged by dynamic computed tomography (CT), magnetic resonance imaging, ultrasound, and positron emission tomography. Endostatin given on this schedule was essentially free of significant drug-related toxicity. Two transient episodes of grade 1 rash were observed. No clinical responses were observed. Endostatin pharmacokinetics were linear with dose, and serum concentrations were achieved that are associated with antitumor activity in preclinical models. No aggregate effect on circulating proangiogenic growth factors were seen, although several patients exhibited persistent declines in vascular endothelial growth factor levels while enrolled in the study. A few patients demonstrated changes in their dynamic CT scans suggestive of a decline in microvessel density, although overall, no consistent effect of endostatin on tumor vasculature was seen. Endostatin given daily as a 1-hour intravenous infusion was well tolerated without dose-limiting toxicity at doses up to 300 mg/m2.


Revisiting Correlation Between Pre Operative High Resolution Computed Tomography and Operative Findings in Attico Antral Disease.
(1) To correlate the findings of high resolution computed tomography (HRCT) scans with operative findings in chronic otitis media (attico antral disease). (2) To assess the role of HRCT in chronic otitis media (attico antral disease). This prospective observational study undertaken at a tertiary level teaching hospital included 50 patients of chronic otitis media (attico antral disease) who underwent pre-operative HRCT scanning and the findings were compared with the operative findings and correlation between the two was assessed with appropriate statistical methods. HRCT findings correlated well for the status of malleus and incus, facial nerve canal, lateral semicircular canal, and sinus plate but were less accurate for stapes and tegmen plate. As for disease extent and prediction of cholesteatoma the degree of correlation was site dependent being greater in mastoid air cell system and epitympanum and lesser in mesotympanum and hypotympanum. HRCT despite of its value in management of chronic otitis media has its drawbacks and limitations. CT's accuracy of prediction in some aspects of the disease varies with the site of pathology and this point must always be kept in mind by the operating surgeon. We suggest that each health care centre should establish their own correlative indices for HRCT temporal bone imaging in COM. HRCT cannot be entirely relied upon in management of chronic otitis media patients. However against the backdrop of improved radiological skills in interpreting temporal bone ct images, improved CT machines and importantly the growing concern over medicolegal issues, the role of pre operative CT scan in COM is much more than what was thought previously. Undoubtedly, it is a very useful 'aid' to management BUT a well-trained, experienced and alert surgeon is the key for an accurate diagnosis and successful management of chronic otitis media (attico-antral disease).


Low-Energy Electron Irradiation Damage in Few-Monolayer Pentacene Films.
Crystalline films of pentacene molecules, two to four monolayers in thickness, are grown via in situ sublimation on silicon substrates in the ultrahigh vacuum chamber of a low-energy electron microscope. It is observed that the diffraction pattern of the pentacene layers fades upon irradiation with low-energy electrons. The damage cross section is found to increase by more than an order of magnitude for electron energies from 0 to 10 eV and by another order of magnitude from 10 to 40 eV. Close to 0 eV, damage is virtually nil. Creation of chemically reactive atomic centers after electron attachment or impact ionization is thought to trigger chemical reactions between neighboring molecules that gradually transform the layer into a disordered carbon nanomembrane. Additionally, diminishing spectroscopic features related to the unoccupied band structure of the layers, accompanied by loss of definition in real-space images, and an increase in the background intensity of diffraction images during irradiation point to chemical changes and formation of a disordered layer.


Dietary Assessment on a Mobile Phone Using Image Processing and Pattern Recognition Techniques: Algorithm Design and System Prototyping.
Dietary assessment, while traditionally based on pen-and-paper, is rapidly moving towards automatic approaches. This study describes an Australian automatic food record method and its prototype for dietary assessment via the use of a mobile phone and techniques of image processing and pattern recognition. Common visual features including scale invariant feature transformation (SIFT), local binary patterns (LBP), and colour are used for describing food images. The popular bag-of-words (BoW) model is employed for recognizing the images taken by a mobile phone for dietary assessment. Technical details are provided together with discussions on the issues and future work.


Characteristics of motor vehicle accidents in the Herzegovina region.
Epidemiological studies around the world point to motor vehicle accidents as being one of the leading causes of death. The objective of our study was to analyze some characteristics of motor vehicle accidents in the region of Herzegovina. The study included 226 patients treated at the Clinic for Surgical Diseases and Urology of the University Clinical Hospital Mostar in 2005. A total of 78.3% or examinees were men and 21.7% women. The majority of patients of both sexes were aged between 20 and 30 years. Most of the accidents occurred during autumn and winter months, on weekends and in sunny-dry weather conditions. As expected, most of the hospitalized patients were injured either while driving an automobile or in a pedestrian versus automobile collisions. Of the 226 patients 92.5% admitted to the emergency department and 60.2% were hospitalized. Most of the injured (72.1%) were not under the influence of alcohol at time of the accident. 82.7% of injured were examined by medical personnel at the site within 30 minutes from injury but only 43.0% of them arrived at the hospital within 30 minutes of injury. As most of the injured patients were not under the influence of alcohol at the time of the injury we are of the opinion that more attention in preventing motor vehicle accidents should be directed to speeding and the state of the motorways and vehicles themselves. We propose a large-scale epidemiological study in the Herzegovina region and a review of current road management practices and emergency protocols.


Culture, immortalization, and characterization of human meibomian gland epithelial cells.
Meibomian gland epithelial cells are essential in maintaining the health and integrity of the ocular surface. However, very little is known about their physiological regulation. In this study, the cellular control mechanisms were explored, first to establish a defined culture system for the maintenance of primary epithelial cells from human meibomian glands and, second, to immortalize these cells, thereby developing a preclinical model that could be used to identify factors that regulate cell activity. Human meibomian glands were removed from lid segments after surgery, enzymatically digested, and dissociated. Isolated epithelial cells were cultured in media with or without serum and/or 3T3 feeder layers. To attempt immortalization, the cells were exposed to retroviral human telomerase reverse transcriptase (hTERT) and/or SV40 large T antigen cDNA vectors, and antibiotic-resistant cells were selected, expanded, and subcultured. Analyses for possible biomarkers, cell proliferation and differentiation, lipid-related enzyme gene expression, and the cellular response to androgen were performed with biochemical, histologic, and molecular biological techniques. It was possible to isolate viable human meibomian gland epithelial cells and to culture them in serum-free medium. These cells proliferated, survived through at least the fifth passage, and contained neutral lipids. Infection with hTERT immortalized these cells, which accumulated neutral lipids during differentiation, expressed multiple genes for lipogenic enzymes, responded to androgen, and continued to proliferate. The results show that human meibomian gland epithelial cells may be isolated, cultured, and immortalized.


Differential diagnosis of lower extremity enlargement in pediatric patients referred with a diagnosis of lymphedema.
There are many causes for a large lower limb in the pediatric age group. These children are often mislabeled as having lymphedema, and incorrect diagnosis can lead to improper treatment. The purpose of this study was to determine the differential diagnosis in pediatric patients referred for lower extremity "lymphedema" and to clarify management. The authors' Vascular Anomalies Center database was reviewed between 1999 and 2010 for patients referred with a diagnosis of lymphedema of the lower extremity. Records were studied to determine the correct cause for the enlarged extremity. Alternative diagnoses, sex, age of onset, and imaging studies were also analyzed. A referral diagnosis of lower extremity lymphedema was given to 170 children; however, the condition was confirmed in only 72.9 percent of patients. Forty-six children (27.1 percent) had another disorder: microcystic/macrocystic lymphatic malformation (19.6 percent), noneponymous combined vascular malformation (13.0 percent), capillary malformation (10.9 percent), Klippel-Trenaunay syndrome (10.9 percent), hemihypertrophy (8.7 percent), posttraumatic swelling (8.7 percent), Parkes Weber syndrome (6.5 percent), lipedema (6.5 percent), venous malformation (4.3 percent), rheumatologic disorder (4.3 percent), infantile hemangioma (2.2 percent), kaposiform hemangioendothelioma (2.2 percent), or lipofibromatosis (2.2 percent). Age of onset in children with lymphedema was older than in patients with another diagnosis (p = 0.027). "Lymphedema" is not a generic term. Approximately one-fourth of pediatric patients with a large lower extremity are misdiagnosed as having lymphedema; the most commonly confused causes are other types of vascular anomalies. History, physical examination, and often radiographic studies are required to differentiate lymphedema from other conditions to ensure the child is managed appropriately.


Structural basis for enzymatic evolution from a dedicated ADP-ribosyl cyclase to a multifunctional NAD hydrolase.
Cyclic ADP-ribose (cADPR) is a universal calcium messenger molecule that regulates many physiological processes. The production and degradation of cADPR are catalyzed by a family of related enzymes, including the ADP-ribosyl cyclase from Aplysia california (ADPRAC) and CD38 from human. Although ADPRC and CD38 share a common evolutionary ancestor, their enzymatic functions toward NAD and cADPR homeostasis have evolved divergently. Thus, ADPRC can only generate cADPR from NAD (cyclase), whereas CD38, in contrast, has multiple activities, i.e. in cADPR production and degradation, as well as NAD hydrolysis (NADase). In this study, we determined a number of ADPRC and CD38 structures bound with various nucleotides. From these complexes, we elucidated the structural features required for the cyclization (cyclase) reaction of ADPRC and the NADase reaction of CD38. Using the structural approach in combination with site-directed mutagenesis, we identified Phe-174 in ADPRC as a critical residue in directing the folding of the substrate during the cyclization reaction. Thus, a point mutation of Phe-174 to glycine can turn ADPRC from a cyclase toward an NADase. The equivalent residue in CD38, Thr-221, is shown to disfavor the cyclizing folding of the substrate, resulting in NADase being the dominant activity. The comprehensive structural comparison of CD38 and APDRC presented in this study thus provides insights into the structural determinants for the functional evolution from a cyclase to a hydrolase.


Endoscopic ultrasound-guided fine needle aspiration of the celiac ganglion: A diagnostic pitfall.
Endoscopic ultrasound guided fine-needle aspiration (EUS-FNA) is now widely used as a primary tool in the evaluation of lymphadenopathy in both the mediastinum and abdomen. A sympathetic ganglion may be mistaken for an enlarged lymph node on endoscopic ultrasound and are rarely sampled as such. A 51-year-old female presented with a history of weight loss, vomiting for several months, and right upper quadrant discomfort. Computed tomography (CT) and magnetic resonance imaging (MRI) scans showed a dilated common bile duct (CBD) with a possible periampullary mass, paraaortic, and pericelial lymph nodes suspicious for metastatic disease. Endosonography revealed a 17 mm oval hypoechoic structure with distinct margins in the para-aortic, celiac axis region suggestive of an enlarged lymph node. An EUS-FNA was done. Cytology revealed ganglion cells with large oval epithelial-like cells with round nuclei and prominent nucleoli consistent with a benign sympathetic ganglion. It is crucial for the cytopathologist to be aware of the fact that the endoscopist might have sampled a celiac ganglion instead of a celiac lymph node and be able to distinguish the cytological features of a benign sympathetic ganglion from a malignant process.


The structures of frataxin oligomers reveal the mechanism for the delivery and detoxification of iron.
Defects in the mitochondrial protein frataxin are responsible for Friedreich ataxia, a neurodegenerative and cardiac disease that affects 1:40,000 children. Here, we present the crystal structures of the iron-free and iron-loaded frataxin trimers, and a single-particle electron microscopy reconstruction of a 24 subunit oligomer. The structures reveal fundamental aspects of the frataxin mechanism. The trimer has a central channel in which one atom of iron binds. Two conformations of the channel with different metal-binding affinities suggest that a gating mechanism controls whether the bound iron is delivered to other proteins or transferred to detoxification sites. The trimer constitutes the basic structural unit of the 24 subunit oligomer. The architecture of this oligomer and several features of the trimer structure demonstrate striking similarities to the iron-storage protein ferritin. The data reveal how stepwise assembly provides frataxin with the structural flexibility to perform two functions: metal delivery and detoxification.


Kinematic analyses including finite helical axes of drop jump landings demonstrate decreased knee control long after anterior cruciate ligament injury.
The purpose was to evaluate the dynamic knee control during a drop jump test following injury of the anterior cruciate ligament injury (ACL) using finite helical axes. Persons injured 17-28 years ago, treated with either physiotherapy (ACLPT, n = 23) or reconstruction and physiotherapy (ACLR, n = 28) and asymptomatic controls (CTRL, n = 22) performed a drop jump test, while kinematics were registered by motion capture. We analysed the Preparation phase (from maximal knee extension during flight until 50 ms post-touchdown) followed by an Action phase (until maximal knee flexion post-touchdown). Range of knee motion (RoM), and the length of each phase (Duration) were computed. The finite knee helical axis was analysed for momentary intervals of ~15° of knee motion by its intersection (ΔAP position) and inclination (ΔAP Inclination) with the knee's Anterior-Posterior (AP) axis. Static knee laxity (KT100) and self-reported knee function (Lysholm score) were also assessed. The results showed that both phases were shorter for the ACL groups compared to controls (CTRL-ACLR: Duration 35±8 ms, p = 0.000, CTRL-ACLPT: 33±9 ms, p = 0.000) and involved less knee flexion (CTRL-ACLR: RoM 6.6±1.9°, p = 0.002, CTRL-ACLR: 7.5 ±2.0°, p = 0.001). Low RoM and Duration correlated significantly with worse knee function according to Lysholm and higher knee laxity according to KT-1000. Three finite helical axes were analysed. The ΔAP position for the first axis was most anterior in ACLPT compared to ACLR (ΔAP position -1, ACLPT-ACLR: 13±3 mm, p = 0.004), with correlations to KT-1000 (rho 0.316, p = 0.008), while the ΔAP inclination for the third axis was smaller in the ACLPT group compared to controls (ΔAP inclination -3 ACLPT-CTRL: -13±5°, p = 0.004) and showed a significant side difference in ACL injured groups during Action (Injured-Non-injured: 8±2.7°, p = 0.006). Small ΔAP inclination -3 correlated with low Lysholm (rho 0.391, p = 0.002) and high KT-1000 (rho -0.450, p = 0.001). Conclusions Compensatory movement strategies seem to be used to protect the injured knee during landing. A decreased ΔAP inclination in injured knees during Action suggests that the dynamic knee control may remain compromised even long after injury.


Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells.
The binding and subsequent intracellular processing of transferrin and transferrin receptors was studied in A431 cells using 125I-transferrin and a monoclonal antibody to the receptor (ATR) labeled with 125I and gold colloid. Using 125I-transferrin we have shown that, whereas at 37 degrees C uptake proceeded linearly for up to 60 min, most of the ligand that was bound was internalized and then rapidly returned to the incubation medium undegraded. At 37 degrees C, the intracellular half-life of the most rapidly recycled transferrin was 7.5 min. 125I-ATR displayed the same kinetics of uptake but following its internalization at 37 degrees C, it was partially degraded. At 22 degrees C and below, the intracellular degradation of 125I-ATR was selectively inhibited and as a result it accumulated intracellularly. Electron microscopy of conventional thin sections and of whole-cell mounts was used to follow the uptake and processing of transferrin receptors labeled with ATR-gold colloid complexes. Using a pulse-chase protocol, the intracellular pathway followed by internalized ATR gold-receptor complexes was outlined in detail. Within 5 min at 22 degrees C the internalized complexes were transferred from coated pits on the cell surface to a system of narrow, branching cisternae within the peripheral cytoplasm. By 15 min they reached larger, more dilated elements that, in thin section, appeared as irregular profiles containing small (30-50-nm diam) vesicles. By 30 min, the gold complexes were located predominantly within typical spherical multivesicular bodies lying in the peripheral cytoplasm, and by 40-60 min, they reached a system of cisternal and multivesicular body elements in the juxtanuclear area. At 22 degrees C, no other compartments became labeled but if they were warmed to 37 degrees C the gold complexes were transferred to lysosome-like elements. Extracting ATR-gold complexes with Triton X after a 30-min chase at 22 degrees C and purifying them on Sepharose-transferrin indicated that the internalized complexes remained bound to the transferrin receptor during their intracellular processing.


A model of phospholipid biosynthesis in tumor in response to an anticancer agent in vivo.
We study, in this paper, a model for the core of the system of the Glycerophospholipid metabolism in the murine cells. It comprises the simple and enzymatic reactions of PhosphatidylEthanolamine and the PhosphatidylCholine. The model's general structure is taken from a number of books and articles. We translate this model into a set of ordinary differential equations (ODEs), to propose a quantitative explanation of the experimental experiences and the observed results. In order to make it usable as a basis for simulations and mathematical analysis we need to make precise the various constants present in the equations but which are usually not directly accessible in the literature. In a first step we considered experimental data of rat's liver cells obtained by NMR spectroscopy: given the values of metabolite concentrations we find appropriate parameter values which allow us to describe the system with ODEs. We have then performed several analyses using the developed model such as stability analysis. A first interesting result is the global stability of the system which was observed by simulation and then proved by mathematical arguments. A second important result is that we observe on the diagrams that the steady state for normal cells is precisely a singular point of order two, whereas tumoral cells present different characteristics; this fact has been proved for PhosphatidylEthanolamine N-Methyl transferase (PEMT), an enzyme which seems to be identified for the first time as a crucial element in the tumoral process. In a second step we applied our model to experimental data of proton HRMAS NMR spectroscopy for solid B16 melanoma and Lewis lung (3LL) 3LL carcinoma cells treated by Chloroethyl Nitrosourea (CENU). We performed a complete comparative analysis of parameters in order to learn the predictive statements to explain increases and decreases which one can observe in concentrations.


Polarimetric multi-mode tilted fiber grating sensors.
The polarimetric sensing characteristics of multi-mode-fiber based tilted fiber Bragg gratings (MMF-TFBG) have been analyzed and experimentally demonstrated. The larger diameter fiber core and graded index core/cladding profile enable the tilted gratings to excite multiple high-order core modes with significantly different polarization dependence and to form a well-defined "comb" of spectrally separated resonances at different wavelengths. Orientation-recognized twist/rotation measurements (-90° to 90°) have been achieved with sensitivity of 0.075 dB/deg by using intensity monitoring of two orthogonally polarized odd core-modes (LP11 and LP12). The proposed sensor is compact, works in reflection (a short section of MMF-TFBG spliced with a leading-in single mode fiber without any offset or tapering), is insensitive to temperature (intensity detection instead of wavelength monitoring) and is immune to unwanted intensity fluctuations (differential intensity measurement). Other TFBG sensing modalities, such as lateral pressure and surrounding refractive index are demonstrated separately with the same device configuration and interrogation principles.


LATS-YAP/TAZ controls lineage specification by regulating TGFβ signaling and Hnf4α expression during liver development.
The Hippo pathway regulates the self-renewal and differentiation of various adult stem cells, but its role in cell fate determination and differentiation during liver development remains unclear. Here we report that the Hippo pathway controls liver cell lineage specification and proliferation separately from Notch signalling, using mice and primary hepatoblasts with liver-specific knockout of Lats1 and Lats2 kinase, the direct upstream regulators of YAP and TAZ. During and after liver development, the activation of YAP/TAZ induced by loss of Lats1/2 forces hepatoblasts or hepatocytes to commit to the biliary epithelial cell (BEC) lineage. It increases BEC and fibroblast proliferation by up-regulating TGFβ signalling, but suppresses hepatoblast to hepatocyte differentiation by repressing Hnf4α expression. Notably, oncogenic YAP/TAZ activation in hepatocytes induces massive p53-dependent cell senescence/death. Together, our results reveal that YAP/TAZ activity levels govern liver cell differentiation and proliferation in a context-dependent manner.


Skeletal muscle cellularity and glycogen distribution in the hypermuscular Compact mice.
The TGF-beta member myostatin acts as a negative regulator of skeletal muscle mass. The Compact mice were selected for high protein content and hypermuscularity, and carry a naturally occurring 12-bp deletion in the propeptide region of the myostatin precursor. We aimed to investigate the cellular characteristics and the glycogen distribution of the Compact tibialis anterior (TA) muscle by quantitative histochemistry and spectrophotometry. We have found that the deficiency in myostatin resulted in significantly increased weight of the investigated hindlimb muscles compared to wild type. Although the average glycogen content of the individual fibers kept unchanged, the total amount of glycogen in the Compact TA muscle increased two-fold, which can be explained by the presence of more fibers in Compact compared to wild type muscle. Moreover, the ratio of the most glycolytic IIB fibers significantly increased in the Compact TA muscle, of which glycogen content was the highest among the fast fibers. In summary, myostatin deficiency caused elevated amount of glycogen in the TA muscle but did not increase the glycogen content of the individual fibers despite the marked glycolytic shift observed in Compact mice.


Magnification of secondary production by kelp detritus in coastal marine ecosystems.
Kelps are highly productive seaweeds found along most temperate latitude coastlines, but the fate and importance of kelp production to nearshore ecosystems are largely unknown. The trophic role of kelp-derived carbon in a wide range of marine organisms was assessed by a natural experiment. Growth rates of benthic suspension feeders were greatly increased in the presence of organic detritus (particulate and dissolved) originating from large benthic seaweeds (kelps). Stable carbon isotope analysis confirmed that kelp-derived carbon is found throughout the nearshore food web.


Promoting notochord fate and repressing muscle development in zebrafish axial mesoderm.
Cell fate decisions in early embryonic cells are controlled by interactions among developmental regulatory genes. Zebrafish floating head mutants lack a notochord; instead, muscle forms under the neural tube. As shown previously, axial mesoderm in floating head mutant gastrulae fails to maintain expression of notochord genes and instead expresses muscle genes. Zebrafish spadetail mutant gastrulae have a nearly opposite phenotype; notochord markers are expressed in a wider domain than in wild-type embryos and muscle marker expression is absent. We examined whether these two phenotypes revealed an antagonistic genetic interaction by constructing the double mutant. Muscle does not form in the spadetail;floating head double mutant midline, indicating that spadetail function is required for floating head mutant axial mesoderm to transfate to muscle. Instead, the midline of spadetail;floating head double mutants is greatly restored compared to that of floating head mutants; the floor plate is almost complete and an anterior notochord develops. In addition, we find that floating head mutant cells can make both anterior and posterior notochord when transplanted into a wild-type host, showing that enviromental signals can override the predisposition of floating head mutant midline cells to make muscle. Taken together, these results suggest that repression of spadetail function by floating head is critical to promote notochord fate and prevent midline muscle development, and that cells can be recruited to the notochord by environmental signals.


A regional multilevel analysis: can skilled birth attendants uniformly decrease neonatal mortality?
Globally 40 % of deaths to children under-five occur in the very first month of life with three-quarters of these deaths occurring during the first week of life. The promotion of delivery with a skilled birth attendant (SBA) is being promoted as a strategy to reduce neonatal mortality. This study explored whether SBAs had a protective effect against neonatal mortality in three different regions of the world. The analysis pooled data from nine diverse countries for which recent Demographic and Health Survey data were available. Multilevel logistic regression was used to understand the influence of skilled delivery on two outcomes-neonatal mortality during the first week of life and during the first day of life. Control variables included age, parity, education, wealth, residence (urban/rural), geographic region (Africa, Asia and Latin America/Caribbean), antenatal care and tetanus immunization. The direction of the effect of skilled delivery on neonatal mortality was dependent on geographic region. While having a SBA at delivery was protective against neonatal mortality in Latin America/Caribbean, in Asia there was only a protective effect for births in the first week of life. In Africa SBAs were associated with higher neonatal mortality for both outcomes, and the same was true for deaths on the first day of life in Asia. Many women in Africa and Asia deliver at home unless a complication occurs, and thus skilled birth attendants may be seeing more women with complications than their unskilled counterparts. In addition there are issues with the definition of a SBA with many attendants in both Africa and Asia not actually having the needed training and equipment to prevent neonatal mortality. Considerable investment is needed in terms of training and health infrastructure to enable these providers to save the youngest lives.


Redox albuminomics: oxidized albumin in human diseases.
Albumin is the major contributor to colloid oncotic pressure and also serves as an important carrier protein of many endogenous and exogenous molecules throughout the body. In blood and extravascular fluids, albumin is susceptible to different oxidative modifications, especially thiol oxidation and carbonylation. Because of its metal-binding properties and the redox properties of its Cys34 thiol, albumin displays an important antioxidant activity. As albumin is the predominant protein in most body fluids, its Cys34 represents the largest fraction of free thiols within body fluids. Evidence that albumin oxidation takes place in vivo has been reported only recently. Different redox proteomic, mass spectrometric, and chromatographic techniques have shown albumin redox modifications in various human pathophysiological conditions. As a whole, most data here presented demonstrate that massive albumin oxidation occurs in vivo in different biological fluids and, to some extent, that this process is correlated to organ dysfunction. Recent reports suggest that the albumin redox state may serve as a global biomarker for the redox state in the body in various human diseases. However, further study is required to elucidate the exact relationship between albumin oxidation and pathology. In addition, it is unknown if some albumin oxidized forms may also have diagnostic uses. Application of specific redox proteomics techniques for the characterization of oxidized albumin forms in screening studies is required. A further challenge will be to analyze how these oxidative albumin modifications are related to real impact to the body.


Dual-spindle formation in zygotes keeps parental genomes apart in early mammalian embryos.
At the beginning of mammalian life, the genetic material from each parent meets when the fertilized egg divides. It was previously thought that a single microtubule spindle is responsible for spatially combining the two genomes and then segregating them to create the two-cell embryo. We used light-sheet microscopy to show that two bipolar spindles form in the zygote and then independently congress the maternal and paternal genomes. These two spindles aligned their poles before anaphase but kept the parental genomes apart during the first cleavage. This spindle assembly mechanism provides a potential rationale for erroneous divisions into more than two blastomeric nuclei observed in mammalian zygotes and reveals the mechanism behind the observation that parental genomes occupy separate nuclear compartments in the two-cell embryo.


Saturable-absorber-based spatial filtering of high-power laser beams.
A saturable-absorber-based technique for spatial filtering of high-average-power laser beams is described. For a focused, radially symmetric beam having its highest intensity at the center, this saturable absorber behaves like a soft aperture with gradually increasing attenuation toward the beam edges, thus selectively transmitting the low divergence components that are confined close to the central axis of the propagating laser beam. This technique has been successfully used to reduce the divergence of a high-power, high-repetition-rate, tunable, narrowband, pulsed dye laser. Our results demonstrate how a judicious choice of operating parameters allows spatial filtering to be achieved with the introduction of a minimum absorption loss of the laser beam in the saturable absorber. Following a time-dependent analysis of a rate equation model describing the propagation and interaction of the laser beam with the saturable absorber, we have also obtained theoretical estimates for the extent of spatial filtering. Our theoretical estimates have been found to be in good agreement with our experimental observations.


Evidence-based palaeopathology: meta-analysis of PubMed-listed scientific studies on ancient Egyptian mummies.
There is a plethora of published scientific studies on ancient Egyptian mummies. Surprisingly, hitherto there is no systematic review of this research, which would help to assess the quality of this vast body of published literature and thus to increase "evidence" in palaeopathological research. The aim of this study was to review all PubMed-listed scientific studies performed on Ancient Egyptian mummies. A total of 131 studies were found in the database for the selected time period, 1977-2005. Our "meta-analysis" showed that the number of publications per year varies enormously. The majority of mummies examined date to the third intermediate and Ptolemaic periods; data from other time periods were lacking. Identification of the cause of death and (14)C-dating of the mummy or funeral goods were rarely addressed. There was a tendency towards an increased use of non-invasive examination methods in more modern times. Our meta-analysis addressed both scientific content (e.g. palaeopathological findings/examination methods) and publication issues (e.g. location of the first author or year of publication) in these studies. Based on our experience, we recommend some minimum publication standards for palaeopathologic studies on ancient mummies, which shall improve evidence-based research in palaeopathology in general.


Vascular perfusion chilling of red meat carcasses - A feasibility study.
Meat carcasses must be chilled to below 7°C before leaving the slaughterhouse. Typically this is done by passing cold air over the surfaces of eviscerated and de-hided carcasses. This surface cooling can take many hours to reduce centre temperatures to below 7°C. In vascular perfusion chilling (VPC), a cold fluid is circulated through the intact vascular system, offering significant reductions in cooling time. This paper describes a small feasibility study to evaluate vascular perfusion techniques for rapid chilling of lamb carcasses using a proprietary Flo-ice(™) system. This produces pumpable ice slurries containing very fine ice particles, suitable for circulating through vascular systems. VPC was found to be capable of rapid initial reduction of carcass temperatures in comparison with air chilling (mean times to 20°C in deep legs were reduced from 2.6 to 1.3h, which was significantly different at P<0.05). In all cases however, uptake of perfusate into the carcasses occurred. This limited the duration of the perfusion treatment and as a result restricted the period of enhanced cooling. Samples from carcasses treated with VPC were lighter (P<0.05, with mean measured L value increasing from 43.4 to 46.8) and more yellow (P<0.05, with mean measured b value increasing from 6.7 to 7.9) than samples from conventionally chilled carcasses, and had lower shear force values when cooked (P<0.05, with mean force reducing from 10.0 to 6.8kg). This was most probably due to the added water in the meat. Microbial quality of the meat was not significantly affected by the perfusion treatments.


Denervation and high-fat diet reduce insulin signaling in T-tubules in skeletal muscle of living mice.
Insulin stimulates muscle glucose transport by translocation of GLUT4 to sarcolemma and T-tubules. Despite muscle glucose uptake playing a major role in insulin resistance and type 2 diabetes, the temporal and spatial changes in insulin signaling and GLUT4 translocation during these conditions are not well described. We used time-lapse confocal imaging of green fluorescent protein (GFP) ADP-ribosylation factor nucleotide-binding site opener (ARNO) (evaluation of phosphatidylinositide 3-kinase activation) and GLUT4-GFP-transfected quadriceps muscle in living, anesthetized mice either muscle denervated or high-fat fed. T-tubules were visualized with sulforhodamine B dye. In incubated muscle, glucose transport was measured by 2-deoxy-D-[(3)H]-glucose uptake, and functional detubulation was carried out by osmotic shock. Muscle fibers were immunostained for insulin receptors. Denervation and high-fat diet reduced insulin-mediated glucose transport. In denervated muscle, insulin-stimulated phosphatidylinositol 3,4,5 P(3) (PIP3) production was abolished in T-tubules, while PIP3 production at sarcolemma was increased 2.6-fold. Correspondingly, GLUT4-GFP translocation to T-tubules was abolished, while translocation to sarcolemma was increased 2.3-fold. In high fat-fed mice, a approximately 65% reduction in both insulin-induced T-tubular PIP3 production and GLUT4-GFP translocation was seen. Sarcolemma was less affected, with reductions of approximately 40% in PIP3 production and approximately 15% in GLUT4-GFP translocation. Access to T-tubules was not compromised, and insulin receptor distribution in sarcolemma and T-tubules was unaffected by denervation or high-fat feeding. Detubulation of normal muscle reduced basal and abolished insulin-induced glucose transport. Our findings demonstrate, for the first time, that impaired insulin signaling and GLUT4 translocation is compartmentalized in muscle and primarily localized to T-tubules and not sarcolemma during insulin resistance.


How a neuropsychiatric brain bank should be run: a consensus paper of Brainnet Europe II.
The development of new molecular and neurobiological methods, computer-assisted quantification techniques and neurobiological investigation methods which can be applied to the human brain, all have evoked an increased demand for post-mortem tissue in research. Psychiatric disorders are considered to be of neurobiological origin. Thus far, however, the etiology and pathophysiology of schizophrenia, depression and dementias are not well understood at the cellular and molecular level. The following will outline the consensus of the working group for neuropsychiatric brain banking organized in the Brainnet Europe II, on ethical guidelines for brain banking, clinical diagnostic criteria, the minimal clinical data set of retrospectively analyzed cases as well as neuropathological standard investigations to perform stageing for neurodegenerative disorders in brain tissue. We will list regions of interest for assessments in psychiatric disorder, propose a dissection scheme and describe preservation and storage conditions of tissue. These guidelines may be of value for future implementations of additional neuropsychiatric brain banks world-wide.


CT coronary angiography in patients with suspected angina due to coronary heart disease (SCOT-HEART): an open-label, parallel-group, multicentre trial.
The benefit of CT coronary angiography (CTCA) in patients presenting with stable chest pain has not been systematically studied. We aimed to assess the effect of CTCA on the diagnosis, management, and outcome of patients referred to the cardiology clinic with suspected angina due to coronary heart disease. In this prospective open-label, parallel-group, multicentre trial, we recruited patients aged 18-75 years referred for the assessment of suspected angina due to coronary heart disease from 12 cardiology chest pain clinics across Scotland. We randomly assigned (1:1) participants to standard care plus CTCA or standard care alone. Randomisation was done with a web-based service to ensure allocation concealment. The primary endpoint was certainty of the diagnosis of angina secondary to coronary heart disease at 6 weeks. All analyses were intention to treat, and patients were analysed in the group they were allocated to, irrespective of compliance with scanning. This study is registered with ClinicalTrials.gov, number NCT01149590. Between Nov 18, 2010, and Sept 24, 2014, we randomly assigned 4146 (42%) of 9849 patients who had been referred for assessment of suspected angina due to coronary heart disease. 47% of participants had a baseline clinic diagnosis of coronary heart disease and 36% had angina due to coronary heart disease. At 6 weeks, CTCA reclassified the diagnosis of coronary heart disease in 558 (27%) patients and the diagnosis of angina due to coronary heart disease in 481 (23%) patients (standard care 22 [1%] and 23 [1%]; p<0·0001). Although both the certainty (relative risk [RR] 2·56, 95% CI 2·33-2·79; p<0·0001) and frequency of coronary heart disease increased (1·09, 1·02-1·17; p=0·0172), the certainty increased (1·79, 1·62-1·96; p<0·0001) and frequency seemed to decrease (0·93, 0·85-1·02; p=0·1289) for the diagnosis of angina due to coronary heart disease. This changed planned investigations (15% vs 1%; p<0·0001) and treatments (23% vs 5%; p<0·0001) but did not affect 6-week symptom severity or subsequent admittances to hospital for chest pain. After 1·7 years, CTCA was associated with a 38% reduction in fatal and non-fatal myocardial infarction (26 vs 42, HR 0·62, 95% CI 0·38-1·01; p=0·0527), but this was not significant. In patients with suspected angina due to coronary heart disease, CTCA clarifies the diagnosis, enables targeting of interventions, and might reduce the future risk of myocardial infarction. The Chief Scientist Office of the Scottish Government Health and Social Care Directorates funded the trial with supplementary awards from Edinburgh and Lothian's Health Foundation Trust and the Heart Diseases Research Fund.


Uncorrected Myopic Refractive Error Increases Microsaccade Amplitude.
Human brain generates miniature eye movements, such as microsaccades, to counteract image fading due to visual adaptation. Generation of microsaccade relies on the amount of retinal error or acuity demand for a desired visual task. The goal of this study was to assess the influence of visual blur, induced by uncorrected refractive error on microsaccades and saccades. Ten subjects with myopia held their gaze on a visual target during two experiment conditions: corrected refractive error and uncorrected refractive error. Eye movements were measured with high-resolution video oculography under binocular viewing conditions during both tasks. Gaze holding function, microsaccades, and visually guided saccades were analyzed and compared during both tasks. We found an increase in the amplitude of microsaccades in the presence of uncorrected refractive error, but the microsaccade frequency and velocity remained unchanged. The microsaccade amplitude systematically increased with an increase in uncorrected refractive error. The main sequence relationship relating the saccade amplitude with respective peak velocity was not significantly different between two conditions. The onset latency, peak velocities, and accuracy of visually guided saccades also were unchanged between the two conditions. These results suggest that visual blur, hence the precision of an image on the fovea, has an important role in calibrating the amplitude of fixational eye movements, such as microsaccades.