Uptake of lactosylated low-density lipoprotein by galactose-specific receptors in rat liver.
The liver contains two types of galactose receptors, specific for Kupffer and parenchymal cells respectively. These receptors are only expressed in the liver, and therefore are attractive targets for the specific delivery of drugs. We provided low-density lipoprotein (LDL), a particle with a diameter of 23 nm in which a variety of drugs can be incorporated, with terminal galactose residues by lactosylation. Radioiodinated LDL, lactosylated to various extents (60-400 mol of lactose/ mol of LDL), was injected into rats. The plasma clearance and hepatic uptake of radioactivity were correlated with the extent of lactosylation. Highly lactosylated LDL (greater than 300 lactose/LDL) is completely cleared from the blood by liver within 10 min. Pre-injection with N-acetylgalactosamine blocks liver uptake, which indicates that the hepatic recognition sites are galactose-specific. The hepatic uptake occurs mainly by parenchymal and Kupffer cells. At a low degree of lactosylation, approx. 60 lactose/LDL, the specific uptake (ng/mg of cell protein) is 28 times higher in Kupffer cells than in parenchymal cells. However, because of their much larger mass, parenchymal cells are the main site of uptake. At high degrees of lactosylation (greater than 300 lactose/LDL), the specific uptake in Kupffer cells is 70-95 times that in parenchymal cells. Under these conditions, Kupffer cells are, despite their much smaller mass, the main site of uptake. Thus not only the size but also the surface density of galactose on lactosylated LDL is important for the balance of uptake between Kupffer and parenchymal cells. This knowledge should allow us to design particulate galactose-bearing carriers for the rapid transport of various drugs to either parenchymal cells or Kupffer cells.


Drug Development Strategy for Type 2 Diabetes: Targeting Positive Energy Balances.
Newer classes of medications have been proven useful in glycemic control in type 2 diabetes (T2D), but many do not appear capable to slow down the progressive loss of ß-cell function, or to improve population-level glycemic control. Positive energy balance, e.g. surplus energy intake over expenditure, is at the core for developing metabolic syndrome and T2D. Currently available glycemic control drugs come to the market based on their 1-2 years risk-benefit profiles, but most of them do not correct positive energy balance and lose efficacy in the long-term. This denouement is destined by a positive energy balance of T2D. There is continuous endeavor/investment in new drugs for T2D. In this review, we compared the effects of commonly used oral hypoglycemic agents on energy balance and discussed several novel therapeutic targets/approaches for T2D that could potentially correct positive energy balance: changing the composition of intestinal host-microbiota to alleviate excess caloric consumption, controlling chylomicron uptake into intestinal lacteals to reduce excessive caloric intake, and activating pyruvate kinase M2 (PKM2) to ameliorate glucose metabolism and increase energy expenditure. We further reviewed how nicotine affects body weight and ameliorates positive energy balance, and ways to encourage people to adopt a more healthy lifestyle by exercising more and/or decreasing caloric intake. These potential targets/approaches may hopefully correct positive energy balance, delay disease progression, reverse some pathophysiological changes, and eventually prevent and/or cure the disease. Drug development strategies applying new insights into T2D process and therapeutic index to correct positive energy balance need to be seriously considered.


Plasma levels of osteopontin identify patients at risk for organ damage in systemic lupus erythematosus.
Osteopontin (OPN) has been implicated as a mediator of Th17 regulation via type I interferon (IFN) receptor signaling and in macrophage activity at sites of tissue repair. This study assessed whether increased circulating plasma OPN (cOPN) precedes development of organ damage in pediatric systemic lupus erythematosus (pSLE) and compared it to circulating plasma neutrophil gelatinase-associated lipocalin (cNGAL), a predictor of increased SLE disease activity. cOPN and cNGAL were measured in prospectively followed pSLE (n=42) and adult SLE (aSLE; n=23) patients and age-matched controls. Time-adjusted cumulative disease activity and disease damage were respectively assessed using adjusted-mean SLE disease activity index (SLEDAI) (AMS) and SLICC/ACR damage index (SDI). Compared to controls, elevated cOPN and cNGAL were observed in pSLE and aSLE. cNGAL preceded worsening SLEDAI by 3-6 months (P=0.04), but was not associated with increased 6-month AMS. High baseline cOPN, which was associated with high IFNalpha activity and expression of autoantibodies to nucleic acids, positively correlated with 6-month AMS (r=0.51 and 0.52, P=0.001 and 0.01 in pSLE and aSLE, respectively) and was associated with SDI increase at 12 months in pSLE (P=0.001). Risk factors for change in SDI in pSLE were cOPN (OR 7.5, 95% CI [2.9-20], P=0.03), but not cNGAL, cumulative prednisone, disease duration, immunosuppression use, gender or ancestry using univariate and multivariate logistic regression. The area under the curve (AUC) when generating the receiver-operating characteristic (ROC) of baseline cOPN sensitivity and specificity for the indication of SLE patients with an increase of SDI over a 12 month period is 0.543 (95% CI 0.347-0.738; positive predictive value 95% and negative predictive value 38%). High circulating OPN levels preceded increased cumulative disease activity and organ damage in SLE patients, especially in pSLE, and its value as a predictor of poor outcome should be further validated in large longitudinal cohorts.


DNA methylation modules associate with incident cardiovascular disease and cumulative risk factor exposure.
Epigenome-wide association studies using DNA methylation have the potential to uncover novel biomarkers and mechanisms of cardiovascular disease (CVD) risk. However, the direction of causation for these associations is not always clear, and investigations to-date have often failed to replicate at the level of individual loci. Here, we undertook module- and region-based DNA methylation analyses of incident CVD in the Women's Health Initiative (WHI) and Framingham Heart Study Offspring Cohort (FHS) in order to find more robust epigenetic biomarkers for cardiovascular risk. We applied weighted gene correlation network analysis (WGCNA) and the Comb-p algorithm to find methylation modules and regions associated with incident CVD in the WHI dataset. We discovered two modules whose activation correlated with CVD risk and replicated across cohorts. One of these modules was enriched for development-related processes and overlaps strongly with epigenetic aging sites. For the other, we showed preliminary evidence for monocyte-specific effects and statistical links to cumulative exposure to traditional cardiovascular risk factors. Additionally, we found three regions (associated with the genes SLC9A1, SLC1A5, and TNRC6C) whose methylation associates with CVD risk. In sum, we present several epigenetic associations with incident CVD which reveal disease mechanisms related to development and monocyte biology. Furthermore, we show that epigenetic modules may act as a molecular readout of cumulative cardiovascular risk factor exposure, with implications for the improvement of clinical risk prediction.


Effect of Selective Thrombus Aspiration on Serum Lipoprotein-Associated Phospholipase A2 in Patients with ST-Elevation Myocardial Infarction Undergoing Primary Percutaneous Coronary Intervention with High Thrombus Burden.
Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a potential therapeutic target in acute coronary syndromes. Although recent evidence does not support the routine use of manual thrombus aspiration (TA) in patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PPCI), the use of TA is associated with a significant improvement in myocardial reperfusion, especially in patients with high thrombus burden (HTB). We hypothesized that TA would reduce the serum Lp-PLA2 levels in STEMI patients undergoing PPCI with HTB. Our study cohort included 320 consecutive STEMI patients undergoing PPCI with HTB who were randomly assigned to receive either TA before PPCI (TA group, n = 160) or PPCI alone (standard PPCI group, n = 160). The baseline characteristics of study participants were well-matched. After 30 ± 2 days, serum Lp-PLA2 levels decreased by 53.9% in the TA group (152.9 ± 58.1 ng/mL) and decreased by 31.2% in the standard PPCI group (84.2 ± 86.6 ng/mL, p < 0.001). The TA group had a significantly lower prevalence of balloon predilatation, number of stents used, total stent length and corrected thrombolysis in myocardial infarction frame count, and a higher percentage of myocardial blush grade ≥ 2 compared with the standard PPCI group (all p < 0.001). No significant difference between the groups was observed in 30 ± 2 days for major adverse cardiovascular and cerebrovascular events (p = 0.702). After 30 ± 2 days of treatment, TA may significantly reduce serum levels of Lp-PLA2 in STEMI patients undergoing PPCI with HTB.


Soluble transferrin receptor, ferritin and soluble transferrin receptor--Ferritin index in assessment of anaemia in rhaeumatoid arthritis.
Anaemia of chronic disease (ACD) is a frequent complication of rheumatoid arthritis (RA). A diagnostic difficulty in RA is the distinction between iron deficiency anaemia (IDA) and ACD. The aim of our study was to evaluate the usefulness of serum soluble transferrin receptor (sTfR) and sTfR/log ferritin (TfR-F) index to diagnose iron deficiency in RA patients with anaemia. Routine laboratory indices of anaemia and sTfR were measured in 20 healthy persons to form the control group, 30 patients with iron deficiency anaemia and 28 RA patients with anaemia. Serum sTfR levels were significantly elevated above the cut-off value in patients with IDA and those in the iron depleted RA subgroup (ferritin < 60 microg/L) compared with those in the control and iron repleted RA subgroup (ferritin > 60 microg/L). The same was observed for TfR-F index. However, five patients in the iron repleted RA sub group had an elevated sTfR level, of which two had increased TfR-F index. Serum sTfR correlated well with the markers of anaemia and not with ESR. Ferritin had no correlation with markers of anaemia but correlated well with ESR. Measurement of sTfR and TfR-F index are good indicators of iron deficiency in RA patients with anaemia. To be cost effective, sTfR can be estimated in RA patients with anaemia when the ferritin level is more than 60 microg/L.


A novel protein targeting domain directs proteins to the anterior cytoplasmic face of the flagellar pocket in African trypanosomes.
The flagellar pocket of African trypanosomes is a critical sorting station for protein and membrane trafficking, and is considered to be an Achilles' heel of this deadly pathogen. Although several proteins, including receptors for host-derived growth factors, are targeted specifically to the flagellar pocket, the signals responsible for this restricted subcellular localization are entirely unknown. Using T lymphocyte triggering factor-green fluorescent protein (TLTF(1)-GFP) fusion proteins, we demonstrate that an internal 144 amino acid domain of TLTF from Trypanosoma brucei is sufficient for directing GFP to the cytoplasmic side of the anterior flagellar pocket. Immuno-gold electron microscopy reveals that the TLTF-GFP fusion protein is located in an electron dense structure that immediately abuts the anterior flagellar pocket membrane. The amino acid sequence of the TLTF targeting domain does not resemble previously characterized protein trafficking signals, and random mutagenesis reveals that flagellar pocket targeting is conferred by a structural motif, rather than a short, contiguous array of amino acids. The aberrant sorting of two mutant proteins into the flagellum, and the targeting of a related human protein to the plus end of the trypanosome's cytoskeletal microtubules, lead us to suggest that flagellar pocket targeting involves interactions with the trypanosome cytoskeleton. The finding that TLTF-GFP is restricted to the anterior, cytoplasmic face of the flagellar pocket membrane, suggests that there is structural heterogeneity in the membrane of this organelle.


Bone fragility in men--where are we?
This is a summary of several aspects of the epidemiology, pathogenesis and treatment arising directly and indirectly from the proceedings of the Third International Osteoporosis in Men meeting held in Genoa in May 2005. Advances in the study of bone fragility in men have taken place, but many challenges remain. Although the epidemiology of hip fractures is well documented, the epidemiology of other non-vertebral fractures is less well defined even though these fractures contribute substantially to the global burden of fractures in men. The epidemiology of vertebral fragility fractures is derived mostly from cross sectional data. The comparable prevalence of vertebral fractures in men and women is likely to be misleading because of traumatic vertebral fractures that arise in young men. Prospective studies are needed to define the proportion of these fractures that are traumatic. After the age of 50 years, the incidence of vertebral fractures in men is about one third to one half of that in women. As in women, most vertebral and non-vertebral fragility fractures occur in persons without osteoporosis. Identifying these individuals is an unmet challenge. The absolute risk for fractures appears no different by sex in men and women of the same age and bone mineral density (BMD) so that the diagnostic threshold for osteoporosis in women can be used in men. Fracture risk varies around the world and is unlikely to be explained solely by variations in BMD, though there are few data comparing men and women of different races. Both the notion that men lose less bone than women from the endosteal envelope and that they gain more on the periosteal envelope during advancing age needs reassessment as recent evidence challenges these observations. Sex differences in the net gain and loss from these surfaces are likely to be site specific, and research is needed to specify this heterogeneity and the reasons for it. The independent and co-dependent effects of sex hormones and the growth hormone/insulin like growth factor 1 axis on periosteal and endosteal modeling and remodeling during growth as well as ageing are poorly defined. The anti-fracture efficacy and safety of androgens and other agents remain incompletely investigated in men. A great deal of research is needed to advance our understanding of bone fragility in men.


The effect of a brief mindfulness-based intervention on personal recovery in people with bipolar disorder: a randomized controlled trial (study protocol).
With the advent of the recovery movement in mental health, a humanistic paradigm shift has occurred, placing the focus on personal recovery (i.e., hope, identity, and life meaning) instead of functional or clinical recovery only (i.e., symptom reduction or increases in physical function). Along the journey of recovery, people with bipolar disorder (BD) struggle to cope with recurring mood fluctuations between depression and mania. Mindfulness-based interventions (MBIs) have the potential to result in improvements in personal recovery outcomes. Thus, this protocol will evaluate the efficacy and mechanisms of a brief MBI for helping individuals with BD with their personal recovery. It is hypothesized that adults with BD randomly assigned to a brief MBI intervention will report greater improvements in personal recovery than those in a waiting list control condition. In addition, it is hypothesized that such benefits will be mediated by improvements in emotion awareness, emotion regulation, and illness acceptance. Moreover, the specific stage of BD is hypothesized to moderate the beneficial effects of the brief MBI, such that those in the early stage of BD will report more benefits regarding emotion awareness and emotion regulation, whereas those in the late stage of BD will report more advantages concerning illness acceptance. One hundred and fifty-four adults with BD will be recruited from hospitals and community settings for this research project. This study will use a mixed methods design. A randomized-controlled trial will be conducted to compare a brief MBI (four sessions in total) group and a waiting list control group. Assessments will be made at baseline, after intervention, and at six-month follow-up. In addition, a qualitative and participatory research method called Photovoice will be employed to further understand the experiences of the participants who receive the brief MBI along their personal recovery journey. If the study hypotheses are supported, the findings from this research project will provide empirical support for an alternative treatment. Moreover, by identifying the mechanisms of the beneficial effects of the brief MBI, the findings will highlight process variables that could be specifically targeted to make MBI treatment even more effective in this population. This study is registered with the Chinese Clinical Trial Registry ( ChiCTR- 1900024658 ). Registered 20th July 2019.


Comparative genomics of clinical Stenotrophomonas maltophilia isolates reveals genetic diversity which correlates with colonization and persistence in vivo.
Stenotrophomonas maltophilia is a Gram-negative emerging opportunistic pathogen often present in people with respiratory diseases such as cystic fibrosis (CF). People with CF (pwCF) experience lifelong polymicrobial infections of the respiratory mucosa. Our prior work showed that Pseudomonas aeruginosa promotes persistence of S. maltophilia in mouse respiratory infections. As is typical for environmental opportunistic pathogens, S. maltophilia has a large genome and a high degree of genetic diversity. In this study, we evaluated the genomic content of S. maltophilia, combining short and long read sequencing to construct nearly complete genomes of 10 clinical isolates. The genomes of these isolates were then compared with all publicly available S. maltophilia genome assemblies, and each isolate was then evaluated for colonization/persistence in vivo, both alone and in coinfection with P. aeruginosa. We found that while the overall genome size and GC content were fairly consistent between strains, there was considerable variability in both genome structure and gene content. Similarly, there was significant variability in S. maltophilia colonization and persistence in experimental mouse respiratory infections in the presence or absence of P. aeruginosa. Ultimately, this study gives us a greater understanding of the genomic diversity of clinical S. maltophilia isolates, and how this genomic diversity relates to both interactions with other pulmonary pathogens and to host disease progression. Identifying the molecular determinants of infection with S. maltophilia can facilitate development of novel antimicrobial strategies for a highly drug-resistant pathogen.


Pregnancy decision making as a significant life event: a commitment approach.
A value-laden basis for commitment to goals and the behavioral and affective sequelae of commitment were examined in the context of a stressful life event. Fifty-seven women who were interviewed during a clinic visit for a pregnancy test (Time 1 [T1]) subsequently received positive test results and were then interviewed 2 days later (Time 2 [T2]) and a month later (Time 3 [T3]). The intentionality and the meaning of the pregnancy were associated with self-reported commitment to the pregnancy at T1. In turn, commitment predicted affective states both prior to (T1) and shortly after (T2) receiving test results. Initial commitment also predicted decisions to continue versus to terminate the pregnancy. The decision to continue the pregnancy appeared to bolster self-reported commitment. Relatedly, those continuing the pregnancy reported smoking fewer cigarettes at T3 than at T1. For those aborting the pregnancy, commitment at T1 was negatively related to adjustment at T3. Initial commitment predicted subsequent depression, guilt, hostility among those who aborted, whereas commitment predicted anxiety among those who continued the pregnancy. Other correlates of commitment (pregnancy concerns, religion, abortion history, and other life goals) were explored.


Electrophysiology of functional subsidiary pacemakers in canine right atrium.
Glass microelectrodes were used to study the electrical activity of the subsidiary atrial pacemaker (SAP) cells that maintain atrial excitation after suppression of the sinoatrial node. Tissues with documented SAP activity were isolated from the canine inferior right atrium and superfused in vitro with Tyrode solution containing norepinephrine (NE, 10(-8)-10(-7) M). SAP action potentials exhibited prominent diastolic depolarization and a significantly lower maximum diastolic potential, take-off potential, overshoot, rate of rise, and amplitude than typical atrial muscle. Withdrawal of NE completely blocked SAP propagation, although SAP automaticity continued at a slower rate. Acetylcholine (ACh, 5 X 10(-8) M) usually produced complete exit block and decreased spontaneous rate. Higher concentrations of ACh (10(-6) M) elicited a prominent hyperpolarization (19.2 +/- 6.6 mV), completely suppressing SAP automaticity. In quiescent preparations exposed to NE greater than or equal to 10(-7) M, external stimuli at short cycle lengths (less than 1,000 ms) elicited action potentials with delayed afterdepolarizations, which frequently caused nondriven repetitive activity. This triggered activity was inhibited by verapamil or withdrawal of NE. These studies identify and characterize the electrical activity of functional subsidiary pacemakers located in a specific region of the inferior right atrium. In addition, fibers within this region display triggered activity. Spontaneous activity generated by fibers within the SAP region may cause atrial dysrhythmias.


Development and characterization of novel microsatellite markers by Next Generation Sequencing for the blue and red shrimp Aristeus antennatus.
The blue and red shrimp, Aristeus antennatus, is a commercially important crustacean, in the Mediterranean Sea, which has been listed as a priority species for fishery management. Hypervariable microsatellite markers could be a useful tool to identify genetic stocks among geographically close fishing grounds. Potential microsatellite markers (97) identified from next-generation sequencing of an individual shrimp using a 454 GS Junior Pyrosequencer were tested on a preliminary panel of 15 individuals representing the four worldwide genetic stocks of the species from which 35 polymorphic loci were identified and used to characterize an additional 20 individuals from the Western Mediterranean Sea. In the Western Mediterranean sample, 32 out of 35 were polymorphic loci and the number of alleles per locus ranged from 2 to 14 and expected heterozygosity ranged from 0.050 to 0.968. No linkage disequilibrium was detected, indicating the independence of the loci. These novel microsatellites provide additional tools to address questions relating to genetic diversity, parentage studies and connectivity patterns of A. antennatus populations and help develop effective strategies to ensure long-term sustainability of this resource.


Plasmacytoid urothelial carcinoma (UC) are luminal tumors with similar CD8+ Tcell density and PD-L1 protein expression on immune cells as compared to conventional UC.
Plasmacytoid urothelial carcinoma (UC) is a rare pathological variant of UC with low chemotherapeutic sensitivity and dismal outcomes. The molecular and immune profiles of such tumors remain poorly investigated. Herein, we investigated the phenotypical features of a cohort of plasmacytoid UC (n=32) by comparison to a control group of conventional high-grade UC with matched clinicopathological characteristics (n=30). Histopathological analysis included the following antibodies: p63, GATA3, CK5/6, CK20 and HER2. In addition, the density of intra-tumor CD8+ lymphocytes, and PD-L1 expression in tumor (TC) and immune cells (IC) were evaluated. Plasmacytoid UC expressed GATA3 (97% vs 86% P=0.18), CK20 (59% vs 36% P=0.08) markers and showed a significantly higher rate of HER2 overexpression (2+ and 3+ score: 25% vs 0%, P<0.01) compared to controls. A significantly lower expression of CK5/6 (22% vs 56%, P<0.05) and p63 (41% vs 80%, P<0.05) was observed in plasmacytoid UC compared to controls. The density of tumor-infiltrating CD8+ cells was similar between plasmacytoid and conventional UC (P=0.9). PD-L1 expression on IC was similar compared to conventional UC (P=0.3). Together, our study demonstrated that plasmacytoid UC belong to the luminal subtype and display a rather inflamed microenvironment similar to conventional UC. These data support the inclusion of plasmacytoid variant of UC in clinical trials evaluating immune checkpoint inhibitors monotherapy or combination immunotherapeutic strategies.


Thyroid hormone effects on androgen receptor messenger RNA expression in rat Sertoli and peritubular cells.
Postnatal Sertoli cell maturation is characterized by a pronounced rise in androgen receptor (AR) expression, which increases several fold between birth and adulthood. Since both 3,3',5-triiodothyronine (T3) and FSH regulate Sertoli cell proliferation and differentiation, we have determined the effects of T3 and FSH on AR mRNA expression in cultured Sertoli cells from 5-day-old rats. These cultures contain 5-9% peritubular cells, which also express AR mRNA. To insure that the observed T3 responses did not result from peritubular cells, we examined T3 effects on AR mRNA expression in cultured 20-day-old Sertoli cells (which contain minimal peritubular contamination) and peritubular cells, and measured thyroid hormone receptor (TR) mRNA expression in both of these cell types. Sertoli cells from 5- and 20-day-old rat testes were grown in serum-free medium alone (controls) or with ovine FSH (100 ng/ml) and/or T3 (100 nM) for 4 days. Peritubular cells purified from 20-day-old rat testes were grown in serum-containing medium for 8 days. These cells were split 1:4, and grown an additional 8 days, the last 4 days in serum-free medium with or without T3. TR and AR mRNA levels in all cultures were determined by Northern blotting. AR mRNA levels in 5- and 20-day-old cultured Sertoli cells were significantly (P < 0.05) increased by both T3 and FSH alone. Furthermore, AR mRNA levels in Sertoli cells treated with T3 and FSH were greater than with either alone. TR mRNA expression was detected in cultured peritubular cells, but TR mRNA levels in these cells were only approximately 30% of that seen in 20-day-old cultured Sertoli cells. In contrast to Sertoli cells, T3 did not affect peritubular AR mRNA expression. These results indicate that T3 is an important regulator of the postnatal Sertoli cell AR mRNA increase. The additive effects of maximally stimulatory doses of FSH and T3 suggest these hormones work through different mechanisms to increase AR mRNA. TR mRNA expression in peritubular cells indicates these cells may be direct T3 targets, though the function of T3 in these cells is unknown.


Comparative chemical composition and variability of biological activity of methanolic extracts from Hypericum perforatum L.
The biovariability of Hypericum perforatum L. (St. John's Wort) grown wild in Calabria and Sardinia (Italy) was reported with the aim to characterize the species through the isolation, detection, and quantitative evaluations of chemical markers (hypericin, quercetin, rutin) by HPLC analysis. Antioxidant activity of the methanolic H. perforatum extracts showed that the Calabrian samples were more active than those from Sardinia. The antibacterial activity evidenced the best performance on the gram positive bacteria with a MIC value of 50 microg/mL. Moreover, antifungal activity of all the extracts was also tested which showed interesting results particularly on the phytopathogene fungus P. ultimum. The variability shown by the samples could be attributed to environmental factors such as chemical-physical properties, composition of the soil, geographical coordinate, altitude, and solar exposure. The phytochemical analysis and the biological activity data suggested a possible use of H. perforatum extracts in the alimentary, cosmetic, and pharmaceutical fields.


Subcellular location and molecular mobility of human cytosolic sulfotransferase 1C1 in living human embryonic kidney 293 cells.
Cytosolic sulfotransferases were first isolated from the hepatic cytosol, and they have been localized in the cytoplasm of formaldehyde-fixed human cell samples. The current work was carried out to determine the subcellular localization and molecular mobility of cytosolic sulfotransferases in living human embryonic kidney (HEK) 293 cells. In this work, the subcellular location of human cytosolic sulfotransferase 1C1 (SULT1C1) was studied in cultured HEK293 cells using confocal laser-scanning microscopy. A green fluorescent protein (GFP)-tagged SULT1C1 protein was localized in the cytoplasm of living HEK293 cells. This is consistent with results from previous studies on several other cytosolic sulfotransferase isoforms. Fluorescence recovery after photobleaching microscopy was performed to assess the molecular mobility of the expressed GFP-SULT1C1 molecules. The results suggested that the expressed recombinant GFP-SULT1C1 molecules in living HEK293 cells may include both mobile and immobile populations. To obtain additional insights into the subcellular location of SULT1C1, two machine learning algorithms, Sequential Minimal Optimization and Multilayer Perceptron, were used to compute the probability distribution for the localization of SULT1C1 in nine selected cellular compartments. The resulting probability distribution suggested that the most likely subcellular location of SULT1C1 is the cytosol.


Construction, substitution, and sorting of metallo-organic structures via subcomponent self-assembly.
Subcomponent self-assembly allows the construction of complex architectures from simple building blocks via formation of covalent bonds around metal templates. Since both covalent and coordinative bonds are formed reversibly, a wealth of rearrangement reactions is possible involving substitution at both intraligand (often C=N) and metal-ligand (N --> metal) bonds. If the possibilities latent within a set of subcomponents and metal ions are understood, one may also select specific structures from among dynamic libraries of products. The parallel preparation of structures from "nonorthogonal" mixtures of subcomponents is also possible, as is the direction of subcomponents to specific sites within product structures.


The relational luring effect: Retrieval of relational information during associative recognition.
Here we argue that semantic relations (e.g., works in: nurse-hospital) have abstract independent representations in long-term memory (LTM) and that the same representation is accessed by all exemplars of a specific relation. We present evidence from 2 associative recognition experiments that uncovered a novel relational luring effect (RLE) in recognition memory. Participants studied word pairs, and then discriminated between intact (old) pairs and recombined lures. In the first experiment participants responded more slowly to lures that were relationally similar (table-cloth) to studied pairs (floor-carpet), in contrast to relationally dissimilar lures (pipe-water). Experiment 2 extended the RLE by showing a continuous effect of relational lure strength on recognition times (RTs), false alarms, and hits. It used a continuous pair recognition task, where each recombined lure or target could be preceded by 0, 1, 2, 3 or 4 different exemplars of the same relation. RTs and false alarms increased linearly with the number of different previously seen relationally similar pairs. Moreover, more typical exemplars of a given relation lead to a stronger RLE. Finally, hits for intact pairs also rose with the number of previously studied different relational instances. These results suggest that semantic relations exist as independent representations in LTM and that during associative recognition these representations can be a spurious source of familiarity. We discuss the implications of the RLE for current models of semantic and episodic memory, unitization in associative recognition, analogical reasoning and retrieval, as well as constructive memory research. (PsycINFO Database Record


The role of patient-clinician information engagement and information seeking from nonmedical channels in fruit and vegetable intake among cancer patients.
Previous research suggests positive effects of health information seeking on prevention behaviors such as diet, exercise, and fruit and vegetable consumption. The present study builds upon this research and strengthens causal claims from it by examining the lagged effect of patient-clinician information engagement on fruit and vegetable consumption as well as the indirect effect on the outcome through seeking information from nonmedical channels. The results are based on data collected from a randomly drawn sample of breast, prostate, and colorectal cancer patients from the Pennsylvania Cancer Registry who completed mail surveys in the Fall of 2006 and 2007. There was a 65% response rate for baseline subjects (resulting n = 2,013); of those, 1,293 were interviewed 1 year later, and 1,257 were available for our analyses. Results show a positive lagged main effect of patient-clinician information engagement at baseline on fruit and vegetable consumption at follow-up (B = 0.26, SE = 0.10, p = .01). The mediation analysis shows that patient-clinician information engagement leads to increased fruit and vegetable consumption among cancer patients, in part through patients' information seeking from nonmedical channels. Implications of these findings for the cancer patient population and for physicians are discussed.


Isoleucine infusion during "simulated" upper gastrointestinal bleeding improves liver and muscle protein synthesis in cirrhotic patients.
Upper gastrointestinal (GI) bleeding in cirrhotic patients has a high incidence of mortality and morbidity. Postbleeding catabolism has been hypothesized to be partly due to the low biological value of hemoglobin, which lacks the essential amino acid isoleucine. The aims were to study the metabolic consequences of a "simulated" upper GI bleed in patients with cirrhosis of the liver and the effects of intravenous infusion of isoleucine. Portal drained viscera, liver, muscle, and kidney protein kinetics were quantified using a multicatheterization technique during routine portography. Sixteen overnight-fasted, metabolically stable patients who received an intragastric infusion of an amino acid solution mimicking hemoglobin every 4 hours were randomized to saline or isoleucine infusion and received a mixture of stable isotopes (L-[ring-2H5]phenylalanine, L-[ring-2H4]tyrosine, and L-[ring-2H2]tyrosine) to determine organ protein kinetics. This simulated bleed resulted in hypoisoleucinemia that was attenuated by isoleucine infusion. Isoleucine infusion during the bleed resulted in a positive net balance of phenylalanine across liver and muscle, whereas renal and portal drained viscera protein kinetics were unaffected. In the control group, no significant effect was shown. The present study investigated hepatic and portal drained viscera protein metabolism selectively in humans. The data show that hepatic and muscle protein synthesis is stimulated by improving the amino acid composition of the upper GI bleed by simultaneous intravenous isoleucine administration.


Violence, homicide and suicide: strong correlation and wide variation across districts.
The clinical assessment and management of the risk of violence and suicide by people with mental illness may have to focus on environmental as well as individual factors. To investigate possible associations between violence, homicide and suicide rates, population density and indices of deprivation, with particular reference to inner-city boroughs. Coroners' statistics in London for homicide and suicide were obtained, with police-reported homicide and violence rates as a validity check. Correlations were made between these data and population density, the Jarman under-privileged areas score, and the Mental Illness Needs Index for each of the 32 London boroughs. Homicide rates had a 14.3-fold range, suicide a 4.4-fold range and interpersonal violence a 6.6-fold range. The variables under study were strongly correlated with each other. Rates were highest in boroughs with high population density and deprivation scores. The associations persisted when covarying for deprivation, age structure or population density. Because violence, homicide and suicide are so closely correlated, they are likely to be valid indices of the differences between adjacent boroughs; this has implications for the delivery of preventive and mental health services and for clinical management of risk.


Comparing Temporospatial Performance During Brisk and Self-Paced Walking by Men With Osteomyoplastic Transfemoral Amputation and Controls Using Pressure and Muscle Activation Peak Times.
The aim of osteomyoplastic transfemoral amputation (OTFA) is to produce sustained, robust prosthetic gait performance by residuum reconstructing. A better understanding of residuum-socket interface pressures (RSI) and residuum muscle activation should uniquely reveal gait stability to better inform long-term rehabilitation goals. The objectives of this study are to characterize RSI pressures and residuum muscle activation in men with OTFA while walking at two speeds and compare temporospatial muscle activation with intact controls. In this study, we observed and compared healthy men with OTFA and controls during 2-min gait trials at brisk and self-paced speeds, two visits, and 1 year apart. RSI pressures and hip adductors, hamstrings, and quadriceps activation were recorded for those with OTFA. OTFA temporospatial muscle activation patterns were compared with the controls. Within the extracted strides, heel-strike and toe-off events and EMG activation peak times were characterized and compared. Peak times for pressure and EMG activity were examined in individual muscles and antagonist muscles of residual and intact limbs. Six men with OTFA exhibited adductor, hamstring, and quadriceps co-contraction within intact and residual limbs, regardless of walking speed or trial. Co-contraction within their intact limb occurred throughout the gait cycle. Within the residuum, co-contraction occurred during weight transference. The 75% most likely RSI peaks occurred during stance. EMG peaks were 75% most likely to occur during early stance, terminal stance-initial swing, and terminal swing. Participants with OTFA demonstrated adductors-hamstrings-quadriceps co-contraction in the intact thigh and residuum with corresponding RSI pressure increase, primarily during transitions between stance and swing, indicating gait instability, demonstrating the need to explicitly address these deficits continuously in rehabilitation and wellness settings.


[Chlorine channels activated by suberyldicholine in mollusk neurons].
Patch-clamp technique in cell-attached configuration was applied to investigate ion permeability induced by suberyldicholine in the neurons of freshwater mollusc Planorbarius corneus. The inward currents through single channels were registered at patch potentials 50-100 mV more negative than resting potential of the cell. When the patch pipette contained suberyldicholine (5 mol/l), single channel currents were recorded which grouped in bursts and bursts of openings could themselves be grouped together in clusters of bursts. The desensitisation was not too pronounced: usually we were able to observe single-channel currents for 20-30 minutes. The unit conductance gamma was found to be about 10 pS.


Comparative Cytogenetics of the Congo African Grey Parrot (Psittacus erithacus).
The Congo African grey parrot (Psittacus erithacus, PER) is an endemic species of Central Africa, valued for its intelligence and listed as vulnerable due to poaching and habitat destruction. Improved knowledge about the P. erithacus genome is needed to address key biological questions and conservation of this species. The P. erithacus genome was studied using conventional and molecular cytogenetic approaches including Zoo-FISH. P. erithacus has a 'typical' parrot karyotype with 2n = 62-64 and 8 pairs of macrochromosomes. A distinct feature was a sharp macro-microchromosome boundary. Telomeric sequences were present at all chromosome ends and interstitially in PER2q, the latter coinciding with a C-band. NORs mapped to 4 pairs of microchromosomes which is in contrast to a single NOR in ancestral type avian karyotypes. Zoo-FISH with chicken macrochromosomes GGA1-9 and Z revealed patterns of conserved synteny similar to many other avian groups, though neighboring synteny combinations of GGA6/7, 8/9, and 1/4 were distinctive only to parrots. Overall, P. erithacus shared more Zoo-FISH patterns with neotropical macaws than Australian species such as cockatiel and budgerigar. The observations suggest that Psittaciformes karyotypes have undergone more extensive evolutionary rearrangements compared to the majority of other avian genomes.


Non-mydriatic fundus photography: a viable alternative to fundoscopy for identification of diabetic retinopathy in an Aboriginal population in rural Western Australia?
To evaluate the Canon CR5-45NM non-mydriatic fundus camera (Canon, Kanagawa, Japan) for identifying retinopathy and the need for laser treatment in a population of Aboriginal patients with diabetes mellitus in rural Western Australia. Diabetic Aboriginal patients were photographed through undilated pupils using a Canon CR5-45NM non-mydriatic fundus camera, after which ophthalmoscopy was performed using indirect ophthalmoscopy through dilated pupils. The examining ophthalmologist recorded the presence of retinopathy and the need for laser treatment. A proportion of patients were rephotographed through dilated pupils. Photographs were reviewed by a second ophthalmologist who evaluated the quality of the image, the presence of retinopathy and the need for laser treatment. Results of fundus photographs and ophthalmoscopy were compared. Three hundred and twenty-eight eyes in 164 Aboriginal patients were examined. The mean patient age was 48.2 years (range 16-81 years) and the mean duration of diabetes was 7.5 years (range 1-35 years). Seventy-four eyes (22.6%) were diagnosed with retinopathy using combined examination techniques, 44 (59.5%) of which were identified by ophthalmoscopy and 55 (74.3%) by photography. Thirty-five eyes were deemed to need treatment, 18 (51.4%) of which were identified by ophthalmoscopy and 30 (85.7%) by photography. Kappa coefficient measurement for agreement for presence of retinopathy and need for referral was 0.41 and 0.53, respectively. Photograph quality was significantly improved following pupil dilation. The Canon CR5-45NM non-mydriatic fundus camera was relatively good at identifying diabetic retinopathy and could usefully be applied within a screening programme for treatable disease within this population.


[Correlation analysis of clock genes and MEN2 medullary thyroid carcinoma].
Objective: To investigate the correlation between CLOCK and BMAL1 genes and MEN2 medullary thyroid carcinoma (MTC). Methods: Thirteen cases with MEN2 MTC and thirteen cases with non-MEN2 MTC were selected who were treated in the Yantai Yuhuangding Hospital between January 2013 and September 2021. Clinical indicators such as blood calcitonin level, tumor diameter and metastatic lymph node of patients were collected. The expression differences of CLOCK and BMAL1 between MEN2 MTC and para-carcinoma tissue as well as between MEN2 MTC and non-MEN2 MTC were detected by immunohistochemistry and qPCR. The correlation between lymph node metastasis and CLOCK or BMAL1 expression was analyzed. Protein-protein interaction (PPI) network analysis combined with qPCR and correlation analysis was used to explore the expression regulation relationship between RET and circadian clock genes. The rhythm disorder of MEN2 cells was verified by lipopolysaccharide cell stimulation experiment after dexamethasone rhythm synchronization. Results: MEN2 MTC exhibited typical RET gene mutation. The mean blood calcitonin level, the tumor diameter and the number of metastatic lymph nodes of patients with MEN2 MTC were higher than those of patients with non-MEN2 MTC (t value was 2.76, 2.53, 2.26, all P<0.05). Immunohistochemical results showed that the expression levels of CLOCK and BMAL1 in MEN2 MTC were higher than those in non-MEN2 MTC, while negatively expressed in para-cancerous thyroid follicle. qPCR displayed that the expression of CLOCK gene in cancer tissues was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.68 and 2.86, all P<0.05); the expression of BMAL1 gene in MEN2 MTC was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.21 and 2.35, all P<0.05). Correlation analysis showed that the expression levels of CLOCK and BMAL1 genes were positively correlated with the number of lymph node metastases in patients with MEN2 MTC (r=0.65, P<0.001; r=0.52, P=0.005). PPI network analysis indicated that the expression of CLOCK gene was positively correlated with the abnormal expression of RET gene (r=0.96, P<0.001). With lipopolysaccharide to stimulate cultured cells in vitro after dexamethasone rhythm synchronization, the expressions of CLOCK and BMAL1 in MEN2 MTC cells (0.47±0.22 and 2.60±1.48) at 12 hours of synchronization were significantly lower than those in para-cancerous tissues (1.70±1.62 and 8.23±2.52), the difference was statistically significant(t=5.04, P=0.007; t=3.34, P=0.029). Conclusion: CLOCK and BMAL1 are correlated with the occurrence and development of MEN2 MTC, and may be potential targets for the development of new therapeutic strategies for MEN2 MTC.


Identification and its vicissitudes as observed in the perversions.
This paper aims to contribute to the clarification of the different meanings of the concept of identification and the different processes involved in the making of an identification, by considering the perversions as disorders of internalization. The different forms of internalization are regarded as lying along a continuum of which identification is at one extreme and involves the incorporation of (part of) the object-representation into the self-representation. The pervert is shown to be unable to carry out the process of identification because he is fixated at an early developmental phase which I refer to as the 'core complex': instead of identification he has to make use of 'stimulation' in which the subject unconsciously models himself on (parts of) the object. The crucial role the body plays in these dynamics is considered. Exploration of these issues highlights how, in order for identification to take place, the issues of the core complex need to be adequately resolved in the course of normal development.


Soluble CD70: a novel immunotherapeutic agent for experimental glioblastoma.
Given the overall poor outcome with current treatment strategies in malignant gliomas, immunotherapy has been considered a promising experimental approach to glioblastoma for more than 2 decades. A cell surface molecule, CD70, may induce potent antitumor immune responses via activation of the costimulatory receptor CD27 expressed on immune effector cells. There is evidence that a soluble form of CD70 (sCD70) may exhibit biological activity, too. A soluble costimulatory ligand is attractive because it may facilitate immune activation and may achieve a superior tissue distribution. To test the antiglioma effect of sCD70, the authors genetically modified SMA-560 mouse glioma cells to secrete the extracellular domain of CD70. They assessed the immunogenicity of the transfected cells in cocultures with immune effector cells by the determination of immune cell proliferation and the release of interferon-gamma. Syngeneic VM/Dk mice were implanted orthotopically with control or sCD70-releasing glioma cells to determine a survival benefit mediated by sCD70. Depletion studies were performed to identify the cellular mediators of prolonged survival of sCD70-releasing glioma-bearing mice. The authors found that ectopic expression of sCD70 enhanced the proliferation and interferon-gamma release of syngeneic splenocytes in vitro. More importantly, sCD70 prolonged the survival of syngeneic VM/Dk mice bearing intracranial SMA-560 gliomas. The survival rate at 60 days increased from 5 to 45%. Antibody-mediated depletion of CD8-positive T cells abrogates the survival advantage conferred by sCD70. These data suggest that sCD70 is a potent stimulator of antiglioma immune responses that depend critically on CD8-positive T cells. Soluble CD70 could be a powerful adjuvant for future immunotherapy trials for glioblastoma.


Kernels for small molecules and the prediction of mutagenicity, toxicity and anti-cancer activity.
Small molecules play a fundamental role in organic chemistry and biology. They can be used to probe biological systems and to discover new drugs and other useful compounds. As increasing numbers of large datasets of small molecules become available, it is necessary to develop computational methods that can deal with molecules of variable size and structure and predict their physical, chemical and biological properties. Here we develop several new classes of kernels for small molecules using their 1D, 2D and 3D representations. In 1D, we consider string kernels based on SMILES strings. In 2D, we introduce several similarity kernels based on conventional or generalized fingerprints. Generalized fingerprints are derived by counting in different ways subpaths contained in the graph of bonds, using depth-first searches. In 3D, we consider similarity measures between histograms of pairwise distances between atom classes. These kernels can be computed efficiently and are applied to problems of classification and prediction of mutagenicity, toxicity and anti-cancer activity on three publicly available datasets. The results derived using cross-validation methods are state-of-the-art. Tradeoffs between various kernels are briefly discussed. Datasets available from http://www.igb.uci.edu/servers/servers.html


Single-layer continuous versus two-layer interrupted intestinal anastomosis: a prospective randomized trial.
To determine the suitability of a single-layer continuous technique for intestinal anastomosis in a surgical training program. Several recent reports have advocated the use of a continuous single-layer technique for intestinal anastomosis. Purported advantages include shorter time for construction, lower cost, and perhaps a lower rate of anastomotic leakage. The authors hypothesized that the single-layer continuous anastomosis could be safely introduced into a surgical training program and that it could be performed in less time and at a lower cost than the two-layer interrupted anastomosis. The study was conducted during a 3-year period ending September 1999. All adult patients requiring intestinal anastomosis were considered eligible. Patients who required anastomosis to the stomach, duodenum, and rectum were excluded. Patients were also excluded if the surgeon did not believe either technique could be used. Patients were randomly assigned to one- or two-layer techniques. Single-layer anastomoses were performed with a continuous 3-0 polypropylene suture. Two-layer anastomoses were constructed using interrupted 3-0 silk Lembert sutures for the outer layer and a continuous 3-0 polyglycolic acid suture for the inner layer. The time for anastomosis began with the placement of the first stitch and ended when the last stitch was cut. Anastomotic leak was defined as radiographic demonstration of a fistula or nonabsorbable material draining from a wound after oral administration, or visible disruption of the suture line during reexploration. Sixty-five single-layer and 67 two-layer anastomoses were performed. The groups were evenly matched according to age, sex, diagnosis, and location of the anastomosis. Two leaks (3.1%) occurred in the single-layer group and one (1.5%) in the two-layer group. Two abscesses (3.0%) occurred in each group. A mean of 20.8 minutes was required to construct a single-layer anastomosis versus 30.7 minutes for the two-layer technique. Mean length of stay was 7.9 days for single-layer patients and 9.9 days for two-layer patients; this difference did not quite reach statistical significance. Cost of materials was $4.61 for the single-layer technique and $35.38 for the two-layer method. A single-layer continuous anastomosis can be constructed in significantly less time and with a similar rate of complications compared with the two-layer technique. It also costs less than any other method and can be incorporated into a surgical training program without a significant increase in complications.


The degrees of freedom problem in human standing posture: collective and component dynamics.
The experiment was setup to investigate the coordination and control of the degrees of freedom (DFs) of human standing posture with particular reference to the identification of the collective and component variables. Subjects stood in 3 postural tasks: feet side by side, single left foot quiet stance and single left foot stance with body rocking at the ankle joint in the sagittal plane. All three postural tasks showed very high coherence (∼ 1) of center of pressure (COP)--center of mass (COM) in the low frequency range. The ankle and hip coherence was mid range (∼.5) with the tasks having different ankle/hip compensatory cophase patterns. The findings support the view that the in-phase relation of the low frequency components of the COP-COM dynamic is the collective variable in the postural tasks investigated. The motions of the individual joints (ankle, knee, hip, neck) and couplings of pair wise joint synergies (e.g., ankle-hip) provide a supporting cooperative role to the preservation of the collective variable in maintaining the COM within the stability region of the base of support (BOS) and minimizing the amount of body motion consistent with the task constraint.


Role of fibroblast growth factor receptors 1 and 2 in the ureteric bud.
Fibroblast growth receptors (FGFRs) consist of four signaling family members. Mice with deletions of fgfr1 or fgfr2 are embryonic lethal prior to the onset of kidney development. To determine roles of FGFR1 and FGFR2 in the ureteric bud, we used a conditional targeting approach. First, we generated transgenic mice using the Hoxb7 promoter to drive cre recombinase and green fluorescent protein expression throughout ureteric bud tissue. We crossed Hoxb7creEGFP mice with mice carrying lox-p sites flanking critical regions of fgfr1 and/or fgfr2. Absence of fgfr1 from the ureteric bud (fgfr1(UB-/-)) results in no apparent renal abnormalities. In contrast, fgfr2(UB-/-) mice have very aberrant ureteric bud branching, thin ureteric bud stalks, and fewer ureteric bud tips. Fgfr2(UB-/-) ureteric bud tips also demonstrate inappropriate regions of apoptosis and reduced proliferation. The nephrogenic mesenchymal lineage in fgfr2(UB-/-) mice develops normal-appearing glomeruli and tubules, and only slightly fewer nephrons than controls. In contrast, fgfr2(UB-/-) kidneys have abnormally thickened subcapsular cortical stromal mesenchyme. Ultimately, fgfr2(UB-/-) adult kidneys are small and abnormally shaped or are hydronephrotic. Finally, there are no additional abnormalities in the fgfr1/2(UB-/-) kidneys versus the fgfr2(UB-/-) kidneys. In conclusion, FGFR2, but not FGFR1, appears crucial for ureteric bud branching morphogenesis and stromal mesenchyme patterning.


A plea to stop using the case-control design in retrospective database studies.
The case-control design is widely used in retrospective database studies, often leading to spectacular findings. However, results of these studies often cannot be replicated, and the advantage of this design over others is questionable. To demonstrate the shortcomings of applications of this design, we replicate two published case-control studies. The first investigates isotretinoin and ulcerative colitis using a simple case-control design. The second focuses on dipeptidyl peptidase-4 inhibitors and acute pancreatitis, using a nested case-control design. We include large sets of negative control exposures (where the true odds ratio is believed to be 1) in both studies. Both replication studies produce effect size estimates consistent with the original studies, but also generate estimates for the negative control exposures showing substantial residual bias. In contrast, applying a self-controlled design to answer the same questions using the same data reveals far less bias. Although the case-control design in general is not at fault, its application in retrospective database studies, where all exposure and covariate data for the entire cohort are available, is unnecessary, as other alternatives such as cohort and self-controlled designs are available. Moreover, by focusing on cases and controls it opens the door to inappropriate comparisons between exposure groups, leading to confounding for which the design has few options to adjust for. We argue that this design should no longer be used in these types of data. At the very least, negative control exposures should be used to prove that the concerns raised here do not apply.


Ipatasertib exhibits anti‑tumorigenic effects and enhances sensitivity to paclitaxel in endometrial cancer in vitro and in vivo.
Endometrial cancer is the most common gynecologic cancer and one of the only cancers for which incidence and mortality is steadily increasing. Although curable with surgery in the early stages, endometrial cancer presents a significant clinical challenge in the metastatic and recurrent setting with few novel treatment strategies emerging in the past fifty years. Ipatasertib (IPAT) is an orally bioavailable pan‑AKT inhibitor, which targets all three AKT isoforms and has demonstrated anti‑tumor activity in pre‑clinical models, with clinical trials emerging for many cancer types. In the present study, the MTT assay was employed to evaluate the therapeutic efficacy of IPAT or IPAT in combination with paclitaxel (PTX) in endometrial cancer cell lines and primary cultures of endometrial cancer. The effect of IPAT and PTX on the growth of endometrial tumors was evaluated in a transgenic mouse model of endometrial cancer. Apoptosis was assessed using cleaved caspase assays and cellular stress was assessed using ROS, JC1 and tetramethylrhodamine ethyl ester assays. The protein expression levels of markers of apoptosis and cellular stress, and DNA damage were evaluated using western blotting and immunohistochemistry. IPAT significantly inhibited cell proliferation, caused cell cycle G1 phase arrest, and induced cellular stress and mitochondrial apoptosis in a dose dependent manner in human endometrial cancer cell lines. Combined treatment with low doses of IPAT and PTX led to synergistic inhibition of cell proliferation and induction of cleaved caspase 3 activity in the human endometrial cancer cell lines and the primary cultures. Furthermore, IPAT effectively reduced tumor growth, accompanied by decreased protein expression levels of Ki67 and phosphorylation of S6 in the Lkb1fl/flp53fl/fl mouse model of endometrioid endometrial cancer. The combination of IPAT and PTX resulted in increased expression of phosphorylated‑H2AX and KIF14, markers of DNA damage and microtubule dysfunction respectively, as compared with IPAT alone, PTX alone or placebo‑treated mice. The results of the present study provide a biological rationale to evaluate IPAT and the combination of IPAT and PTX in future clinical trials for endometrial cancer.


Cortical regulation of striatal medium spiny neuron dendritic remodeling in parkinsonism: modulation of glutamate release reverses dopamine depletion-induced dendritic spine loss.
Striatal medium spiny neurons (MSNs) receive glutamatergic afferents from the cerebral cortex and dopaminergic inputs from the substantia nigra (SN). Striatal dopamine loss decreases the number of MSN dendritic spines. This loss of spines has been suggested to reflect the removal of tonic dopamine inhibitory control over corticostriatal glutamatergic drive, with increased glutamate release culminating in MSN spine loss. We tested this hypothesis in two ways. We first determined in vivo if decortication reverses or prevents dopamine depletion-induced spine loss by placing motor cortex lesions 4 weeks after, or at the time of, 6-hydroxydopamine lesions of the SN. Animals were sacrificed 4 weeks after cortical lesions. Motor cortex lesions significantly reversed the loss of MSN spines elicited by dopamine denervation; a similar effect was observed in the prevention experiment. We then determined if modulating glutamate release in organotypic cocultures prevented spine loss. Treatment of the cultures with the mGluR2/3 agonist LY379268 to suppress corticostriatal glutamate release completely blocked spine loss in dopamine-denervated cultures. These studies provide the first evidence to show that MSN spine loss associated with parkinsonism can be reversed and point to suppression of corticostriatal glutamate release as a means of slowing progression in Parkinson's disease.


Decoupling brain activations of muscle-caused activations and mental intention-cause activations using the general linear model: A functional near-infrared spectroscopy study.
Integrating a brain-computer interface into a lower-limb medical rehabilitation assistive device can enhance rehabilitation efficiency. The latest research in the field focuses on the decoding performance of different motions. However, the difference between muscle-caused primitive activation and mental intention-caused activation has not been fully investigated. Thus, our study tried to decouple these two kinds of cerebral activation using a general linear model (GLM). Nine healthy and right-handed subjects were recruited for a two-section experiment. They were asked to extend or flex their knees while seated in the first section of the experiment or standing in the second section. Functional near-infrared spectroscopy (fNIR) was adopted to monitor their hemodynamic changes. Two groups of paradigms (one for circle-wise analysis, the other for full-section analysis) were constructed from the experimental paradigm. Each group consisted of three (the first intention, the second intention, and the muscle activation). The constructed paradigms were fed to the Balloon model for six desired hemodynamic responses (dHRFs). The regressor of GLM consisted of three dHRFs and the corresponding motion artifacts and drifts. The simulated physiological noises were included in the structured background matrix. The results showed that all subjects had similar cerebral activation patterns for the intention to extend or flex knees. The activation during musclecaused activation was less intense than that caused by both intentions. This finding can help further research on more efficient motion intention detection and the possibility of multiple motions decoding.


The views of key stakeholders in Zimbabwe on the introduction of postgraduate family medicine training: A qualitative study.
Strengthening primary health care (PHC) is a priority for all effective health systems, and family physicians are seen as a key member of the PHC team. Zimbabwe has joined a number of African countries that are seriously considering the introduction of postgraduate family medicine training. Implementation of training, however, has not yet happened. To explore the views of key stakeholders on the introduction of postgraduate family medicine training. Key academic, governmental and professional stakeholders in Zimbabwean health and higher education systems. Twelve semi-structured interviews were conducted with purposively selected key stakeholders. Data were recorded, transcribed and analysed using the framework method. Anticipated benefits: More effective functioning of PHC and district health services with reduced referrals, improved access to more comprehensive services and improved clinical outcomes. Opportunities: International trend towards family medicine training, government support, availability of a small group of local trainers, need to revise PHC policy. Anticipated barriers: Family medicine is unattractive as a career choice because it is largely unknown to newly qualified doctors and may not be recognised in private sector. There is concern that advocacy is mainly coming from the private sector. Threats: Economic conditions, poor remuneration, lack of funding for resources and new initiatives, resistance from other specialists in private sector. Stakeholders anticipated significant benefits from the introduction of family medicine training and identified a number of opportunities that support this, but also recognised the existence of major barriers and threats to successful implementation.


Effect of first, second, and third trimester placental factors on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes.
To investigate effect of first, second, and third trimester placental factors (PF) on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes (PBLs), and to explore their influence on human immunodeficiency virus (HIV) vertical transmission. Human peripheral blood mononuclear cells (PBMCs) were treated with first, second, and third trimester PF (concentration 25%) respectively for 24 hours. The expression of CD4, CCR5, and CXCR4 in PBLs, and the percentages of CCR5(+), CXCR4(+)ìand CCR5(+)CXCR4(+) cells in peripheral blood CD4(+) lymphocytes were determined with flow cytometry. All trimester PFs reduced CCR5 expression in PBLs. The efficiency of the first trimester PF was higher than that of the second and third trimester PF. The percentage of CCR5(+) cells in peripheral blood CD4(+) lymphocytes of PF groups was significantly lower than that of the control group, and the percentage of CCR5(+) cells in peripheral blood CD4(+) lymphocytes of the first trimester PF group was significantly lower than that of the second and third trimester group. The percentages of CCR5(+)CXCR4(+) cells in peripheral blood CD4(+) lymphocytes of PF groups were significantly decreased as compared with the control group, and the percentage of CCR5(+)CXCR4(+) cells in peripheral blood CD4(+) lymphocytes of the first trimester PF group was significantly lower than that of the third trimester PF group. PF can reduce the expression of CCR5 in human PBLs and peripheral blood CD4(+) lymphocytes, indicating that PF might reduce R5 virus infection via preventing HIV entry, and might play an important role in reducing R5 virus intrauterine infection.


Clinical synovitis in a particular joint is associated with progression of erosions and joint space narrowing in that same joint, but not in patients initially treated with infliximab.
To assess the relationship between joint tenderness, swelling and joint damage progression in individual joints and to evaluate the influence of treatment on these relationships. First-year data of the Behandel Strategieën (BeSt) study were used, in which patients recently diagnosed as having rheumatoid arthritis (RA) were randomly assigned into four different treatment strategies. Baseline and 1-year x-rays of the hands and feet were assessed using the Sharp-van der Heijde score (SHS). With generalised estimating equations, 3-monthly assessments of tender and swollen joints of year 1 were related to erosion progression, joint space narrowing (JSN) progression and total SHS progression at the individual joint level (definition > 0.5 SHS units) in year 1, corrected for potential confounders and within-patient correlation for multiple joints per patient. During year 1, 59% of all 13 959 joints analysed were ever tender and 45% ever swollen, 2.1% showed erosion progression, 1.9% JSN progression and 3.6% SHS progression. Swelling and tenderness were both independently associated with erosion and JSN progression with comparable OR, although with higher OR in the hands than in the feet. Local swelling and tenderness were not associated with local damage progression in patients initially treated with infliximab. Clinical signs of synovitis are associated with erosion and JSN progression in individual joints after 1 year in RA. A disconnect between synovitis and joint damage progression was observed at joint level in patients who were treated with methotrexate and infliximab as initial treatment, confirming the disconnect between synovitis and the development of joint damage in tumour necrosis factor blockers seen at patient level.


[Thromboembolic diseases in pregnancy].
In pregnancy, thromboembolic complications are six times more frequent than in nonpregnant women. Maternal age, idiopathic or secondary thrombosis in the patients history, the mode of delivery, bed rest and / or obesity as well as the thrombophilic defects are considerable factors for an increased risk of deep vein thrombosis (DVT) and pulmonary embolism (PE). Compared to unfractionated heparins (UFHs) prophylactic treatment relies mainly on low molecular weight heparins (LMWHs) which safety is now well established in pregnant women. In view of their multiple side effects oral anticoagulants such as cumarine or aspirin are contraindicated or of subordinate importance. The procedures to exclude DVT follow the usual diagnostic algorithms considering maximum radiation protection in case of further apparative diagnostics. Although there are reported good experiences with LMWHs in the treatment of DVT, UFHs are still the preferred drugs in this clinical condition. Insufficient heparin treatment or impending postthrombotic complications may be reasons for additional therapies like thrombolysis or operative thrombectomy.


Cardiovascular magnetic resonance normal values in children for biventricular wall thickness and mass.
Pediatric patients are becoming increasingly referred for cardiovascular magnetic resonance (CMR). Measurement of ventricular wall thickness is typically part of the assessment and can be of diagnostic importance, e.g. in arterial hypertension. However, normal values for left ventricular (LV) and right ventricular (RV) wall thickness in pediatric patients are lacking. The aim of this study was to establish pediatric centile charts for segmental LV and RV myocardial thickness in a retrospective multicenter CMR study. CMR was performed in 161 healthy children and adolescents with an age range between 6 and 18 years from two centers in the UK and Germany as well as from a previously published CMR project of the German Competence Network for Congenital Heart Defects. LV myocardial thickness of 16 segments was measured on the short axis stack using the American Heart Association segmentation model. In addition, the thickness of the RV inferior and anterior free wall as well as biventricular mass was measured. The mean age (standard deviation) of the subjects was 13.6 (2.9) years, 64 (39.7%) were female. Myocardial thickness of the basal septum (basal antero- and inferoseptal wall) was 5.2 (1.1) mm, and the basal lateral wall (basal antero- and inferolateral) measured 5.1 (1.2) mm. Mid-ventricular septum (antero- and inferoseptal wall) measured 5.5 (1.2) mm, and mid-ventricular lateral wall (antero- and inferolateral wall) was 4.7 (1.2) mm. Separate centile charts for boys and girls for all myocardial segments and myocardial mass were created because gender was significantly correlated with LV myocardial thickness (p < 0.001 at basal level, p = 0.001 at midventricular level and p = 0.005 at the apex) and biventricular mass (LV, p < 0.001; RV, p < 0.001). We established CMR normal values of segmental myocardial thickness and biventricular mass in children and adolescents. Our data are of use for the detection of abnormal myocardial properties and can serve as a reference in future studies and clinical practice.


A sacrificial process for fabrication of biodegradable polymer membranes with submicron thickness.
A new sacrificial molding process using a single mask has been developed to fabricate ultrathin 2-dimensional membranes from several biocompatible polymeric materials. The fabrication process is similar to a sacrificial microelectromechanical systems (MEMS) process flow, where a mold is created from a material that can be coated with a biodegradable polymer and subsequently etched away, leaving behind a very thin polymer membrane. In this work, two different sacrificial mold materials, silicon dioxide (SiO2 ) and Liftoff Resist (LOR) were used. Three different biodegradable materials; polycaprolactone (PCL), poly(lactic-co-glycolic acid) (PLGA), and polyglycidyl methacrylate (PGMA), were chosen as model polymers. We demonstrate that this process is capable of fabricating 200-500 nm thin, through-hole polymer membranes with various geometries, pore-sizes and spatial features approaching 2.5 µm using a mold fabricated via a single contact photolithography exposure. In addition, the membranes can be mounted to support rings made from either SU8 or PCL for easy handling after release. Cell culture compatibility of the fabricated membranes was evaluated with human dermal microvascular endothelial cells (HDMECs) seeded onto the ultrathin porous membranes, where the cells grew and formed confluent layers with well-established cell-cell contacts. Furthermore, human trabecular meshwork cells (HTMCs) cultured on these scaffolds showed similar proliferation as on flat PCL substrates, further validating its compatibility. All together, these results demonstrated the feasibility of our sacrificial fabrication process to produce biocompatible, ultra-thin membranes with defined microstructures (i.e., pores) with the potential to be used as substrates for tissue engineering applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1192-1201, 2016.


Midwives on the move: comparing the requirements for practice and integration contexts for internationally educated midwives in Canada with the U.S., U.K. and Australia.
Midwives have long been internationally mobile, but there has been relatively little attention paid to the requirements that internationally educated midwives (IEMs) must meet to practice and become integrated into the health-care systems of their destination country. This paper examines from a comparative perspective the policy context and integration procedures that IEMs must follow in order to practice in Canada and how this compares with the U.S., the U.K., and Australia. The data upon which this paper is based are largely derived from an analysis of documents and websites for key organisations involved in the integration process supplemented with interviews with key informants influential in the assessment and integration of IEMs. What these data reveal is that the challenges for IEMs derive in large part from the differences in entry to practice requirements-midwifery and nursing training in three of the cases, baccalaureate training in one. Another critical factor is whether bridging or adaptation programmes are available (rarely in the U.S.), and whether they focus more on orientation objectives (as they do in the U.K. and Australia) or also the upgrading of skills (as they do in Canada) critical for IEM professional integration. These different approaches to the integration of IEMs have important implications for the 'brain drain' and 'brain waste' of much needed midwifery skills in both source and destination countries.


Murine Model of Tuberculosis. In vitro and in vivo Study.
Animal models that mimic the whole spectrum of susceptibility and resistance of human populations to tuberculosis would be useful for studying pathogenesis of tuberculosis, mechanisms of the host defense, and for testing prospective prophylactic and therapeutic tuberculosis vaccines and drugs. Inbred mouse strains are broadly and successfully used to model human tuberculosis and study mechanisms of host-mycobacteria interaction. Here we consider numerous aspects of murine models of tuberculosis. We analyze the course of tuberculosis depending on route and dosage of mycobacterial infection, and discuss the conformity of these approaches to the natural route of human infection. The advantage of the mouse model is provided by the availability of hundreds of inbred strains with different genetic background. It allows a genetic analysis of the mouse susceptibility/resistance to tuberculosis. Mouse strains with specific properties such as congenic, recombinant, recombinant inbred, recombinant congenic, mutant and usage of whole genome screening led to mapping loci controlling these traits. Transgenic mouse strains with gene insertion or targeted mutation (knock-out of a gene) are very effective tools for studying the role of specific genes controlling the anti-tuberculosis immunity. Mice with knock-out genes for cytokines, chemokines, cell subpopulations, cell receptors, and enzymes are broadly used to identify the mechanisms of tuberculosis control. In this review we examine a mouse model for anti-tuberculosis vaccine and drug testing. In addition, we present some important phenomena of tuberculosis immunity - latent tuberculosis/reactivation and immunological memory to tuberculosis.


The effects of Nigella sativa oil, thymoquinone, propolis, and caffeic acid phenethyl ester on radiation-induced cataract.
The aim of this study was to investigate the antioxidant and radioprotective effects of propolis, caffeic acid phenethyl ester (CAPE), Nigella sativa oil (NSO), and thymoquinone (TQ) against ionizing radiation-induced cataracts in lens after total cranium irradiation of rats with single dose of 5-Gy cobalt-60 gamma rays. A total of 74 Sprague-Dawley rats were divided into 8 groups to test the radioprotective effectiveness of Nigella sativa oil, thymoquine, propolis, or caffeic acid phenethyl ester administered by either orogastric tube or intraperitoneal injection. Appropriate control groups were also studied. Chylack's cataract classification was used in the study. At the end of the tenth day, cataracts developed in 80 % of the rats in the radiotherapy group. After irradiation, cataract rate dropped to 20 % in NSO, 30 % in propolis, 40 % in CAPE, and 50 % in TQ groups and was limited to grade 1 and grade 2. Cataract formation was observed the least in NSO group and the most in TQ group. In the irradiated (IR) group, superoxide dismutase activity was lower, while glutathione peroxidase and xanthine oxidase activities and malondialdehyde level were higher compared with the other groups. Total superoxide scavenger activity and nonenzymatic superoxide scavenger activity were not statistically significant in IR group compared with the other groups. The findings obtained in the study might suggest that propolis, CAPE, NSO, and TQ could prevent cataractogenesis in ionizing radiation-induced cataracts in the lenses of rats, wherein propolis and NSO were found to be more potent.


Understanding the influence of all nodes in a network.
Centrality measures such as the degree, k-shell, or eigenvalue centrality can identify a network's most influential nodes, but are rarely usefully accurate in quantifying the spreading power of the vast majority of nodes which are not highly influential. The spreading power of all network nodes is better explained by considering, from a continuous-time epidemiological perspective, the distribution of the force of infection each node generates. The resulting metric, the expected force, accurately quantifies node spreading power under all primary epidemiological models across a wide range of archetypical human contact networks. When node power is low, influence is a function of neighbor degree. As power increases, a node's own degree becomes more important. The strength of this relationship is modulated by network structure, being more pronounced in narrow, dense networks typical of social networking and weakening in broader, looser association networks such as the Internet. The expected force can be computed independently for individual nodes, making it applicable for networks whose adjacency matrix is dynamic, not well specified, or overwhelmingly large.


Evaluation of the mouse model for acute otitis media.
Various animal models have been employed for otitis media research. The mouse has been studied less, in spite of its many advantages. To better understand the suitability of the mouse for studies of otitis media, an evaluation was made of its middle ear inflammatory processes following inoculation with heat-killed Streptococcus pneumoniae (strain 6A), one of the three most common bacteria to cause otitis media in the human. A total of 94 BALB/c mice were injected transtympanically with three concentrations of heat-killed bacteria (10(4), 10(6), and 10(9) organisms per ml) and inflammation evaluated with both histologic examination and auditory brainstem response audiometry. Dose-related measures of the time course of inflammation showed it was maximal at 3 days. PBS-injected control mice also demonstrated some degree of middle ear inflammation. Therefore, inflammation measures from PBS injected mice were used as the threshold above which histologic inflammatory changes would be considered a response to bacteria. These quantitative comparisons of bacterial and PBS inoculations revealed the most significant middle ear measures of inflammation were amount of fluid in the middle ear, tympanic membrane thickness, and number of inflammatory cells. The induction of middle ear inflammation in the mouse demonstrated the applicability of this model for investigations of otitis media.


Cancer patients hospitalised in the last week of life risk insufficient care quality - a population-based study from the Swedish Register of Palliative Care.
One-quarter of all cancer deaths in Sweden occur in hospitals. If the place of death affects the quality of end-of-life (EOL) is largely unknown. This population-based, retrospective study included all adults cancer deaths reported to the Swedish Register of Palliative Care in 2011-2013 (N = 41,729). Hospital deaths were compared to deaths occurring in general or specialised palliative care, or in nursing homes with respect to care quality indicators in the last week of life. Odds ratios (OR) with 95% confidence intervals (CI) were calculated with specialised palliative home care as reference. Preferred place of death was unknown for 63% of hospitalised patients and consistent with the actual place of death in 25% compared to 97% in palliative home care. Hospitalised patients were less likely to be informed when death was imminent (OR: 0.3; CI: 0.28-0.33) as were their families (OR: 0.51; CI: 0.46-0.57). Validated screening tools were less often used in hospitals for assessment of pain (OR: 0.32; CI: 0.30-0.34) or other symptoms (OR: 0.31; CI: 0.28-0.34) despite similar levels of EOL symptoms. Prescriptions of as needed drugs against anxiety (OR: 0.27; CI: 0.24-0.30), nausea (OR: 0.19; CI: 0.17-0.21), or pulmonary secretions (OR: 0.29; CI: 0.26-0.32) were less prevalent in hospitals. Bereavement support was offered after 57% of hospital deaths compared to 87-97% in palliative care units and 72% in nursing homes. Dying in hospital was associated with inferior end-of-life care quality among cancer patients in Sweden.


Manager traits and quality-of-care performance in hospitals.
This paper aims to assess the impact of the leadership traits of chief executive officers (CEOs) on hospital performance in the USA. The effectiveness and efficiency of the CEO is of critical importance to the performance of any organization, including hospitals. Management systems and manager behaviours (traits) are of crucial importance to any organization because of their connection with organizational performance. To identify key factors associated with the quality of care delivered by hospitals, the authors gathered perceptions of manager traits from chief executive officers (CEOs) and followers in three groups of US hospitals delivering different levels of quality of care performance. Three high- and three low-performing hospitals were selected from the top and bottom 20th percentiles, respectively, using a national hospital ranking system based on standard quality of care performance measures. Three lean hospitals delivering intermediate performance were also selected. A survey was used to gather perceptions of manager traits (providing a modern or lean management system inclination) from CEOs and their followers in the three groups, which were compared. Four traits were found to be significantly different (alpha < 0.05) between lean (intermediate-) and low-performing hospitals. The different perceptions between these two hospital groups were all held by followers in the low-performing hospitals and not the CEOs, and all had a modern management inclination. No differences were found between lean (intermediate-) and high-performing hospitals, or between high- and low-performing hospitals. These findings support a need for hospital managers to acquire appropriate traits to achieve lean transformation, support a benefit of measuring manager traits to assess progress towards lean transformation and lend weight to improved quality of care that can be delivered by hospitals adopting a lean system of management.


The Use of Cardiac Autonomic Responses to Aerobic Exercise in Elderly Stroke Patients: Functional Rehabilitation as a Public Health Policy.
The development of public policies must be guided by full knowledge of the health-disease process of the population. Aerobic exercises are recommended for rehabilitation in stroke patients, and have been shown to improve heart rate variability (HRV). Our aim was to compare the cardiac autonomic modulation of elderly stroke patients with that of healthy elderly people during and after an acute bout of aerobic exercise. A total of 60 elderly people participated in the study (30 in the control group, mean age of 67 ± 4 years; 30 in the stroke group, mean age of 69 ± 3 years). HRV was analyzed in rest-10 min of rest in supine position; exercise-the 30 min of peak exercise; and recovery-30 min in supine position post-exercise. Taking rest and exercises together, for SDNN, RMSSD, pNN50, RRTri, and TINN, there was no difference between the stroke and control groups (p = 0.062; p = 0.601; p = 0.166; p = 0.224, and p = 0.059, respectively). The HF (ms2) was higher and the LF/HF ratio was lower for the stroke group than the control group (p < 0.001 and p = 0.007, respectively). The SD2 was lower for the stroke group than for the control group (p = 0.041). Stroke patients present reduced variability at rest, sympathetic predominance during exercise, and do not return to baseline after the 30 min of recovery, with similar responses found in the healthy elderly group.


Health-related quality of life in long-term survivors with localised prostate cancer by therapy-Results from a population-based study.
Several therapies for localised prostate cancer (PC) are available; all yield similar survival rates. However, each therapy has significant side effects that can influence patients' health-related quality of life (HRQoL) in the long run. The study sample included 911 survivors with localised PC, 5-15 years post-diagnosis who were identified from the population-based CAESAR + study in Germany. HRQoL was assessed using the EORTC QLQ-C30 and EORTC QLQ-PR25 questionnaires. The association between type of therapy and HRQoL was assessed with multivariable linear regression and global F-test adjusting for age, time since diagnosis and comorbidities. Overall, survivors treated with radical prostatectomy (RP) or radiotherapy (RT) alone reported the best HRQoL and the lowest symptom burden. Conversely, survivors treated with androgen deprivation therapy (ADT) (& RP/RT) or RP & RT (in combination) reported the worst HRQoL and the highest symptom burden. Significant differences among treatment groups in HRQoL were found for global health status (p = 0.041), social functioning (p = 0.007), urinary symptoms (p = 0.035), bowel symptoms (p = 0.017) and hormonal treatment-related symptoms (p < 0.001) among other symptoms. Long-term localised PC survivors formerly treated with a combination of RP and RT or with ADT report poorer HRQoL and more symptoms than patients treated with either RP or RT alone.


Chemical composition and properties of ashes from combustion plants using Miscanthus as fuel.
Miscanthus giganteus is one of the energy crops considered to show potential for a substantial contribution to sustainable energy production. In the literature there is little data available about the chemical composition of ashes from the combustion of Miscanthus and practically no data about their physical properties. However, for handling, treatment and utilization of the ashes this information is important. In this study ashes from two biomass combustion plants using Miscanthus as fuel were investigated. The density of the ashes was 2230±35kg/m3, which was similar to the density of ashes from straw combustion. Also the bulk densities were close to those reported for straw ashes. The flowability of the ashes was a little worse than the flowability of ashes from wood combustion. The measured heavy metal concentrations were below the usual limits for utilization of the ashes as soil conditioner. The concentrations in the bottom ash were similar to those reported for ash from forest residue combustion plants. In comparison with cyclone fly ashes from forest residue combustion the measured heavy metal concentrations in the cyclone fly ash were considerably lower. Cl-, S and Zn were enriched in the cyclone fly ash which is also known for ashes from wood combustion. In comparison with literature data obtained from Miscanthus plant material the concentrations of K, Cl- and S were lower. This can be attributed to the fact that the finest fly ash is not collected by the cyclone de-dusting system of the Miscanthus combustion plants.


Transcriptional responses in a Drosophila defensive symbiosis.
Inherited symbionts are ubiquitous in insects and can have important consequences for the fitness of their hosts. Many inherited symbionts defend their hosts against parasites or other natural enemies; however, the means by which most symbionts confer protection is virtually unknown. We examine the mechanisms of defence in a recently discovered case of symbiont-mediated protection, where the bacterial symbiont Spiroplasma defends the fly Drosophila neotestacea from a virulent nematode parasite, Howardula aoronymphium. Using quantitative PCR of Spiroplasma infection intensities and whole transcriptome sequencing, we attempt to distinguish between the following modes of defence: symbiont-parasite competition, host immune priming and the production of toxic factors by Spiroplasma. Our findings do not support a model of exploitative competition between Howardula and Spiroplasma to mediate defence, nor do we find strong support for host immune priming during Spiroplasma infection. Interestingly, we recovered sequence for putative toxins encoded by Spiroplasma, including a novel putative ribosome-inactivating protein, transcripts of which are up-regulated in response to nematode exposure. Protection via the production of toxins may be a widely used and important mechanism in heritable defensive symbioses in insects.


Casting preforms for microstructured polymer optical fibre fabrication.
A monolithic structured polymer preform was formed by in-situ chemical polymerization of high-purity MMA monomer in a home-made mould. The conditions for fabrication of the preforms were optimized and the preform was drawn to microstructured polymer optical fibre. The optical properties of the resultant elliptical-core fibre were measured. This technique provides advantages over alternative preform fabrication methods such as drilling and capillary stacking, which are less suitable for mass production.


Nutrition, insulin, IGF-1 metabolism and cancer risk: a summary of epidemiological evidence.
A Western lifestyle--characterized by low physical activity, and high dietary intake, animal protein, saturated fats and rapidly digestible carbohydrates--is associated with increased risks of many cancers. It has been postulated that, at least in part, these increases may be mediated by alterations in the metabolism of insulin and insulin-like growth factors (IGFs), related to nutritional lifestyle. Insulin regulates energy metabolism and increases the bio-activity of IGF-1 by enhancing its synthesis and by decreasing several of its binding proteins (IGFBP-1 and 2). Insulin and IGF-1 both stimulate anabolic processes as a function of available energy and elementary substrates (e.g. amino acids). The anabolic signals by insulin or IGF-1 can promote tumour development by inhibiting apoptosis, and by stimulating cell proliferation. Epidemiological evidence is accumulating and suggests that the risk of cancers of the colon, pancreas, endometrium, breast and prostate are related to circulating levels of insulin, IGF-1, or both. Nutritional energy balance, macronutrient composition of the diet and physical activity levels appear to be major determinants of IGF-1 bioactivity.


Localization of SV40 genes within supercoiled loop domains.
Recent studies indicate that eukaryotic DNA is organized into supercoiled loop domains. These loops appear to be anchored at their bases to an insoluble nuclear skeleton or matrix. Most of the DNA in the loops can be released from the matrix by nuclease digestion; the residual DNA remaining with the nuclear matrix represents sequences at the base of the loops, and possibly other sequences which are intimately associated with the nuclear matrix for other reasons. Using a quantitative application of the Southern blotting technique, we have found this residual DNA from SV40 infected 3T3 cells to be enriched in SV40 sequences, indicating that they reside near matrix-DNA attachment points. An enrichment of 3-7 fold relative to total cellular DNA, was found in each of three different lines of SV40 infected 3T3 cells. Control experiments with globin genes showed no such enrichment in this residual matrix DNA. This sequence specificity suggests that the spatial organization of DNA sequences within loops may be related to the functionality of these sequences within the cell.


Assessment of left ventricular mass in hypertrophic cardiomyopathy by real-time three-dimensional echocardiography using single-beat capture image.
Left ventricular (LV) mass is an important prognostic indicator in hypertrophic cardiomyopathy. Although LV mass can be easily calculated using conventional echocardiography, it is based on geometric assumptions and has inherent limitations in asymmetric left ventricles. Real-time three-dimensional echocardiographic (RT3DE) imaging with single-beat capture provides an opportunity for the accurate estimation of LV mass. The aim of this study was to validate this new technique for LV mass measurement in patients with hypertrophic cardiomyopathy. Sixty-nine patients with adequate two-dimensional (2D) and three-dimensional echocardiographic image quality underwent cardiac magnetic resonance (CMR) imaging and echocardiography on the same day. Real-time three-dimensional echocardiographic images were acquired using an Acuson SC2000 system, and CMR-determined LV mass was considered the reference standard. Left ventricular mass was derived using the formula of the American Society of Echocardiography (M-mode mass), the 2D-based truncated ellipsoid method (2D mass), and the RT3DE technique (RT3DE mass). The mean time for RT3DE analysis was 5.85 ± 1.81 min. Intraclass correlation analysis showed a close relationship between RT3DE and CMR LV mass (r = 0.86, P < .0001). However, LV mass by the M-mode or 2D technique showed a smaller intraclass correlation coefficient compared with CMR-determined mass (r = 0.48, P = .01, and r = 0.71, P < .001, respectively). Bland-Altman analysis showed reasonable limits of agreement between LV mass by RT3DE imaging and by CMR, with a smaller positive bias (19.5 g [9.1%]) compared with that by the M-mode and 2D methods (-35.1 g [-20.2%] and 30.6 g [17.6%], respectively). RT3DE measurement of LV mass using the single-beat capture technique is practical and more accurate than 2D or M-mode LV mass in patients with hypertrophic cardiomyopathy.


The Effects of Repeated Sprints on the Kinematics of 3-Point Shooting in Basketball.
Fatigue modifies the kinematics of various sports-related movements. Basketball induces fatigue, however, the effects of fatigue on the kinematics of shooting have never been studied. This study analysed the effects of fatigue induced by repeated sprints on the kinematics of 3-point shooting (3PS) in young, elite basketball players (U18 level). 3D joint angles were calculated at the maximum and minimum heights of the centre of mass during 3PS, using inertial measurement units (Biomech system, Xsens Technologies BV, Enschede, The Netherlands). Height, velocity and the angle of the ball at the time of release were extrapolated from the wrist joint angles. All players performed four 3PS actions in dynamic conditions before and after a fatigue protocol at 70% of their maximal exercise capacity. The fatigue protocol consisted of a shuttle test with repeated 20-m sprints interspersed with sets of 5 jumps. There was no change in the kinematics of 3PS (p > 0.05), or the ball release variables (p > 0.05) following the fatigue protocol. This suggests that elite basketball players are able to cope with physical fatigue while performing coordinated movements such as 3PS.