Outcome of implant therapy involving localised lateral alveolar ridge and/or sinus floor augmentation: a clinical and radiographic retrospective 1-year study.
To retrospectively evaluate the 1-year outcome of implant therapy involving various types of localised lateral alveolar ridge and/or sinus floor augmentation procedures performed before implant placement. Forty-seven patients were consecutively treated with oral implants on average 6.5 months after bone grafting. A total of 82 regions were augmented and 151 implants were inserted. All regions were augmented with autogenous bone harvested intraorally. Block grafts (13 regions, 16%), particulate grafts (25 regions, 30%), and a combination of block and particulate grafts (44 regions, 54%) were used. The particulate grafts were sometimes mixed with less than 25% anorganic porous bovine-derived bone mineral (28 regions, 34%). Finally, the grafted areas were in most cases covered by either an expanded polytetrafluoroethylene (e-PTFE) membrane (58 regions, 71%) or a resorbable collagen membrane (11 regions, 13%). The selection of the performed procedure was based upon the experience of the surgeon and thorough evaluation of each case. All surgical procedures were performed by two oral and maxillofacial surgeons with extensive clinical experience with regenerative procedures. The outcome measures were complications related to the augmentation procedure, prosthesis survival, implant survival and peri-implant marginal bone loss. No dropouts occurred. Complications occurred in eight patients (17%) after bone harvesting: temporary paraesthesia (two patients, 4%), membrane exposure (five patients, 11%) and acute sinusitis (one patient, 2%). Membrane exposure was exclusively observed when an e-PTFE membrane was used. However, the subsequent implant placement was not compromised. Regarding the patient with acute sinusitis, the graft was removed and implant/prosthesis placement was not possible. No further prosthesis failures occurred during the 1-year follow-up period. Thus, the prosthesis survival was 99%. The implant survival involving sinus lift, lateral augmentation, and combined sinus lift and lateral augmentation was 91%, 97%, and 100%, respectively. A total of four patients (9%) experienced implant failure. The mean peri-implant marginal bone loss at patient level involving sinus lift, lateral augmentation, and combined sinus lift and lateral augmentation was 0.60 mm (range: 0.31-1.25), 0.31 mm (range: 0.00-0.75) and 0.41 mm (range: 0.00-1.25), respectively. No technical complications were observed. Moreover, no episodes of peri-implant mucositis or peri-implantitis were registered. Localised lateral alveolar ridge and/or sinus floor augmentation performed before implant placement seems to be associated with few complications after 1 year. However, it should be emphasised that all surgical procedures were selected and performed by oral and maxillofacial surgeons with extensive clinical experience in the field of regenerative procedures.


Coming of Age With Cancer: Physical, Social, and Financial Barriers to Independence Among Emerging Adult Survivors
To explore the transition to self-care among a sample of emerging adult cancer survivors after hematopoietic cell transplantation (HCT). . 18 HCT survivors who were aged 18-29 years at the time of HCT for a primary hematologic malignancy and were 8-60 months post-HCT participated in the study. The study took place in the hematology outpatient setting at City of Hope National Medical Center. . The authors conducted in-depth semistructured interviews and analyzed interview transcripts using grounded theory methodology. . Health-related setbacks following HCT disrupted not only participants' journey toward self-care, but also their overarching developmental trajectory toward adulthood. Physically, participants struggled with lack of personal space around caregivers, but felt unready to live on their own. Socially, they relied on multiple caregivers to avoid relying too much on any one person. Financially, participants worried about prolonged dependence and increased needs in the future. . Nurses can support the transition to self-care among emerging adults after HCT by recognizing the broader developmental impact of their cancer experience.


Impact of a nationwide public campaign on delays and outcome in Swiss patients with acute coronary syndrome.
To improve the outcome in patients with acute coronary syndrome (ACS), public campaigns have been launched aimed at shortening presentation delays and allowing more efficient treatment. Their impact is uncertain. Pre-hospital delays and outcome in patients with ACS included in the Swiss National AMIS Plus registry were assessed prospectively before and after a nationwide multimedia campaign ('HELP') by the Swiss Heart Foundation in 2007. The campaign aimed at better awareness of symptoms, increasing knowledge in laymen of resuscitation and more rapid access to medical services. The primary study endpoint was the time between onset of symptoms and hospital admission. Secondary endpoints were successful out-of-hospital resuscitation, symptoms upon admission, and outcome. Between January 2005 and December 2008, 8906 ACS patients (61% ST-segment elevation myocardial infarction (STEMI), 39% non-ST-segment elevation myocardial infarction/unstable angina pectoris (NSTEMI/UAP), mean age 65 ± 13 years, 75% males) admitted within 24 hours after onset of symptoms were enrolled. The median pre-hospital delay was reduced from 197 minutes during the pre-intervention period to 180 minutes during the post-intervention period (reduction 10% (95% confidence interval (CI) 6-14%); P < 0.001), in STEMI (reduction 10% (95% CI 5-14%); P < 0.001) and NSTEMI patients (reduction 11% (95% CI 4-17%); P = 0.001), due to pronounced effects in males (reduction 12% (95% CI 7-16%); P < 0.001) and in patients ≤75 years (reduction 12% (95% CI 8-16%); P < 0.001). Out-of-hospital resuscitation increased (odds ratio (OR) 1.26 (95% CI 1.06-1.54); P = 0.02). Overall outcome remained unchanged, however, the rate of re-infarction showed a decrease (OR 0.58 (95% CI 0.36-0.91); P = 0.021). After a nationwide educational campaign, shorter pre-hospital delays were observed, and more patients were able to be treated promptly. These results may be useful in planning future health strategies to improve management and outcome of patients with ACS, especially in female and elderly patients.


Multivariate modeling of proteins related to trapezius myalgia, a comparative study of female cleaners with or without pain.
The prevalence of chronic trapezius myalgia is high in women with high exposure to awkward working positions, repetitive movements and movements with high precision demands. The mechanisms behind chronic trapezius myalgia are not fully understood. The purpose of this study was to explore the differences in protein content between healthy and myalgic trapezius muscle using proteomics. Muscle biopsies from 12 female cleaners with work-related trapezius myalgia and 12 pain free female cleaners were obtained from the descending part of the trapezius. Proteins were separated with two-dimensional differential gel electrophoresis (2D-DIGE) and selected proteins were identified with mass spectrometry. In order to discriminate the two groups, quantified proteins were fitted to a multivariate analysis: partial least square discriminate analysis. The model separated 28 unique proteins which were related to glycolysis, the tricaboxylic acid cycle, to the contractile apparatus, the cytoskeleton and to acute response proteins. The results suggest altered metabolism, a higher abundance of proteins related to inflammation in myalgic cleaners compared to healthy, and a possible alteration of the contractile apparatus. This explorative proteomic screening of proteins related to chronic pain in the trapezius muscle provides new important aspects of the pathophysiology behind chronic trapezius myalgia.


Which factors are important for the successful development and implementation of clinical pathways? A qualitative study.
Clinical pathways (CPs) are detailed longitudinal care plans delineating measures to be conducted during a patient's treatment. Although positive effects on resource consumption and quality of care have been shown, CPs are still underutilised in many clinical settings because their development and implementation are difficult. Evidence underpinning successful development and implementation is sparse. The authors conducted semistructured face-to-face interviews with key staff members involved in the design and implementation of CPs in a large surgery department. Interviewees were asked to provide opinions on various issues, which were previously identified as potentially important in CP development and implementation. The transcribed text was read and coded independently by two researchers. Respondents highlighted the importance of a multidisciplinary participatory approach for CP design and implementation. There was a strong initial fear of losing individual freedom of treatment, which subsided after people worked with CPs in clinical everyday life. It was appreciated that the project originated from people at different levels of the department's hierarchy. Likewise, it was felt that CP implementation granted more autonomy to lower-level staff. The structured qualitative approach of this study provides information on what issues are considered important by staff members for CP design and implementation. Whereas some concepts such as the importance of a multidisciplinary approach or continuous feedback of results are known from theories, others such as strengthening the authority especially of lower-level health professionals through CPs have not been described so far. Many of the findings point towards strong interactions between factors important for CP implementation and a department's organisational structure.


A framework to rank genomic alterations as targets for cancer precision medicine: the ESMO Scale for Clinical Actionability of molecular Targets (ESCAT).
In order to facilitate implementation of precision medicine in clinical management of cancer, there is a need to harmonise and standardise the reporting and interpretation of clinically relevant genomics data. The European Society for Medical Oncology (ESMO) Translational Research and Precision Medicine Working Group (TR and PM WG) launched a collaborative project to propose a classification system for molecular aberrations based on the evidence available supporting their value as clinical targets. A group of experts from several institutions was assembled to review available evidence, reach a consensus on grading criteria and present a classification system. This was then reviewed, amended and finally approved by the ESMO TR and PM WG and the ESMO leadership. This first version of the ESMO Scale of Clinical Actionability for molecular Targets (ESCAT) defines six levels of clinical evidence for molecular targets according to the implications for patient management: tier I, targets ready for implementation in routine clinical decisions; tier II, investigational targets that likely define a patient population that benefits from a targeted drug but additional data are needed; tier III, clinical benefit previously demonstrated in other tumour types or for similar molecular targets; tier IV, preclinical evidence of actionability; tier V, evidence supporting co-targeting approaches; and tier X, lack of evidence for actionability. The ESCAT defines clinical evidence-based criteria to prioritise genomic alterations as markers to select patients for targeted therapies. This classification system aims to offer a common language for all the relevant stakeholders in cancer medicine and drug development.


Detection of paroxysmal nocturnal hemoglobinuria (PNH) in bone marrow aspirates☆.
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired condition in which, due to a mutation of the phosphatidylinositol glycan class A gene, hematopoietic cells lack proteins that are normally anchored to the cell surface by glycosylphosphatidylinositol (GPI). Thus, PNH cells show poor expression of surface proteins, such as CD55 and CD59, and dim or absent binding of fluorescently labeled modified aerolysin (FLAER). In clinical diagnostic laboratories, the detection and quantitation of PNH is currently performed by flow cytometry (FC) analysis of peripheral blood (PB) samples. Although PB remains the preferred source of cells for PNH detection, we and other authors have shown that a careful FC analysis of bone marrow (BM) aspirates can provide results for PNH detection and quantitation equivalent to those obtained with PB. Here, we review studies delineating the expression of GPI-anchored proteins and FLAER binding in normal BM cells, and summarize published findings demonstrating the feasibility of identifying and quantitating PNH cells in BM using FLAER as well as antibodies against GPI-anchored proteins. Detection of PNH cells in BM should be useful in patients with unsuspected PNH or with severe cytopenia in whom PB FC analysis may be challenging.


Neural-network-based approximate output regulation of discrete-time nonlinear systems.
The existing approaches to the discrete-time nonlinear output regulation problem rely on the offline solution of a set of mixed nonlinear functional equations known as discrete regulator equations. For complex nonlinear systems, it is difficult to solve the discrete regulator equations even approximately. Moreover, for systems with uncertainty, these approaches cannot offer a reliable solution. By combining the approximation capability of the feedforward neural networks (NNs) with an online parameter optimization mechanism, we develop an approach to solving the discrete nonlinear output regulation problem without solving the discrete regulator equations explicitly. The approach of this paper can be viewed as a discrete counterpart of our previous paper on approximately solving the continuous-time nonlinear output regulation problem.


Phospholipase D1 Acts through Akt/TopBP1 and RB1 to Regulate the E2F1-Dependent Apoptotic Program in Cancer Cells.
The RB1/E2F1 signaling pathway is frequently deregulated in colorectal cancer and has been suggested to intersect with Wnt/β-catenin and PI3K/Akt pathways, but molecular evidence for this link is lacking. In this study, we demonstrate that phospholipase D1 (PLD1), a transcriptional target of β-catenin/TCF4, orchestrates functional interactions between these pathways during intestinal tumor development. Overexpression of PLD1 in intestinal epithelial cells protected cells from apoptosis induced by PLD1 ablation in the Apcmin/+ mouse model of intestinal tumorigenesis. Mechanistic investigations revealed that genetic and pharmacologic targeting of PLD1 promote the E2F1-dependent apoptotic program via both miR-192/4465-mediated downregulation of RB1 and inhibition of Akt-TopBP1 pathways. Moreover, the miRNA-RB1 axis and Akt pathway also contributed to the PLD1-mediated self-renewal capacity of colon cancer-initiating cells. Finally, PLD1-driven E2F1 target gene expression positively correlated with tumor stage in patients with colorectal cancer. Overall, our findings suggest that PLD1 mediates cross-talk between multiple major signaling pathways to promote the survival and malignancy of colon cancer cells and may therefore represent an ideal signaling node for therapeutic targeting. Cancer Res; 77(1); 142-52. ©2016 AACR.


Tubulin chaperone E binds microtubules and proteasomes and protects against misfolded protein stress.
Mutation of tubulin chaperone E (TBCE) underlies hypoparathyroidism, retardation, and dysmorphism (HRD) syndrome with defective microtubule (MT) cytoskeleton. TBCE/yeast Pac2 comprises CAP-Gly, LRR (leucine-rich region), and UbL (ubiquitin-like) domains. TBCE folds alpha-tubulin and promotes alpha/beta dimerization. We show that Pac2 functions in MT dynamics: the CAP-Gly domain binds alpha-tubulin and MTs, and functions in suppression of benomyl sensitivity of pac2Delta mutants. Pac2 binds proteasomes: the LRR binds Rpn1, and the UbL binds Rpn10; the latter interaction mediates Pac2 turnover. The UbL also binds the Skp1-Cdc53-F-box (SCF) ubiquitin ligase complex; these competing interactions for the UbL may impact on MT dynamics. pac2Delta mutants are sensitive to misfolded protein stress. This is suppressed by ectopic PAC2 with both the CAP-Gly and UbL domains being essential. We propose a novel role for Pac2 in the misfolded protein stress response based on its ability to interact with both the MT cytoskeleton and the proteasomes.


Mineralocorticoid Receptor Antagonists for Treatment of Hypertension and Heart Failure.
Spironolactone and eplerenone are both mineralocorticoid-receptor antagonists. These compounds block both the epithelial and nonepithelial actions of aldosterone, with the latter assuming increasing clinical relevance. Spironolactone and eplerenone both affect reductions in blood pressure either as mono- or add-on therapy; moreover, they each afford survival benefits in diverse circumstances of heart failure and the probability of renal protection in proteinuric chronic kidney disease. However, as use of mineralocorticoid-blocking agents has expanded, the hazards inherent in taking such drugs have become more apparent. Whereas the endocrine side effects of spironolactone are in most cases little more than a cosmetic annoyance, the potassium-sparing effects of both spironolactone and eplerenone can prove disastrous, even fatal, if sufficient degrees of hyperkalemia emerge. For most patients, however, the risk of developing hyperkalemia in and of itself should not discourage the sensible clinician from bringing these compounds into play. Hyperkalemia should always be considered a possibility in patients receiving either of these medications; therefore, anticipatory steps should be taken to minimize the likelihood of its occurrence if long-term therapy of these agents is being considered.


Transplantation of an allogeneic vein bioengineered with autologous stem cells: a proof-of-concept study.
Extrahepatic portal vein obstruction can have severe health consequences. Variceal bleeding associated with this disorder causes upper gastrointestinal bleeding, leading to substantial morbidity and mortality. We report the clinical transplantation of a deceased donor iliac vein graft repopulated with recipient autologous stem cells in a patient with extrahepatic portal vein obstruction. A 10 year old girl with extrahepatic portal vein obstruction was admitted to the Sahlgrenska University Hospital in Gothenburg, Sweden, for a bypass procedure between the superior mesenteric vein and the intrahepatic left portal vein (meso Rex bypass). A 9 cm segment of allogeneic donor iliac vein was decellularised and subsequently recellularised with endothelial and smooth muscle cells differentiated from stem cells obtained from the bone marrow of the recipient. This graft was used because the patient's umbilical vein was not suitable and other strategies (eg, liver transplantation) require lifelong immunosuppression. The graft immediately provided the recipient with a functional blood supply (25-30 cm/s in the portal vein and 40 mL/s in the artery was measured intraoperatively and confirmed with ultrasound). The patient had normal laboratory values for 9 months. However, at 1 year the blood flow was low and, on exploration, the shunt was patent but too narrow due to mechanical obstruction of tissue in the mesocolon. Once the tissue causing the compression was removed the graft dilated. We therefore used a second stem-cell populated vein graft to lengthen the previous graft. After this second operation, the portal pressure was reduced from 20 mm Hg to 13 mm Hg and blood flow was 25-40 cm/s in the portal vein. With restored portal circulation the patient has substantially improved physical and mental function and growth. The patient has no anti-endothelial cell antibodies and is receiving no immunosuppressive drugs. An acellularised deceased donor vein graft recellularised with autologous stem cells can be considered for patients in need of vascular vein shunts without the need for immunosuppression. Swedish Government.


Interoceptive awareness is associated with acute alcohol-induced changes in subjective effects.
Interoception, the sensing of bodily signals, is related to emotional reactivity and may contribute to the pathophysiology of addiction. Evidence is accumulating that individuals with alcohol use disorders and other substance-dependences show altered interoceptive processing, however little is known about the acute effects of alcohol on interoception and how this may influence the perception of drug induced effects. In a double-blind design, fifty (30 females) healthy young participants were given a beverage containing either a low (0.4 g/kg, n = 18) or high (0.6 g/kg, n = 15) alcohol dose or a placebo (n = 17). After alcohol administration, participants completed two interoceptive paradigms, the heart-beat tracking and heart-beat discrimination tasks, both assessing different accuracy and metacognitive measures of interoception. Subjective feelings elicited by alcohol administration were also measured. Participants under the low alcohol dose had lower metacognitive interoceptive awareness on the discrimination task compared to placebo. Participants under alcohol experienced feelings of light-headedness, which were positively associated with increased interoceptive awareness in the cardiac discrimination task. These results provide evidence for a relationship between interoceptive processing and the perception of drug-induced mood changes. This finding, showing how interoceptive awareness of cardiac discrimination contributes to the appraisal of subjective light-headedness generated by alcohol administration, brings novel perspectives to the understanding of drug discrimination and reinforcement mechanisms.


The barley immune receptor Mla recognizes multiple pathogens and contributes to host range dynamics.
Crop losses caused by plant pathogens are a primary threat to stable food production. Stripe rust (Puccinia striiformis) is a fungal pathogen of cereal crops that causes significant, persistent yield loss. Stripe rust exhibits host species specificity, with lineages that have adapted to infect wheat and barley. While wheat stripe rust and barley stripe rust are commonly restricted to their corresponding hosts, the genes underlying this host specificity remain unknown. Here, we show that three resistance genes, Rps6, Rps7, and Rps8, contribute to immunity in barley to wheat stripe rust. Rps7 cosegregates with barley powdery mildew resistance at the Mla locus. Using transgenic complementation of different Mla alleles, we confirm allele-specific recognition of wheat stripe rust by Mla. Our results show that major resistance genes contribute to the host species specificity of wheat stripe rust on barley and that a shared genetic architecture underlies resistance to the adapted pathogen barley powdery mildew and non-adapted pathogen wheat stripe rust.


Multiscale Investigation of the Depth-Dependent Mechanical Anisotropy of the Human Corneal Stroma.
To investigate the depth-dependent mechanical anisotropy of the human corneal stroma at the tissue (stroma) and molecular (collagen) level by using atomic force microscopy (AFM). Eleven human donor corneas were dissected at different stromal depths by using a microkeratome. Mechanical measurements were performed in 15% dextran on the surface of the exposed stroma of each sample by using a custom-built AFM in force spectroscopy mode using both microspherical (38-μm diameter) and nanoconical (10-nm radius of curvature) indenters at 2-μm/s and 15-μm/s indentation rates. Young's modulus was determined by fitting force curve data using the Hertz and Hertz-Sneddon models for a spherical and a conical indenter, respectively. The depth-dependent anisotropy of stromal elasticity was correlated with images of the corneal stroma acquired by two-photon microscopy. The force curves were obtained at stromal depths ranging from 59 to 218 μm. At the tissue level, Young's modulus (ES) showed a steep decrease at approximately 140-μm stromal depth (from 0.8 MPa to 0.3 MPa; P = 0.03) and then was stable in the posterior stroma. At the molecular level, Young's modulus (EC) was significantly greater than at the tissue level; EC decreased nonlinearly with increasing stromal depth from 3.9 to 2.6 MPa (P = 0.04). The variation of microstructure through the thickness correlated highly with a nonconstant profile of the mechanical properties in the stroma. The corneal stroma exhibits unique anisotropic elastic behavior at the tissue and molecular levels. This knowledge may benefit modeling of corneal behavior and help in the development of biomimetic materials.


The ARF family of transcription factors and their role in plant hormone-responsive transcription.
Auxin response factors or ARFs are a recently discovered family of transcription factors that bind with specificity to auxin response elements (AuxREs) in promoters of primary or early auxin-responsive genes. ARFs have an amino-terminal DNA-binding domain related to the carboxyl-terminal DNA-binding domain in the maize transactivator VIVIPAROUS1. All but one ARF identified to date contain a carboxyl-terminal protein-protein interaction domain that forms a putative amphipathic alpha-helix. A similar carboxyl-terminal protein-protein interaction domain is found in the Aux/IAA class of auxin-inducible proteins. Some ARFs contain transcriptional activation domains, while others contain repression domains. ARFs appear to play a pivotal role in auxin-regulated gene expression of primary response genes.


Bioeconomic modelling of raccoon rabies spread management impacts in Quebec, Canada.
Beginning in 2006, point infection control operations and aerial distribution of oral rabies vaccines along the US border were performed in Quebec, Canada, to control the potential spread of raccoon rabies. A benefit-cost analysis assessed the economic efficiency of this rabies control programme into the future. In this study, a mathematical simulation model was used to determine the potential spread of raccoon rabies from the 2006 index case, and incidence rates of human post-exposure prophylaxis (PEP), animal testing and human exposure investigations were calculated. Benefits were calculated as the potential savings from reduced numbers of human PEP, animal testing and human exposure investigations owing to control, which ranged from $47 million to $53 million. Programme cost scenarios were based on projections of total expenditures, which ranged from $33 million to $49 million. Economic efficiency was indicated for approximately half of the modelled scenarios, with the greatest benefit-cost ratios resulting from reduced future programme costs.


Observer variation in the diagnosis of follicular variant of papillary thyroid carcinoma.
The histopathologic diagnosis of follicular variant of papillary thyroid carcinoma (FVPCA) can be difficult. Recent reports have suggested that this neoplasm may be frequently overdiagnosed by pathologists. We examined the observer variation in the diagnosis of FVPCA in 87 tumors by 10 experienced thyroid pathologists. The criteria that the reviewers considered most helpful for making a diagnosis of FVPCA were also assessed. A concordant diagnosis of FVPCA was made by all 10 reviewers with a cumulative frequency of 39%. In this series, 24.1% of the patients had metastatic disease (n = 21). In the cases with metastatic disease, a diagnosis of FVPCA was made by all 10 reviewers with a cumulative frequency of 66.7%, and 7 of the reviewers made a diagnosis of FVPCA with a cumulative frequency of 100%. The most important criteria used to diagnose FVPCA included the presence of cytoplasmic invaginations into the nucleus (pseudo-inclusions), abundant nuclear grooves, and ground glass nuclei. These results suggest that although the diagnosis of FVPCA is variable even among experienced thyroid pathologists, most reviewers agreed on this diagnosis for patients with metastatic disease. The use of well-defined histopathologic features should improve the consistency in diagnosing FVPCA. Since most cases with metastatic disease had obvious invasion, caution should be used in making a diagnosis of FVPCA in the absence of the major histopathologic features or clear-cut invasive growth.


Onset of Juvenile Court Involvement: Exploring Gender-Specific Associations with Maltreatment and Poverty.
Despite increased attention to gender differences in youthful offending, no known studies have examined the relative impact of poverty, maltreatment, and their combination on gender-specific patterns of offending. This research addresses the question of the differential impact of maltreatment and poverty on the onset of status and delinquent petitions for girls compared to boys. A sample of youth born in 1982-1986 in the Midwest was examined. The independent variables were poverty, maltreatment, and both. The risks of delinquent petition and status petition were analyzed using separate Cox proportional hazards models by gender. A second set of analyses were conducted on a subset of youth reported for maltreatment. There was an increase in the likelihood of juvenile court petition based on the combination of poverty and maltreatment risk factors compared to maltreatment only. This increase in risk held true only for the boys in the maltreatment subsample. Thus, the notion of these risk factors being additive is supported with males, but only for females when a non-maltreatment comparison group exists. The gender-specific nature of these relationships supports conceptual propositions that girls' pathways to the juvenile justice system are distinct from boys'. Implications for theory, research, and practice are discussed.


A hybrid NRPS-PKS gene cluster related to the bleomycin family of antitumor antibiotics in Alteromonas macleodii strains.
Although numerous marine bacteria are known to produce antibiotics via hybrid NRPS-PKS gene clusters, none have been previously described in an Alteromonas species. In this study, we describe in detail a novel hybrid NRPS-PKS cluster identified in the plasmid of the Alteromonasmacleodii strain AltDE1 and analyze its relatedness to other similar gene clusters in a sequence-based characterization. This is a mobile cluster, flanked by transposase-like genes, that has even been found inserted into the chromosome of some Alteromonasmacleodii strains. The cluster contains separate genes for NRPS and PKS activity. The sole PKS gene appears to carry a novel acyltransferase domain, quite divergent from those currently characterized. The predicted specificities of the adenylation domains of the NRPS genes suggest that the final compound has a backbone very similar to bleomycin related compounds. However, the lack of genes involved in sugar biosynthesis indicates that the final product is not a glycopeptide. Even in the absence of these genes, the presence of the cluster appears to confer complete or partial resistance to phleomycin, which may be attributed to a bleomycin-resistance-like protein identified within the cluster. This also suggests that the compound still shares significant structural similarity to bleomycin. Moreover, transcriptomic evidence indicates that the NRPS-PKS cluster is expressed. Such sequence-based approaches will be crucial to fully explore and analyze the diversity and potential of secondary metabolite production, especially from increasingly important sources like marine microbes.


PKM2 tyrosine phosphorylation and glutamine metabolism signal a different view of the Warburg effect.
New evidence suggests that the receptor tyrosine kinase FGFR1 (fibroblast growth factor receptor 1) directly phosphorylates pyruvate kinase M2 (PKM2), resulting in reduced conversion of phosphoenolpyruvate to pyruvate, which is further catabolized to lactate by lactate dehydrogenase A. Mutation of the critical tyrosine Tyr(105) to Phe rendered PKM2 more active but was associated with decreased cellular lactate production, increased oxygen consumption, and decreased hypoxic cell proliferation relative to wild-type PKM2. The apparent paradoxical effect of growth signaling through tyrosine phosphorylation, which decreases rather than increases PKM2 activity, stimulates a revised perspective of the Warburg effect. This effect, which describes the propensity for cancer cells to convert glucose to lactate at a high rate, must now accommodate links among glycolysis, the tricarboxylic acid cycle, and glutamine metabolism in cancer cells.


Where You Live Matters: Structural Correlates of HIV Risk Behavior Among Young Men Who Have Sex with Men in Metro Detroit.
Structural characteristics are linked to HIV/STI risks, yet few studies have examined the mechanisms through which structural characteristics influence the HIV/STI risk of young men who have sex with men (YMSM). Using data from a cross-sectional survey of YMSM (ages 18-29) living in Detroit Metro (N = 328; 9 % HIV-positive; 49 % Black, 27 % White, 15 % Latino, 9 % Other race), we used multilevel modeling to examine the association between community-level characteristics (e.g., socioeconomic disadvantage; distance to LGBT-affirming institutions) and YMSM's HIV testing behavior and likelihood of engaging in unprotected anal intercourse with serodiscordant partner(s). We accounted for individual-level factors (race/ethnicity, poverty, homelessness, alcohol and marijuana use) and contextual factors (community acceptance and stigma regarding same-sex sexuality). YMSM in neighborhoods with greater disadvantage and nearer to an AIDS Service Organization were more likely to have tested for HIV and less likely to report serodiscordant partners. Community acceptance was associated with having tested for HIV. Efforts to address YMSM's exposure to structural barriers in Detroit Metro are needed to inform HIV prevention strategies from a socioecological perspective.


IFN-γ extends the immune functions of Guanylate Binding Proteins to inflammasome-independent antibacterial activities during Francisella novicida infection.
Guanylate binding proteins (GBPs) are interferon-inducible proteins involved in the cell-intrinsic immunity against numerous intracellular pathogens. The molecular mechanisms underlying the potent antibacterial activity of GBPs are still unclear. GBPs have been functionally linked to the NLRP3, the AIM2 and the caspase-11 inflammasomes. Two opposing models are currently proposed to explain the GBPs-inflammasome link: i) GBPs would target intracellular bacteria or bacteria-containing vacuoles to increase cytosolic PAMPs release ii) GBPs would directly facilitate inflammasome complex assembly. Using Francisella novicida infection, we investigated the functional interactions between GBPs and the inflammasome. GBPs, induced in a type I IFN-dependent manner, are required for the F. novicida-mediated AIM2-inflammasome pathway. Here, we demonstrate that GBPs action is not restricted to the AIM2 inflammasome, but controls in a hierarchical manner the activation of different inflammasomes complexes and apoptotic caspases. IFN-γ induces a quantitative switch in GBPs levels and redirects pyroptotic and apoptotic pathways under the control of GBPs. Furthermore, upon IFN-γ priming, F. novicida-infected macrophages restrict cytosolic bacterial replication in a GBP-dependent and inflammasome-independent manner. Finally, in a mouse model of tularemia, we demonstrate that the inflammasome and the GBPs are two key immune pathways functioning largely independently to control F. novicida infection. Altogether, our results indicate that GBPs are the master effectors of IFN-γ-mediated responses against F. novicida to control antibacterial immune responses in inflammasome-dependent and independent manners.


Return to Sport Rates in Physically Active Individuals 6 Months After Arthroscopy for Femoroacetabular Impingement Syndrome.
Six months is the expected time frame to return to sport (RTS) after hip arthroscopy for femoroacetabular impingement syndrome (FAIS). The primary aims of this study were to (1) report the RTS rate of physically active individuals 6 months after arthroscopic surgery for FAIS and (2) compare the self-reported hip function between those who were able to RTS against those who had not. Cohort study. The Ohio State University Wexner Medical Center. A total of 42 physically active individuals scheduled for hip arthroscopy for FAIS. Self-reported function, including preinjury and current activity levels and ability to participate in sport, were collected on 42 individuals with FAIS prior to surgery and at 6-month follow-up. Participants were allocated into 2 groups based on self-reported RTS status at 6-month follow-up. Separate 2-way analyses of variance were used to test the interaction of groups (those who did/did not RTS). Main effects were reported in the absence of statistically significant interactions (P ≤ .05). Altogether, 28 of 42 participants (66%) returned for 6-month follow-up; 5 of the 14 participants (35.7%) lost to follow-up underwent contralateral hip surgery within the study time frame. At the 6-month follow-up, 16 of the remaining 28 participants (57.1%) reported that they had returned to sport, 5 of whom returned at their prior level of participation. There was no statistically significant interaction of group and time for either hip outcome score subscale (P ≥ .20). Self-reported hip function improved over time, regardless of group (P ≤ .001). Participants in the yRTS group demonstrated higher hip outcome score-sport scores than did the nRTS group, regardless of time (P = .04). Though just over half of participants returned to sport 6 months after hip arthroscopy for FAIS, only 18% returned to their previous level of sports participation. Participants who returned to sport reported better function than those who did not, but self-reported hip function improved over time regardless of group. The most commonly reported reasons for not returning to sport were weakness (69.6%), fear (65.2%), and pain (56.5%).


Simultaneous oscillations of cytoplasmic free Ca2+ concentration and Ins(1,4,5)P3 concentration in mouse pancreatic beta-cells.
Changes in the cytoplasmic free Ca2+ concentration ([Ca2+]i) in stimulated cells are often oscillatory, but the mechanisms that drive these oscillations are still a matter of controversy: different models of the generation of these [Ca2+]i oscillations make different assumptions as to whether oscillations in Ins(1,4,5)P3 concentration are necessary for this process. We have looked for changes in inositol polyphosphate levels that might occur in suspensions of murine pancreatic beta-cells when these cells are induced to display synchronized oscillations in [Ca2+]i by the sequential addition of glucose, an alpha 2-adrenergic stimulus and extracellular Ca2+. The intracellular level of Ins(1,4,5)P3 oscillated in a manner approximately in synchrony with changes in [Ca2+]i. Oscillations in the levels of Ins(1,4,5)P3 metabolites [Ins(1,3,4)P3 and inositol bisphosphates] were slightly delayed relative to the Ins(1,4,5)P3 oscillations, and the concentration of Ins(1,3,4,5,6)P5 remained approximately constant during the [Ca2+]i oscillations. These results demonstrate that [Ins(1,4,5)P3] and [Ca2+]i oscillate in synchrony in at least one type of cell. Whether such oscillations in intracellular [Ins(1,4,5)P3] provide a primary driving force for [Ca2+]i oscillations either in beta-cells or in other stimulated cells remains to be determined. Even if they do not, the [Ins(1,4,5)P3] oscillations will at least provide an amplifying influence on the [Ca2+]i changes.


External ecological niche for Candida albicans within reducing, oxygen-limited zones of wetlands.
Candida albicans within the human host is well studied; however, identifying environmental reservoirs of pathogens is epidemiologically valuable for disease management. Oxygen-limited, carbohydrate-rich zones of wetlands, to which sewage-borne C. albicans is often exposed, are characteristically similar to the gastrointestinal reservoir. Consequently, using quantitative real-time PCR (qRT-PCR) and gas chromatography-mass spectrometry (GC-MS), we demonstrated that oxygen-limited zones in polluted wetlands may act as potential reservoirs of C. albicans.


Laparoscopic uterosacral nerve ablation for alleviating chronic pelvic pain: a randomized controlled trial.
Chronic pelvic pain is a common condition with a major effect on health-related quality of life, work productivity, and health care use. Operative interruption of nerve trunks in the uterosacral ligaments by laparoscopic uterosacral nerve ablation (LUNA) is a treatment option for patients with chronic pelvic pain. To assess the effectiveness of LUNA in patients with chronic pelvic pain. Randomized controlled trial of 487 women with chronic pelvic pain lasting longer than 6 months without or with minimal endometriosis, adhesions, or pelvic inflammatory disease, who were recruited to the study by consultant gynecological surgeons from 18 UK hospitals between February 1998 and December 2005. Follow-up was conducted by questionnaires mailed at 3 and 6 months and at 1, 2, 3, and 5 years. Bilateral LUNA or laparoscopy without pelvic denervation (no LUNA); participants were blinded to the treatment allocation. The primary outcome was pain, which was assessed by a visual analogue scale. Data concerning the 3 types of pain (noncyclical pain, dysmenorrhea, and dyspareunia) were analyzed separately as was the worst pain level experienced from any of these 3 types of pain. The secondary outcome was health-related quality of life, which was measured using a generic instrument (EuroQoL EQ-5D and EQ-VAS). After a median follow-up of 69 months, there were no significant differences reported on the visual analogue pain scales for the worst pain (mean difference between the LUNA group and the no LUNA group, -0.04 cm [95% confidence interval {CI}, -0.33 to 0.25 cm]; P = .80), noncyclical pain (-0.11 cm [95% CI, -0.50 to 0.29 cm]; P = .60), dysmenorrhea (-0.09 cm [95% CI, -0.49 to 0.30 cm]; P = .60), or dyspareunia (0.18 cm [95% CI, -0.22 to 0.62 cm]; P = .40). No differences were observed between the LUNA group and the no LUNA group for quality of life. Among women with chronic pelvic pain, LUNA did not result in improvements in pain, dysmenorrhea, dyspareunia, or quality of life compared with laparoscopy without pelvic denervation. controlled-trials.com Identifier: ISRCTN41196151.


Common pathogenic mechanism in development progression of liver injury caused by non-alcoholic or alcoholic steatohepatitis.
This review showed the common pathogenic mechanism in the development of non-alcoholic or alcoholic steatohepatitis. In particular, we describe the role of innate immune system and oxidative stress caused by gut-derived endotoxin. Gut-derived endotoxin plays an important role in alcoholic liver injury. It was reported that acute ethanol administration reduced activation of Kupffer cells. It is therefore possible that alcohol-induced hepatocellular damage occurs as a result of bacterial or endotoxin translocation under a reduction of the reticuloendothelial system (RES) function in alcoholic liver disease (ALD). On the other hand, recently, attention has been directed toward the effect of ethanol ingestion on Kupffer cell function, which is stimulated by gut-derived endotoxin via mechanisms dependent on increased gut permeability and the possible relationship between Kupffer cells and alcohol-induced liver injury. It is generally accepted that activation of the innate immune system and increased release of proinflammatory cytokines and other mediators plays an important role in the development of ALD. It was shown that Kupffer cells activation by endotoxin via Toll-like receptor (TLR-4) is involved in alcohol-induced liver injury and that ethanol-induced oxidative stress is important in the regulation of transcription factor NF-kappaB activation and that cytokine production by Kupffer cells. TNF-alpha and free radicals are produced in early alcohol-induced liver injury. In support of this finding, the pathology caused by alcohol was blocked nearly completely in TNF-alpha receptor 1. Many pathways have been suggested to contribute to the ability of ethanol to induce a state of oxidative stress. One central pathway appears to be the induction of the CYP2E1 form of cytochrome P450 enzymes by ethanol. Initial efforts to clarify the mechanisms that promote the progression from steatosis to steatohepatitis somewhat artificially divides disease mechanisms into "first and second" hit. The best candidates for these second hits were considered to be oxidative stress (CYP2E1 induction) and associated lipid peroxidation and cyokines, principally, TNF-alpha. Some of the most definitive data on the importance of the innate immune system or oxidative stress in the pathogenesis of liver disease come from studies of alcoholic and non-alcoholic steatohepatitis in animals.


N-nitrosamine formation by monochloramine, free chlorine, and peracetic acid disinfection with presence of amine precursors in drinking water system.
In this study, the formation of eight N-nitrosamines, N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine, N-nitrosomethylamine, N-nitrosodi-n-propylamine, N-nitrosodi-n-butylamine, N-Nitrosopiperidine, N-Nitrosopyrrolidine, N-Nitrosomorpholine, were systematically evaluated with respect to seven N-nitrosamine precursors (dimethylamine, trimethylamine, 3-(dimethylaminomethyl)indole, 4-dimethylaminoantipyrine, ethylmethylamine, diethylamine, dipropylamine) and three disinfectants (monochloramine, free chlorine, peracetic acid) under variable dosages, exposure times, and pH in a drinking water system. Without the presence of the seven selected N-nitrosamine precursors N-nitrosamine formation was not observed under any tested condition except very low levels of N-Nitrosopyrrolidine under some conditions. With selected N-nitrosamine precursors present N-nitrosamines formed at different levels under different conditions. The highest N-nitrosamine formation was NDMA with a maximum concentration of 1180 ng/L by monochloramine disinfection with precursors present; much lower levels of N-nitrosamines were formed by free chlorine disinfection; and no detectable level of N-nitrosamines were observed by peracetic acid disinfection except low level of N-Nitrosodi-n-propylamine under some conditions. NDMA formation was not affected by pH while four other N-nitrosamine formations were slightly affected by sample pH tested between 7 and 9, with formation decreasing with increasing pH. Monochloramine exposure time study displayed fast formation of N-nitrosamines, largely formed in four hours of exposure and maximized after seven days. This was a systematic study on the N-nitrosamine formation with the seven major N-nitrosamine precursors presence and absence under different conditions, including peracetic acid disinfection which has not been studied elsewhere.


Dynamics of microring resonator modulators.
A dynamic model for the transmission of a microring modulator based on changes in the refractive index, loss, or waveguide-ring coupling strength is derived to investigate the limitations to the intensity modulation bandwidth. Modulation bandwidths approaching the free spectral range frequency are possible if the waveguide-ring coupling strength is varied, rather than the refractive index or loss of the ring. The results illustrate that via controlled coupling, resonant modulators with high quality factors can be designed to operate at frequencies much larger than the resonator linewidth.


Role of phenylthiocarbamide as a genetic marker in predicting the predisposition of disease traits in humans.
The main objective of this study is to find out the genetic variation and predisposition of overweight/obese, smoking/alcoholism and thyroid disease traits among tasters and non-tasters in Mysore population, South India. Bitter-taste perception for phenylthiocarbamide (PTC) is a classically variable trait both within and between human populations. Many studies have reported that in world population, approximately 30% of them are PTC non-tasters and 70% are tasters. This investigation was conducted during the year 2009-2010 involving a total 1352 study subjects and divided into three different groups, considering the age ranging from 13 to 50 years. Phenylthiocarbamide taste sensitivity was measured by administering a freshly prepared 0.025% of phenylthiocarbamide solution using the Harris and Kalmus method with a slight modification and the results were recorded. In the first group of 100 obese/overweight children, 28% are taster and 72% are non-taster and among 100 control group 67% are tasters and 43% are non-tasters. In second group, out of 1152 individuals 710 (61.63%) are tasters and 442 (38.37%) are non-tasters including both males and females. In the third group, out of each 100 thyroid patients and the control group, tasters are significantly more frequent (61.41%) than the non-tasters (38.58%) in the control group, but a higher proportion of non-tasters are recorded among individuals with thyroid problems (68%) compared to tasters (32%). There is a significant higher incidence of PTC tasters than non-tasters among general population in this study. As phenotypic variation in PTC sensitivity is genetic in origin, this may represent a surrogate risk factor for the development of multifactorial disease and disorders.


[Effect of GMCSF-absence on neovascularization during wound healing].
To study the effect of GMCSF-absence on the rate of wound healing and neovascularization during wound repair. 30 wild type (WT) mice and 30 GMCSF- absence mice (GMCSF-KO) were obtained. They were received full thickness skin wound (0.8 cm x 0.8 cm) in each side of midline after deeply anesthesia. In the different post-injury time points, the wound sites were digitally photographed to calculate the percentage of wound closure by using computer image analyses software. The wound specimens were also obtained dynamically for immunohistological analysis of CD31 at wound site. The analysis of the wound closure showed that wound healing in GMCSF-KO mice was delayed significantly comparing with that in WT mice from the day 3 post-wounding. At days 7 and 10 after wounding significantly more numbers of blood vessels were formed in the WT controls compared to the GMCSF-KO mice. GMCSF-absence inhibits neovascularization during wound repair and leads to the delay of wound healing.


Short-term toxicological effects of cadmium on the woodlouse, Porcellio laevis (Crustacea, Isopoda).
Terrestrial isopods have been used in a number of studies as test organisms concerning the effects of chemicals in a (eco-)toxicological context. They are saprophages, which are in direct contact with polluted plant material. Cadmium is a metal with no known biological function and is toxic to life. There are a variety of anthropogenic sources of cadmium in the environment. The aims of this study were to determine the LC50 for cadmium sulfate for Porcellio laevis and the acute effect of this metal salt on the mass of this species and also to determine if P. laevis could distinguish and avoid cadmium sulfate-contaminated leaves. Acute toxicity tests and behavioral tests were conducted using concentrations ranging from 10,000 to 40,000mg/kg cadmium sulfate. The LC50 obtained of 26,700 mg/kg cadmium sulfate is an indication of P. laevis' tolerance to cadmium contamination. P. laevis also exhibited the ability to distinguish and avoid leaves contaminated with high cadmium sulfate levels.


Using Survey Questions to Measure Preferences: Lessons from an Experimental Validation in Kenya.
Can a short survey instrument reliably measure a range of fundamental economic preferences across diverse settings? We focus on survey questions that systematically predict behavior in incentivized experimental tasks among German university students (Becker et al. 2016) and were implemented among representative samples across the globe (Falk et al. 2018). This paper presents results of an experimental validation conducted among low-income individuals in Nairobi, Kenya. We find that quantitative survey measures -- hypothetical versions of experimental tasks -- of time preference, attitude to risk and altruism are good predictors of choices in incentivized experiments, suggesting these measures are broadly experimentally valid. At the same time, we find that qualitative questions -- self-assessments -- do not correlate with the experimental measures of preferences in the Kenyan sample. Thus, caution is needed before treating self-assessments as proxies of preferences in new contexts.


BET bromodomain proteins are required for glioblastoma cell proliferation.
Epigenetic proteins have recently emerged as novel anticancer targets. Among these, bromodomain and extra terminal domain (BET) proteins recognize lysine-acetylated histones, thereby regulating gene expression. Newly described small molecules that inhibit BET proteins BRD2, BRD3, and BRD4 reduce proliferation of NUT (nuclear protein in testis)-midline carcinoma, multiple myeloma, and leukemia cells in vitro and in vivo. These findings prompted us to determine whether BET proteins may be therapeutic targets in the most common primary adult brain tumor, glioblastoma (GBM). We performed NanoString analysis of GBM tumor samples and controls to identify novel therapeutic targets. Several cell proliferation assays of GBM cell lines and stem cells were used to analyze the efficacy of the drug I-BET151 relative to temozolomide (TMZ) or cell cycle inhibitors. Lastly, we performed xenograft experiments to determine the efficacy of I-BET151 in vivo. We demonstrate that BRD2 and BRD4 RNA are significantly overexpressed in GBM, suggesting that BET protein inhibition may be an effective means of reducing GBM cell proliferation. Disruption of BRD4 expression in glioblastoma cells reduced cell cycle progression. Similarly, treatment with the BET protein inhibitor I-BET151 reduced GBM cell proliferation in vitro and in vivo. I-BET151 treatment enriched cells at the G1/S cell cycle transition. Importantly, I-BET151 is as potent at inhibiting GBM cell proliferation as TMZ, the current chemotherapy treatment administered to GBM patients. Since I-BET151 inhibits GBM cell proliferation by arresting cell cycle progression, we propose that BET protein inhibition may be a viable therapeutic option for GBM patients suffering from TMZ resistant tumors.


Which Performance Tests Best Define the Special Judo Fitness Test Classification in Elite Judo Athletes?
The normative values of the Special Judo Fitness Test (SJFT) are used to evaluate judo athletes, and the question arises of which performance tests from crucial motor abilities best define the SJFT classification in elite judo athletes. This study aimed to investigate the relationship between elite judo athletes' physical performance and the evaluation using SJFT index norms. Nineteen judo athletes (11 females) (22.8 ± 2.5 years old) from the senior judo national team voluntarily participated in this study. Body composition, reaction time, balance, flexibility, agility, hand grip strength, 20 m sprint, vertical jump, SJFT, and Wingate tests were performed by athletes on four separate days at one-day intervals. Athletes were classified as regular and above (≥regular) or poor and below (≤poor) according to their SJFT index scores. Simple logistic regression analysis was used to evaluate this classification's consistency with performance test results. Odds ratios (OR) and 95% confidence intervals related to each possible factor and Wald test statistics were calculated. The SJFT index classification was associated with Wingate peak and mean power, vertical jump, and sprint performance results (p < 0.05), whereas it was not associated with body fat percentage, agility, reaction time, hand grip strength, flexibility, and balance performances (p > 0.05). SJFT index classificatory norms are mainly related to athletes' anaerobic power. Higher anaerobic power increases athletes' possibility of being classified as ≥regular.


Quality of life and outcomes after pancreaticoduodenectomy.
To assess the quality of life (QOL) and functional outcome of patients after pancreaticoduodenectomy. Pancreaticoduodenectomy is gaining acceptance and is being performed in increasing numbers for various malignant and benign diseases of the pancreas and periampullary region. There is a general impression that pancreaticoduodenectomy can severely impair QOL and alter normal activities. Only a few small studies have evaluated QOL after pancreaticoduodenectomy. A standard QOL questionnaire was sent to 323 patients surviving pancreaticoduodenectomy who had undergone surgery at The Johns Hopkins Hospital between 1981 and 1997. Thirty items on a visual analog scale were categorized into three domains: physical (15 items), psychological (10 items), and social (5 items). Scores are reported as a percentile, with 100% being the highest possible score. The same QOL questionnaire was also sent to laparoscopic cholecystectomy patients and healthy controls. A separate component of the questionnaire asked about functional outcomes and disabilities. Overall QOL scores for the 192 responding pancreaticoduodenectomy patients in the three domains (physical, psychological, social) were 78%, 79%, and 81%, respectively. These QOL scores were comparable to those of the 37 laparoscopic cholecystectomy patients and the 31 healthy controls. The pancreaticoduodenectomy patients were subgrouped into chronic pancreatitis, other benign disease, pancreatic adenocarcinoma, and other cancers. Patients who underwent resection for chronic pancreatitis and pancreatic adenocarcinoma had significantly lower QOL scores in the physical and psychological domains compared with the laparoscopic cholecystectomy patients and the healthy controls. Common problems after pancreaticoduodenectomy were weight loss, abdominal pain, fatigue, foul stools, and diabetes. This is the largest single-institution experience assessing QOL after pancreaticoduodenectomy. These data demonstrate that as a group, patients who survive pancreaticoduodenectomy have near-normal QOL scores. Many patients report weight loss and symptoms consistent with pancreatic exocrine and endocrine insufficiency. Most patients have QOL scores comparable to those of control patients and can function independently in daily activities.


Advanced analytical methods to assess physical activity behaviour using accelerometer raw time series data: a protocol for a scoping review.
Physical activity (PA) is a complex multidimensional human behaviour. Currently, there is no standardised approach for measuring PA using wearable accelerometers in health research. The total volume of PA is an important variable because it includes the frequency, intensity and duration of activity bouts, but it reduces them down to a single summary variable. Therefore, analytical approaches using accelerometer raw time series data taking into account the way PA are accumulated over time may provide more clinically relevant features of physical behaviour. Advances on these fields are highly needed in the context of the rapid development of digital health studies using connected trackers and smartwatches. The objective of this review will be to map advanced analytical approaches and their multidimensional summary variables used to provide a comprehensive picture of PA behaviour. This scoping review will be guided by the Arksey and O'Malley methodological framework. A search for relevant publications will be undertaken in MEDLINE (PubMed), Embase and Web of Science databases. The selection of articles will be limited to studies published in English from January 2010 onwards. Studies including analytical methods that go beyond total PA volume, average daily acceleration and the conventional cut-point approaches, involving tri-axial accelerometer data will be included. Two reviewers will independently screen all citations, full-text articles and extract data. The data will be collated, stored and charted to provide a descriptive summary of the analytical methods and outputs, their strengths and limitations and their association with different health outcomes. This protocol describes a systematic method to identify, map and synthesise advanced analytical approaches and their multidimensional summary variables used to investigate PA behaviour and identify potentially clinically relevant features. The results of this review will be useful to guide future research related to analysing PA patterns, investigate their association with health conditions and suggest appropriate recommendations for changes in PA behaviour. The results may be of interest to sports scientists, clinical researchers, epidemiologists and smartphone application developers in the field of PA assessment. This protocol has been registered with the Open Science Framework (OSF): https://osf.io/yxgmb .


Inconsistency between direct and indirect comparisons of competing interventions: meta-epidemiological study.
To investigate the agreement between direct and indirect comparisons of competing healthcare interventions. Meta-epidemiological study based on sample of meta-analyses of randomised controlled trials. Data sources Cochrane Database of Systematic Reviews and PubMed. Inclusion criteria Systematic reviews that provided sufficient data for both direct comparison and independent indirect comparisons of two interventions on the basis of a common comparator and in which the odds ratio could be used as the outcome statistic. Inconsistency measured by the difference in the log odds ratio between the direct and indirect methods. The study included 112 independent trial networks (including 1552 trials with 478,775 patients in total) that allowed both direct and indirect comparison of two interventions. Indirect comparison had already been explicitly done in only 13 of the 85 Cochrane reviews included. The inconsistency between the direct and indirect comparison was statistically significant in 16 cases (14%, 95% confidence interval 9% to 22%). The statistically significant inconsistency was associated with fewer trials, subjectively assessed outcomes, and statistically significant effects of treatment in either direct or indirect comparisons. Owing to considerable inconsistency, many (14/39) of the statistically significant effects by direct comparison became non-significant when the direct and indirect estimates were combined. Significant inconsistency between direct and indirect comparisons may be more prevalent than previously observed. Direct and indirect estimates should be combined in mixed treatment comparisons only after adequate assessment of the consistency of the evidence.


[Dietary patterns and correlates in adults living in Ribeirão Preto, São Paulo State, Brazil: the OBEDIARP Project].
This study aimed to identify and analyze correlates of dietary patterns in residents of Ribeirão Preto, São Paulo State, Brazil. This was a cross-sectional study including both men and women aged > 30 years (n = 930, weighted sample = 2,197). Factor analysis was applied to identify food consumption patterns, using a semi-quantitative questionnaire. Four patterns were identified: (a) obesogenic: associated with more physical activity, more schooling, and age < 40 years; (b) healthy: associated with female gender, individuals without overweight, older adults, central obesity, more physical activity, and higher socioeconomic status; (c) mixed: without overweight, female gender, and younger adults; and (d) popular: associated with absence of hypercholesterolemia and lower family income. The results highlight the need to encourage eating healthy foods in order to prevent chronic non-communicable diseases.


Identification of new inhibitors for low molecular weight protein tyrosine phosphatase isoform B.
The National Cancer Institute Diversity Set II (1356 compounds) and Diversity Set III (1597 compounds) were screened via in silico methods as potential inhibitors of low molecular weight protein tyrosine phosphatase (LWM-PTP) isoform B (EC 18.104.22.168). Those candidates that demonstrated comparable or better docking scores than that of pyridoxal 5'-phosphate (PLP), one of the most potent known inhibitors of LMW-PTP with a competitive inhibitor dissociation constant (Kis) of 7.6μM (pH 5.0), were analyzed via in vitro kinetic assays against LMW-PTP isoform B. While none of the compounds tested in vitro was significantly better that PLP, five compounds showed comparable inhibition. These five compounds are very diverse in structure and represent new therapeutic leads for inhibition of this isozyme.


Autoimmunity and the immunotherapy of cancer: targeting the "self" to destroy the "other".
It is increasingly clear that immunity to "self"-antigens may result in tumor destruction in mouse and man. But which antigens should be targeted with therapeutic cancer vaccines? In the case of melanoma, recognition of melanocyte differentiation antigens (MDA) can be associated with autoimmune depigmentation (vitiligo). We propose that intersection of protein transport to melanosomes and endosomes allows for the loading of MDA-derived peptides on MHC class II molecules, resulting in the activation of MDA-specific CD4+ "helper" T cells that aid the induction of melanoma-specific CD8+ T cells. Thus, the immunogenicity of MDA may be a consequence of their unique cell biology. Studies of MDA-based vaccines can provide new insight into the development of more effective cancer vaccines.


Cutaneous leishmaniasis vector control perspectives using lambdacyhalothrin residual house spraying in El Ingenio, Miranda State, Venezuela.
An indoor vector control trial was carried out between December 1996 and February 1997, with the aim of reducing the population densities of Lutzomyia ovallesi, the proven vector of cutaneous leishmaniasis (CL) in El Ingenio, Miranda State, Venezuela. Houses were matched according to their structure ("bahareque" [mud and straw], concrete, and wood) and randomly assigned to a control group (n = 19) or a treated group (n = 20) that was sprayed with 25 mg/m(2) of lambdacyhalothrin. This dose was selected on the basis of a laboratory susceptibility test of Lu. ovallesi to pyrethroids. Sandfly abundance was measured using CDC light traps 7-79 d post-intervention, by which time the sandfly population in control houses had declined to very low levels, at what was the end of the transmission season. Catches of total females, fed females, and males were significantly lower in sprayed than in control houses immediately after treatment. Catches of total females and males in sprayed houses recovered, reaching the level of control houses after 7 and 11 weeks, respectively. Bioassays following WHO protocols using contact cones on a sandfly laboratory colony showed a short residual effect of the insecticide explaining these results. In contrast, there was no tendency for catches of fed females in the sprayed houses to recover during the course of the trial. Previous studies of adult population dynamics showed a short season of high abundance. Combined with these results, we believe that 2 indoor sprayings with lambdacyhalothrin, the first at the beginning of November and the second at the beginning of January, would considerably reduce the Lu. ovallesi population, and control CL transmission in this and other foci where this sandfly is considered to be the main vector.


International Germ Cell Consensus Classification: a prognostic factor-based staging system for metastatic germ cell cancers. International Germ Cell Cancer Collaborative Group.
Cisplatin-containing chemotherapy has dramatically improved the outlook for patients with metastatic germ cell tumors (GCT), and overall cure rates now exceed 80%. To make appropriate risk-based decisions about therapy and to facilitate collaborative trials, a simple prognostic factor-based staging classification is required. Collaborative groups from 10 countries provided clinical data on patients with metastatic GCT treated with cisplatin-containing chemotherapy. Multivariate analyses of prognostic factors for progression and survival were performed and models were validated on an independent data set. Data were available on 5,202 patients with nonseminomatous GCT (NSGCT) and 660 patients with seminoma. Median follow-up time was 5 years. For NSGCT the following independent adverse factors were identified: mediastinal primary site; degree of elevation of alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and lactic dehydrogenase (LDH); and presence of nonpulmonary visceral metastases (NPVM), such as liver, bone, and brain. For seminoma, the predominant adverse feature was the presence of NPVM. Integration of these factors produced the following groupings: good prognosis, comprising 60% of GCT with a 91% (89% to 93%) 5-year survival rate; intermediate prognosis, comprising 26% of GCT with a 79% (75% to 83%) 5-year survival rate; and poor prognosis, comprising 14% of GCT (all with NSGCT) with a 48% (42% to 54%) 5-year survival rate. An easily applicable, clinically based, prognostic classification for GCT has been agreed on between all the major clinical trial groups who are presently active worldwide. This should be used in clinical practice and in the design and reporting of clinical trials to aid international collaboration and understanding.


Exercise for low-income patients with diabetes: a continuous quality improvement project.
The specific aim of this project was to form a partnership between a community health center (CHC) to improve access to exercise for low-income patients with type 2 diabetes. Eligible participants were members of the Siouxland Community Health Center (SCHC), 19 years of age or older, with a diagnosis of type 2 diabetes. Patients were medically waived and received an invitation to exercise at the Siouxland YMCA. An exercise coach (pharmacist, registered nurse, or medical assistant) met with each patient and scheduled and conducted exercise visits for participating patients 2 times each week. Patients' baseline clinical variables were taken and assessed quarterly over a 12-month period. This collaboration established an ongoing exercise program with a total of 1297 exercise encounters in a 12-month period. Forty-eight of 130 eligible patients (36.9%) visited the YMCA on at least 1 occasion. Patients were categorized as experimental (56.3%), involved (25%), and regular (18.8%) exercisers. Nine (18.8%) of the 48 patients attended supplementary YMCA exercise encounters. Five (11%) of the 48 patients renewed and purchased a membership at the YMCA and adopted a self-management approach to regular exercise. The creation of a partnership between a community health center and an exercise organization permitted disadvantaged patients with diabetes an affordable exercise program and contributed to improved clinical measurements. The exercise program can be used as a model for other community health centers and health professionals who desire to improve the implementation of an exercise program for patients with diabetes.


Plasma phthalate and bisphenol a levels and oxidant-antioxidant status in autistic children.
Phthalates and bisphenol A (BPA) are endocrine disruting chemicals (EDCs) that are suggested to exert neurotoxic effects. This study aimed to determine plasma phthalates and BPA levels along with oxidant/antioxidant status in autistic children [n=51; including 12 children were diagnosed with "Pervasive Developmental Disorder-Not Otherwise Specified (PDD-NOS)]. Plasma levels of BPA, di (2-ethylhexyl)-phthalate (DEHP) and its main metabolite mono (2-ethylhexyl)-phthalate (MEHP); thiobarbituric acid reactive substance (TBARS) and carbonyl groups; erythrocyte glutathione peroxidase (GPx1), thioredoxin reductase (TrxR), catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR) activities and glutathione (GSH) and selenium levels were measured. Plasma BPA levels of children with PDD-NOS were significantly higher than both classic autistic children and controls (n=50). Carbonyl, selenium concentrations and GPx1, SOD and GR activities were higher (p<0.05); CAT activity was markedly lower in study group. BPA exposure might be associated with PDD-NOS. Intracellular imbalance between oxidant and antioxidant status might facilitate its neurotoxicity.


Effect of Early-life Gut Mucosal Compromise on Disease Progression in NOD Mice.
Disease expression in spontaneous nonobese diabetic (NOD) mice depends on environmental stimuli such as stress, diet, and gut microbiota composition. We evaluated a brief, early-life gut intervention in which pups were weaned to low-dose dextran sulfate sodium (DSS). We hypothesized that the mucus-reducing effect of this compound and subsequent increased host-bacterial contact would delay disease onset and decrease insulitis due to enhanced oral tolerance. However, disease incidence did not differ between groups, although median survival (time point when 50% of the mice are still alive) of the control group was 184 d compared with 205 d for DSS-treated mice. Mean age at disease onset (that is, blood glucose of at least 12 mmol/L) was 164 d for control mice and 159 d for DSS-treated mice. In addition, 62.5% of control mice reached a blood glucose of 12 mmol/L before 30 wk of age compared with 59% in DSS-treated mice, which had a significant transient increase in serum insulin in week 4. No changes were found in immune cells collected from spleen, pancreatic lymph nodes, and mesenteric lymph nodes. Although mice received a low dose of DSS, the subsequent reduction in the diversity of the microbiota during weeks 4 through 6 led to increased cecal length and weight and, in week 13, a tendency toward decreased colon length, with increased leakage of LPS to the blood. We conclude that mucus reduction and subsequent increased host-bacterial contact did not affect overall disease progression in NOD mice.


Step Duration Effects on Blood Loss in Simulated Designs of Tourniquet Use Procedure.
We sought new knowledge by further developing a model of using calculations in the simulation of a first-aid task. The purpose of this study was to develop the model to investigate the performance of tourniquet use in its component steps. We aimed to design an experiment on a desktop computer by mathematically manipulating simulated data in tourniquet use. A time factor of tourniquet use was ranged widely through time challenges in five degrees from ideal to worst performances. Redesigning the task was assessed by time costs and blood losses. The step of tourniquet application took 17% of the trial time and securing the tourniquet after bleeding control took the longest amount of the trial time, 31%. A minority of the time (48% [17% + 31%] to apply tourniquet plus secure it) was spent after the tourniquet touched the patient, whereas most of the time (52%) was spent before the tourniquet touched the patient. The step of tourniquet application lost 14% of the total blood lost, whereas no blood was lost during securing the tourniquet, because that was the moment of bleeding control despite securing the tourniquet taking much time (31%). Most (86%) of blood lost occurred before the tourniquet touched the patient. But blood losses differed 10-fold, with a maximum of 2,434mL, which, when added to a pretask indication blood loss of 177mL, summed to 2,611mL. Before redesigning the task, costs of donning gloves and calling 9-1-1 included uncontrolled bleeding, but gloving mitigated risk of spreading pathogens among people. By step and person, redesigns of the task altered the risk-benefit profile. The model was useful because it simulated where most of the bleeding occurred before the tourniquet touched the patient. Modeling simulated redesigns of the task, which showed changes in the task's risk-benefit profile by step and among persons. The model generated hypotheses for future research, including the capability to screen candidate ideas among task designs.


A reference map of potential determinants for the human serum metabolome.
The serum metabolome contains a plethora of biomarkers and causative agents of various diseases, some of which are endogenously produced and some that have been taken up from the environment1. The origins of specific compounds are known, including metabolites that are highly heritable2,3, or those that are influenced by the gut microbiome4, by lifestyle choices such as smoking5, or by diet6. However, the key determinants of most metabolites are still poorly understood. Here we measured the levels of 1,251 metabolites in serum samples from a unique and deeply phenotyped healthy human cohort of 491 individuals. We applied machine-learning algorithms to predict metabolite levels in held-out individuals on the basis of host genetics, gut microbiome, clinical parameters, diet, lifestyle and anthropometric measurements, and obtained statistically significant predictions for more than 76% of the profiled metabolites. Diet and microbiome had the strongest predictive power, and each explained hundreds of metabolites-in some cases, explaining more than 50% of the observed variance. We further validated microbiome-related predictions by showing a high replication rate in two geographically independent cohorts7,8 that were not available to us when we trained the algorithms. We used feature attribution analysis9 to reveal specific dietary and bacterial interactions. We further demonstrate that some of these interactions might be causal, as some metabolites that we predicted to be positively associated with bread were found to increase after a randomized clinical trial of bread intervention. Overall, our results reveal potential determinants of more than 800 metabolites, paving the way towards a mechanistic understanding of alterations in metabolites under different conditions and to designing interventions for manipulating the levels of circulating metabolites.


Statistical alignment with a sequence evolution model allowing rate heterogeneity along the sequence.
We present a stochastic sequence evolution model to obtain alignments and estimate mutation rates between two homologous sequences. The model allows two possible evolutionary behaviors along a DNA sequence in order to determine conserved regions and take its heterogeneity into account. In our model, the sequence is divided into slow and fast evolution regions. The boundaries between these sections are not known. It is our aim to detect them. The evolution model is based on a fragment insertion and deletion process working on fast regions only and on a substitution process working on fast and slow regions with different rates. This model induces a pair hidden Markov structure at the level of alignments, thus making efficient statistical alignment algorithms possible. We propose two complementary estimation methods, namely, a Gibbs sampler for Bayesian estimation and a stochastic version of the EM algorithm for maximum likelihood estimation. Both algorithms involve the sampling of alignments. We propose a partial alignment sampler, which is computationally less expensive than the typical whole alignment sampler. We show the convergence of the two estimation algorithms when used with this partial sampler. Our algorithms provide consistent estimates for the mutation rates and plausible alignments and sequence segmentations on both simulated and real data.


Effect of pulmonary hypertension on gas exchange.
This paper reviews the effects of pulmonary artery hypertension on gas exchange by exploring three different issues, namely: 1) how does gas exchange behave in diseases characterized by increased vascular tone (primary pulmonary hypertension (PPH), chronic obstructive pulmonary disease (COPD) and interstitial pulmonary fibrosis (IPF)) or decreased vascular tone ("hepatopulmonary syndrome"); 2) how does exercise, as a non-pharmacological tool of increasing pulmonary blood flow, modify gas exchange in these diseases; and 3) how do several drugs that lower (vasodilators) or increase (almitrine) the active component of pulmonary hypertension interact with gas exchange. Available data show that: 1) in PPH a high pulmonary vascular tone enhances gas exchange and when it is lowered, either by oxygen or vasodilators, ventilation perfusion (VA/Q) distributions deteriorate; 2) in COPD a lowered (vasodilators) or augmented (almitrine) active vascular tone is almost invariably paralleled by a deterioration or enhancement of ventilation-perfusion matching, respectively; 3) in IPF an adequate active response of the pulmonary vasculature is essential to maintain gas exchange, both at rest and during exercise; and 4) in patients with liver cirrhosis a low pulmonary vascular tone induces an abnormal VA/Q distribution. In summary, these data show that any situation and/or therapeutic intervention that lowers the active vascular tone deteriorates VA/Q relationships and vice versa. The final effect of pulmonary vascular tone on arterial oxygen tension (PaO2) is less predictable. The reason for this uncertainty is that the actual PaO2 value depends on the interplay of the intra- and extrapulmonary factors that control gas exchange in humans, and not only on the degree of VA/Q mismatching.


Serum xanthine oxidase: origin, regulation, and contribution to control of trypanosome parasitemia.
African trypanosomiasis is caused by Salivarian trypanosomes, tsetse fly-transmitted protozoa that inhabit the blood plasma, lymph and interstitial fluids, and, in the case of Trypanosoma brucei species, also the cerebrospinal fluid of mammal hosts. Trypanosomiasis in people and domestic animals manifests as recurring waves of parasites in the blood and is typically fatal. In contrast, trypanosomiasis in Cape buffaloes, which are naturally selected to resist the disease, is characterized by the development of only one or a few waves of parasitemia, after which the infection becomes cryptic, being maintained by the presence of 1-20 mammal-infective organisms/ml of blood. The control of the acute phase of parasitemia in Cape buffaloes correlates with a decline in blood catalase activity and the generation of trypanocidal H(2)O(2) in serum during the catabolism of endogenous purine by xanthine oxidase. Here we review features of this response, and of trypanosome metabolism, that facilitate H(2)O(2)-mediated killing of the parasites with minimal damage to the host. We also discuss the origin and regulation of serum xanthine oxidase and catalase, and show how recovery of serum catalase in infected Cape buffaloes precludes a role for H(2)O(2) in the long-term, stable suppression of trypanosome parasitemia.


Sustained proliferation in cancer: Mechanisms and novel therapeutic targets.
Proliferation is an important part of cancer development and progression. This is manifest by altered expression and/or activity of cell cycle related proteins. Constitutive activation of many signal transduction pathways also stimulates cell growth. Early steps in tumor development are associated with a fibrogenic response and the development of a hypoxic environment which favors the survival and proliferation of cancer stem cells. Part of the survival strategy of cancer stem cells may manifested by alterations in cell metabolism. Once tumors appear, growth and metastasis may be supported by overproduction of appropriate hormones (in hormonally dependent cancers), by promoting angiogenesis, by undergoing epithelial to mesenchymal transition, by triggering autophagy, and by taking cues from surrounding stromal cells. A number of natural compounds (e.g., curcumin, resveratrol, indole-3-carbinol, brassinin, sulforaphane, epigallocatechin-3-gallate, genistein, ellagitannins, lycopene and quercetin) have been found to inhibit one or more pathways that contribute to proliferation (e.g., hypoxia inducible factor 1, nuclear factor kappa B, phosphoinositide 3 kinase/Akt, insulin-like growth factor receptor 1, Wnt, cell cycle associated proteins, as well as androgen and estrogen receptor signaling). These data, in combination with bioinformatics analyses, will be very important for identifying signaling pathways and molecular targets that may provide early diagnostic markers and/or critical targets for the development of new drugs or drug combinations that block tumor formation and progression.


Induction and prepatterning of the zebrafish pectoral fin bud requires axial retinoic acid signaling.
Vertebrate forelimbs arise as bilateral appendages from the lateral plate mesoderm (LPM). Mutants in aldh1a2 (raldh2), an embryonically expressed gene encoding a retinoic acid (RA)-synthesizing enzyme, have been used to show that limb development and patterning of the limb bud are crucially dependent on RA signaling. However, the timing and cellular origin of RA signaling in these processes have remained poorly resolved. We have used genetics and chemical modulators of RA signaling to resolve these issues in the zebrafish. By rescuing pectoral fin induction in the aldh1a2/neckless mutant with exogenous RA and by blocking RA signaling in wild-type embryos, we find that RA acts as a permissive signal that is required during the six- to eight-somite stages for pectoral fin induction. Cell-transplantation experiments show that RA production is not only crucially required from flanking somites, but is sufficient to permit fin bud initiation when the trunk mesoderm is genetically ablated. Under the latter condition, intermediate mesoderm alone cannot induce the pectoral fin field in the LPM. We further show that induction of the fin field is directly followed by a continued requirement for somite-derived RA signaling to establish a prepattern of anteroposterior fates in the condensing fin mesenchyme. This process is mediated by the maintained expression of the transcription factor hand2, through which the fin field is continuously posteriorized, and lasts up to several hours prior to limb-budding. Thus, RA signaling from flanking somites plays a dual early role in the condensing limb bud mesenchyme.


Integrating clinical research into clinical decision making.
Evidence-based medicine has placed a general priority on knowledge gained from clinical research for clinical decision making. However, knowledge derived from empiric, population-based research, while valued for its ability to limit bias, is not directly applicable to the care of individual patients. The gap between clinical research and individual patient care centers on the fact that empiric research is not generally designed to answer questions of direct relevance to individual patients. Clinicians must utilize other forms of medical knowledge, including pathophysiologic rationale and clinical experience, in order to arrive at the best medical decision for a particular patient. In addition, clinicians must also elucidate and account for the goals and values of individual patients as well as barriers and facilitators of care inherent in the system in which they practice. Evidence-based guidelines and protocols, then, can never be prescriptive. Clinicians must continue to rely on clinical judgment, negotiating potentially conflicting warrants for action, in an effort to arrive at the best decision for a particular patient.


Dissecting adipose tissue lipolysis: molecular regulation and implications for metabolic disease.
Lipolysis is the process by which triglycerides (TGs) are hydrolyzed to free fatty acids (FFAs) and glycerol. In adipocytes, this is achieved by sequential action of adipose TG lipase (ATGL), hormone-sensitive lipase (HSL), and monoglyceride lipase. The activity in the lipolytic pathway is tightly regulated by hormonal and nutritional factors. Under conditions of negative energy balance such as fasting and exercise, stimulation of lipolysis results in a profound increase in FFA release from adipose tissue (AT). This response is crucial in order to provide the organism with a sufficient supply of substrate for oxidative metabolism. However, failure to efficiently suppress lipolysis when FFA demands are low can have serious metabolic consequences and is believed to be a key mechanism in the development of type 2 diabetes in obesity. As the discovery of ATGL in 2004, substantial progress has been made in the delineation of the remarkable complexity of the regulatory network controlling adipocyte lipolysis. Notably, regulatory mechanisms have been identified on multiple levels of the lipolytic pathway, including gene transcription and translation, post-translational modifications, intracellular localization, protein-protein interactions, and protein stability/degradation. Here, we provide an overview of the recent advances in the field of AT lipolysis with particular focus on the molecular regulation of the two main lipases, ATGL and HSL, and the intracellular and extracellular signals affecting their activity.


Loss of protein tyrosine phosphatase, non-receptor type 2 is associated with activation of AKT and tamoxifen resistance in breast cancer.
Breast cancer is a heterogeneous disease and new clinical markers are needed to individualise disease management and therapy further. Alterations in the PI3K/AKT pathway, mainly PIK3CA mutations, have been shown frequently especially in the luminal breast cancer subtypes, suggesting a cross-talk between ER and PI3K/AKT. Aberrant PI3K/AKT signalling has been connected to poor response to anti-oestrogen therapies. In vitro studies have shown protein tyrosine phosphatase, non-receptor type 2 (PTPN2) as a previously unknown negative regulator of the PI3K/AKT pathway. Here, we evaluate possible genomic alterations in the PTPN2 gene and its potential as a new prognostic and treatment predictive marker for endocrine therapy benefit in breast cancer. PTPN2 gene copy number was assessed by real-time PCR in 215 tumour samples from a treatment randomised study consisting of postmenopausal patients diagnosed with stage II breast cancer 1976-1990. Corresponding mRNA expression levels of PTPN2 were evaluated in 86 available samples by the same methodology. Gene copy loss of PTPN2 was detected in 16% (34/215) of the tumours and this was significantly correlated with lower levels of PTPN2 mRNA. PTPN2 gene loss and lower mRNA levels were associated with activation of AKT and a poor prognosis. Furthermore, PTPN2 gene loss was a significant predictive marker of poor benefit from tamoxifen treatment. In conclusion, genomic loss of PTPN2 may be a previously unknown mechanism of PI3K/AKT upregulation in breast cancer. PTPN2 status is a potential new clinical marker of endocrine treatment benefit which could guide further individualised therapies in breast cancer.


What we should know about portal vein thrombosis in cirrhotic patients: a changing perspective.
Portal vein thrombosis (PVT) is one of the most common complications occurring during the natural course of liver cirrhosis. Even though PVT is often asymptomatic, the worsening of liver function, an unexpected episode of gastrointestinal bleeding or ascitic decompensation may be landmarks of PVT development. Beyond these clinical manifestations, it is debated whether PVT really has an impact on liver cirrhosis natural history or rather represents only one of its consequences. Probably PVT development should not only be considered as a matter of impaired blood flow or pro-coagulation tendency. On one hand, PVT seems a consequence of the worsening in portal vein outflow due to the increased hepatic resistance in cirrhotic livers. On the other hand, vascular microthrombosis secondary to necroinflammation may cause liver ischemia and infarction, with loss of hepatic tissue (parenchymal extinction) which is replaced by fibrotic tissue. Therefore, PVT might also be considered as the overt manifestation of the liver fibrosing process evolution and anticoagulant therapy may thus have microscopic indirect effects also on the progression of liver disease. At present, a connection between PVT development and the progression of liver fibrosis/cirrhosis has not yet been demonstrated. Nevertheless, it is not clear if PVT development may worsen cirrhotic patients' outcome by itself. Some authors tried to assess liver transplant benefit in PVT cirrhotic patients but data are contrasting. In this review, we will try to answer these questions, providing a critical analysis of data reported in literature.


Sequence analysis and expression of the bacterial dichloromethane dehalogenase structural gene, a member of the glutathione S-transferase supergene family.
The nucleotide sequence of a cloned 2.8-kilobase-pair BamHI-PstI fragment containing dcmA, the dichloromethane dehalogenase structural gene from Methylobacterium sp. strain DM4, was determined. An open reading frame with a coding capacity of 287 amino acids (molecular weight, 37,430) was identified as dcmA by its agreement with the N-terminal amino acid sequence, the total amino acid composition, and the subunit size of the purified enzyme. Alignment of the deduced dichloromethane dehalogenase amino acid sequence with amino acid sequences of the functionally related eucaryotic glutathione S-transferases revealed three regions containing highly conserved amino acid residues and indicated that dcmA is a member of the glutathione S-transferase supergene family. The 5' terminus of in vivo dcmA transcripts was determined by nuclease S1 mapping to be 82 base pairs upstream of the GTG initiation codon of dcmA. Despite a putative promoter sequence with high resemblance to the Escherichia coli -10 and -35 consensus sequences, located at an appropriate distance from the transcription start point, dcmA was only marginally expressed in E. coli. The strong induction of dichloromethane dehalogenase in Methylobacterium sp. by dichloromethane was abolished by deleting the 1.3-kilobase-pair upstream region of dcmA. Plasmid constructs devoid of this region directed expression of dichloromethane dehalogenase at a constitutively induced level.


Must we press on until a young mother dies? Remifentanil patient controlled analgesia in labour may not be suited as a "poor man's epidural".
The epidural route is still considered the gold standard for labour analgesia, although it is not without serious consequences when incorrect placement goes unrecognized, e.g. in case of intravascular, intrathecal and subdural placements. Until now there has not been a viable alternative to epidural analgesia especially in view of the neonatal outcome and the need for respiratory support when long-acting opioids are used via the parenteral route. Pethidine and meptazinol are far from ideal having been described as providing rather sedation than analgesia, affecting the cardiotocograph (CTG), causing fetal acidosis and having active metabolites with prolonged half-lives especially in the neonate. Despite these obvious shortcomings, intramuscular and intravenously administered pethidine and comparable substances are still frequently used in delivery units. Since the end of the 90 ths remifentanil administered in a patient-controlled mode (PCA) had been reported as a useful alternative for labour analgesia in those women who either don't want, can't have or don't need epidural analgesia. In view of the need for conversion to central neuraxial blocks and the analgesic effect remifentanil has been demonstrated to be superior to pethidine. Despite being less effective in terms of the resulting pain scores, clinical studies suggest that the satisfaction with analgesia may be comparable to that obtained with epidural analgesia. Owing to this fact, remifentanil has gained a place in modern labour analgesia in many institutions. However, the fact that remifentanil may cause harm should not be forgotten when the use of this potent mu-agonist is considered for the use in labouring women. In the setting of one-to-one midwifery care, appropriate monitoring and providing that enough experience exists with this potent opioid and the treatment of potential complications, remifentanil PCA is a useful option in addition to epidural analgesia and other central neuraxial blocks. Already described serious consequences should remind us not refer to remifentanil PCA as a "poor man's epidural" and to safely administer remifentanil with an appropriate indication. Therefore, the authors conclude that economic considerations and potential cost-savings in conjunction with remifentanil PCA may not be appropriate main endpoints when studying this valuable method for labour analgesia.


Evaluation of a single injection of 99mTc-labeled diethylenetriaminepentaacetic acid for measuring glomerular filtration rate in horses.
Glomerular filtration rate (GFR) was measured in 12 clinically normal horses, using the standard inulin clearance method, and values were compared with values for 2 methods, using a single rapid IV injection of 99mTc-labeled diethylenetriaminepentaacetic acid (99mTc-DTPA). The first 99mTc-DTPA method used a 2-compartment model to calculate GFR blood clearance of the tracer. The second method used sequential digital gamma camera images of the kidneys to determine fractional accumulation of the total dose of the tracer in the kidneys (percentage of injected dose, gamma camera) from 0 to 10 minutes after radionuclide administration. Linear correlation among the 3 methods was determined. Mean (+/- SD) GFR, using the inulin clearance method, was 154.67 +/- 42.28 ml/min/100 kg of body weight. Mean GFR, using the 2-compartment blood clearance curve, was 146.92 +/- 27.49 ml/min/100 kg. Mean GFR, using percentage of injected dose (gamma camera method) was 154.7 +/- 22.00 ml/min/100 kg. The percentage of injected dose (gamma camera method) did not correlate significantly to the inulin clearance results. However, a significant (r = 0.666, P less than 0.018) correlation was observed between the inulin method and the 99mTc-DTPA blood clearance method. Significant (P less than 0.0001) difference also was observed in the split function of the equine kidneys, with GFR of the right kidney contributing 60.1 +/- 9.12% of the total function, as determined by 99mTc-DTPA gamma camera imaging. Because the 99mTc-DTPA blood clearance method does not require urine collection, it may be a more practical procedure to measure GFR in the horse.