Disorders of cancer metabolism: The therapeutic potential of cannabinoids.
Abnormal energy metabolism, as one of the important hallmarks of cancer, was induced by multiple carcinogenic factors and tumor-specific microenvironments. It comprises aerobic glycolysis, de novo lipid biosynthesis, and glutamine-dependent anaplerosis. Considering that metabolic reprogramming provides various nutrients for tumor survival and development, it has been considered a potential target for cancer therapy. Cannabinoids have been shown to exhibit a variety of anticancer activities by unclear mechanisms. This paper first reviews the recent progress of related signaling pathways (reactive oxygen species (ROS), AMP-activated protein kinase (AMPK), mitogen-activated protein kinases (MAPK), phosphoinositide 3-kinase (PI3K), hypoxia-inducible factor-1alpha (HIF-1α), and p53) mediating the reprogramming of cancer metabolism (including glucose metabolism, lipid metabolism, and amino acid metabolism). Then we comprehensively explore the latest discoveries and possible mechanisms of the anticancer effects of cannabinoids through the regulation of the above-mentioned related signaling pathways, to provide new targets and insights for cancer prevention and treatment.


Interplay between quantum interference and conformational fluctuations in single-molecule break junctions.
We theoretically explored the combined role of conformational fluctuations and quantum interference in determining the electrical conductance of single-molecule break junctions. In particular we computed the conductance of a family of methylsulfide-functionalized trans-α,ω-diphenyloligoene molecules, with terminal phenyl rings containing meta or para linkages, for which (at least in the absence of fluctuations) destructive interference in the former is expected to decrease their electrical conductance compared with the latter. We compared the predictions of density functional theory (DFT), in which fluctuational effects are absent, with results for the conformationally-averaged conductance obtained from an ensemble of conformations obtained from classical molecular dynamics. We found that junctions formed from these molecules exhibit distinct transport regimes during junction evolution and the signatures of quantum interference in these molecules survive the effect of conformational fluctuations. Furthermore, the agreement between theory and experiment is significantly improved by including conformational averaging.


Early sexual debut: Voluntary or coerced? Evidence from longitudinal data in South Africa--the Birth to Twenty Plus study.
Early sexual debut, voluntary or coerced, increases risks to sexual and reproductive health. Sexual coercion is increasingly receiving attention as an important public health issue owing to its association with adverse health and social outcomes. To describe voluntary and coerced experience at sexual debut. A longitudinal perspective among 2,216 adolescents (1,149 females, 1,067 males) in a birth cohort study in South Africa, analysing data collected on six occasions between 11 and 18 years. The median age of sexual debut was 16 years for females and 15 for males. Reported coerced sexual debut included children < 11 years of age. Males reported earlier sexual debut with both voluntary and coerced sexual experience, than females (p < 0.0001). Sexual coercion at early sexual debut among both male and female adolescents occurred mostly through sexual intercourse with older adolescents and partners of the same age. The identified time periods and age groups need to be targeted for interventions to delay sexual debut and prevent sexual coercion among young people. More research is needed to understand underlying predisposing risk factors for sexual coercion at sexual debut, both early and not early.


Surgical resident perceptions of trauma surgery as a specialty.
Presenting the opinions of surgical residents about the appeal of trauma surgery as a specialty may influence current reform. Survey study. Academic research. General surgery residents (postgraduate years 1-5 and recent graduates) registered with the American College of Surgeons. A 22-item survey. Career plans and perceptions about trauma surgery as a specialty. Of 6006 mailed surveys, we had a 20.1% response rate. Midlevel residents comprised most of the respondents, and most were undecided about their career choice or planned to enter general surgical private practice. The typical residency programs represented were academic (81.7%), urban (90.6%), and level I trauma centers (78.7%), and included more than 6 months of trauma experience (77.6%). Most respondents (70.6%) thought that trauma surgery was unappealing. The most important deterrents to entering the field were lifestyle, poor reimbursement, and limited operating room exposure, while increased surgical critical care was not seen as a restriction. When questioned about the future of trauma surgery, they believed that trauma surgeons should perform elective (86.8%) and nontrauma emergency (91.5%) cases and would benefit from active association with an outpatient clinic (76.0%). Intellectual challenge and exciting nature of the field were listed as the most appealing aspects, and ideal practice characteristics included guaranteed salary and time away from work. As demand for trauma surgeons increases, resident interest has dwindled. As a specialty, trauma surgery must undergo changes that reflect the needs of the incoming generation. We present a sampling of current surgical resident opinion and offer these data to assist the changing discipline and the evolving field of acute care surgery.


Synthesis and Evaluation of a Radioiodinated Tracer with Specificity for Poly(ADP-ribose) Polymerase-1 (PARP-1) in Vivo.
Interest in nuclear imaging of poly(ADP-ribose) polymerase-1 (PARP-1) has grown in recent years due to the ability of PARP-1 to act as a biomarker for glioblastoma and increased clinical use of PARP-1 inhibitors. This study reports the identification of a lead iodinated analog 5 of the clinical PARP-1 inhibitor olaparib as a potential single-photon emission computed tomography (SPECT) imaging agent. Compound 5 was shown to be a potent PARP-1 inhibitor in cell-free and cellular assays, and it exhibited mouse plasma stability but approximately 3-fold greater intrinsic clearance when compared to olaparib. An (123)I-labeled version of 5 was generated using solid state halogen exchange methodology. Ex vivo biodistribution studies of [(123)I]5 in mice bearing subcutaneous glioblastoma xenografts revealed that the tracer had the ability to be retained in tumor tissue and bind to PARP-1 with specificity. These findings support further investigations of [(123)I]5 as a noninvasive PARP-1 SPECT imaging agent.


Evidence for the involvement of testicular protein CRISP2 in mouse sperm-egg fusion.
CRISP2, originally known as Tpx-1, is a cysteine-rich secretory protein specifically expressed in male haploid germ cells. Although likely to be involved in gamete interaction, evidence for a functional role of CRISP2 in fertilization still remains poor. In the present study, we used a mouse model to examine the subcellular localization of CRISP2 in sperm and its involvement in the different stages of fertilization. Results from indirect immunofluorescence and protein extraction experiments indicated that mouse CRISP2 is an intraacrosomal component that remains associated with sperm after capacitation and the acrosome reaction (AR). In vitro fertilization assays using zona pellucida-intact mouse eggs showed that an antibody against the protein significantly decreased the percentage of penetrated eggs, with a coincident accumulation of perivitelline sperm. The failure to inhibit zona pellucida penetration excludes a detrimental effect of the antibody on sperm motility or the AR, supporting a specific participation of CRISP2 at the sperm-egg fusion step. In agreement with this evidence, recombinant mouse CRISP2 (recCRISP2) specifically bound to the fusogenic area of mouse eggs, as previously reported for rat CRISP1, an epididymal protein involved in gamete fusion. In vitro competition investigations showed that incubation of mouse zona-free eggs with a fixed concentration of recCRISP2 and increasing amounts of rat CRISP1 reduced the binding of recCRISP2 to the egg, suggesting that the proteins interact with common complementary sites on the egg surface. Our findings indicate that testicular CRISP2, as observed for epididymal CRISP1, is involved in sperm-egg fusion through its binding to complementary sites on the egg surface, supporting the idea of functional cooperation between homologous molecules to ensure the success of fertilization.


A heat shock-responsive domain of human HSF1 that regulates transcription activation domain function.
Human heat shock factor 1 (HSF1) stimulates transcription from heat shock protein genes following stress. We have used chimeric proteins containing the GAL4 DNA binding domain to identify the transcriptional activation domains of HSF1 and a separate domain that is capable of regulating activation domain function. This regulatory domain conferred heat shock inducibility to chimeric proteins containing the activation domains. The regulatory domain is located between the transcriptional activation domains and the DNA binding domain of HSF1 and is conserved between mammalian and chicken HSF1 but is not found in HSF2 or HSF3. The regulatory domain was found to be functionally homologous between chicken and human HSF1. This domain does not affect DNA binding by the chimeric proteins and does not contain any of the sequences previously postulated to regulate DNA binding of HSF1. Thus, we suggest that activation of HSF1 by stress in humans is controlled by two regulatory mechanisms that separately confer heat shock-induced DNA binding and transcriptional stimulation.


Meta-review of the barriers and facilitators to women accessing perinatal mental healthcare.
Perinatal mental health (PMH) problems are common and can have an adverse impact on women and their families. However, research suggests that a substantial proportion of women with PMH problems do not access care. To synthesise the results from previous systematic reviews of barriers and facilitators to women to seeking help, accessing help, and engaging in PMH care, and to suggest recommendations for clinical practice and policy. A meta-review of systematic reviews. Seven databases were searched and reviewed using a Preferred Reporting Items for Systematic Reviews and Meta Analyses search strategy. Studies that focused on the views of women seeking help and accessing PMH care were included. Data were analysed using thematic synthesis. Assessing the Methodological Quality of Systematic Reviews-2 was used to assess review methodology. To improve validity of results, a qualitative sensitivity analysis was conducted to assess whether themes remained consistent across all reviews, regardless of their quality rating. A total of 32 reviews were included. A wide range of barriers and facilitators to women accessing PMH care were identified. These mapped across a multilevel model of influential factors (individual, healthcare professional, interpersonal, organisational, political and societal) and across the care pathway (from decision to consult to receiving care). Evidence-based recommendations to support the design and delivery of PMH care were produced based on identified barriers and facilitators. The identified barriers and facilitators point to a complex interplay of many factors, highlighting the need for an international effort to increase awareness of PMH problems, reduce mental health stigma, and provide woman-centred, flexible care, delivered by well trained and culturally sensitive primary care, maternity, and psychiatric health professionals. CRD42019142854.


Flexible regression models for ROC and risk analysis, with or without a gold standard.
A novel semiparametric regression model is developed for evaluating the covariate-specific accuracy of a continuous medical test or biomarker. Ideally, studies designed to estimate or compare medical test accuracy will use a separate, flawless gold-standard procedure to determine the true disease status of sampled individuals. We treat this as a special case of the more complicated and increasingly common scenario in which disease status is unknown because a gold-standard procedure does not exist or is too costly or invasive for widespread use. To compensate for missing data on disease status, covariate information is used to discriminate between diseased and healthy units. We thus model the probability of disease as a function of 'disease covariates'. In addition, we model test/biomarker outcome data to depend on 'test covariates', which provides researchers the opportunity to quantify the impact of covariates on the accuracy of a medical test. We further model the distributions of test outcomes using flexible semiparametric classes. An important new theoretical result demonstrating model identifiability under mild conditions is presented. The modeling framework can be used to obtain inferences about covariate-specific test accuracy and the probability of disease based on subject-specific disease and test covariate information. The value of the model is illustrated using multiple simulation studies and data on the age-adjusted ability of soluble epidermal growth factor receptor - a ubiquitous serum protein - to serve as a biomarker of lung cancer in men. SAS code for fitting the model is provided. Copyright © 2015 John Wiley & Sons, Ltd.


aP2-Cre-mediated inactivation of estrogen receptor alpha causes hydrometra.
In this study we describe the reproductive phenotypes of a novel mouse model in which Cre-mediated deletion of ERα is regulated by the aP2 (fatty acid binding protein 4) promoter. ERα-floxed mice were crossed with transgenic mice expressing Cre-recombinase under the control of the aP2 promoter to generate aP2-Cre/ERα(flox/flox) mice. As expected, ERα mRNA levels were reduced in adipose tissue, but in addition we also detected an 80% reduction of ERα levels in the hypothalamus of aP2-Cre/ERα(flox/flox) mice. Phenotypic analysis revealed that aP2-Cre/ERα(flox/flox) female mice were infertile. In line with this, aP2-Cre/ERα(flox/flox) female mice did not cycle and presented 3.8-fold elevated estrogen levels. That elevated estrogen levels were associated with increased estrogen signaling was evidenced by increased mRNA levels of the estrogen-regulated genes lactoferrin and aquaporin 5 in the uterus. Furthermore, aP2-Cre/ERα(flox/flox) female mice showed an accumulation of intra-uterine fluid, hydrometra, without overt indications for causative anatomical anomalies. However, the vagina and cervix displayed advanced keratosis with abnormal quantities of accumulating squamous epithelial cells suggesting functional obstruction by keratin plugs. Importantly, treatment of aP2-Cre/ERα(flox/flox) mice with the aromatase inhibitor Letrozole caused regression of the hydrometra phenotype linking increased estrogen levels to the observed phenotype. We propose that in aP2-Cre/ERα(flox/flox) mice, increased serum estrogen levels cause over-stimulation in the uterus and genital tracts resulting in hydrometra and vaginal obstruction.


Characterization of a novel rat epididymal cell line to study epididymal function.
The epididymis is an androgen-dependent organ that allows spermatozoa to become fully functional as they pass through this tissue. The specialized functions of the epididymis are mediated by interactions between epididymal epithelial cells and between epididymal cells and spermatozoa. Although the critical role of the epididymis in sperm maturation is well established, the mechanisms regulating cell-cell interactions remain poorly understood because of the lack of appropriate cell line models. We now report the characterization of a novel rat caput epididymal cell line (RCE) that was immortalized by transfecting primary cultures of rat epididymal cells with the simian virus 40 large T antigen. At the electron microscope level, the cell line was composed of epithelial principal cells with characteristics of in vivo cells; principal cells had well-developed Golgi apparatus, abundant endoplasmic reticulum cisternae, and few endosomes. RCE cells expressed the mRNAs coding for the androgen receptor, estrogen receptor alpha, and 4-ene-steroid-5-alpha-reductase types 1 and 2 as well as epididymal-specific markers Crisp-1 and epididymal retinoic acid binding protein. Epididymal retinoic acid binding protein expression was significantly induced with dihydrotestosterone, although this effect was not blocked by flutamide, suggesting that RCE cells are not androgen responsive. Neighboring cells formed tight and gap junctions characteristic of epididymal cells in vivo and expressed tight (occludin and claudin-1, -3, and -4) and gap junctional proteins (connexin-26, -30.3, -32, and -43). The RCE cell line displays many characteristics of epithelial principal cells, thus providing a model for studying epididymal cell functions.


Tularaemia seroprevalence of captured and wild animals in Germany: the fox (Vulpes vulpes) as a biological indicator.
A total of 2475 animals from Germany, both captive and wild, were tested for antibodies against Francisella tularensis to obtain more knowledge about the presence of this pathogen in Germany. An indirect and a competitive ELISA served as screening methods, positive and inconclusive samples were confirmed by Western blot. Of the zoo animals sampled between 1992 and 2007 (n = 1122), three (0·3%) were seropositive. The seroconversion of a hippopotamus in Berlin Zoo was documented. From 1353 serum samples of wild foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and wild boars (Sus scrofa), collected between 2005 and 2009 in the federal state of Brandenburg (surrounding Berlin), a total of 101 (7·5%) tested positive for antibodies to F. tularensis lipopolysaccharide. Our results indicate a higher seroprevalence of F. tularensis in wildlife in eastern Germany than commonly assumed. Furthermore, we found foxes and raccoon dogs to be biological indicators for tularaemia.


International Union of Pharmacology. XXVIII. Proteinase-activated receptors.
Proteinase-activated receptors (PARs) represent a unique subclass of G-protein-coupled receptors of which four family members have now been cloned from a number of species. The novel mechanism of receptor activation involves the proteolytic unmasking of a cryptic N-terminal receptor sequence that, remaining tethered, binds to and triggers receptor function. In addition, short (five to six amino acids) synthetic peptides, based on the proteolytically revealed motif, can activate PARs without the unmasking of the tethered ligand. This article summarizes the experiments leading to the pharmacological characterization and cloning of the four PAR family members and provides a rationale for their designation by the acronym "PAR". The ability to distinguish among the PARs pharmacologically 1) with selective proteinase activators, 2) with receptor-selective peptide agonists, and 3) with peptide and nonpeptide antagonists is discussed, as are the molecular mechanisms of receptor activation and desensitization/internalization. Finally, the potential physiological roles of the PARs, which are widely distributed in many organs in the settings of tissue injury, repair, and remodeling, including embryogenesis and oncogenesis are discussed, and the newly appreciated roles of proteinases as signaling molecules that can act as either functional agonists or antagonists are highlighted.


Endogenous opioids may mediate secondary damage after experimental brain injury.
Although endogenous opioids have been implicated in the pathophysiology of spinal cord injury and brain ischemia, the role of specific opioid peptides and opiate receptors in the pathophysiology of traumatic brain injury remains unexplored. This study examined regional changes in brain opioid immunoreactivity and cerebral blood flow (CBF) after fluid-percussion brain injury in the cat and compared the effect of an opiate antagonist (Win 44,441-3 [Win-(-)]) with its dextroisomer Win 44,441-2 [Win-(+)] (which is inactive at opiate receptors) in the treatment of brain injury. Dynorphin A immunoreactivity (Dyn A-IR) but not leucine-enkephalin-like immunoreactivity accumulated in injury regions after traumatic injury; Dyn-IR increases also occurred predominantly in those areas showing significant decreases in regional CBF. Administration of Win-(-) but not Win-(+) or saline at 15 min after injury significantly improved mean arterial pressure, electroencephalographic amplitude, and regional CBF and reduced the severity and incidence of hemorrhage. Win-(-) also significantly improved survival after brain injury. Taken together, these findings suggest that dynorphin, through actions at opiate receptors, may contribute to the pathophysiology of secondary brain injury after head trauma and indicate that selective opiate-receptor antagonists may be useful in treatment of traumatic brain injury.


Inherently chiral cone-calix[4]arenes via a subsequent upper rim ring-closing/opening methodology.
Access to chiral calix[4]arenes can unlock novel supramolecular architectures for enantioselective catalysis and molecular recognition. However, accessibility to these structures has been significantly hindered so far. We report herein the synthesis and characterization of di- and trifunctionalized cone-calix[4]arenes featuring a lactone moiety spanning the distal positions at the upper rim. The lactones force the whole skeleton to assume pinched-cone conformations. The ring-closure is favored by the high conformational flexibility of the calixarene scaffold. The new lactones are remarkably stable in the solid state, while a quick hydrolysis to restore the parent carboxylic acids occurs in solution under acidic/basic conditions. Slow aminolyses of lactones 2-3 yield inherently chiral products featuring three different functionalities at the upper rim, at room temperature. The subsequent ring-closing/opening methodology presented here highlights the versatility of these lactones as powerful synthons for the preparation of a variety of threefold upper rim functionalized, inherently chiral calix[4]arenes fixed in the cone structure.


Antibody-based exosite inhibitors of ADAMTS-5 (aggrecanase-2).
Adamalysin-like metalloproteinases with thrombospondin (TS) motifs (ADAMTS)-5 is the multi-domain metalloproteinase that most potently degrades aggrecan proteoglycan in the cartilage and its activity is implicated in the development of osteoarthritis (OA). To generate specific exosite inhibitors for it, we screened a phage display antibody library in the presence of the zinc-chelating active site-directed inhibitor GM6001 (Ilomastat) and isolated four highly selective inhibitory antibodies. Two antibodies were mapped to react with exosites in the catalytic/disintegrin domains (Cat/Dis) of the enzyme, one in the TS domain and one in the spacer domain (Sp). The antibody reacting with the Sp blocked the enzyme action only when aggrecan or the Escherichia coli-expressed aggrecan core protein were substrates, but not against a peptide substrate. The study with this antibody revealed the importance of the Sp for effective aggrecanolytic activity of ADAMTS-5 and that this domain does not interact with sulfated glycosaminoglycans (GAGs) but with the protein moiety of the proteoglycan. An antibody directed against the Cat/Dis of ADAMTS-5 was effective in a cell-based model of aggrecan degradation; however, the anti-Sp antibody was ineffective. Western blot analysis of endogenous ADAMTS-5 expressed by human chondrocytes showed the presence largely of truncated forms of ADAMTS-5, thus explaining the lack of efficacy of the anti-Sp antibody. The possibility of ADAMTS-5 truncation must then be taken into account when considering developing anti-ancillary domain antibodies for therapeutic purposes.


Removal of luminal content protects the small intestine during hemorrhagic shock but is not sufficient to prevent lung injury.
The small intestine plays a key role in the pathogenesis of multiple organ failure following circulatory shock. Current results show that reduced perfusion of the small intestine compromises the mucosal epithelial barrier, and the intestinal contents (including pancreatic digestive enzymes and partially digested food) can enter the intestinal wall and transport through the circulation or mesenteric lymph to other organs such as the lung. The extent to which the luminal contents of the small intestine mediate tissue damage in the intestine and lung is poorly understood in shock. Therefore, rats were assigned to three groups: No-hemorrhagic shock (HS) control and HS with or without a flushed intestine. HS was induced by reducing the mean arterial pressure (30 mmHg; 90 min) followed by return of shed blood and observation (3 h). The small intestine and lung were analyzed for hemorrhage, neutrophil accumulation, and cellular membrane protein degradation. After HS, animals with luminal contents had increased neutrophil accumulation, bleeding, and destruction of E-cadherin in the intestine. Serine protease activity was elevated in mesenteric lymph fluid collected from a separate group of animals subjected to intestinal ischemia/reperfusion. Serine protease activity was elevated in the plasma after HS but was detected in lungs only in animals with nonflushed lumens. Despite removal of the luminal contents, lung injury occurred in both groups as determined by elevated neutrophil accumulation, permeability, and lung protein destruction. In conclusion, luminal contents significantly increase intestinal damage during experimental HS, suggesting transport of luminal contents across the intestinal wall should be minimized.


Effect of blue-blocking glasses in major depressive disorder with sleep onset insomnia: A randomized, double-blind, placebo-controlled study.
Blue wavelengths form the portion of the visible electromagnetic spectrum that most potently regulates circadian rhythm. We hypothesized that wearing blue-blocking (BB) glasses in the evening may influence circadian rhythm disturbances in patients with major depressive disorder (MDD), resulting in improved sleep and mood. We used a randomized placebo-controlled double-blinded design. Patients with MDD with sleep onset insomnia were randomly assigned to wearing either BB glasses or clear glasses (placebo). Patients were instructed to wear the glasses from 20:00 hours until bedtime for 2 weeks. We assessed sleep state (sleep quality on a visual analog scale, the Morningness-Eveningness Questionnaire [MEQ], and a sleep diary) and depressive symptoms at baseline and after 2 weeks. Data were analyzed with a full analysis set. In total, 20 patients were randomly assigned to the BB and placebo groups (BB group, n = 10; placebo group, n = 10). There were three dropouts (BB group, n = 1; placebo group, n = 2). At baseline, sleep quality, sleep latency (assessed via a sleep diary), and antipsychotics use differed between the groups. To take account of these differences, the baseline sleep state or depressive symptoms and antipsychotics use were used as covariates in the later analysis. The change scores for sleep quality did not show a significant improvement in the BB group compared with the placebo group (mean [standard deviation, SD] scores for BB versus placebo: 36.1 [31.7] versus 16.2 [15.1], p = 0.43), although half of the BB group showed a clear improvement in sleep quality. The change in MEQ scores did not significantly differ between the groups (p = 0.14), although there was a trend of a shift to morning type in the BB group (3.10 [4.95] points) and to evening type in the placebo group (0.50 [3.89] points). There were no statistically significant changes in depressive symptoms in either group. Across both groups, 40% of the participants reported pain or discomfort from wearing the glasses, which were available in only one size. Thus, the failure to find significant differences may have resulted from the glasses used in this study. Glasses fitted to individual patients may improve efficacy and safety. Replication of the study with a larger sample size and size-adjustable glasses is needed.


Urgent need for "EBMM" in pediatric oncology: Evidence based medical marijuana.
Marijuana has been used by many different civilizations for numerous different purposes, including its use for medical indications. Recently, there has been significant media coverage of the efficacy of medical marijuana in the treatment of seizures in children with Dravet syndrome, and this has led many to search for other possible pediatric indications for cannabinoids, including many different indications in pediatric cancer. However, there is very little evidence on safety or efficacy of cannabinoids in children being treated with cancer. This commentary accompanies a recent paper by a group in Israel who have published their experience of medical marijuana in 50 children and adolescents with cancer, showing excellent satisfaction and better symptom control, and without significant adverse drug reactions. This study from Israel is an excellent first step, but prospective well-designed trials of medical marijuana in pediatric oncology are urgently needed.


Hearing loss and cognition: the role of hearing AIDS, social isolation and depression.
Hearing loss is associated with poor cognitive performance and incident dementia and may contribute to cognitive decline. Treating hearing loss with hearing aids may ameliorate cognitive decline. The purpose of this study was to test whether use of hearing aids was associated with better cognitive performance, and if this relationship was mediated via social isolation and/or depression. Structural equation modelling of associations between hearing loss, cognitive performance, social isolation, depression and hearing aid use was carried out with a subsample of the UK Biobank data set (n = 164,770) of UK adults aged 40 to 69 years who completed a hearing test. Age, sex, general health and socioeconomic status were controlled for as potential confounders. Hearing aid use was associated with better cognition, independently of social isolation and depression. This finding was consistent with the hypothesis that hearing aids may improve cognitive performance, although if hearing aids do have a positive effect on cognition it is not likely to be via reduction of the adverse effects of hearing loss on social isolation or depression. We suggest that any positive effects of hearing aid use on cognition may be via improvement in audibility or associated increases in self-efficacy. Alternatively, positive associations between hearing aid use and cognition may be accounted for by more cognitively able people seeking and using hearing aids. Further research is required to determine the direction of association, if there is any direct causal relationship between hearing aid use and better cognition, and whether hearing aid use results in reduction in rates of cognitive decline measured longitudinally.


The use of electrophysiological monitoring in the intraoperative management of intracranial aneurysms.
Somatosensory evoked potentials (SSEPs) and brainstem auditory evoked potentials (BAEPs) have been increasingly utilised during surgery for intracranial aneurysms to identify cerebral ischaemia. Between July 1994 and April 1996, we surgically treated 70 aneurysms in 49 consecutive patients (58 operations) with the aid of intraoperative evoked potential monitoring. This study sought to evaluate the usefulness of SSEP and BAEP monitoring during intracranial aneurysm surgery. Mean patient age was 51.9 (range 18-79) years. The sizes of the aneurysms were 3-4 mm (15), 5-9 mm (26), 10-14 mm (11), 15-19 mm (seven), 20-24 mm (six), and >25 mm (five). SSEPs were monitored in 58 procedures (100%) and BAEPs in 15 (26%). The neurological status of the patients was evaluated before and after surgery. Thirteen of the 58 procedures (22%) monitored with SSEPs had SSEP changes (12 transient, one persistent); 45 (78%) had no SSEP changes. Three of 15 patients (20%) monitored with BAEPs had changes (two transient, one persistent); 12 (80%) had no BAEP changes. Of the 14 patients with transient SSEP or BAEP changes, these changes resolved with adjustment or removal of aneurysm clips (nine), elevating MAP (four), or retractor adjustment (one). Mean time from precipitating event to electrophysiological change was 8.9 minutes (range 3-32), and the mean time for recovery of potentials in patients with transient changes was 20.2 minutes (range 3-60). Clinical outcome was excellent in 39 patients, good in five, and poor in three (two patients died), and was largely related to pretreatment grade. SSEPs and BAEPs are useful in preventing clinical neurological injury during surgery for intracranial aneurysms and in predicting which patients will have unfavourable outcomes.


Novel bright-emission small-molecule NIR-II fluorophores for in vivo tumor imaging and image-guided surgery.
Though high brightness and biocompatible small NIR-II dyes are highly desirable in clinical or translational cancer research, their fluorescent cores are relatively limited and their synthetic processes are somewhat complicated. Herein, we have explored the design and synthesis of novel NIR-II fluorescent materials (H1) without tedious chromatographic isolation with improved fluorescence performance (QY ≈ 2%) by introducing 2-amino 9,9-dialkyl-substituted fluorene as a donor into the backbone. Several types of water-soluble and biocompatible NIR-II probes: SXH, SDH, and H1 NPs were constructed via different chemical strategies based on H1, and then their potential to be used in in vivo tumor imaging and image-guided surgery in the NIR-II region was explored. High levels of uptake were obtained for both passive and active tumor targeting probes SXH and SDH. Furthermore, high resolution imaging of blood vessels on tumors and the whole body of living mice using H1 NPs for the first time has demonstrated precise NIR-II image-guided sentinel lymph node (SLN) surgery.


A modified Bohlman technique using a novel implant for treatment of high-grade spondylolisthesis.
Several surgical options for managing high-grade spondylolisthesis have been described in the literature and range from posterior-only in situ fusion to circumferential fusion with complete reduction of the dislocation. The level of evidence supporting any one technique is weak, and to date there is no Level I or II evidence supporting any current surgical treatment option. Techniques have evolved as implant technology has advanced and surgeons have gained experience with deformity correction. Still, the paucity of cases at any one institution limits the ability to perform clinical studies in a prospective and randomized fashion. To the authors' knowledge, the use of the AxiaLif bolt in a modified Bohlman technique has not been described. In the setting of a case of symptomatic high-grade spondylolisthesis refractory to nonoperative management, the authors describe a modified Bohlman technique in which they used the AxiaLif bolt rather than the fibula graft that was originally described. They then supplemented this with pedicle screw instrumentation and an iliac crest autograft. At the 2-year follow-up exam, the patient exhibited relief of his preoperative back and leg pain and he had returned to all activities. The latest radiographs demonstrated successful fusion. A single-stage, posterior instrumented fusion in which the AxiaLif bolt is used in lieu of fibula autograft or allograft in a modified Bohlman technique is technically less demanding, does not have the morbidity associated with harvesting a fibula autograft, and carries no risk of disease transmission associated with the use of allograft.


The burden of Alzheimer's disease and other types of dementia in the Middle East and North Africa region, 1990-2019.
Alzheimer's disease (AD) is the most common cause of dementia and this progressive neurological disorder is associated with substantial mortality and morbidity. We aimed to report the burden of AD and other types of dementia in the Middle East and North Africa (MENA) region, by age, sex and sociodemographic index (SDI), for the period 1990-2019. publicly accessible data on the prevalence, death and disability-adjusted life years (DALYs) because of AD, and other types of dementia, were retrieved from the global burden of disease 2019 project for all MENA countries from 1990 to 2019. in 2019, the age-standardised point prevalence of dementia was 777.6 per 100,000 populations in MENA, which was 3.0% higher than in 1990. The age-standardised death and DALY rates of dementia were 25.5 and 387.0 per 100,000, respectively. In 2019, the highest DALY rate was observed in Afghanistan and the lowest rate was in Egypt. That same year, the age-standardised point prevalence, death and DALY rates increased with advancing age and were higher for females of all age groups. From 1990 to 2019, the DALY rate of dementia decreased with increasing SDI up to 0.4, then slightly increased up to an SDI of 0.75, followed by a decrease for the remaining SDI levels. the point prevalence of AD and other types of dementia has increased over the past three decades, and in 2019, the corresponding regional burden was higher than the global average.


Hydrogen sulfide ameliorates aging-associated changes in the kidney.
Aging is associated with replacement of normal kidney parenchyma by fibrosis. Because hydrogen sulfide (H2S) ameliorates kidney fibrosis in disease models, we examined its status in the aging kidney. In the first study, we examined kidney cortical H2S metabolism and signaling pathways related to synthesis of proteins including matrix proteins in young and old male C57BL/6 mice. In old mice, increase in renal cortical content of matrix protein involved in fibrosis was associated with decreased H2S generation and AMPK activity, and activation of insulin receptor (IR)/IRS-2-Akt-mTORC1-mRNA translation signaling axis that can lead to increase in protein synthesis. In the second study, we randomized 18-19 month-old male C57BL/6 mice to receive 30 μmol/L sodium hydrosulfide (NaHS) in drinking water vs. water alone (control) for 5 months. Administration of NaHS increased plasma free sulfide levels. NaHS inhibited the increase in kidney cortical content of matrix proteins involved in fibrosis and ameliorated glomerulosclerosis. NaHS restored AMPK activity and inhibited activation of IR/IRS-2-Akt-mTORC1-mRNA translation axis. NaHS inhibited age-related increase in kidney cortical content of p21, IL-1β, and IL-6, components of the senescence-associated secretory phenotype. NaHS abolished increase in urinary albumin excretion seen in control mice and reduced serum cystatin C levels suggesting improved glomerular clearance function. We conclude that aging-induced changes in the kidney are associated with H2S deficiency. Administration of H2S ameliorates aging-induced kidney changes probably by inhibiting signaling pathways leading to matrix protein synthesis.


Mucin gene expression in normal, preneoplastic, and neoplastic human gastric epithelium.
Mucins synthesized by malignant cells may contribute (via decreased cellular adhesion and immune recognition) to cancer invasion and metastases. Human mucins are derived from a heterogeneous family of genes, labeled MUC1-6. Our aim was to determine the pattern of mucin gene expression in normal, preneoplastic (intestinal metaplasia), and malignant gastric specimens. Probes and antibodies for specific mucin tandem repeat sequences were used for RNA and immunohistochemical analysis. Normal stomach mucosa was characterized by expression of MUC1, MUC5, and MUC6 mRNA and immunoreactive protein, without MUC2, MUC3, and MUC4 gene expression. In contrast, high levels of MUC2 and MUC3 mucin mRNA and immunoreactive protein were found in specimens with intestinal metaplasia. Gastric cancers exhibited markedly altered secretory mucin mRNA levels compared with adjacent normal mucosa, with decreased levels of MUC5 and MUC6 mRNA and increased levels of MUC3 and MUC4 mRNA. Overall, immunoreactive MUC1 mucin was detected in 72% of 33 gastric cancers, and secretory mucin core peptides were expressed in 34% (MUC2), 45% (MUC3), 19% (MUC5), and 57% (MUC6) of these specimens. Coexpression of multiple (three or more) mucin core proteins occurred in 15 of 25 (60%) advanced (stages III and IV) cancers compared with 1 of 8 (12.5%) early (stages I and II) cancers (P < 0.048). We conclude that human gastric epithelium has a unique mucin gene pattern, which becomes markedly altered in preneoplastic and neoplastic specimens. Increased mucin gene heterogeneity in gastric adenocarcinomas is associated with advanced cancer stage.


High-energy femtosecond stretched-pulse fiber laser with a nonlinear optical loop mirror.
A stretched-pulse fiber laser with a nonlinear optical loop mirror (NOLM) that produces 100-fs pulses with 1-nJ energy is demonstrated. These results constitute a 30-fold increase in pulse energy over previously reported femtosecond fiber lasers with a NOLM. Compared with previous stretched-pulse lasers, this laser offers a cleaner spectrum and improved stability, with comparable pulse duration and energy. Implications for the construction of truly environmentally stable lasers are discussed.


From Neuron Biophysics to Orientation Selectivity in Electrically Coupled Networks of Neocortical L2/3 Large Basket Cells.
In the neocortex, inhibitory interneurons of the same subtype are electrically coupled with each other via dendritic gap junctions (GJs). The impact of multiple GJs on the biophysical properties of interneurons and thus on their input processing is unclear. The present experimentally based theoretical study examined GJs in L2/3 large basket cells (L2/3 LBCs) with 3 goals in mind: (1) To evaluate the errors due to GJs in estimating the cable properties of individual L2/3 LBCs and suggest ways to correct these errors when modeling these cells and the networks they form; (2) to bracket the GJ conductance value (0.05-0.25 nS) and membrane resistivity (10 000-40 000 Ω cm(2)) of L2/3 LBCs; these estimates are tightly constrained by in vitro input resistance (131 ± 18.5 MΩ) and the coupling coefficient (1-3.5%) of these cells; and (3) to explore the functional implications of GJs, and show that GJs: (i) dynamically modulate the effective time window for synaptic integration; (ii) improve the axon's capability to encode rapid changes in synaptic inputs; and (iii) reduce the orientation selectivity, linearity index, and phase difference of L2/3 LBCs. Our study provides new insights into the role of GJs and calls for caution when using in vitro measurements for modeling electrically coupled neuronal networks.


Economic feasibility of no-tillage and manure for soil carbon sequestration in corn production in northeastern Kansas.
This study examined the economic potential of no-tillage versus conventional tillage to sequester soil carbon by using two rates of commercial N fertilizer or beef cattle manure for continuous corn (Zea mays L.) production. Yields, input rates, field operations, and prices from an experiment were used to simulate a distribution of net returns for eight production systems. Carbon release values from direct, embodied, and feedstock energies were estimated for each system, and were used with soil carbon sequestration rates from soil tests to determine the amount of net carbon sequestered by each system. The values of carbon credits that provide an incentive for managers to adopt production systems that sequester carbon at greater rates were derived. No-till systems had greater annual soil carbon gains, net carbon gains, and net returns than conventional tillage systems. Systems that used beef cattle manure had greater soil carbon gains and net carbon gains, but lower net returns, than systems that used commercial N fertilizer. Carbon credits would be needed to encourage the use of manure-fertilized cropping systems.


Highly Stretchable, Weavable, and Washable Piezoresistive Microfiber Sensors.
A key challenge in electronic textiles is to develop an intrinsically conductive thread of sufficient robustness and sensitivity. Here, we demonstrate an elastomeric functionalized microfiber sensor suitable for smart textile and wearable electronics. Unlike conventional conductive threads, our microfiber is highly flexible and stretchable up to 120% strain and possesses excellent piezoresistive characteristics. The microfiber is functionalized by enclosing a conductive liquid metallic alloy within the elastomeric microtube. This embodiment allows shape reconfigurability and robustness, while maintaining an excellent electrical conductivity of 3.27 ± 0.08 MS/m. By producing microfibers the size of cotton threads (160 μm in diameter), a plurality of stretchable tubular elastic piezoresistive microfibers may be woven seamlessly into a fabric to determine the force location and directionality. As a proof of concept, the conductive microfibers woven into a fabric glove were used to obtain physiological measurements from the wrist, elbow pit, and less accessible body parts, such as the neck and foot instep. Importantly, the elastomeric layer protects the sensing element from degradation. Experiments showed that our microfibers suffered minimal electrical drift even after repeated stretching and machine washing. These advantages highlight the unique propositions of our wearable electronics for flexible display, electronic textile, soft robotics, and consumer healthcare applications.


Oestrogen receptor in the granulosa cell during postnatal development of the rat ovary.
Numbers of granulosa cells obtained from follicles of immature rats increased from 1.6 X 10(5) cells/ovary on day 8 to 7.1 X 10(6) cells/ovary on day 40 of age, the day of vaginal opening and first pro-oestrus. Very high levels of cytosol oestrogen receptor were found on day 8 (175,000 sites/cell) but by day 19 20,000 sites/cell were found. Nuclear receptor concentrations were highest on day 12 (5400 +/- 1470 (S.D.)sites/cell) and again on day 21 (5400 +/- 2300 sites/cell). After day 21 both cytosol and nuclear oestrogen receptor concentrations fell and remained low until nuclear concentrations rose at day 40. Two consecutive daily injections of FSH/LH (5 i.u.) increased cell number over control in animals killed on day 22, gave no significant alteration in animals killed on day 26 or 28 but decreased numbers in animals aged 32 and 35 days. Only on day 22 was the increase in cell number associated with an increase in nuclear oestrogen receptor concentrations. Indeed on days 32 and 35 increased nuclear receptor concentrations were associated with a decreased cell number.


Influence of polymerisation method and type of fibre on fracture strength of endodontically treated teeth.
The aim of this study was to investigate (i) the effect of direct or indirect polymerisation of adhesive-impregnated ribbon fibre under 4-mm bulk-filled composites on fracture strength; (ii) to compare polyethylene ribbon fibre-reinforcement composites with short fibre-reinforced composite; and (iii) the effectiveness of polyethylene ribbon fibre according to the restorative materials used (low-viscous bulk-fill composite, high-viscous flowable composite or conventional paste composite). Seventy molars were divided into seven groups; (groups 1-2) Ribbond-reinforced Surefil-SDR; (group 3) Ribbond-reinforced G Aenial Flo; (group 4) Ribbond-reinforced G Aenial Posterior; (group 5) short fibre-reinforced composite everX Posterior; (group 6) unfilled cavity; (group 7) intact teeth. Ribbond was adopted to cavity walls by impregnating an adhesive and using a flowable composite. In group 1, adhesive-impregnated Ribbond was polymerised directly using a light-curing-unit, and indirectly in group 2 under 4-mm bulk-filled composite. Direct or indirect polymerisation of adhesive impregnated ribbon fibre under 4-mm bulk-filled composite did not change the fracture strength results. Polyethylene ribbon fibre-reinforced groups (groups 1-4) and short fibre-reinforced composite group (group 5) displayed similar results. Polyethylene ribbon fibre can be used safely under 4-mm bulk-filled composites. Ribbond-reinforced low-viscous bulk-fill, high-viscous flowable, and conventional paste composite exhibited similar fracture strength results.


Carotid-cavernous sinus fistulas and venous thrombosis.
Radiographic signs of cavernous sinus thrombosis were found in eight consecutive patients with an angiographic diagnosis of carotid-cavernous sinus fistula; six were of the dural type and the ninth case was of a shunt from a cerebral hemisphere vascular malformation. Diagnostic features consisted of filling defects within the cavernous sinus and its tributaries, an abnormal shape of the cavernous sinus, an atypical pattern of venous drainage, and venous stasis. Progression of thrombosis was demonstrated in five patients who underwent follow-up angiography. Because of a high incidence of spontaneous resolution, patients with dural-cavernous sinus fistulas who show signs of venous thrombosis at angiography should be followed conservatively.


Inhibition of myostatin signaling through Notch activation following acute resistance exercise.
Myostatin is a TGFβ family member and negative regulator of muscle size. Due to the complexity of the molecular pathway between myostatin mRNA/protein and changes in transcription, it has been difficult to understand whether myostatin plays a role in resistance exercise-induced skeletal muscle hypertrophy. To circumvent this problem, we determined the expression of a unique myostatin target gene, Mighty, following resistance exercise. Mighty mRNA increased by 6 h (82.9 ± 24.21%) and remained high out to 48 h (56.5 ± 19.67%) after resistance exercise. Further examination of the soleus, plantaris and tibialis anterior muscles showed that the change in Mighty mRNA at 6 h correlated with the increase in muscle size associated with this protocol (R(2) = 0.9996). The increase in Mighty mRNA occurred both independent of Smad2 phosphorylation and in spite of an increase in myostatin mRNA (341.8 ± 147.14% at 3 h). The myostatin inhibitor SKI remained unchanged. However, activated Notch, another potential inhibitor of TGFβ signaling, increased immediately following resistance exercise (83 ± 11.2%) and stayed elevated out to 6 h (78 ± 16.6%). Electroportion of the Notch intracellular domain into the tibialis anterior resulted in an increase in Mighty mRNA (63 ± 13.4%) that was equivalent to the canonical Notch target HES-1 (94.4 ± 7.32%). These data suggest that acute resistance exercise decreases myostatin signaling through the activation of the TGFβ inhibitor Notch resulting in a decrease in myostatin transcriptional activity that correlates well with muscle hypertrophy.


Incidence and clinical significance of immunogenicity to infliximab in Crohn's disease: a critical systematic review.
Infliximab (IFX) is a chimeric (mouse/human) anti-TNF-alpha monoclonal antibody approved for the treatment of refractory luminal and fistulizing Crohn's disease (CD). It is a source of potential immunogenicity for humans, with the occurrence of anti-infliximab antibodies (ATIs), which are thought to interfere with the pharmacodynamics and/or pharmacokinetics of the compound. It remains unclear whether ATIs have any clinical importance for drug efficacy or safety. We review studies specifically evaluating the incidence of ATIs in CD and their impact on the efficacy and safety of IFX. A systematic review was undertaken by electronic searches of the PubMed and SCOPUS databases from earliest records to October 2008, as well as reference lists of all relevant articles and relevant abstracts from meetings. The biological and clinical mechanisms of ATI development are poorly understood. The incidence of ATIs in vivo depends on multiple analytical and clinical factors, both patient- and treatment-related. The presence of ATIs is weakly and variably associated with clinical response or infusion reactions, but not with reactions relevant to clinical decision-making. Enormous variation in the methods of reporting ATIs and immunogenicity of IFX make almost any interpretation possible from different studies, but few have clinical relevance. There is no clear evidence that ATIs have an impact on efficacy or safety, nor a need to measure or prevent them in clinical practice. Circulating drug concentration may be a more relevant measure of immunogenicity.


Self-assembly of inverse patchy colloids with tunable patch coverage.
We report a simple and scalable technique for the preparation of patchy particles with tunable patch coverage. These particles are categorized as inverse patchy colloidal particles (IPCs) as the patches repel each other while patch and non-patch surfaces attract. We demonstrate the effect of patch coverage, concentration of electrolyte and concentration of particles on the self-assembly of IPC particles. The study identifies various clustering zones such as (1) finite-sized clusters, (2) chain-like assemblies and (3) irregular amorphous aggregates. The linear assemblies are observed over a wide range of particle concentrations and salt concentrations. The anisotropic electrostatic interaction controls the formation of chain-like assemblies. In an extended study, we use negatively charged isotropic colloidal (NCIC) particles to tune the self-assembly of IPC particles. Interestingly, we observe significant improvement in the clustering efficiency of IPC particles leading to the formation of co-polymeric, flexible branched chains. Depending on the number ratio of NCIC particles with respect to IPC particles, the clustering process is classified into three different phases such as (1) finite-sized, (2) linear and (3) dispersed state. Using a quantitative analysis we show that such evolution of structures is attributed to seeding and crowding effects caused by the addition of NCIC particles. The use of NCIC particles thus control the self-assembly of inverse patchy colloids and tune the number and shape of the self-assembled structures.


Verbal abilities in children of mothers with epilepsy: Association to maternal folate status.
To examine the effect of maternal folic acid supplementation and maternal plasma folate and antiepileptic drug (AED) concentrations on language delay in AED-exposed children of mothers with epilepsy. Children of mothers with and without epilepsy enrolled from 1999 to 2008 in the Norwegian Mother and Child Cohort study were included. Information on medical history, AED use, and folic acid supplementation during pregnancy was collected from parent-completed questionnaires. Maternal plasma folate and maternal plasma and umbilical cord AED concentrations were measured in blood samples from gestational weeks 17 to 19 and immediately after birth, respectively. Language development at 18 and 36 months was evaluated by the Ages and Stages Questionnaires. A total of 335 AED-exposed children of mothers with epilepsy and 104,222 children of mothers without epilepsy were surveyed. For those with no maternal periconceptional folic acid supplementation, the fully adjusted odds ratio (OR) for language delay in AED-exposed children compared to the controls at 18 months was 3.9 (95% confidence interval [CI] 1.9-7.8, p < 0.001) and at 36 months was 4.7 (95% CI 2.0-10.6, p < 0.001). When folic supplementation was used, the corresponding ORs for language delay were 1.7 (95% CI 1.2-2.6, p = 0.01) and 1.7 (95% CI 0.9-3.2, p = 0.13), respectively. The positive effect of folic acid supplement use on language delay in AED-exposed children was significant only when supplement was used in the period from 4 weeks before the pregnancy and until the end of the first trimester. Folic acid use early in pregnancy may have a preventive effect on language delay associated with in utero AED exposure.


Acute appendicitis during pregnancy: case series of 20 pregnant women.
Acute appendicitis (AA) is the most common cause of acute abdomen during pregnancy. Most of the signs of appendicitis are also found during normal pregnancy period, however, and diagnosis of appendicitis during pregnancy remains challenging. The aim of the current study was to report our clinical experience of AA during pregnancy and investigate optimal management of this difficult situation. Records of 20 pregnant women with diagnosis of AA who underwent appendectomy between 2005 and 2015 were included in this study. Data were collected retrospectively. Patients were evaluated according to age, signs and symptoms, gestational age, physical findings, serum white blood cell count, ultrasound (US) findings, pathology reports, surgical technique, operation time, and complications. Of 20 patients, 16 (80%) underwent open appendectomy and 4 (20%) underwent laparoscopic appendectomy. Mean age of patients was 29.6±5.6 years. Most common symptom was abdominal pain (95%). Six (30%) patients were in first trimester, 9 (45%) patients were in second trimester and 5 (25%) patients in were in third trimester. US findings consistent with AA were found in 12 (60%) patients. Negative appendectomy rate was 30%. Maternal complication was seen in only 1 (5%) patient. No fetal complication was observed. Accurate diagnosis and prompt surgical treatment of AA in pregnant women should be performed due to high rates of maternal and fetal complications.


Understanding trends in C-H bond activation in heterogeneous catalysis.
While the search for catalysts capable of directly converting methane to higher value commodity chemicals and liquid fuels has been active for over a century, a viable industrial process for selective methane activation has yet to be developed. Electronic structure calculations are playing an increasingly relevant role in this search, but large-scale materials screening efforts are hindered by computationally expensive transition state barrier calculations. The purpose of the present letter is twofold. First, we show that, for the wide range of catalysts that proceed via a radical intermediate, a unifying framework for predicting C-H activation barriers using a single universal descriptor can be established. Second, we combine this scaling approach with a thermodynamic analysis of active site formation to provide a map of methane activation rates. Our model successfully rationalizes the available empirical data and lays the foundation for future catalyst design strategies that transcend different catalyst classes.


Mammalian cell transformation by a murine retrovirus vector containing the avian erythroblastosis virus erbB gene.
A recombinant murine retrovirus vector containing the v-erbB gene of avian erythroblastosis virus was constructed to investigate v-erbB as a transforming gene for mammalian cells. A restriction fragment containing the v-erbB sequences from a molecular clone of avian erythroblastosis virus was inserted into a Moloney murine leukemia virus vector. The construct, designated MuLV/erbB, transformed NIH 3T3 cells at a high efficiency in the DNA transfection assay. Individual MuLV/erbB transfectants grew in soft agar and were tumorigenic. The transfectants contained v-erbB DNA sequences, expressed v-erbB-specific transcripts, and synthesized v-erbB-related glycoproteins. The majority of transfectants produced two major v-erbB gene products of 58 and 66 kilodaltons. However, some transfectants produced much smaller v-erbB-specific proteins. Tunicamycin experiments revealed that the size heterogeneity observed between different transfectants was not due to variations in glycoprotein processing, implying that, in some cases, alterations in the MuLV/erbB genome occurred during the transfection process. These findings indicate that expression of the complete v-erbB gene product is not required for transformation of NIH 3T3 cells. A transmissible murine v-erbB (M-erbB) virus was generated by infection of nonproducer transfectants with amphotrophic murine leukemia virus. Transmission of the rescued M-erbB virus was confirmed by DNA, RNA, and protein analyses. The introduction of a transforming v-erbB gene into mammalian cells by virus infection provides a means of analyzing the mechanism by which this epidermal growth factor receptor-related gene alters the growth and differentiation of cells from various lineages.


Genetic Disruption of the Multifunctional CD98/LAT1 Complex Demonstrates the Key Role of Essential Amino Acid Transport in the Control of mTORC1 and Tumor Growth.
The CD98/LAT1 complex is overexpressed in aggressive human cancers and is thereby described as a potential therapeutic target. This complex promotes tumorigenesis with CD98 (4F2hc) engaging β-integrin signaling while LAT1 (SLC7A5) imports essential amino acids (EAA) and promotes mTORC1 activity. However, it is unclear as to which member of the heterodimer carries the most prevalent protumoral action. To answer this question, we explored the tumoral potential of each member by gene disruption of CD98, LAT1, or both and by inhibition of LAT1 with the selective inhibitor (JPH203) in six human cancer cell lines from colon, lung, and kidney. Each knockout respectively ablated 90% (CD98 KO: ) and 100% (LAT1 KO: ) of Na(+)-independent leucine transport activity. LAT1 KO: or JPH203-treated cells presented an amino acid stress response with ATF4, GCN2 activation, mTORC1 inhibition, and severe in vitro and in vivo tumor growth arrest. We show that this severe growth phenotype is independent of the level of expression of CD98 in the six tumor cell lines. Surprisingly, CD98 KO: cells with only 10% EAA transport activity displayed a normal growth phenotype, with mTORC1 activity and tumor growth rate undistinguishable from wild-type cells. However, CD98 KO: cells became extremely sensitive to inhibition or genetic disruption of LAT1 (CD98 KO: /LAT1 KO: ). This finding demonstrates that the tumoral potential of CD98 KO: cells is due to residual LAT1 transport activity. Therefore, these findings clearly establish that LAT1 transport activity is the key growth-limiting step of the heterodimer and advocate the pharmacology development of LAT1 transporter inhibitors as a very promising anticancer target. Cancer Res; 76(15); 4481-92. ©2016 AACR.


A meta-analysis of research on sensory integration treatment.
The purpose of this study was to find whether existing studies of treatment using sensory integration approaches support the efficacy of these approaches. With meta-analysis, the results of sensory integration efficacy research studies published from 1972 to the present were synthesized and analyzed. Sixteen studies were used to compare sensory integration effect with no treatment (SI/NT), and 16 were used to compare sensory integration effect with alternative treatments (SI/ALT). Overall average effect sizes, comparisons of the effect sizes for different dependent variables, and secondary factors associated with effect size variation were examined. The weighted average effect size of SI/NT studies was .29. However, there was a significant difference between the average effect sizes of the earlier studies (.60) and the more recent studies (.03). Of the outcome measures, larger effect sizes were found in the psychoeducational category (.39) and motor category (.40). Of SI/ALT studies, the average effect size was .09, not significantly different from zero. Three central conclusions can be made. First, in the SI/NT comparison, a significant effect was replicated for sensory integration treatment effects in earlier studies, but more recent studies did not show overall positive effects. Second, larger effect sizes were found in psychoeducational and motor categories. Third, sensory integration treatment methods were found to be as effective as various alternative treatment methods.


Assessing the Genotypic Differences between Strains of Corynebacterium pseudotuberculosis biovar equi through Comparative Genomics.
Seven genomes of Corynebacterium pseudotuberculosis biovar equi were sequenced on the Ion Torrent PGM platform, generating high-quality scaffolds over 2.35 Mbp. This bacterium is the causative agent of disease known as "pigeon fever" which commonly affects horses worldwide. The pangenome of biovar equi was calculated and two phylogenomic approaches were used to identify clustering patterns within Corynebacterium genus. Furthermore, other comparative analyses were performed including the prediction of genomic islands and prophages, and SNP-based phylogeny. In the phylogenomic tree, C. pseudotuberculosis was divided into two distinct clades, one formed by nitrate non-reducing species (biovar ovis) and another formed by nitrate-reducing species (biovar equi). In the latter group, the strains isolated from California were more related to each other, while the strains CIP 52.97 and 1/06-A formed the outermost clade of the biovar equi. A total of 1,355 core genes were identified, corresponding to 42.5% of the pangenome. This pangenome has one of the smallest core genomes described in the literature, suggesting a high genetic variability of biovar equi of C. pseudotuberculosis. The analysis of the similarity between the resistance islands identified a higher proximity between the strains that caused more severe infectious conditions (infection in the internal organs). Pathogenicity islands were largely conserved between strains. Several genes that modulate the pathogenicity of C. pseudotuberculosis were described including peptidases, recombination enzymes, micoside synthesis enzymes, bacteriocins with antimicrobial activity and several others. Finally, no genotypic differences were observed between the strains that caused the three different types of infection (external abscess formation, infection with abscess formation in the internal organs, and ulcerative lymphangitis). Instead, it was noted that there is a higher phenetic correlation between strains isolated at California compared to the other strains. Additionally, high variability of resistance islands suggests gene acquisition through several events of horizontal gene transfer.


Continent-wide risk assessment for the establishment of nonindigenous species in Antarctica.
Invasive alien species are among the primary causes of biodiversity change globally, with the risks thereof broadly understood for most regions of the world. They are similarly thought to be among the most significant conservation threats to Antarctica, especially as climate change proceeds in the region. However, no comprehensive, continent-wide evaluation of the risks to Antarctica posed by such species has been undertaken. Here we do so by sampling, identifying, and mapping the vascular plant propagules carried by all categories of visitors to Antarctica during the International Polar Year's first season (2007-2008) and assessing propagule establishment likelihood based on their identity and origins and on spatial variation in Antarctica's climate. For an evaluation of the situation in 2100, we use modeled climates based on the Intergovernmental Panel on Climate Change's Special Report on Emissions Scenarios Scenario A1B [Nakićenović N, Swart R, eds (2000) Special Report on Emissions Scenarios: A Special Report of Working Group III of the Intergovernmental Panel on Climate Change (Cambridge University Press, Cambridge, UK)]. Visitors carrying seeds average 9.5 seeds per person, although as vectors, scientists carry greater propagule loads than tourists. Annual tourist numbers (∼33,054) are higher than those of scientists (∼7,085), thus tempering these differences in propagule load. Alien species establishment is currently most likely for the Western Antarctic Peninsula. Recent founder populations of several alien species in this area corroborate these findings. With climate change, risks will grow in the Antarctic Peninsula, Ross Sea, and East Antarctic coastal regions. Our evidence-based assessment demonstrates which parts of Antarctica are at growing risk from alien species that may become invasive and provides the means to mitigate this threat now and into the future as the continent's climate changes.


Huntington's disease blood and brain show a common gene expression pattern and share an immune signature with Alzheimer's disease.
There is widespread transcriptional dysregulation in Huntington's disease (HD) brain, but analysis is inevitably limited by advanced disease and postmortem changes. However, mutant HTT is ubiquitously expressed and acts systemically, meaning blood, which is readily available and contains cells that are dysfunctional in HD, could act as a surrogate for brain tissue. We conducted an RNA-Seq transcriptomic analysis using whole blood from two HD cohorts, and performed gene set enrichment analysis using public databases and weighted correlation network analysis modules from HD and control brain datasets. We identified dysregulated gene sets in blood that replicated in the independent cohorts, correlated with disease severity, corresponded to the most significantly dysregulated modules in the HD caudate, the most prominently affected brain region, and significantly overlapped with the transcriptional signature of HD myeloid cells. High-throughput sequencing technologies and use of gene sets likely surmounted the limitations of previously inconsistent HD blood expression studies. Our results suggest transcription is disrupted in peripheral cells in HD through mechanisms that parallel those in brain. Immune upregulation in HD overlapped with Alzheimer's disease, suggesting a common pathogenic mechanism involving macrophage phagocytosis and microglial synaptic pruning, and raises the potential for shared therapeutic approaches.


Rare case of Killian-Pallister syndrome associated with idiopathic short stature detected with fluorescent in situ hybridization on buccal smear.
Killian-Pallister syndrome (KPS) is a rare form of chromosomal mosaicism and is defined by the existence of an extra chromosome 12 in some cell lines in one individual. The degree of mosaicism varies among tissues and dictates the clinical presentation of the syndrome. The clinical features of Killian-Pallister syndrome include mental retardation, typical facial dysmorphism and pigmentation defects. We present a rare case of Killian-Pallister syndrome with severe form of the disease associated with isolated growth hormone deficiency and low-rate mosaicism on buccal smear. The absence of a marker chromosome 12p in lymphocyte cultures and the low degree of mosaicism lead to frequent misdiagnosis of this condition. The selection of tissue sampling is crucial in establishing the diagnosis of Killian-Pallister syndrome. Fluorescent in situ hybridisation on buccal smear remains the golden standard as a screening method if a suspicion of the syndrome exists.


Green tea polyphenol EGCG suppresses cigarette smoke condensate-induced NF-kappaB activation in normal human bronchial epithelial cells.
Cigarette smoke is a powerful inducer of inflammatory responses resulting in disruption of major cellular pathways with transcriptional and genomic alterations driving the cells towards carcinogenesis. Cell culture and animal model studies indicate that (-)-epigallocatechin-3-gallate (EGCG), the major polyphenol present in green tea, possesses potent anti-inflammatory and antiproliferative activity capable of selectively inhibiting cell growth and inducing apoptosis in cancer cells without adversely affecting normal cells. Here, we demonstrate that EGCG pretreatment (20-80 microM) of normal human bronchial epithelial cells (NHBE) resulted in significant inhibition of cigarette smoke condensate (CSC)-induced cell proliferation. Nuclear factor-kappaB (NF-kappaB) controls the transcription of genes involved in immune and inflammatory responses. In most cells, NF-kappaB prevents apoptosis by mediating cell survival signals. Pretreatment of NHBE cells with EGCG suppressed CSC-induced phosphorylation of IkappaBalpha, and activation and nuclear translocation of NF-kappaB/p65. NHBE cells transfected with a luciferase reporter plasmid containing an NF-kappaB-inducible promoter sequence showed an increased reporter activity after CSC exposure that was specifically inhibited by EGCG pretreatment. Immunoblot analysis showed that pretreatment of NHBE cells with EGCG resulted in a significant downregulation of NF-kappaB-regulated proteins cyclin D1, MMP-9, IL-8 and iNOS. EGCG pretreatment further inhibited CSC-induced phosphorylation of ERK1/2, JNK and p38 MAPKs and resulted in a decreased expression of PI3K, AKT and mTOR signaling molecules. Taken together, our data indicate that EGCG can suppress NF-kappaB activation as well as other pro-survival pathways such as PI3K/AKT/mTOR and MAPKs in NHBE cells, which may contribute to its ability to suppress inflammation, proliferation and angiogenesis induced by cigarette smoke.


Mesenchymal stem cells do not exert direct beneficial effects on CNS remyelination in the absence of the peripheral immune system.
Remyelination is the natural repair mechanism in demyelinating disorders such as multiple sclerosis (MS) and it was proposed that it might protect from axonal loss. For unknown reasons, remyelination is often incomplete or fails in MS lesions and therapeutic treatments to enhance remyelination are not available. Recently, the transplantation of exogenous mesenchymal stem cells (MSC) has emerged as a promising tool to enhance repair processes. This included the animal model experimental autoimmune encephalomyelitis (EAE), a commonly used model for the autoimmune mechanisms of MS. However, in EAE it is not clear if the beneficial effect of MSC derives from a direct influence on brain resident cells or if this is an indirect phenomenon via modulation of the peripheral immune system. The aim of this study was to determine potential regenerative functions of MSC in the toxic cuprizone model of demyelination that allows studying direct effects on de- and remyelination without the influence of the peripheral immune system. MSC from three different species (human, murine, canine) were transplanted either intraventricularly into the cerebrospinal fluid or directly into the lesion of the corpus callosum at two time points: at the onset of oligodendrocyte progenitor cell (OPC) proliferation or the peak of OPC proliferation during cuprizone induced demyelination. Our results show that MSC did not exert any regenerative effects after cuprizone induced demyelination and oligodendrocyte loss. During remyelination, MSC did not influence the dynamics of OPC proliferation and myelin formation. In conclusion, MSC did not exert direct regenerative functions in a mouse model where peripheral immune cells and especially T lymphocytes do not play a role. We thus suggest that the peripheral immune system is required for MSC to exert their effects and this is independent from a direct influence of the central nervous system.


Targeting Nrf2 signaling improves bacterial clearance by alveolar macrophages in patients with COPD and in a mouse model.
Patients with chronic obstructive pulmonary disease (COPD) have innate immune dysfunction in the lung largely due to defective macrophage phagocytosis. This deficiency results in periodic bacterial infections that cause acute exacerbations of COPD, a major source of morbidity and mortality. Recent studies indicate that a decrease in Nrf2 (nuclear erythroid-related factor 2) signaling in patients with COPD may hamper their ability to defend against oxidative stress, although the role of Nrf2 in COPD exacerbations has not been determined. Here, we test whether activation of Nrf2 by the phytochemical sulforaphane restores phagocytosis of clinical isolates of nontypeable Haemophilus influenza (NTHI) and Pseudomonas aeruginosa (PA) by alveolar macrophages from patients with COPD. Sulforaphane treatment restored bacteria recognition and phagocytosis in alveolar macrophages from COPD patients. Furthermore, sulforaphane treatment enhanced pulmonary bacterial clearance by alveolar macrophages and reduced inflammation in wild-type mice but not in Nrf2-deficient mice exposed to cigarette smoke for 6 months. Gene expression and promoter analysis revealed that Nrf2 increased phagocytic ability of macrophages by direct transcriptional up-regulation of the scavenger receptor MARCO. Disruption of Nrf2 or MARCO abrogated sulforaphane-mediated bacterial phagocytosis by COPD alveolar macrophages. Our findings demonstrate the importance of Nrf2 and its downstream target MARCO in improving antibacterial defenses and provide a rationale for targeting this pathway, via pharmacological agents such as sulforaphane, to prevent exacerbations of COPD caused by bacterial infection.


Electric-field-switchable stratified volume holograms in photorefractive polymers.
Two- and four-layer stratified volume holograms have been fabricated from the photorefractive polymeric material poly(methyl methacrylate): (1,3-dimethyl-2,2-tetramethylene-5-nitrobenzimidazoline):C(60) and characterized by holographic four-wave mixing experiments. Coherent addition of diffracted fields from the individual layers is observed, leading to a diffraction efficiency that increases with the square of the active layer thickness. Electricfield switching of the diffraction efficiencies of individual layers is demonstrated. The angular selectivity of the diffraction efficiency is also characterized for one, two, and four active layers. The angular width of the peaks narrows with increasing total structure thickness in agreement with theory.


Monogene and polygene therapy for the treatment of experimental prostate cancers by use of apoptotic genes bax and bad driven by the prostate-specific promoter ARR(2)PB.
We have shown that adenovirus-mediated manipulation of apoptotic genes such as bax could be a therapeutic option for prostate cancer. Unfortunately, the response of experimental prostate tumors to a single therapeutic gene of the apoptotic pathway is short-lived, and most of these tumors relapse after a short period of time. In this investigation we present data generated with adenovirus AvARR(2)PB-Bad, in which the apoptotic gene bad was placed under the control of the dihydrotestosterone (DHT)-inducible third-generation probasin-derived promoter ARR(2)PB. This therapeutic virus was given alone or in combination with other therapeutic viruses to a variety of in vitro and in vivo experimental models of prostate cancer. On infection with AvARR(2)PB-Bad, DHT-induced Bad overexpression occurred specifically in androgen receptor-positive (AR(+)) cells of prostatic derivation. The apoptotic effect of AvARR(2)PB-Bad (group 1) was compared with that of AvARR(2)PB-Bax (which overexpresses the apoptotic protein Bax) (group 2), with that of the combination AvARR(2)PB-Bad plus AvARR(2)PB-Bax (group 3), and with that of the control virus AvARR(2)PB-CAT (group 4) in the cell line LNCaP. In addition to identifying the modality of apoptosis induction by overexpressed Bad, the results suggested that group 3 contained more apoptotic cells than any other group. In additional studies, AR(+) androgen-dependent LNCaP cells or AR(+) and androgen-independent C4-2 cells were injected subcutaneously into nude mice. Four groups of six LNCaP or C4-2 tumors were treated with the same combinations of viruses discussed above for groups 1, 2, 3, and 4. Treatment resulted in decreased tumor size in groups 1, 2, and 3 compared with group 4. There was a better response in group 3 compared with group 2, and in group 2 compared with group 1. A better response in group 3 was confirmed during a 8-week follow-up period, in which no treatment was administered. Two LNCaP and C4-2 tumors of group 3 disappeared at the end of treatment and did not recur after an 8-week follow-up period. The data suggest that polygene therapy with apoptotic molecules is more effective in experimental models of androgen-dependent or -independent prostate cancer than monogene therapy.


Replication regions from plant-pathogenic Pseudomonas syringae plasmids are similar to ColE2-related replicons.
Many strains of the phytopathogen Pseudomonas syringae contain mutually compatible plasmids that share extensive regions of sequence homology and essential replication determinants. The replication regions of two compatible large plasmids involved in virulence or pathogenicity, pPT23A from P. syringae pv. tomato strain PT23 and pAV505 from P. syringae pv. phaseolicola strain HRI1302A, were isolated. DNA sequencing of the origins of replication revealed homologous ORFs, designated ORF-Pto and ORF-Pph, respectively. Both ORFs are 1311 bp long and encode peptides of 437 amino acids with predicted molecular masses of 48259 (Pto) and 48334 (Pph) Da. Expression of the two ORFs in Escherichia coli produced peptides of 50 kDa (Pto) and 56 kDa (Pph). The predicted peptides showed an overall identity of 897 %, being highly conserved from residues 1 to 373, but showing considerable variation in their C-terminal regions (50% identity over the last 64 aa). The two ORFs had significant similarity with the putative replication protein from plasmid pTiK12 of Thiobacillus intermedius and other CoIE2-related plasmids. However, both peptides were 100 residues longer than any of the known CoIE2-related rep sequences. Subcloning of fragments from the replication region of pPT23A revealed the presence of at least three incompatibility determinants, designated IncA, IncB and IncC. Partial sequencing of the region downstream of ORF-Pto revealed homology to the ru/AB genes, involved in UV resistance, from plasmid pPSR1. It is proposed that the replication origin of pPT23A serves as the prototype of a family of related plasmids.


Effects of different surface conditioning methods on the bond strength of composite resin to amalgam.
Repairing amalgam restorations with composite resins using surface conditioning methods is a conservative treatment approach. This study investigated the effects of different conditioning methods that could be used for repair of amalgam fractures. Amalgam (N=96) was condensed into cavities within autopolymerizing polymethylmethacrylate (PMMA), and the exposed surface of each specimen (diameter, 6 mm; thickness, 2 mm) was ground finished. The specimens were randomly divided into nine experimental groups (n=12 per group), depending on the conditioning method used. The control group had natural central incisors with amalgam (n=12). The combination of the following conditioning methods was tested: silicacoating (Sc), sandblasting (Sb), metal primers, coupling agents, fiber (Fb) application, and opaquers (O). Five types of silanes, metal primers, or adhesives (Visiobond [V], Porcelain Photobond [PP], Alloy Primer [AP], Unibond sealer [Us], ESPE-Sil [ES]), and four opaquers, namely, Clearfil St Opaquer (CstO), Sinfony (S), Miris (M), and an experimental Opaquer (EO-Cavex), were used. The groups were as follows: group 1, Sc+ES+S+V; group 2, Sc+ES+CstO+V; group 3, Sc+ES+M+V; group 4, Sc+ES+EO+V; group 5, Sb+AP+S; group 6, Sb+AP+PP+CstO; group 7, Sc+ES+S+Fb+V+Fb; group 8-control, SC+ES+V; and group 9, Etch+Sc+ES+S+Us. One repair composite was used for all groups (Clearfil Photo Bond Posterior, Kuraray, Tokyo, Japan). Shear bond strengths (SBSs) (MPa ± SD) were evaluated after 5 weeks of water storage (analysis of variance [ANOVA], Tukey honestly significant differences [HSD], α=0.05). Group 1 exhibited significantly higher values (35.5 ± 4.1) than were seen in group 4 (19.4 ± 8.9), group 6 (19.1 ± 7.8), and group 8 (20.1 ± 4.1) (p<0.05). Group 9 exhibited significantly lower values (8.3 ± 3.4) than were noted in groups 1 to 3 (35.5 ± 4.1; 27 ± 12.5; 24.4 ± 5.1, respectively) (p<0.05). Group 7 (16.4 ± 5.9) showed significantly lower values than were observed in group 1 (35.5 ± 4.1) (p<0.05). Surface conditioning techniques affected the bond strengths of composite adhesion to amalgam. Experimental opaquer exhibited lower values. Leaving a small border of enamel around the restoration decreased the bond strength.


Technology assessment No. 7: Hysteroscopy.
Hysteroscopy is performed to view and treat pathology within the uterine cavity and endocervix. Diagnostic hysteroscopy allows visualization of the endocervical canal, endometrial cavity, and fallopian tube ostia. Operative hysteroscopy incorporates the use of mechanical, electrosurgical, or laser instruments to treat intracavitary pathology and perform hysteroscopic sterilization procedures. Selection of a distending medium requires consideration of the advantages, disadvantages, and risks associated with various media as well as their compatibility with electrosurgical or laser energy. A preoperative consultation allows the patient and physician to discuss the hysteroscopic procedure, weigh its inherent risks and benefits, review the patient's medical history for any comorbid conditions, and exclude pregnancy. Known pregnancy, genital tract infections, and active herpetic infection are contraindications to hysteroscopy. The most common perioperative complications associated with operative hysteroscopy are hemorrhage, uterine perforation, and cervical laceration. The procedure is minimally invasive and can be used with a high degree of safety.


A high-throughput functional complementation assay for classification of BRCA1 missense variants.
Mutations in BRCA1 and BRCA2 account for the majority of hereditary breast and ovarian cancers, and therefore sequence analysis of both genes is routinely conducted in patients with early-onset breast cancer. Besides mutations that clearly abolish protein function or are known to increase cancer risk, a large number of sequence variants of uncertain significance (VUS) have been identified. Although several functional assays for BRCA1 VUSs have been described, thus far it has not been possible to conduct a high-throughput analysis in the context of the full-length protein. We have developed a relatively fast and easy cDNA-based functional assay to classify BRCA1 VUSs based on their ability to functionally complement BRCA1-deficient mouse embryonic stem cells. Using this assay, we have analyzed 74 unclassified BRCA1 missense mutants for which all predicted pathogenic variants are confined to the BRCA1 RING and BRCT domains. BRCA1 VUSs are frequently found in patients with hereditary breast or ovarian cancer and present a serious problem for clinical geneticists. This article describes the generation, validation, and application of a reliable high-throughput assay for the functional classification of BRCA1 sequence variants of uncertain significance.


A systematic review finds prediction models for chronic kidney disease were poorly reported and often developed using inappropriate methods.
Chronic kidney disease (CKD) is a global health concern that is increasing mainly as the result of increasing incidences of diabetes and hypertension. Furthermore, if left untreated, individuals with CKD may progress to end-stage kidney failure. Identifying individuals with undiagnosed CKD or those who are at an increased risk of developing CKD or progressing to end-stage kidney disease (ESKD) is therefore an important challenge. We sought to systematically review and critically assess the conduct and reporting of methods used to develop risk prediction models for predicting the risk of having undiagnosed (prevalent) or future risk of developing (incident) CKD or end-stage kidney failure in adults. We conducted a systematic search of PubMed database to identify studies published up until September 2011 that describe the development of models combining two or more variables to predict the risk of prevalent or incident CKD or ESKD. We extracted key information that describes aspects of developing a prediction model, including the study design, data quality, sample size and number of events, outcome definition, risk predictor selection and coding, missing data, model-building strategies, and aspects of performance. Eleven studies describing the development of 14 prediction models were included. Eight studies reported the development of 11 models to predict incident CKD or ESKD, whereas 3 studies developed models for prevalent CKD. A total of 97 candidate risk predictors were considered, and 43 different risk predictors featured in the 14 prediction models. A method, not recommended to select risk predictors for inclusion in the multivariate model, using statistical significance from univariate screening was carried out in six studies. Missing data were frequently poorly handled and reported with no mention of missing data in four studies; 4 studies explicitly excluded individuals with missing data, and only 2 studies used multiple imputation to replace missing values. We found that prediction models for chronic kidney were often developed using inappropriate methods and were generally poorly reported. Using poor methods can affect the predictive ability of the models, whereas inadequate reporting hinders an objective evaluation of the potential usefulness of the model.


The role of high-energy phosphate in norepinephrine-induced acute renal failure in the dog.
Previous studies have demonstrated that pretreatment with mannitol, furosemide, or bradykinin can attenuate the severity of norepinephrine-induced renal functional impairment. The present studies were designed to evaluate the possibility that these agents are protective, in part, by preserving cellular metabolic integrity. The renal cortex was repetitively biopsied during the course of this study, and high-pressure liquid chromatography was used to analyze the tissue content of adenine nucleotides (expressed in nanomoles per gram of wet tissue). The adenine nucleotide charge ratio (CR) and total adenine nucleotide (TAN) content were calculated as indices of cellular metabolic integrity. In addition to the above-established protective agents, phenoxybenzamine was used to evaluate a direct toxic effect of norepinephrine on renal tissue. Inulin clearance at 3 hours post infusion (expressed as a percent of control) was 7% with norepinephrine alone and, in the protected groups, 36% with bradykinin, 61% with furosemide, 51% with mannitol, and 100% with phenoxybenzamine. There was no change in CR or TAN with phenoxybenzamine. In contrast, during norepinephrine administration CR fell significantly in all other groups. Three hours after stopping norepinephrine, CR had returned toward control values and the level of CR was significantly better in all protected groups when compared with norepinephrine alone. Similarly, the levels of TAN were significantly diminished in the norepinephrine-alone group when compared to all protected groups, and there was significantly more tubular necrosis as well. The maintenance of higher levels of TAN and the preserved ability to regenerate adenosine triphosphate in the protected groups, when compared to the norepinephrine-alone group, support the contention that these agents offer protection, at least in part, by preserving cellular metabolic integrity.


Distribution and functional significance of somatostatin receptors in malignant melanoma.
Malignant melanoma is a neuroendocrine tumor that contains somatostatin receptors (SSTRs). Adjuvant therapy for melanoma is limited. Because melanomas arise from neural crest cells, we sought to evaluate the distribution of SSTR subtypes found in these tumors and their functional significance by imaging with 111In-pentetreotide scintigraphy (OctreoScan). Octreotide binds with greatest affinity to SSTR2 and SSTR5. Studying the expression of SSTRs in melanoma may demonstrate a potential role for octreotide in the treatment of melanoma. A series of 23 melanomas from 17 patients who underwent resection of regional or distant metastases were evaluated for the presence of SSTRs by the reverse transcriptase-polymerase chain reaction (RT-PCR) using primers specific for SSTR1 through SSTR5. Identity of RT-PCR products was confirmed by Southern blot analysis. Sixteen patients underwent preoperative OctreoScan. SSTR1 was expressed in 96% of tumors, SSTR2 in 83%, SSTR3 in 61%, SSTR4 in 57%, and SSTR5 in 9%. OctreoScan imaged 63% of tumors. There was no correlation between SSTR subtype expression and OctreoScan result. Most of the melanomas expressed mRNA for SSTR1 and SSTR2, with approximately half expressing SSTR3 and SSTR4. The SSTR mRNA for SSTR2 appears to be transcribed into functional protein that binds 111In-pentetreotide in more than half of these patients. Although OctreoScan has limited sensitivity for localizing melanomas, tumors that can be imaged by OctreoScan may be amenable to adjuvant therapy with octreotide or targeted therapy with high-energy radioisotope-labeled octreotide. These studies clearly define melanoma as a neuroendocrine tumor, which may open new avenues for tumor control.


10 years outcome from childhood traumatic brain injury.
Childhood traumatic brain injury (TBI) is a common, acquired disability, which has significant implications for subsequent development, and for later quality of life. To date few studies have documented outcomes in these children into adolescence, when academic, social and personal demands increase. The objective of this study was to document functional outcomes at 10 years post-injury, and to identify predictors of outcome including injury, socio-demographic and pre-injury characteristics. The study employed consecutive recruitment to a prospective, longitudinal study. Children with a diagnosis of TBI between 2 and 12 years were initially recruited and divided according to injury severity (mild, moderate, severe). The sample was reviewed at 10 years post-injury and intellectual, adaptive, executive and social domains were investigated. Results indicated that, at 10 years post child TBI, survivors' functional abilities fell overall within the low average to average range regardless of injury severity, suggesting no extreme impairments at a group level for any of the domains under investigation. Significant group differences were identified, though, for adaptive abilities and for speed of processing, with more severe injury associated with poorer performances in each instance. Further, a similar trend was identified for measures of intellectual ability and executive functions. Individual rates of impairment were considerably higher than population expectations across all severity groups for these domains. Although rates of social impairment were also elevated, they were less related to injury factors, suggesting that cognitive/adaptive outcomes and social consequences of TBI may have different bases.


Potential impact of exclusion criteria on results of hypertension trials.
Recent trials of antihypertensive therapy, including the Veterans Administration trials, the Hypertension Detection and Follow-up Program, the Multiple Risk Factor Intervention Trial, the Australian Mild Hypertension Trial, and the British Medical Research Council Trial, are reviewed with a particular emphasis on the criteria leading to the exclusion of potentially eligible participants. The observation of all-cause and cause-specific mortality rates in the group ultimately selected to participate in the trial is suggested as an index to the general applicability of trial results. Because end-point rates are fundamental for determining sample size, substantial reduction in these end-point rates by patient exclusion should be taken into account by the trial design. Some recent trials may have generated end-point-event rates so low that the power of the trial to detect reasonable treatment effects was substantially reduced. Future trials should attempt to take this important factor into account at the design stage.


Susceptibility and aversion of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Cry1F Bt maize and considerations for insect resistance management.
Bacillus thuringiensis (Bt) maize was developed primarily for North American pests such as European corn borer (Ostrinia nubilalis (Hübner)). However, most Bt maize products are also cultivated outside of North America, where the primary pests may be different and may have lower susceptibility to Bt toxins. Fall armyworm (Spodoptera frugiperda JE Smith) is an important pest and primary target of Bt maize in Central and South America. S. frugiperda susceptibility to Cry1F (expressed in event TC1507) is an example of a pest-by-toxin interaction that does not meet the high-dose definition. In this study, the behavioral and toxic response of S. frugiperda to Cry1F maize was investigated by measuring the percentage of time naive third instars spent feeding during a 3-min exposure. S. frugiperda also were exposed as third instars to Cry1F maize for 14 d to measure weight gain and survival. S. frugiperda demonstrated an initial, postingestive aversive response to Cry1F maize, and few larvae survived the 14 d exposure. The role of susceptibility and avoidance are discussed in the context of global IRM refuge strategy development for Bt products.


Bond characteristics of porcelain fused to milled titanium.
A bonding agent has recently been introduced that prepares the surface of milled titanium copings for bonding to low fusing dental porcelains. This study evaluated the effectiveness of the bonding agent by comparing the shear and three-point bending strength of specimens made with three combinations of materials: 1) milled titanium/porcelain with bonding agent, 2) the same milled titanium/porcelain without bonding agent, and 3) cast high palladium/conventional porcelain. Shear specimens consisting of porcelain cylinders 6 mm in diameter and 8 mm in length were fired to the ends of metal cylinders 6 mm in diameter and 13 mm in length. Three-point bend specimens made of 25 x 3 x 0.5 mm metal bars were veneered with 8 x 3 x 1 mm of porcelain in the bar's center. Specimens were tested in shear and bending in a universal testing machine. The data were analyzed using one-way ANOVA and Newman-Kuels post hoc tests (p = 0.05). When a titanium bonding agent was used, porcelain to titanium bond strength was slightly but statistically significantly greater than the porcelain to high palladium bond strength. The result was the same when measured by both shear and three-point bending tests. Without the bonding agent, the shear strength of porcelain to titanium was significantly lower than that of the bonding agent and high palladium groups. The use of a bonding agent improves the bond strength of porcelain-fused-to-milled titanium.