17 beta-Estradiol augments endothelium-dependent contractions to arachidonic acid in rabbit aorta.
Experiments were designed to determine the effects of chronic estrogen treatment on endothelium-dependent responses to arachidonic acid in the aorta of the rabbit. Ovariectomized rabbits were treated with either placebo or 17 beta-estradiol for 14 days. The aortas were removed, and rings cut from each blood vessel were suspended for measurement of isometric force. In rings contracted with a submaximal concentration of norepinephrine, endothelium-dependent contractions to arachidonic acid were augmented by estrogen treatment. Prostaglandin F2 alpha, E2, the thromboxane analogue U 46619, and prostacyclin contracted aortic smooth muscle. Only the contractions to prostacyclin were augmented by estrogen treatment. Estrogen treatment enhanced the contractions to norepinephrine in aortas with, but not in those without, endothelium; these contractions were not affected by inhibition of uptake of the neurotransmitter. Indomethacin inhibited contractions to norepinephrine in rings with endothelium from estrogen-treated rabbits. These results indicate that chronic treatment with estrogens can affect arterial reactivity to norepinephrine through an endothelium-dependent mechanism that may involve the metabolism of arachidonic acid by cyclooxygenase. The altered sensitivity of the smooth muscle to prostacyclin may contribute in part to the augmented contractions to arachidonic acid with estrogen treatment.


In Vivo Inhibition of Proteasome Activity and Tumour Growth by Murraya koenigii Leaf Extract in Breast Cancer Xenografts and by Its Active Flavonoids in Breast Cancer Cells.
Inhibition of the 26S proteasome is an attractive approach for anticancer therapy. Proteasome inhibitors are known to selectively target cancer cells and make them more sensitive to chemotherapeutic agents. Murraya koenigii is a medicinally important herb of Asian origin and a rich source of bioactive compounds such as flavonoids and alkaloids. In the present study, we investigated the proteasome inhibitory and apoptotic effect of M. koenigii leaf extract in vivo in a xenograft tumor mouse model, and also assessed the toxicity if any in normal mice. M. koenigii extract did not lead to any toxicity in mice. Analysis of extract revealed the presence of flavonoid compounds which act as proteasome inhibitors. Quercetin treatment led to the decrease in the cell viability and arrest of cells in G2/M phase. Quercetin, Apigenin, Kaempferol and Rutin; flavonoids present in the leaf extract, dose-dependently inhibited the endogenous 26S proteasome activity in MDA-MB-231 cells. Reduction in tumor growth was associated with a decrease in proteasomal enzyme activities in the treated groups. Increased caspase-3 activity and TUNEL-positive cells indicated enhanced apoptosis with Murraya leaf extract treatment. Decreased expression of angiogenic and anti-apoptotic gene markers is indicative of inhibition of angiogenesis and promotion of apoptosis in the leaf extract treated tumors.


beta-cell transcription factors and diabetes: no evidence for diabetes-associated mutations in the gene encoding the basic helix-loop-helix transcription factor neurogenic differentiation 4 (NEUROD4) in Japanese patients with MODY.
The basic helix-loop-helix (bHLH) family of transcription factors plays an important role in the normal development and function of the endocrine pancreas. Heterozygous mutations in the gene encoding one member of this family, NeuroD1/BETA2, are associated with a monogenic form of diabetes that resembles maturity-onset diabetes of the young (MODY) in many respects. This result prompted us to screen the genes encoding related bHLH transcription factors that are also expressed in pancreatic islets for diabetes-associated mutations. We have screened 57 unrelated Japanese subjects with a clinical diagnosis of MODY for mutations in the NeuroD4/Math-3/ATH-3 gene (NEUROD4). This analysis revealed seven frequent polymorphisms that were not associated with MODY, including five in the 5'-untranslated region (UTR) (-477G/A, -436delA, -324delT, -107insTTTT, and -104T/C [cDNA sequences]) and two in the 3'-UTR (1027C/T and 1076C/A). A missense mutation, K68T (203A/C), was found in a heterozygous state in one MODY subject and two nondiabetic subjects. The results of our study suggest that genetic variation in NEUROD4 is not a common cause of MODY in Japanese.


Multiple sites and actions of gabapentin-induced relief of ongoing experimental neuropathic pain.
Gabapentin (GBP) is a first-line therapy for neuropathic pain, but its mechanisms and sites of action remain uncertain. We investigated GBP-induced modulation of neuropathic pain following spinal nerve ligation (SNL) in rats. Intravenous or intrathecal GBP reversed evoked mechanical hypersensitivity and produced conditioned place preference (CPP) and dopamine (DA) release in the nucleus accumbens (NAc) selectively in SNL rats. Spinal GBP also significantly inhibited dorsal horn wide-dynamic-range neuronal responses to a range of evoked stimuli in SNL rats. By contrast, GBP microinjected bilaterally into the rostral anterior cingulate cortex (rACC), produced CPP, and elicited NAc DA release selectively in SNL rats but did not reverse tactile allodynia and had marginal effects on wide-dynamic-range neuronal activity. Moreover, blockade of endogenous opioid signaling in the rACC prevented intravenous GBP-induced CPP and NAc DA release but failed to block its inhibition of tactile allodynia. Gabapentin, therefore, can potentially act to produce its pain relieving effects by (a) inhibition of injury-induced spinal neuronal excitability, evoked hypersensitivity, and ongoing pain and (b) selective supraspinal modulation of affective qualities of pain, without alteration of reflexive behaviors. Consistent with previous findings of pain relief from nonopioid analgesics, GBP requires engagement of rACC endogenous opioid circuits and downstream activation of mesolimbic reward circuits reflected in learned pain-motivated behaviors. These findings support the partial separation of sensory and affective dimensions of pain in this experimental model and suggest that modulation of affective-motivational qualities of pain may be the preferential mechanism of GBP's analgesic effects in patients.


Angiotensin-converting enzyme inhibition studies by natural leech inhibitors by capillary electrophoresis and competition assay.
A protocol to follow the processing of angiotensin I into angiotensin II by rabbit angiotensin-converting enzyme (ACE) and its inhibition by a novel natural antagonist, the leech osmoregulator factor (LORF) using capillary zonal electrophoresis is described. The experiment was carried out using the Beckman PACE system and steps were taken to determine (a) the migration profiles of angiotensin and its yielded peptides, (b) the minimal amount of angiotensin II detected, (c) the use of different electrolytes and (d) the concentration of inhibitor. We demonstrated that LORF (IPEPYVWD), a neuropeptide previously found in leech brain, is able to inhibit rabbit ACE with an IC(50) of 19.8 micro m. Interestingly, its cleavage product, IPEP exhibits an IC(50) of 11.5 micro m. A competition assay using p-benzoylglycylglycylglycine and insect ACE established that LORF and IPEP fragments are natural inhibitors for invertebrate ACE. Fifty-four percent of insect ACE activity is inhibited with 50 micro m IPEP and 35% inhibition with LORF (25 mm). Extending the peptide at both N- and C-terminus (GWEIPEPYVWDES) and the cleavage of IPEP in IP abolished the inhibitory activity of both peptides. Immunocytochemical data obtained with antisera raised against LORF and leech ACE showed a colocalization between the enzyme and its inhibitor in the same neurons. These results showed that capillary zonal electrophoresis is a useful technique for following enzymatic processes with small amounts of products and constitutes the first evidence of a natural ACE inhibitor in invertebrates.


Rugby World Cup 2011: International Rugby Board injury surveillance study.
To determine the frequency and nature of injuries sustained during the IRB 2011 Rugby World Cup. A prospective, whole population survey. 615 international rugby players representing 20 teams competing at the IRB 2011 Rugby World Cup in New Zealand. The study was implemented according to the international consensus statement for epidemiological studies in rugby union; the main measures included the players' age (years), stature (cm) and body mass (Kg) and the incidence (number of injuries/1000 player-hours), mean and median severity (days absence), location (%), type (%) and cause (%) of match and training injuries. The incidences of injuries were 89.1/1000 player-match-hours (forwards: 85.0; backs: 93.8) and 2.2/1000 player-training-hours (forwards: 2.7; backs: 1.7). The mean severity of injuries was 23.6 days (forwards: 21.2; backs: 26.2) during matches and 26.9 (forwards: 33.4; backs: 14.3) during training. During matches, lower-limb muscle/tendon (31.6%) and ligament (15.8%) and, during training, lower-limb muscle/tendon (51.4%) and trunk muscle/tendon (11.4%) injuries were the most common injuries. The most common cause of injury during matches was the tackle (forwards: 43.6%, backs: 45.2%), and during training was full and semicontact skills activities. The results confirm that rugby, like other full-contact sports, has a high incidence of injury: the results from IRB Rugby World Cup (RWC) 2011 were similar to those reported for RWC 2007.


Medication Management Models for Polymedicated Home-Dwelling Older Adults With Multiple Chronic Conditions: Protocol of a Systematic Review.
Older adults with multiple chronic diseases commonly require complex medication regimes. When combined with frailty, cognitive impairment, and changing pharmacological prescriptions, older adults' polymedication regimes increase the risk of medication-related problems (MRPs) and hospitalization. Effective, well-organized medication management could avoid MRPs and their clinical outcomes. Identify medication management models and analyze their impact on managing and preventing MRPs for polymedicated, home-dwelling older adults. We will conduct a systematic review of published articles in relevant professional scientific journals from inception until March 31, 2019, in the following electronic databases,: Embase; Medline OvidSP; PubMed (NOT Medline[sb]); Cumulative Index to Nursing and Allied Health Literature (CINAHL) EBSCO; PsycINFO OvidSP; Cochrane Library, Wiley; and Web of Science. We will also hand search the bibliographies of all the relevant articles found and search for unpublished studies. We will consider publications in English, French, German, Spanish, Italian, and Portuguese. Retrieved articles will be screened for eligibility. Statistical analyses will be conducted following the recommendations of the Cochrane Handbook for Systematic Reviews of Interventions, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), and Meta-analysis Of Observational Studies in Epidemiology (MOOSE) statements. Data will be analyzed using SPSS Statistics for Windows, version 25.0 (IBM Corp), and Review Manager, version 5.5 (The Nordic Cochrane Centre, The Cochrane Collaboration). A preliminary search in Embase delivered 3272 references. This preliminary search allows us to complete our research strategy with equation development and to search the other databases. Relevant articles identified will allow for searching the reference lists for unpublished studies. The inclusion and exclusion criteria will be rigorously respected in the study selection. The entire study is expected to be completed by January 2020. This review will provide an exhaustive view of medication management models that could be effective for polymedicated, home-dwelling older adults and will allow us to analyze their impact on managing and preventing MRPs. PROSPERO CRD42018117287; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=117287 (Archived by WebCite at http://www.webcitation.org/77fCfbCjT). DERR1-10.2196/13582.


Expert Consensus Guidelines for Stocking of Antidotes in Hospitals That Provide Emergency Care.
We provide recommendations for stocking of antidotes used in emergency departments (EDs). An expert panel representing diverse perspectives (clinical pharmacology, medical toxicology, critical care medicine, hematology/oncology, hospital pharmacy, emergency medicine, emergency medical services, pediatric emergency medicine, pediatric critical care medicine, poison centers, hospital administration, and public health) was formed to create recommendations for antidote stocking. Using a standardized summary of the medical literature, the primary reviewer for each antidote proposed guidelines for antidote stocking to the full panel. The panel used a formal iterative process to reach their recommendation for both the quantity of antidote that should be stocked and the acceptable timeframe for its delivery. The panel recommended consideration of 45 antidotes; 44 were recommended for stocking, of which 23 should be immediately available. In most hospitals, this timeframe requires that the antidote be stocked in a location that allows immediate availability. Another 14 antidotes were recommended for availability within 1 hour of the decision to administer, allowing the antidote to be stocked in the hospital pharmacy if the hospital has a mechanism for prompt delivery of antidotes. The panel recommended that each hospital perform a formal antidote hazard vulnerability assessment to determine its specific need for antidote stocking. Antidote administration is an important part of emergency care. These expert recommendations provide a tool for hospitals that offer emergency care to provide appropriate care of poisoned patients.


Minimal homozygous endothelial deletion of Eng with VEGF stimulation is sufficient to cause cerebrovascular dysplasia in the adult mouse.
Brain arteriovenous malformations (bAVMs) represent a high risk for hemorrhagic stroke, leading to significant neurological morbidity and mortality in young adults. The etiopathogenesis of bAVM remains unclear. Research progress has been hampered by the lack of animal models. Hereditary Hemorrhagic Telangiectasia (HHT) patients with haploinsufficiency of endoglin (ENG, HHT1) or activin receptor-like kinase 1 (ALK1, HHT2) have a higher incidence of bAVM than the general population. We previously induced cerebrovascular dysplasia in the adult mouse that resembles human bAVM through Alk1 deletion plus vascular endothelial growth factor (VEGF) stimulation. We hypothesized that Eng deletion plus VEGF stimulation would induce a similar degree of cerebrovascular dysplasia as the Alk1-deleted brain. Ad-Cre (an adenoviral vector expressing Cre recombinase) and AAV-VEGF (an adeno-associated viral vector expressing VEGF) were co-injected into the basal ganglia of 8- to 10-week-old Eng(2f/2f) (exons 5 and 6 flanked by loxP sequences), Alk1(2f/2f) (exons 4-6 flanked by loxP sequences) and wild-type (WT) mice. Vascular density, dysplasia index, and gene deletion efficiency were analyzed 8 weeks later. AAV-VEGF induced a similar degree of angiogenesis in the brain with or without Alk1- or Eng-deletion. Abnormally patterned and dilated dysplastic vessels were found in the viral vector-injected region of Alk1(2f/2f) and Eng(2f/2f) brain sections, but not in WT. Alk1(2f/2f) mice had about 1.8-fold higher dysplasia index than Eng(2f/2f) mice (4.6 ± 1.9 vs. 2.5 ± 1.1, p < 0.05). However, after normalization of the dysplasia index with the gene deletion efficiency (Alk1(2f/2f): 16% and Eng(2f/2f): 1%), we found that about 8-fold higher dysplasia was induced per copy of Eng deletion (2.5) than that of Alk1 deletion (0.3). ENG-negative endothelial cells were detected in the Ad-Cre-treated brain of Eng(2f/2f) mice, suggesting homozygous deletion of Eng in the cells. VEGF induced more severe vascular dysplasia in the Ad-Cre-treated brain of Eng(2f/2f) mice than that of Eng(+/-) mice. (1) Deletion of Eng induces more severe cerebrovascular dysplasia per copy than that of Alk1 upon VEGF stimulation. (2) Homozygous deletion of Eng with angiogenic stimulation may be a promising strategy for development of a bAVM mouse model. (3) The endothelial cells that have homozygous causal gene deletion in AVM could be crucial for lesion development.


Breast tumour angiogenesis.
The central importance of tumour neovascularization has been emphasized by clinical trials using antiangiogenic therapy in breast cancer. This review gives a background to breast tumour neovascularization in in situ and invasive breast cancer, outlines the mechanisms by which this is achieved and discusses the influence of the microenvironment, focusing on hypoxia. The regulation of angiogenesis and the antivascular agents that are used in an antiangiogenic dosing schedule, both novel and conventional, are also summarized.


Wnt-dependent beta-catenin signaling is activated after unilateral ureteral obstruction, and recombinant secreted frizzled-related protein 4 alters the progression of renal fibrosis.
beta-Catenin functions as a transducer of Wnt signals to the nucleus, where it interacts with the T cell factor (TCF) family of DNA binding proteins to regulate gene expression. On the basis of the genes regulated by beta-catenin and TCF in various biologic settings, two predicted functions of beta-catenin/TCF-dependent transcription are to mediate the loss of epithelial polarity and to promote fibroblast activities, such as the increased synthesis of fibronectin during chronic renal disease. These predictions were tested by determination of the expression and function of an inhibitor of Wnt signaling, secreted frizzled-related protein 4 (sFRP4), during renal tubular epithelial injury initiated by unilateral ureteral obstruction (UUO). Despite increased sFRP4 gene expression in perivascular regions of injured kidneys, total sFRP4 protein levels decreased after injury. The decreased sFRP4 protein levels after UUO accompanied increased Wnt-dependent beta-catenin signaling in tubular epithelial and interstitial cells, along with increased expression of markers of fibrosis. Administration of recombinant sFRP4 protein caused a reduction in tubular epithelial beta-catenin signaling and suppressed the progression of renal fibrosis, as evidenced by a partial maintenance of E-cadherin mRNA expression and a reduction in the amount of fibronectin and alpha-smooth muscle actin proteins. Furthermore, recombinant sFRP4 reduced the number of myofibroblasts, a central mediator of fibrosis. It is concluded that beta-catenin signaling is activated in tubular epithelial and interstitial cells after renal injury, and recombinant sFRP4 can interfere with epithelial de-differentiation and with fibroblast differentiation and function during progression of renal fibrosis.


Altered placental expression of kisspeptin and its receptor in pre-eclampsia.
Kisspeptin, originally identified as metastatin, important in preventing cancer metastasis, has more recently been shown to be important in pregnancy. Roles indicated for kisspeptin in pregnancy include regulating trophoblast invasion and migration during placentation. The pregnancy-specific disorder pre-eclampsia (PE) is now accepted to begin with inadequate trophoblast invasion and the current study therefore sets out to characterise placental expression of both kisspeptin (KISS1) and its receptor (KISS1R) throughout pregnancy and in PE. Placental tissue was obtained from women undergoing elective surgical termination of early pregnancy (n=10) and from women following Caesarean section at term in normal pregnancy (n=10) and with PE (n=10). Immunohistochemistry of paraffin embedded sections and western immunoblotting were performed to assess protein localisation and expression. Quantitative real-time PCR was carried out to evaluate mRNA expression of both KISS1 and KISS1R. Protein and mRNA expression was found to mirror each other with KISS1 expression found to be reduced in PE compared with that in normal term pregnancy. Interestingly, KISS1R expression at both the mRNA and protein levels was found to be increased in PE compared with that in normal term pregnancy. The current findings of increased KISS1R expression may represent a mechanism by which functional activity of KISS1 is higher in PE than in normal pregnancy. Higher levels of activity of KISS1R may be involved in inhibition of trophoblast invasion and angiogenesis, which are associated with PE.


Giant cell angioblastoma in an adult: a unique presentation.
Giant cell angioblastoma is a very rare, locally destructive vascular tumor of intermediate malignancy without metastatic potential. There are only a few cases reported in the literature exclusively in the soft tissue of children. For the first time, we report on an adult patient with a giant cell angioblastoma in the popliteal fossa. The therapy included tumor resection with favorable clinical, oncological and functional outcome. Due to its locally destructive nature, surgery remains the mainstay of treatment. Histologically, giant cell angioblastoma is comprised of nodular aggregates of histiocytoid cells arranged around bland angiomatous spaces. Because of insufficient available data in regard to the definition of the entity, diagnostic criteria and its biological potential, it is not included in the new World Health Organization classification of tumors of soft tissue and bone. The differential diagnosis includes plexiform fibrohistiocytic tumor, myofibroma and giant cell fibroblastoma.


The clinical, endoscopic and histological spectrum of the solitary rectal ulcer syndrome: a single-center experience of 116 cases.
Solitary rectal ulcer syndrome (SRUS) is an uncommon although benign defecation disorder. The aim of this study was to evaluate the variable endoscopic manifestations of SRUS and its association with other diseases. All the patients diagnosed with SRUS histologically from January 1990 to February 2011 at The Aga Khan University, Karachi were included in the study. The medical records were reviewed retrospectively to evaluate the clinical spectrum of the patients along with the endoscopic and histological findings. A total of 116 patients were evaluated. The mean age was 37.4 ± 16.6 (range: 13-80) years, 61 (53%) of the patients were male. Bleeding per rectum was present in 82%, abdominal pain in 49%, constipation in 23% and diarrhea in 22%. Endoscopically, solitary and multiple lesions were present in 79 (68%) and 33 (28%) patients respectively; ulcerative lesions in 90 (78%), polypoidal in 29 (25%), erythematous patches in 3 (2.5%) and petechial spots in one patient. Associated underlying conditions were hemorrhoids in 7 (6%), hyperplastic polyps in 4 (3.5%), adenomatous polyps in 2(2%), history of ulcerative colitis in 3 (2.5%) while adenocarcinoma of colon was observed in two patients. One patient had previous surgery for colonic carcinoma. SRUS may manifest on endoscopy as multiple ulcers, polypoidal growth and erythematous patches and has shown to share clinicopathological features with rectal prolapse, proctitis cystica profunda (PCP) and inflammatory cloacogenic polyp; therefore collectively grouped as mucosal prolapse syndrome. This may be associated with underlying conditions such as polyps, ulcerative colitis, hemorrhoids and malignancy. High index of suspicion is required to diagnose potentially serious disease by repeated endoscopies with biopsies to look for potentially serious underlying conditions associated with SRUS.


Data Analytics and Modeling for Appointment No-show in Community Health Centers.
Using predictive modeling techniques, we developed and compared appointment no-show prediction models to better understand appointment adherence in underserved populations. We collected electronic health record (EHR) data and appointment data including patient, provider and clinical visit characteristics over a 3-year period. All patient data came from an urban system of community health centers (CHCs) with 10 facilities. We sought to identify critical variables through logistic regression, artificial neural network, and naïve Bayes classifier models to predict missed appointments. We used 10-fold cross-validation to assess the models' ability to identify patients missing their appointments. Following data preprocessing and cleaning, the final dataset included 73811 unique appointments with 12,392 missed appointments. Predictors of missed appointments versus attended appointments included lead time (time between scheduling and the appointment), patient prior missed appointments, cell phone ownership, tobacco use and the number of days since last appointment. Models had a relatively high area under the curve for all 3 models (e.g., 0.86 for naïve Bayes classifier). Patient appointment adherence varies across clinics within a healthcare system. Data analytics results demonstrate the value of existing clinical and operational data to address important operational and management issues. EHR data including patient and scheduling information predicted the missed appointments of underserved populations in urban CHCs. Our application of predictive modeling techniques helped prioritize the design and implementation of interventions that may improve efficiency in community health centers for more timely access to care. CHCs would benefit from investing in the technical resources needed to make these data readily available as a means to inform important operational and policy questions.


The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product.
Mitogen-activated protein (MAP) kinases, also known as extracellular signal-regulated kinases (ERKs), are thought to act at an integration point for multiple biochemical signals because they are activated by a wide variety of extracellular signals, rapidly phosphorylated on threonine and tyrosine, and highly conserved. A critical protein kinase lies upstream of MAP kinase and stimulates the enzymatic activity of MAP kinase. The structure of this protein kinase, denoted MEK1, for MAP kinase or ERK kinase, was elucidated from a complementary DNA sequence and shown to be a protein of 393 amino acids (43,500 daltons) that is related most closely in size and sequence to the product encoded by the Schizosaccharomyces pombe byr1 gene. The MEK gene was highly expressed in murine brain, and the product expressed in bacteria phosphorylated the ERK gene product.


Phosphorylation-dependent cleavage regulates von Hippel Lindau proteostasis and function.
Loss of von Hippel Lindau (VHL) protein function is a key driver of VHL diseases, including sporadic and inherited clear cell renal cell carcinoma. Modulation of the proteostasis of VHL, especially missense point-mutated VHL, is a promising approach to augmenting VHL levels and function. VHL proteostasis is regulated by multiple mechanisms including folding, chaperone binding, complex formation and phosphorylation. Nevertheless, many details underlying the regulations of VHL proteostasis are unknown. VHL is expressed as two variants, VHL30 and VHL19. Furthermore, the long-form variant of VHL was often detected as multiple bands by western blotting. However, how these multiple species of VHL are generated and whether the process regulates VHL proteostasis and function are unknown. We hypothesized that the two major species are generated by VHL protein cleavage, and the cleavage regulates VHL proteostasis and subsequent function. We characterized VHL species using genetical and pharmacological approaches and showed that VHL was first cleaved at the N-terminus by chymotrypsin C before being directed for proteasomal degradation. Casein kinase 2-mediated phosphorylation at VHL N-terminus was required for the cleavage. Furthermore, inhibition of cleavage stabilized VHL protein and thereby promoted HIF downregulation. Our study reveals a novel mechanism regulating VHL proteostasis and function, which is significant for identifying new drug targets and developing new therapeutic approaches targeting VHL deficiency in VHL diseases.


Spread of Ebola virus disease based on the density of roads in West Africa.
On March 23rd 2014 the World Health Organization announced that a new Ebola outbreak had appeared in West Africa involving three countries. The objective of this study was to show how a road density index (RDI) could be constructed and a study of its association with Ebola cases during the outbreak. The study was carried out at the district level across the affected countries. RDI was calculated by km2 of territory as a proxy for the mobility of the population. To calculate this index, the number of km of road constructed in each district was estimated and subsequently divided by the area of each district expressed in km2. The accumulated incidence of Ebola was calculated per district. A generalised linear model with a Poisson distribution was used. The RDI varied from 0.12 to 0.84 between the districts. An RDI increase of 0.01 indicates a 3% increase in Ebola infection risk (RR is 1.03; CI 1.03-1.04). The density of the road network can influence the increased incidence of Ebola cases in the affected zone. An exhaustive mapping of the area could help the relevant organisations to manage another outbreak in the future and it could help the distribution of resources in an emergency situation.


Lack of confidence among trainee doctors in the management of diabetes: the Trainees Own Perception of Delivery of Care (TOPDOC) Diabetes Study.
There is an increased prevalence of diabetes. Doctors in training, irrespective of specialty, will have patients with diabetes under their care. To determine levels of confidence of doctors in training in the management of diabetes and establish their training needs in this area of clinical practice. A national online survey of trainee doctors in the UK using a pre-validated questionnaire. A four-point confidence rating scale was used to rate confidence in the management of diabetes and comparators. A six-point scale was used to quantify how often trainees would contribute to the management of patients with diabetes and trainees were asked about their training in managing diabetes. A total of 2149 doctors completed the survey. The percentage 'fully confident' in diagnosing diabetes was 27%, diagnosing and managing hypoglycaemia 55%, diagnosing and managing diabetic ketoacidosis 43%, managing intravenous (IV) insulin 27%, prescribing IV fluids for patients with diabetes 39% and altering diabetes therapy prior to surgery/other procedure 18%. In comparison, 66% and 65% were 'fully confident' in the management of angina and asthma, respectively (P < 0.05). Forty-one percent would take the initiative to optimize glycaemic control for patients under their care >80% of the time. Respectively, 19% and 35% of respondents reported that their undergraduate and postgraduate training had prepared them adequately to optimize treatment of diabetes. The majority (>70%) wanted further training in managing all aspects of diabetes care. Trainee doctors in the UK lack confidence in the management of diabetes, are unlikely to take the initiative to optimize glycaemic control and report a need for further training.


Neck dissection with simultaneous carotid endarterectomy.
Patients with metastatic neck disease from upper aerodigestive tract carcinomas have an extensive history of tobacco and alcohol abuse. These patients are predisposed to develop atherosclerotic vascular disease. An increased incidence and severity of carotid stenosis in patients receiving radiotherapy for head and neck cancers is known. Management of patients with severe carotid stenosis who require surgical treatment of their neck disease has not been described. The authors describe our experience with simultaneous carotid endarterectomy and neck dissection. Prospective data collection. From 1991 to 1997 at West Virginia University Hospitals, Morgantown, West Virginia, and State University of New York (SUNY) at Buffalo, three patients with severe carotid stenosis required surgery for metastatic neck disease. Preoperative evaluation revealed a bilateral carotid stenosis greater than 90% in all patients. All patients underwent modified radical neck dissections and simultaneous carotid endarterectomies with saphenous vein grafting. Two patients, one undergoing partial pharyngectomy and laryngectomy and the other a laryngectomy and neck dissection, had coverage of the carotid artery with the myogenous component of a pectoral major graft. One patient had only a neck dissection. Two patients healed with no local morbidity, no neck recurrence, and a patent carotid artery by Doppler. No strokes were encountered. One patient died of a myocardial infarction. Severe carotid stenosis that requires revascularization may have endarterectomy performed simultaneously with treatment of head and neck primary with no increase in morbidity.


Maternal death after oocyte donation at high maternal age: case report.
The percentage of women giving birth after the age of 35 increased in many western countries. The number of women remaining childless also increased, mostly due to aging oocytes. The method of oocyte donation offers the possibility for infertile older women to become pregnant. Gestation after oocyte-donation-IVF, however, is not without risks for the mother, especially at advanced age. An infertile woman went abroad for oocyte-donation-IVF, since this treatment is not offered in The Netherlands after the age of 45. The first oocyte donation treatment resulted in multiple gestation, but was ended by induced abortion: the woman could not cope with the idea of being pregnant with twins. During the second pregnancy after oocyte donation, at the age of 50, she was mentally more stable. The pregnancy, again a multiple gestation, was uneventful until delivery. Immediately after delivery the woman had hypertension with nausea and vomiting. A few hours later she had an eclamptic fit. HELLP-syndrome was diagnosed. She died due to cerebral haemorrhage. In The Netherlands, the age limit for women receiving donor oocytes is 45 years and commercial oocyte donation is forbidden by law. In other countries there is no age limit, the reason why some women are going abroad to receive the treatment of their choice. Advanced age, IVF and twin pregnancy are all risk factors for pre-eclampsia, the leading cause of maternal death in The Netherlands.Patient autonomy is an important ethical principle, but doctors are also bound to the principle of 'not doing harm', and do have the right to refuse medical treatment such as IVF-treatment. The discussion whether women above 50 should have children is still not closed. If the decision is made to offer this treatment to a woman at advanced age, the doctor should counsel her intensively about the risks before treatment is started.


CD44/CD24 immunophenotypes on clinicopathologic features of salivary glands malignant neoplasms.
Salivary glands malignant neoplasms (SGMNs) account for 3-6% of head and neck cancers and 0.3% of all cancers. Tumor cells that express CD44 and CD24 exhibit a stem-cell-like behavior. CD44 is the binding site for hyaluronic acid, and CD24 is a receptor that interacts with P-selectin to induce metastasis and tumor progression. The present study aims to evaluate the expression of CD44 and CD24 on SGMNs and correlated these data with several clinicopathologic features. Immunohistochemical stains for CD44 and CD24 were performed on tissue microarrays containing SGMN samples from 69 patients. The CD44, CD24 and CD44/CD24 expression phenotypes were correlated to patient clinicopathologic features and outcome. CD44 expression was associated with the primary site of neoplasm (p = 0.046). CD24 was associated with clinical stage III/IV (p = 0.008), T stage (p = 0,27) and lymph node (p = 0,001). The CD44/CD24 profiles were associated with the primary site of injury (p = 0.005), lymph node (p = 0.011) and T stage (p = 0.023). Univariate analysis showed a significant relationship between clinical staging and disease- free survival (p = 0.009), and the overall survival presents relation with male gender (p = 0.011) and metastasis (p = 0.027). In summary, our investigation confirms that the clinical stage, in accordance with the literature, is the main prognostic factor for SGMN. Additionally, we have presented some evidence that the analysis of isolated CD44 and CD24 immunoexpression or the two combined markers could give prognostic information associated to clinicopathologic features in SGMN. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1284611098470676.


Anti-inflammatory effect of cinnamaldehyde and linalool from the leaf essential oil of Cinnamomum osmophloeum Kanehira in endotoxin-induced mice.
Cinnamomum osmophloeum Kanehira is a Taiwan native plant that belongs to genus Cinnamomum and is also known as pseudocinnamomum or indigenous cinnamon. Its leaf is traditionally used by local people in cooking and as folk therapy. We previously demonstrated the chemical composition and anti-inflammatory effect of leaf essential oil of Cinnamomum osmophloeum Kanehira of linalool chemotype in streptozotocin-induced diabetic rats and on endotoxin-injected mice. The aim of the present study is to evaluate whether cinnamaldehyde and linalool the active anti-inflammatory compounds in leaf essential oil of Cinnamomum osmophloeum Kanehira. Before the injection of endotoxin, C57BL/6 mice of the experimental groups were administered cinnamaldehyde (0.45 or 0.9 mg/kg body weight) or linalool (2.6 or 5.2 mg/kg body weight), mice of the positive control group were administered the leaf essential oil (13 mg/kg body weight), and mice of the negative group were administered vehicle (corn oil, 4 mL/kg body weight) by gavage every other day for two weeks. All mice received endotoxin (i.p. 10 mg/mL/kg body weight) the next day after the final administration and were killed 12 h after the injection. Normal control mice were pretreated with vehicle followed by the injection with saline. None of the treatment found to affect body weight or food or water intake of mice before the injection of endotoxin. Cinnamaldehyde and linalool were found significantly reversed endotoxin-induced body weight loss and lymphoid organ enlargement compared with vehicle (P < 0.05). Both compounds also significantly lowered endotoxin-induced levels of peripheral nitrate/nitrite, interleukin (IL)-1β, IL-18, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and High-mobility group box 1 protein (HMGB-1), and levels of nitrate/nitrite, IL-1β, TNF-α, and IFN-γ in spleen and mesenteric lymph nodes (MLNs) (P < 0.05). Endotoxin-induced expression of toll-like receptor 4 (TLR4), Myeloid differentiation primary response gene 88 (MyD88), myeloid differentiation protein 2 (MD2), Nod-like receptor family, pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC), and caspase-1 in spleen and mesenteric lymph nodes (MLNs) were inhibited by all tested doses of cinnamaldehyde and linalool (P < 0.05). Subsequently, the activation of nuclear factor (NF)-κB and the activity of caspase-1 in spleen and MLNs were also suppressed by these two compounds (P < 0.05). In addition, cinnamaldehyde and linalool at the dose equivalent to their corresponding content in the tested dose of the leaf essential oil, which was 0.9 mg/kg and 5.2 mg/kg, respectively, showed similar or slightly less inhibitory activity for most of these inflammatory parameters compared with that of the leaf essential oil. Our data confirmed the potential use of leaf essential oil of Cinnamomum osmophloeum Kanehira as an anti-inflammatory natural product and provide evidence for cinnamaldehyde and linalool as two potent agents for prophylactic use in health problems associated with inflammations that being attributed to over-activated TLR4 and/or NLRP3 signaling pathways.


Chromatin Immunoprecipitation (ChIP) to Assess Histone Marks in Auxin-treated Arabidopsis thaliana Inflorescence Tissue.
Chromatin Immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) or high-throughput sequencing (ChIP-seq) has become the gold standard for the identification of binding sites of DNA binding proteins and the localization of histone modification on a locus-specific or genome-wide scale, respectively. ChIP experiments can be divided into seven critical steps: (A) sample collection, (B) crosslinking of proteins to DNA, (C) nuclear extraction, (D) chromatin isolation and fragmentation by sonication, (E) immunoprecipitation of histone marks by appropriate antibodies, (F) DNA recovery, and (G) identification of precipitated protein-associated DNA by qPCR or high-throughput sequencing. Here, we describe a time-efficient protocol that can be used for ChIP-qPCR experiments to study the localization of histone modifications in young inflorescences of the model plants Arabidopsis thaliana.


Central nervous system infections in patients with severe burns.
Central nervous system (CNS) infections develop in 3-9% of neurosurgical ICU patients and 0.4-2% of all patients hospitalized with head trauma. CNS infection incidence in burn patients is unknown and this study sets out to identify the incidence and risk factors associated with CNS infections. A retrospective electronic chart review was performed from 1 July 2003 to 30 June 2008 evaluating inpatient medical records along with cerebrospinal fluid (CSF) microbiological results for the presence of CNS infection. The presence of facial and head injuries and burns, along with intracranial interventions were reviewed for association with CNS infections. There were 1964 admissions with 2 patients (0.1%) found to have CNS infection; 1 each with MRSA and Acinetobacter baumannii. Both patients had facial burns and trauma to their head that required intracranial surgery. Of note, both patients had bacteremia with the same microorganisms isolated from their CSF and both survived. Of all patients, 29% had head or neck trauma and burns; 0.35% of those had a CNS infection. Scalp harvest for grafts or debridement of burned scalp was performed on 125 patients of which 9 had an invasive surgical procedure that involved penetration of the skull. The 2 infected patients were from these 9 intracranial surgical patients revealing a 22% infection rate. The incidence of CNS infections in patients with severe burns is extremely low at 0.1%. This rate was low even with head and face burns with trauma unless the patient underwent an intracranial procedure.


Lesions of the fornix but not the entorhinal or perirhinal cortex interfere with contextual fear conditioning.
The effects of entorhinal cortex lesions, combined entorhinal and perirhinal cortex lesions, and fornix lesions on the conditioning of fear responses (freezing) to contextual stimuli were examined using a conditioning procedure known to produce hippocampal-dependent contextual conditioning. Lesions of the entorhinal and or entorhinal plus perirhinal cortex did not disrupt contextual conditioning, but lesions of the fornix did. None of the lesions affected conditioning to an explicit conditioned stimulus. Given that the entorhinal cortex is the primary linkage between the neocortex and the hippocampus and that the fornix is the primary linkage with subcortical structures, subcortical inputs to and outputs of the hippocampus appear to be sufficient to mediate contextual fear conditioning. As a result, the presumption that neocortical information is required for contextual fear conditioning, and perhaps other hippocampal-dependent functions, should be reevaluated.


Pharmacology of L-754,142, a highly potent, orally active, nonpeptidyl endothelin antagonist.
L-754,142, (-)-N-(4-iso-propylbenzenesulfonyl)-alpha-(4-carboxyl-2-n-propy lphenoxy)-3,4- methylenedioxyphenylacetamide, is a potent nonpeptidyl endothelin antagonist (e.g., Ki: cloned human ETA = 0.062 nM: cloned human ETB = 2.25 nM), with high specificity for endothelin receptors. In vitro, L-754,142 is a potent antagonist of ET-1-induced phosphatidyl inositol hydrolysis in Chinese hamster ovary cells expressing cloned human endothelin receptors (IC50: hETA = 0.35 nM; hETB = 26 nM) and of ET-1 induced contractions in rabbit iliac artery rings (pA2 = 7.74) and rat aortic rings (pA2 = 8.7). In vivo, L-754,142 is a potent and specific antagonist of exogenously administered ET-1 or big ET-1, L-754,142 fully protects against ET-1-induced lethality in mice (AD50 = 0.26 mg/kg i.v.). The pressor response to big ET-1 in the anesthetized ferret is blocked by this compound with an ED50 value of 0.019 mg/kg i.v. L-754,142 also blocks the pressor response to big ET-1 in the conscious rat with ED50 values of 0.30 mg/kg i.v. and 0.56 mg/kg p.o. The duration of action of L-754,142 in this rat model is more than 12 hr after an oral dose of 3 mg/kg. In summary, L-754,142 is a potent, orally active ET antagonist with a long duration of action in several in vivo models.


Inhibition of calcium oxalate crystal growth in vitro by uropontin: another member of the aspartic acid-rich protein superfamily.
The majority of human urinary stones are primarily composed of calcium salts. Although normal urine is frequently supersaturated with respect to calcium oxalate, most humans do not form stones. Inhibitors are among the multiple factors that may influence the complex process of urinary stone formation. We have isolated an inhibitor of calcium oxalate crystal growth from human urine by monoclonal antibody immunoaffinity chromatography. The N-terminal amino acid sequence and acidic amino acid content of this aspartic acid-rich protein, uropontin, are similar to those of other pontin proteins from bone, plasma, breast milk, and cells. The inhibitory effect of uropontin on calcium oxalate crystal growth in vitro supports the concept that pontins may have a regulatory role. This function would be analogous to that of other members of the aspartic acid-rich protein superfamily, which stereospecifically regulate the mineralization fronts of calcium-containing crystals.


Cutoff values of serum ferritin and TIBC saturation for the evaluation of gastrointestinal neoplasms in adult anemic patients.
We conducted a retrospective study to find out the optimum values of serum ferritin and other hematologic indices in adult anemic patients who should be referred for thorough gastrointestinal (GI) endoscopic evaluation for GI neoplasms. 544 adult anemic patients were stratified into three groups according to the results of GI endoscopy: benign versus premalignant versus malignant. As compared to non-malignant groups, malignant group demonstrated statistically significant differences in terms of median values of ferritin and total iron-binding capacity (TIBC) saturation. By receiver operating characteristics curve analyses to find out optimum cut-off points of the serum ferritin and TIBC saturation which distinguish between non-malignant diseases and malignant diseases, the cut-off ferritin value of 44.33 ng/mL in male had 72.73 % sensitivity and 70.95 % specificity. The cut-off TIBC saturation value of 9.13 % in male had 73.33 % sensitivity and 70.92 % specificity. The cut-off TIBC saturation value of 6.16 % in female had 69.57 % sensitivity and 65.13 % specificity. It is recommended that adult male patients with anemia undergo thorough endoscopic evaluation to detect GI neoplasms when their serum ferritin levels are ≤44 ng/mL or TIBC saturation values are ≤9 %. For adult female, only TIBC saturation values less than 6 % may contribute to determining whether they undergo GI endoscopic evaluation.


Mass spectrometry of atmospheric aerosols--recent developments and applications. Part II: On-line mass spectrometry techniques.
Many of the significant advances in our understanding of atmospheric particles can be attributed to the application of mass spectrometry. Mass spectrometry provides high sensitivity with fast response time to probe chemically complex particles. This review focuses on recent developments and applications in the field of mass spectrometry of atmospheric aerosols. In Part II of this two-part review, we concentrate on real-time mass spectrometry techniques, which provide high time resolution for insight into brief events and diurnal changes while eliminating the potential artifacts acquired during long-term filter sampling. In particular, real-time mass spectrometry has been shown recently to provide the ability to probe the chemical composition of ambient individual particles <30 nm in diameter to further our understanding of how particles are formed through nucleation in the atmosphere. Further, transportable real-time mass spectrometry techniques are now used frequently on ground-, ship-, and aircraft-based studies around the globe to further our understanding of the spatial distribution of atmospheric aerosols. In addition, coupling aerosol mass spectrometry techniques with other measurements in series has allowed the in situ determination of chemically resolved particle effective density, refractive index, volatility, and cloud activation properties.


One new piperazinedione isolated from a mangrove-derived fungus Aspergillus niger JX-5.
One new piperazinedione derivative, nigerpiperazine A (1), along with six known compounds (2-7) were isolated from the fungus Aspergillus niger JX-5 from mangrove Ceriops tagal. Nigerpiperazine A (1) was elucidated by spectroscopic analyses and 13C NMR chemical shift calculations, compounds 4 and 5 were determined by X-ray and the CD spectra. The absolute configuration of 4 was identified by X-ray for the first time. Compounds 1 and 4 showed inhibitory activities against Helicoverpa armigera Hubner with the IC50 values of 200 and 100 µg/mL, respectively.


Taura syndrome virus from Belize represents a unique variant.
A Taura syndrome virus (TSV) isolate from cultured Penaeus vannamei grown in Belize, Central America was characterized and shown to be a unique isolate. Mortality rates in laboratory infections of specific pathogen-free (SPF) P. vannamei, reactivity of the virus with monoclonal antibody (MAb) 1A1 and phylogenetic analysis demonstrated that the Belize isolate (BLZ02TSV) is a new valiant of TSV. The Hawaiian 1994 TSV isolate (HI94TSV, GenBank AF277675) was used as the reference isolate for these studies. Laboratory infections of SPF P. vannamei with BLZ02TSV demonstrated higher mortalities and earlier onset of mortalities compared to infections with HI94TSV. Shrimp tissues infected with BLZ02TSV reacted with a TSV-specific gene probe by in situ hybridization and were positive by RT-PCR using TSV diagnostic primers, thus indicating that the isolate was TSV. However, Western blot analysis and immunohistochemistry using MAb 1A1 demonstrated that BLZ02TSV did not react with the antibody, suggestive of changes in the VP1 region of the genome that codes for the polypeptide to which MAb 1A1 binds. Phylogenetic analysis of a 1.3 kbp fragment of the TSV VP1 capsid region revealed that BLZ02TSV represents a distinct group among more than 29 isolates of TSV studied thus far. This research demonstrates that BLZ02TSV is a unique isolate of TSV and reiterates a problem related to the use of MAb 1A1 for detection of TSV in clinical specimens.


Screening mammography in women less than age 50 years.
For women aged 40-49 years, to describe the benefits and harms of performing screening mammography, accuracy of digital mammography, and new evidence on the effectiveness of risk-based screening. New data support a 15% reduction in breast cancer mortality for women aged 40-49 years after 10 years of screening; however, the absolute benefit is small and not outweighed by important harms. Digital mammography is more sensitive, but less specific than film mammography in women aged 40-49 years. Risk-based screening that identifies and screens women aged 40-49 years with breast cancer risk similar to an average-risk woman aged 50-59 years results in similar benefits and harms of screening these high-risk women as screening average-risk 50-year-old women. Practitioners should discuss with women aged 40-49 years the benefits and harms of undergoing screening mammography before offering them screening. If women elect to undergo screening mammography, they should undergo biennial screening with digital mammography. Targeting screening for those women aged 40-49 years with risk factors that substantially increase the risk of breast cancer, such as high breast density, family history of breast cancer, and history of benign breast biopsy, could maximize the benefits and minimizes the harms of screening this age group.


A catalytically active high-Mr form of human cathepsin B from sputum.
A cysteine proteinase from purulent sputum was partially purified by a method involving affinity chromatography on Sepharose-aminohexanoylphenylalanylglycinaldehyde semicarbazone. It was immunologically related to lysosomal cathepsin B from human liver and was similar in many, but not all, other aspects. It was catalytically active, as demonstrated by active-site-directed radioiodination, and hydrolysed three cathepsin B substrates, two with Km values similar to those of lysosomal cathepsin B. In addition, the rates of inactivation of the sputum and lysosomal forms of the enzyme by L-3-carboxy-2,3-transepoxypropionyl-leucylamido(4-guanidino) butane (Compound E-64) were very similar. However, the sputum enzyme differed from lysosomal cathepsin B in the following respects. Inhibition by chicken cystatin was much weaker for sputum cathepsin B than for the lysosomal enzyme. Sputum cathepsin B had greater stability at pH 7.5 and a higher apparent Mr, even after deglycosylation, than lysosomal cathepsin B. We conclude that the form of cathepsin B found in sputum is probably a truncated form of human procathepsin B, with some differences in properties that could be of physiological importance.


Oral tranexamic acid (TA) in the treatment of melasma: A retrospective analysis.
Melasma is a common pigmentary disorder among Asians and treatment is challenging. Oral tranexamic acid (TA) has emerged as a potential treatment for refractory melasma. Large-scale studies on its use, outcomes, and safety are limited. We sought to evaluate treatment outcomes and adverse effects of oral TA in melasma in an Asian population. We conducted a retrospective analysis of patients who received oral TA for melasma in a tertiary dermatologic center from January 2010 to June 2014. In all, 561 patients (91.4% female, 8.6% male) were enrolled. Median duration of treatment was 4 months. The majority (503 [89.7%]) improved, 56 (10.0%) had no improvement, and 2 (0.4%) worsened. Patients without family history of melasma had better response rates than those with family history (90.6% vs 60.0%, P = .01). Of the 503 who improved, response was seen within 2 months of TA initiation, with a relapse rate of 27.2%. Adverse events occurred in 40 (7.1%). Most were transient, but 1 developed deep vein thrombosis requiring prompt discontinuation. She was later given the diagnosis of familial protein S deficiency. This was a retrospective study. Oral TA may be an effective adjunct for refractory melasma. Careful screening for personal and familial risk factors for thromboembolism should be done before initiation.


A higher plant extracellular vitronectin-like adhesion protein is related to the translational elongation factor-1 alpha.
Higher plant proteins immunologically related to the animal substrate adhesion molecule vitronectin have recently been observed and implicated in a variety of biological processes, such as plasma membrane-cell wall adhesion, pollen tube extension, and bacterium-plant interaction. We provide evidence that, similar to vitronectin, one of these proteins, PVN1 (plant vitronectin-like 1), isolated from 428 mM NaCl-adapted tobacco cells binds to glass surfaces an heparin. PVN1 was isolated by glass bead affinity chromatography. Isolated PVN1 has adhesive activity based on results from a baby hamster kidney cell-spreading assay. This plant adhesion protein was detected in all tissues examined but was most abundant in roots and salt-adapted cultured cells. Immunogold labeling indicated that PVN1 is localized in the cell wall of cortical and transmitting tissue cells of pollinated mature styles. A partial amino acid sequence of PVN1 revealed no similarity with vitronectin but, instead, was nearly identical to the translational elongation factor-1 alpha (EF-1 alpha). A clone isolated by screening a tobacco cDNA expression library with anti-PVN1 encoded a protein with greater than 93% identity to sequences of EF-1 alpha from plants of numerous species. Immunological cross-reactivity between tobacco PVN1 and EF-1 alpha as well as the reaction between the EF-1 alpha antibody and the 65- and 75-kD vitronectin-like proteins of a fucoidal alga supported the conclusion that the plant extracellular adhesion protein PVN1 is related to EF-1 alpha.


SleepSIM: Conditional GAN-based non-REM sleep EEG Signal Generator.
Synthetic data generation has become increasingly popular with the increasing use of generative networks. Recently, Generative Adversarial Network (GAN) architectures have produced exceptional results in synthetic image generation. However, time series generation still needs to be studied. This paper proposes a Conditional GAN-based system to generate unique samples of non-REM sleep electroencephalographic (EEG) signals. The CGAN model had a 1-D Convolution Neural Network based architecture. The model was trained using real EEG from healthy controls. The trained model can generate an artificial 30-second epoch of non-REM sleep whose power spectrum is identical to that of a real sleep EEG.Clinical relevance- Sleep EEG simulation can be used to train and enhance the skillset of fellows and technicians in the sleep medicine field. Variations in EEG signals can be highly complex to model mathematically; however, here, we harness the power of deep learning, using generative models such as CGANs to train, model complex data distributions, and generate diverse and artificial but realistic EEG signals during non-REM sleep.


CNTF Gene Therapy Confers Lifelong Neuroprotection in a Mouse Model of Human Retinitis Pigmentosa.
The long-term outcome of neuroprotection as a therapeutic strategy for preventing cell death in neurodegenerative disorders remains unknown, primarily due to slow disease progression and the inherent difficulty of assessing neuronal survival in vivo. Employing a murine model of retinal disease, we demonstrate that ciliary neurotrophic factor (CNTF) confers life-long protection against photoreceptor degeneration. Repetitive retinal imaging allowed the survival of intrinsically fluorescent cone photoreceptors to be quantified in vivo. Imaging of the visual cortex and assessment of visually-evoked behavioral responses demonstrated that surviving cones retain function and signal correctly to the brain. The mechanisms underlying CNTF-mediated neuroprotection were explored through transcriptome analysis, revealing widespread upregulation of proteolysis inhibitors, which may prevent cellular/extracellular matrix degradation and complement activation in neurodegenerative diseases. These findings provide insights into potential novel therapeutic avenues for diseases such as retinitis pigmentosa and amyotrophic lateral sclerosis, for which CNTF has been evaluated unsuccessfully in clinical trials.


Arsenic trioxide enhances the radiation sensitivity of androgen-dependent and -independent human prostate cancer cells.
Prostate cancer is the most common malignancy in men. In the present study, LNCaP (androgen-sensitive human prostate cancer cells) and PC-3 cells (androgen-independent human prostate cancer cells) were used to investigate the anti-cancer effects of ionizing radiation (IR) combined with arsenic trioxide (ATO) and to determine the underlying mechanisms in vitro and in vivo. We found that IR combined with ATO increases the therapeutic efficacy compared to individual treatments in LNCaP and PC-3 human prostate cancer cells. In addition, combined treatment showed enhanced reactive oxygen species (ROS) generation compared to treatment with ATO or IR alone in PC-3 cells. Combined treatment induced autophagy and apoptosis in LNCaP cells, and mainly induced autophagy in PC-3 cells. The cell death that was induced by the combined treatment was primarily the result of inhibition of the Akt/mTOR signaling pathways. Furthermore, we found that the combined treatment of cells pre-treated with 3-MA resulted in a significant change in AO-positive cells and cytotoxicity. In an in vivo study, the combination treatment had anti-tumor growth effects. These novel findings suggest that combined treatment is a potential therapeutic strategy not only for androgen-dependent prostate cancer but also for androgen-independent prostate cancer.


Opposite Roles of Salicylic Acid Receptors NPR1 and NPR3/NPR4 in Transcriptional Regulation of Plant Immunity.
Salicylic acid (SA) is a plant defense hormone required for immunity. Arabidopsis NPR1 and NPR3/NPR4 were previously shown to bind SA and all three proteins were proposed as SA receptors. NPR1 functions as a transcriptional co-activator, whereas NPR3/NPR4 were suggested to function as E3 ligases that promote NPR1 degradation. Here we report that NPR3/NPR4 function as transcriptional co-repressors and SA inhibits their activities to promote the expression of downstream immune regulators. npr4-4D, a gain-of-function npr4 allele that renders NPR4 unable to bind SA, constitutively represses SA-induced immune responses. In contrast, the equivalent mutation in NPR1 abolishes its ability to bind SA and promote SA-induced defense gene expression. Further analysis revealed that NPR3/NPR4 and NPR1 function independently to regulate SA-induced immune responses. Our study indicates that both NPR1 and NPR3/NPR4 are bona fide SA receptors, but play opposite roles in transcriptional regulation of SA-induced defense gene expression.


A Multi-Sensor Fusion MAV State Estimation from Long-Range Stereo, IMU, GPS and Barometric Sensors.
State estimation is the most critical capability for MAV (Micro-Aerial Vehicle) localization, autonomous obstacle avoidance, robust flight control and 3D environmental mapping. There are three main challenges for MAV state estimation: (1) it can deal with aggressive 6 DOF (Degree Of Freedom) motion; (2) it should be robust to intermittent GPS (Global Positioning System) (even GPS-denied) situations; (3) it should work well both for low- and high-altitude flight. In this paper, we present a state estimation technique by fusing long-range stereo visual odometry, GPS, barometric and IMU (Inertial Measurement Unit) measurements. The new estimation system has two main parts, a stochastic cloning EKF (Extended Kalman Filter) estimator that loosely fuses both absolute state measurements (GPS, barometer) and the relative state measurements (IMU, visual odometry), and is derived and discussed in detail. A long-range stereo visual odometry is proposed for high-altitude MAV odometry calculation by using both multi-view stereo triangulation and a multi-view stereo inverse depth filter. The odometry takes the EKF information (IMU integral) for robust camera pose tracking and image feature matching, and the stereo odometry output serves as the relative measurements for the update of the state estimation. Experimental results on a benchmark dataset and our real flight dataset show the effectiveness of the proposed state estimation system, especially for the aggressive, intermittent GPS and high-altitude MAV flight.


Long-term exposure to road traffic noise and stroke incidence: a Danish Nurse Cohort study.
Road traffic noise has been linked to increased risk of ischemic heart disease, yet evidence on stroke shows mixed results. We examine the association between long-term exposure to road traffic noise and incidence of stroke, overall and by subtype (ischemic or hemorrhagic), after adjustment for air pollution. Twenty-five thousand six hundred and sixty female nurses from the Danish Nurse Cohort recruited in 1993 or 1999 were followed for stroke-related first-ever hospital contact until December 31st, 2014. Full residential address histories since 1970 were obtained and annual means of road traffic noise (Lden [dB]) and air pollutants (particulate matter with diameter < 2.5 μm and < 10 μm [PM2.5 and PM10], nitrogen dioxide [NO2], nitrogen oxides [NOx]) were determined using validated models. Time-varying Cox regression models were used to estimate hazard ratios (HR) (95% confidence intervals [CI]) for the associations of one-, three-, and 23-year running means of Lden preceding stroke (all, ischemic or hemorrhagic), adjusting for stroke risk factors and air pollutants. The World Health Organization and the Danish government's maximum exposure recommendations of 53 and 58 dB, respectively, were explored as potential Lden thresholds. Of 25,660 nurses, 1237 developed their first stroke (1089 ischemic, 148 hemorrhagic) during 16 years mean follow-up. For associations between a 1-year mean of Lden and overall stroke incidence, the estimated HR (95% CI) in the fully adjusted model was 1.06 (0.98-1.14) per 10 dB, which attenuated to 1.01 (0.93-1.09) and 1.00 (0.91-1.09) in models further adjusted for PM2.5 or NO2, respectively. Associations for other exposure periods or separately for ischemic or hemorrhagic stroke were similar. There was no evidence of a threshold association between Lden and stroke. Long-term exposure to road traffic noise was suggestively positively associated with the risk of overall stroke, although not after adjusting for air pollution.


Are long chain acyl CoAs responsible for suppression of mitochondrial metabolism in hibernating 13-lined ground squirrels?
Hibernation in 13-lined ground squirrels (Ictidomys tridecemlineatus) is associated with a substantial suppression of whole-animal metabolism. We compared the metabolism of liver mitochondria isolated from torpid ground squirrels with those from interbout euthermic (IBE; recently aroused from torpor) and summer euthermic conspecifics. Succinate-fuelled state 3 respiration, calculated relative to mitochondrial protein, was suppressed in torpor by 48% and 44% compared with IBE and summer, respectively. This suppression remains when respiration is expressed relative to cytochrome c oxidase activity. We hypothesized that this suppression was caused by inhibition of succinate transport at the dicarboxylate transporter (DCT) by long-chain fatty acyl CoAs that may accumulate during torpor. We predicted, therefore, that exogenous palmitoyl CoA would inhibit respiration in IBE more than in torpor. Palmitoyl CoA inhibited respiration ~70%, in both torpor and IBE. The addition of carnitine, predicted to reverse palmitoyl CoA suppression by facilitating its transport into the mitochondrial matrix, did not rescue the respiration rates in IBE or torpor. Liver mitochondrial activities of carnitine palmitoyl transferase did not differ among IBE, torpor and summer animals. Although palmitoyl CoA inhibits succinate-fuelled respiration, this suppression is likely not related exclusively to inhibition of the DCT, and may involve additional mitochondrial transporters such as the adenine-nucleotide transporter.


Selective attention modulates the direction of audio-visual temporal recalibration.
Temporal recalibration of cross-modal synchrony has been proposed as a mechanism to compensate for timing differences between sensory modalities. However, far from the rich complexity of everyday life sensory environments, most studies to date have examined recalibration on isolated cross-modal pairings. Here, we hypothesize that selective attention might provide an effective filter to help resolve which stimuli are selected when multiple events compete for recalibration. We addressed this question by testing audio-visual recalibration following an adaptation phase where two opposing audio-visual asynchronies were present. The direction of voluntary visual attention, and therefore to one of the two possible asynchronies (flash leading or flash lagging), was manipulated using colour as a selection criterion. We found a shift in the point of subjective audio-visual simultaneity as a function of whether the observer had focused attention to audio-then-flash or to flash-then-audio groupings during the adaptation phase. A baseline adaptation condition revealed that this effect of endogenous attention was only effective toward the lagging flash. This hints at the role of exogenous capture and/or additional endogenous effects producing an asymmetry toward the leading flash. We conclude that selective attention helps promote selected audio-visual pairings to be combined and subsequently adjusted in time but, stimulus organization exerts a strong impact on recalibration. We tentatively hypothesize that the resolution of recalibration in complex scenarios involves the orchestration of top-down selection mechanisms and stimulus-driven processes.


Partial versus radical nephrectomy for 4 to 7 cm renal cortical tumors.
Recent observations suggest that partial nephrectomy for small renal tumors may be associated with improved survival compared with radical nephrectomy. We evaluated survival in patients with 4 to 7 cm renal tumors in a bi-institutional collaboration. By combining institutional databases from Mayo Clinic and Memorial Sloan-Kettering Cancer Center we identified 1,159 patients with 4.1 to 7.0 cm sporadic, unilateral, solitary, localized renal masses who underwent radical or partial nephrectomy between 1989 and 2006. Patient outcome was compared using Cox proportional hazards regression models. Of the 1,159 patients 873 (75%) and 286 (25%) were treated with radical and partial nephrectomy, respectively. Patients treated with partial vs radical nephrectomy were significantly more likely to have a solitary kidney (10% vs 0.2%) and chronic kidney disease (15% vs 7%, each p <0.001). Median followup in survivors was 4.8 years (range 0 to 19). There was no significant difference in overall survival in patients treated with radical vs partial nephrectomy (p = 0.8). Of 943 patients with renal cell carcinoma those treated with radical nephrectomy were significantly more likely to die of renal cell carcinoma than those treated with partial nephrectomy (HR 2.16, 95% CI 1.04-4.50, p = 0.039) but this only approached statistical significance on multivariate analysis (HR 1.97, 95% CI 0.92-4.20, p = 0.079). Results suggest that overall and cancer specific survival is not compromised when partial nephrectomy is done for 4 to 7 cm renal cortical tumors. With the benefit of preserving renal function our results support partial nephrectomy when technically feasible for renal tumors up to 7 cm.


Chemopreventive effect of apple and berry fruits against colon cancer.
Colon cancer arises due to the conversion of precancerous polyps (benign) found in the inner lining of the colon. Prevention is better than cure, and this is very true with respect to colon cancer. Various epidemiologic studies have linked colorectal cancer with food intake. Apple and berry juices are widely consumed among various ethnicities because of their nutritious values. In this review article, chemopreventive effects of these fruit juices against colon cancer are discussed. Studies dealing with bioavailability, in vitro and in vivo effects of apple and berry juices are emphasized in this article. A thorough literature survey indicated that various phenolic phytochemicals present in these fruit juices have the innate potential to inhibit colon cancer cell lines. This review proposes the need for more preclinical evidence for the effects of fruit juices against different colon cancer cells, and also strives to facilitate clinical studies using these juices in humans in large trials. The conclusion of the review is that these apple and berry juices will be possible candidates in the campaign against colon cancer.


A memory efficient method for structure-based RNA multiple alignment.
Structure-based RNA multiple alignment is particularly challenging because covarying mutations make sequence information alone insufficient. Existing tools for RNA multiple alignment first generate pairwise RNA structure alignments and then build the multiple alignment using only sequence information. Here we present PMFastR, an algorithm which iteratively uses a sequence-structure alignment procedure to build a structure-based RNA multiple alignment from one sequence with known structure and a database of sequences from the same family. PMFastR also has low memory consumption allowing for the alignment of large sequences such as 16S and 23S rRNA. The algorithm also provides a method to utilize a multicore environment. We present results on benchmark data sets from BRAliBase, which shows PMFastR performs comparably to other state-of-the-art programs. Finally, we regenerate 607 Rfam seed alignments and show that our automated process creates multiple alignments similar to the manually curated Rfam seed alignments. Thus, the techniques presented in this paper allow for the generation of multiple alignments using sequence-structure guidance, while limiting memory consumption. As a result, multiple alignments of long RNA sequences, such as 16S and 23S rRNAs, can easily be generated locally on a personal computer. The software and supplementary data are available at http://genome.ucf.edu/PMFastR.


Alcohol Increases Delay and Probability Discounting of Condom-Protected Sex: A Novel Vector for Alcohol-Related HIV Transmission.
Alcohol use, especially at binge levels, is associated with sexual HIV risk behavior, but the mechanisms through which alcohol increases sexual risk taking are not well-examined. Delay discounting, that is, devaluation of future consequences as a function of delay to their occurrence, has been implicated in a variety of problem behaviors, including risky sexual behavior. Probability discounting is studied with a similar framework as delay discounting, but is a distinct process in which a consequence is devalued because it is uncertain or probabilistic. Twenty-three, nondependent alcohol users (13 male, 10 female; mean age = 25.3 years old) orally consumed alcohol (1 g/kg) or placebo in 2 separate experimental sessions. During sessions, participants completed tasks examining delay and probability discounting of hypothetical condom-protected sex (Sexual Delay Discounting Task, Sexual Probability Discounting Task) and of hypothetical and real money. Alcohol decreased the likelihood that participants would wait to have condom-protected sex versus having immediate, unprotected sex. Alcohol also decreased the likelihood that participants would use an immediately available condom given a specified level of sexually transmitted infection (STI) risk. Alcohol did not affect delay discounting of money, but it did increase participants' preferences for larger, probabilistic monetary rewards over smaller, certain rewards. Acute, binge-level alcohol intoxication may increase sexual HIV risk by decreasing willingness to delay sex in order to acquire a condom in situations where one is not immediately available, and by decreasing sensitivity to perceived risk of STI contraction. These findings suggest that delay and probability discounting are critical, but heretofore unrecognized, processes that may mediate the relations between alcohol use and HIV risk.


An ultrasensitive molecularly-imprinted human cardiac troponin sensor.
Cardiac troponin T (TnT) is a highly sensitive cardiac biomarker for myocardial infarction. In this study, the fabrication and characterisation of a novel sensor for human TnT based on a molecularly-imprinted electrosynthesised polymer is reported. A TnT sensitive layer was prepared by electropolymerisation of o-phenylenediamine (o-PD) on a gold electrode in the presence of TnT as a template. To develop the molecularly imprinted polymer (MIP), the template molecules were removed from the modified electrode surface by washing with alkaline ethanol. Electrochemical methods were used to monitor the processes of electropolymerisation, template removal and binding. The imprinted layer was characterised by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM). The incubation of the MIP-modified electrode with respect to TnT concentration resulted in a suppression of the ferro/ferricyanide redox process. Experimental conditions were optimised and a linear relationship was observed between the peak current of [Fe(CN)6](3-)/[Fe(CN)6](4-) and the concentration of TnT in buffer over the range 0.009-0.8 ng/mL, with a detection limit of 9 pg/mL. The TnT MIP sensor was shown to have a high affinity to TnT in comparison with non-imprinted polymer (NIP) electrodes in both buffer and blood serum.


[Social inequality in health and the utilization of health services among the elderly in Latin America].
To learn if there is social inequality in health and in the utilization of health services among the elderly in six cities of Latin America: Buenos Aires, Argentina; São Paulo, Brazil; Santiago, Chile; Havana, Cuba; Mexico City, Mexico; and Montevideo, Uruguay. This study used data from a project called Health, Well-being, and Aging in Latin America and the Caribbean (known as the "SABE project"). To investigate the presence of social inequality in health an ordinal probit model was used, with health status indicated by the following dependent variables: activities of daily living, instrumental activities of daily living, physical mobility, and self-rated health. To measure social inequality in the utilization of health services two characteristics were assessed: one for outpatient medical services (whether or not individuals received outpatient care, and how many times), and one for hospital admissions (whether or not individuals were hospitalized). For outpatient services, we estimated a negative binomial hurdle model, and for hospital admissions, a logit model was constructed. Our main results suggest the presence of social inequality in health in all the cities, with better-off socioeconomic groups having better health. The difference in health was less marked in Havana, Buenos Aires, and Montevideo, and it was more pronounced in São Paulo and Mexico City. With respect to the utilization of health services we found inequalities in the use of outpatient services in Santiago, Mexico City, and São Paulo. In Santiago and Mexico City, more schooling was associated with a higher number of expected outpatient medical visits; however, the opposite was found in São Paulo. For hospital admissions, inequality was found only in São Paulo, with individuals with more years of schooling being much more likely to be hospitalized. To a certain extent, our results reflect the socioeconomic and demographic characteristics of the countries in which each of the six cities is located. The cities in the countries with the worst social indicators (high income inequality and low human development index) tended to have the greatest inequalities in health and in the utilization of health services.


Viral infection increases glucocorticoid-induced interleukin-10 production through ERK-mediated phosphorylation of the glucocorticoid receptor in dendritic cells: potential clinical implications.
The hypothalamic-pituitary-adrenal axis plays a central role in the adaptive response to stress including infection of pathogens through glucocorticoids. Physical and/or mental stress alter susceptibility to viral infection possibly by affecting this regulatory system, thus we explored potential cellular targets and mechanisms that underlie this phenomenon in key immune components dendritic cells (DCs). Dexamethasone (DEX) treatment and subsequent Newcastle disease virus (NDV) infection most significantly and cooperatively stimulated mRNA expression of the interleukin (IL)-10 in murine bone marrow-derived DCs among 89 genes involved in the Toll-like receptor signaling pathways. NDV increased DEX-induced IL-10 mRNA and protein expression by 7- and 3-fold, respectively, which was observed from 3 hours after infection. Conventional DCs (cDCs), but not plasmacytoid DCs (pDCs) were major sources of IL-10 in bone marrow-derived DCs treated with DEX and/or infected with NDV. Murine cytomegalovirus and DEX increased serum IL-10 cooperatively in female mice. Pre-treatment of DCs with the extracellular signal-regulated kinase (ERK) inhibitor U0126 abolished cooperative induction of IL-10 by DEX and NDV. Further, ERK overexpression increased IL-10 promoter activity stimulated by wild-type human GR but not by its mutant defective in serine 203, whereas ERK knockdown abolished NDV/DEX cooperation on IL-10 mRNA and phosphorylation of the mouse GR at serine 213. NDV also increased DEX-induced mRNA expression of three known glucocorticoid-responsive genes unrelated to the Toll-like receptor signaling pathways in DCs. These results indicate that virus and glucocorticoids cooperatively increase production of anti-inflammatory cytokine IL-10 by potentiating the transcriptional activity of GR in DCs, through which virus appears to facilitate its own propagation in infected hosts. The results may further underlie in part known exacerbation of IL-10/T helper-2-related allergic disorders by stress and viral infection.


Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity.
Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe therapeutics. Moringa oleifera is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism of pharmacological actions, antioxidant properties of the Moringa oleifera leaf extracts were tested in two stages of maturity using standard in vitro models. The successive aqueous extract of Moringa oleifera exhibited strong scavenging effect on 2, 2-diphenyl-2-picryl hydrazyl (DPPH) free radical, superoxide, nitric oxide radical and inhibition of lipid per oxidation. The free radical scavenging effect of Moringa oleifera leaf extract was comparable with that of the reference antioxidants. The data obtained in the present study suggests that the extracts of Moringa oleifera both mature and tender leaves have potent antioxidant activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage.


Regulation of Coronary Blood Flow.
The heart is uniquely responsible for providing its own blood supply through the coronary circulation. Regulation of coronary blood flow is quite complex and, after over 100 years of dedicated research, is understood to be dictated through multiple mechanisms that include extravascular compressive forces (tissue pressure), coronary perfusion pressure, myogenic, local metabolic, endothelial as well as neural and hormonal influences. While each of these determinants can have profound influence over myocardial perfusion, largely through effects on end-effector ion channels, these mechanisms collectively modulate coronary vascular resistance and act to ensure that the myocardial requirements for oxygen and substrates are adequately provided by the coronary circulation. The purpose of this series of Comprehensive Physiology is to highlight current knowledge regarding the physiologic regulation of coronary blood flow, with emphasis on functional anatomy and the interplay between the physical and biological determinants of myocardial oxygen delivery. © 2017 American Physiological Society. Compr Physiol 7:321-382, 2017.


Glucose tolerance status is a better predictor of diabetes and cardiovascular outcomes than metabolic syndrome: a prospective cohort study.
To evaluate the importance of oral glucose tolerance test (OGTT) in predicting diabetes and cardiovascular disease in patients with and without Metabolic Syndrome from a population treated in a primary care unit. A prospective cohort study was conducted with subjects regularly attending the primary care unit of Hospital de Clínicas de Porto Alegre. Participants underwent a 75 g OGTT. Metabolic syndrome definition was based on the criteria of IDF/AHA/NHLBI-2010. Participants mean age was 61 ± 12 years (males: 38%; whites: 67%). Of the 148 subjects included, 127 (86%) were followed for 36 ± 14 months, 21 (14%) were lost. Subjects were classified into four groups based on baseline OGTT: 29% normal (n = 43), 28% impaired fasting glucose (IFG; n = 42), 26% impaired glucose tolerance (IGT; n = 38), and 17% diabetes (n = 25). Metabolic syndrome prevalence was lower in normal group (28%), intermediate in IFG (62%) and IGT (65%) groups, and higher among subjects with diabetes (92%; P <0.001). Incidence of diabetes increased along with the stages of glucose metabolism disturbance (normal: 0%, IFG: 16%, IGT: 28%; P = 0.004). No patient with normal OGTT developed diabetes, regardless metabolic syndrome presence. Diabetes at baseline was the major determinant of cardiovascular disease occurrence (normal: 0%, IFG: 4%, IGT: 0%, diabetes: 24%; P = 0.001). In Cox-regression analysis, only the 2 h OGTT results were associated with diabetes (OR = 1.03; 95%CI 1.01-1.06; P <0.001) and cardiovascular disease development (OR = 1.013; 95%CI 1.002-1.025; P = 0.024). In this sample of subjects undergoing diabetes screening, the OGTT predicted diabetes and cardiovascular disease more effectively than the metabolic syndrome status.


Survival from breast cancer among South Asian and non-South Asian women resident in South East England.
Ethnic differences in breast cancer survival have been observed in the USA but have not been examined in Britain. We aimed to investigate such differences between South Asian (i.e. those with family roots in the Indian subcontinent) and non-South Asian (essentially British-native) women in England. Primary breast cancer cases incident in 1986 -1993 and resident in South East England were ascertained through the Thames Cancer and Registry and followed up to the end of 1997. Cases of South Asian ethnicity were identified on the basis of their names by using a previously validated computer algorithm. A total of 1037 South Asian and 50 201 non-South Asian breast cancer cases were included in the analysis; 30% of the South Asian (n=312) and 44% (n=22 201) of the non-South Asian cases died during follow-up. South Asian cases had a higher relative survival than non-South Asians throughout the follow-up period. The 10-year relative survival rates were 72.6% (95% confidence interval: 69.0, 75.9%) and 65.2% (64.5, 65.8%) for South Asians and non-South Asians, respectively. The excess mortality rates experienced by South Asians were 82% (72, 94%) of those experienced by non-South Asians (P=0.004). The magnitude of this effect was slightly reduced with adjustment for differences in age at diagnosis, but was strengthened with further adjustment for differences in stage at presentation and socioeconomic deprivation (excess mortality rates in South Asians relative to non-South Asians=72% (63, 82%), P&<0.001). These findings indicate that the higher survival from breast cancer in the first 10 years after diagnosis among South Asian was not due to differences in age at diagnosis, socioeconomic deprivation or disease stage at presentation.


Exercise Performance and Corticospinal Excitability during Action Observation.
Observation of a model performing fast exercise improves simultaneous exercise performance; however, the precise mechanism underpinning this effect is unknown. The aim of the present study was to investigate whether the speed of the observed exercise influenced both upper body exercise performance and the activation of a cortical action observation network (AON). In Experiment 1, 10 participants completed a 5 km time trial on an arm-crank ergometer whilst observing a blank screen (no-video) and a model performing exercise at both a typical (i.e., individual mean cadence during baseline time trial) and 15% faster than typical speed. In Experiment 2, 11 participants performed arm crank exercise whilst observing exercise at typical speed, 15% slower and 15% faster than typical speed. In Experiment 3, 11 participants observed the typical, slow and fast exercise, and a no-video, whilst corticospinal excitability was assessed using transcranial magnetic stimulation. In Experiment 1, performance time decreased and mean power increased, during observation of the fast exercise compared to the no-video condition. In Experiment 2, cadence and power increased during observation of the fast exercise compared to the typical speed exercise but there was no effect of observation of slow exercise on exercise behavior. In Experiment 3, observation of exercise increased corticospinal excitability; however, there was no difference between the exercise speeds. Observation of fast exercise improves simultaneous upper-body exercise performance. However, because there was no effect of exercise speed on corticospinal excitability, these results suggest that these improvements are not solely due to changes in the activity of the AON.


The Effects of Age, Distraction, and Simulated Central Vision Impairment on Hazard Detection in a Driving Simulator.
Despite similar levels of visual acuity and contrast sensitivity reductions, simulated central vision impairment increased response times to a much greater extent in older than in younger participants. Driving is crucial for maintaining independence in older age, but age-related vision impairments and in-vehicle auditory distractions may impair driving safety. We investigated the effects of age, simulated central vision impairment, and auditory distraction on detection of pedestrian hazards. Thirty-two normally sighted participants (16 younger and 16 older) completed four highway drives in a simulator and pressed the horn whenever they saw a pedestrian. Pedestrians ran toward the road on a collision course with the approaching vehicle. Simulated central vision impairment was achieved by attaching diffusing filters to a pair of laboratory goggles, which reduced visual acuity to 20/80 and contrast sensitivity by 0.35 log units. For drives with distraction, subjects listened to an audiobook and repeated out loud target words. Simulated central vision impairment had a greater effect on reaction times (660-millisecond increase) than age (350-millisecond increase) and distraction (160-millisecond increase) and had a greater effect on older than younger subjects (828- and 492-millisecond increase, respectively). Simulated central vision impairment decreased safe response rates from 94.7 to 78.3%. Distraction did not, however, affect safety because older subjects drove more slowly when distracted (but did not drive more slowly with vision impairment), suggesting that they might have perceived greater threat from the auditory distraction than the vision impairment. Older participants drove more slowly in response to auditory distraction. However, neither older nor younger participants adapted their speed in response to simulated vision impairment, resulting in unsafe detections. These results underline the importance of evaluating safety of responses to hazards as well as reaction times in a paradigm that flexibly allows participants to modify their driving behaviors.


Simultaneous Measurement of 92 Serum Protein Biomarkers for the Development of a Multiprotein Classifier for Ovarian Cancer Detection.
The best known ovarian cancer biomarker, CA125, is neither adequately sensitive nor specific for screening the general population. By using a combination of proteins for screening, it may be possible to increase the sensitivity and specificity over CA125 alone. In this study, we used Proseek Multiplex Oncology II plates to simultaneously measure the expression of 92 cancer-related proteins in serum using proximity extension assays. This technology combines the sensitivity of the PCR with the specificity of antibody-based detection methods, allowing multiplex biomarker detection and high-throughput quantification. We analyzed 1 μL of sera from each of 61 women with ovarian cancer and compared the values obtained with those from 88 age-matched healthy women. Principle component analysis and unsupervised hierarchical clustering separated the ovarian cancer patients from the healthy, with minimal misclassification. Data from the Proseek plates for CA125 levels exhibited a strong correlation with clinical values for CA125. We identified 52 proteins that differed significantly (P < 0.006) between ovarian cancer and healthy samples, several of which are novel serum biomarkers for ovarian cancer. In total, 40 proteins had an estimated area under the ROC curve of 0.70 or greater, suggesting their potential to serve as biomarkers for ovarian cancer. CA125 alone achieved a sensitivity of 93.4% at a specificity of 98%. By adding the Oncology II values for five proteins to CA125 in a multiprotein classifier, we increased the assay sensitivity to 98.4% at a specificity of 98%, thereby improving the sensitivity and specificity of CA125 alone.


Molecular characterization of the surface layer proteins from Clostridium difficile.
Many bacteria express a surface-exposed proteinaceous layer, termed the S-layer, which forms a regular two-dimensional array visible by electron microscopy. Clostridium difficile is unusual in expressing two S-layer proteins (SLPs), which are of varying size in a number of strains. In an approach combining molecular biology with mass spectrometric sequencing strategies, we have identified the structural gene (slpA) for the S-layer from three strains of C. difficile. Both proteins are derived from a common precursor, and processing involves the removal of a signal peptide and a second cleavage to release the two mature SLPs. To our knowledge, this is the first example in which two SLPs have been shown to derive from a single gene product through post-translational processing, rather than from the expression of separate genes. The higher molecular weight (MW) SLP is highly conserved among the three strains, whereas the lower MW SLP shows considerable sequence diversity, reflecting the results from Western blotting. The high-MW SLP shows weak homology to N-acetyl muramoyl-L-alanine amidase from Bacillus subtilis, and both the native SLP from C. difficile and a recombinant protein expressed in Escherichia coli were found to display amidase activity by zymography. The high-MW SLPs showed evidence of glycosylation, whereas the lower MW proteins did not. A family of genes with sequence homology to the amidase domain of the high-MW SLP was identified in the C. difficile strain 630 genome, some of which are located in the same region of the genome as slpA and were shown by reverse transcription-polymerase chain reaction (RT-PCR) analysis to be transcribed.


In vitro antibacterial activity of DU-6859a, a new fluoroquinolone.
The in vitro antibacterial activity of DU-6859a, a new fluoroquinolone, against a wide variety of clinical isolates was evaluated and compared with those of tosufloxacin, ofloxacin, ciprofloxacin, and sparfloxacin. DU-6859a showed potent broad-spectrum activity against gram-positive, gram-negative, and anaerobic bacteria, and its activity was greater than those of the control quinolones. By comparison of MICs at which 90% of strains are inhibited, DU-6859a had potent activity against bacteria resistant to the control quinolones. The time-killing curves of quinolones showed that the number of viable cells decreased rapidly during 2 to 4 of incubation, and regrowth was not seen even after 8 h incubation. At a concentration of four times the MIC, the frequencies of appearance of spontaneous mutants of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa resistant to DU-6859a were < or = 4.0 x 10(-9) to 1.9 x 10(-8). The 50% inhibitory concentrations of DU-6859a were 0.86 and 1.05 micrograms/ml for the supercoiling activities of DNA gyrases isolated from E. coli and P. aeruginosa, respectively. The rank order of the 50% inhibitory concentrations observed for both DNA gyrases roughly paralleled the MICs.


Ion Effect and Metal-Coordinated Cross-Linking for Multiscale Design of Nereis Jaw Inspired Mechanomutable Materials.
The Nvjp-1 protein is a key component in the jaws of Nereis virens, a species of marine worm. It contains over 25 mol % of histidine, which is believed to play a key role in the metal-coordinated cross-linking responsible for the structural stability and exceptional mechanical performance of the worm jaw. Understanding the nanoscale mechanism behind this cross-linking and its pathway in affecting the macroscopic mechanical behavior of the material is crucial to develop bioinspired mechanomutable materials based on Nvjp-1. Here, we use a combination of multiscale modeling and experimental synthesis to understand the behavior of this heterologous-expressed protein from the nano- to the macroscale. We have built a bottom-up molecular-based model, which includes electronic-based density functional theory calculations, atomistic simulation of the nanoscale properties with replica exchange molecular dynamics, and an elastic network model for describing the macroscale behavior at different pHs. This multiscale modeling supports the experimental synthesis of a photo-cross-linked Nvjp-1 hydrogel by proving both the nanoscale mechanisms and mechanical behavior predictions. Our theoretical results agree well with the experimental observations, showing that Nvjp-1 forms a more compact structure in the presence of Zn2+ ions with a suitable pH environment, leading to the formation of more stable intramolecular metal-coordinated cross-links. These metal-coordinated cross-links induce nanoscale aggregation of Nvjp-1, which is responsible for the hydrogel contraction observed in experiments and predicted by the model.