Measurement of extracellular space in the rabbit AV node.
We used quantitative histochemistry to measure the size of the extracellular space (ESC) in various regions of the rabbit heart. When inulin, sucrose, and sorbitol were used as ECS markers, the ECS of the AV-nodal tissue was found to be, respectively, 2.4, 2.2, and 2.5 times larger than that of left ventricular muscle. Glucose was also measured over a 50-fold serum concentration range as an extracellular marker for AV-nodal tissue, left ventricular muscle, and Purkinje fibers. Measurements with glucose also revealed that the ECS of the AV node was 2.5-2.8 times larger than that of ventricular muscle. In contrast, the ECS of the AV node was the same as that of Purkinje fibers when glucose was used as an extracellular marker. ATP content, measured as an intracellular marker, was similar in both AV-nodal and contractile tissue. Collectively, the data obtained with all extracellular markers indicate that the ECS of the AV-nodal region is approximately 2.5 times larger than that of adjacent contractile tissue. Differences in the size of the ECS in various regions of the heart probably have functional significance and should be considered appropriately during the interpretation of data obtained by biochemical and densitometric approaches.


Novel therapeutic approaches toward Hantaan virus and its clinical features' similarity with COVID-19.
Zoonotic virus spill over in human community has been an intensive area of viral pathogenesis and the outbreak of Hantaan virus and severe acute respiratory syndrome coronavirus 2 (SARS CoV2) after late December 2019 caused a global threat. Hantaan virus is second to the COVID-19 outbreak in China with seven cases positive and one death. Both RNA viruses have opposite sense as in (-) for Hantaan virus and (+) for SARS CoV2 but have similarity in the pathogenesis and relevant clinical features including dry cough, high fever, shortness of breath, and SARS associated with pneumonia and certain reported cases with multiple organ failure. Although COVID-19 has global impact with high death toll, Hantaan virus has varyingly high mortality rate between 1% and 40%. Hence, there is a need to explore novel therapeutic targets in Hantaan virus due to its rapid evolution rate in its genetic makeup which governs virulence and target host cells. This review emphasizes the importance of structural and nonstructural proteins of Hantaan virus with relevant insight from SARS CoV2. The envelope glycoproteins such as Gn, Gc, and nucleocapsid protein (N) direct the viral assembly and replication in host cells. Therapeutic treatment has similarity in using ribavirin and extracorporeal membrane oxygenation but lack of efficacious treatment in both cases of SARAS CoV2 and Hantaan virus. Therefore, potential features regarding therapeutic targets for drug discovery for Hantaan viruses are discussed herewith. The conclusive description highlights that N protein is substantially involved in evoking immune response and induces symptoms and could be precursive target for drug discovery studies.


[Sudden bilateral hearing loss with vertigo due to vertebral artery occlusion].
We report a case of sudden bilateral hearing loss associated with an occlusion of the right vertebral artery. A 66-year-old man was admitted to hospital suffering from sudden onset vertigo, deafness, and vomiting. He could not walk due to truncal ataxia. There was positional nystagmus to the right; spontaneous and gaze-evoked nystagmus were absent. He had no facial nerve palsy, dysarthria, pyramidal tract signs, limb ataxia, and sensory impairment. Pure tone audiometry demonstrated a profound sensorineural deafness of both ears. A tentative diagnosis of sudden deafness was made. He was treated with intravenous infusion of corticosteroid; hearing loss of the left ear slightly improved. Cranial MR imaging demonstrated fresh small infarcts in the right cerebellar tonsil, the left cerebellar medulla, and the left middle cerebellar peduncle. MR angiography demonstrated an occlusion of the right vertebral artery. It is possible that reduced perfusion of the anterior inferior cerebellar arteries and internal auditory arteries on both sides resulted in multiple "border zone" infarcts and damage to the inner ear. Clinicians should be aware of the possibility of vertebrobasilar occlusive disease in case of sudden bilateral hearing impairment, even when brainstem or cerebellar signs are absent.


5 Beta,14 beta-androstane-3 beta,14-diol binds to the digitalis receptor site on Na/K-ATPase.
5 beta,14 beta-Androstane-3 beta,-14-diol, the lead (minimum) structure in digitalis compounds, shows the same characteristics of interaction with Na/K-ATPase as ordinary digitalis compounds judged by the following six criteria: (I) shape of the concentration-inhibition curves, (II) species differences in affinity for the enzyme, (III) apparent competition with K+, (IV) competition with digitoxigenin for binding to the enzyme, (V) stabilization of phosphoenzyme formed from ATP, and (VI) enhancement of phosphorylation from orthophosphate.


Thrombin and the thrombin-thrombomodulin complex interaction with plasminogen activator inhibitor type-1.
Thrombin, the final enzyme of the coagulation system, also influences profibrinolytic activity by several mechanisms. These include cellular release of tissue plasminogen activator, activated protein C-induced fibrinolysis, and inactivation of plasminogen activator inhibitor, type 1 (PAI-1). In this report, the role of thrombin in the regulation of PAI-1 is investigated. Our studies demonstrate that thrombin inactivation of PAI-1 occurs via an enzymatic mechanism rather than an enzyme-inhibitor complex mechanism. Evidence to support this conclusion is: (1) concomitant analysis of PAI-1 and thrombin activities demonstrate decreased PAI-1 activity but no loss of thrombin activity; (2) no visible thrombin--PAI-1 complexes by SDS-PAGE analysis; and (3) lack of formation of 125I-thrombin-PAI-1 complexes. Thrombomodulin, a thrombin binding cofactor that modifies thrombin's functions, did not influence the inactivation of PAI-1 by thrombin. We propose that thrombin enzymatically inactivates PAI-1 without forming a stable enzyme-inhibitor complex. The reaction is not affected by thrombomodulin. Overall this reaction occurs so slowly that it is not physiologically relevant without some modifying factor(s).


Item response theory-based validation of a short form of the Eating Behavior Scale for Japanese adults.
Obesity has become a serious social problem in industrialized countries in recent years. Clinically, although the evaluation of dietary behavior abnormalities is as important as any method of risk assessment for obesity, almost all the existing scales with many items may have numerous practical clinical difficulties. In this study, we aimed to prepare a short questionnaire to assess the dietary behavior abnormalities related to obesity. A total of 1032 individuals aged 20 to 59 years participated in the present study. Using item response theory (IRT), we selected the items for a short version from among 30 items of Sakata Eating Behavior Scale (EBS), which is widely used in Japan. As a result of the IRT-based analysis on the original 30-item version, 7 items were adopted as the short version. The correlation between the total score of the original EBS and the EBS short form was extremely high (r = 0.93, P = .001). In examining the criterion validity, for all participants (n = 1032), male (n = 516), and female (n = 516), the correlation coefficients between the total score of the EBS short form and body mass index (BMI) were r = 0.26, r = 0.28, and r = 0.28, respectively. The results of the receiver operating characteristic analysis was performed with obesity BMI > 25 kg/m as a dependent variable, the value of the area under the curve in the ROC was significantly higher in the 7-item version than in the total score of the original items (P = .0005). In conclusion, the 7-item EBS short form was created. Furthermore, it was found that the EBS short form is a reliable and valid measure that can be used as an indicator of obesity in both clinical and research settings.


The search for the uremic toxin: the case for metabolic acidosis.
Much effort has been expended on determining which compound, hormone or metabolic condition causes the uremic syndrome. Byproducts of protein metabolism that can cause uremic symptoms, including loss of lean body mass, have been a focus of research but specific toxins have been difficult to identify. Evidence is provided that implicates metabolic acidosis as the prime signal initiating muscle wasting in uremia since it activates branched-chain ketoacid dehydrogenase and the ubiquitin proteasome pathway. These responses degrade the essential branched chain amino acids and protein in muscle, leading to loss of muscle mass. Correction of the metabolic acidosis with sodium bicarbonate supplements has significant therapeutic implications for uremic patients with even mild degrees of metabolic acidosis.


Widespread circulation of West Nile virus, but not Zika virus in southern Iran.
West Nile virus (WNV) and Zika virus (ZIKV) are mosquito-borne viral infections. Over the past few decades, WNV has been associated with several outbreaks involving high numbers of neuroinvasive diseases among humans. The recent re-emergence of ZIKV has been associated with congenital malformation and also with Guillain-Barre syndrome in adults. The geographic range of arthropod-borne viruses has been rapidly increasing in recent years. The objectives of this study were to determine the presence of IgG specific antibodies and the genome of WNV and ZIKV in human samples, as well as WNV and ZIKV genomes in wild-caught mosquitoes in urban and rural areas of the Hormozgan province, in southern Iran. A total of 494 serum samples were tested for the presence of WNV and ZIKV IgG antibodies using ELISA assays. One hundred and two (20.6%) samples were reactive for WNV IgG antibodies. All serum samples were negative for ZIKV IgG antibodies. Using the multivariable logistic analysis, age (45+ vs. 1-25; OR = 3.4, 95% C.I.: 1.8-6.3), occupation (mostly outdoor vs. mostly indoor; OR = 2.4, 95% C.I.: 1.1-5.2), and skin type(type I/II vs. type III/IV and type V/VI; OR = 4.3, 95% C.I.: 1.7-10.8 and OR = 2.7, 95% C.I.: 1.3-5.5 respectively, skin types based on Fitzpatrick scale) showed significant association with WNV seroreactivity. We collected 2,015 mosquitoes in 136 pools belonging to 5 genera and 14 species. Three pools of Culex pipiens complex were positive for WNV RNA using real-time reverse transcription polymerase chain reaction (rtRT-PCR). ZIKV RNA was not detected in any of the pools. All WNV ELISA reactive serum samples were negative for WNV RNA. In conclusion, we provided evidence of the establishment of WNV in southern Iran and no proof of ZIKV in serum samples or in mosquito vectors. The establishment of an organized arbovirus surveillance system and active case finding strategies seems to be necessary.


Changes in tree community structure in defaunated forests are not driven only by dispersal limitation.
Bushmeat hunting has reduced population sizes of large frugivorous vertebrates throughout the tropics, thereby reducing the dispersal of seeds. This is believed to affect tree population dynamics, and therefore community composition, because the seed dispersal of large-seeded trees depends upon large-bodied vertebrates.We report on a long-running study of the effect of defaunation on a tropical tree community. In three censuses over 11 years, we compared sapling recruitment between a hunted and a nonhunted site, which are nearby and comparable to one another, to determine the extent to which species composition has changed through time following defaunation. We expected to find a reduced abundance of tree species that rely on large frugivores for dispersal at the hunted site and altered community structure as a consequence.Although community composition at the hunted site diverged from that at the nonhunted site, the changes were independent of dispersal syndrome, with no trend toward a decline in species that are dispersed by large, hunted vertebrates. Moreover, the loss of large-bodied dispersers did not generate the changes in tree community composition that we hypothesized. Some species presumed to rely on large-bodied frugivores for dispersal are effectively recruiting despite the absence of their dispersers.Synthesis: The presumption that forests depleted of large-bodied dispersers will experience rapid, directional compositional change is not fully supported by our results. Altered species composition in the sapling layer at the hunted site, however, indicates that defaunation may be connected with changes to the tree community, but that the nature of these changes is not unidirectional as previously assumed. It remains difficult to predict how defaunation will affect tree community composition without a deeper understanding of the driving mechanisms at play.


A nationwide epidemiological study of spinal cord injuries in Taiwan from July 1992 to June 1996.
This prospective epidemiological survey of spinal cord injury (SCI) in Taiwan was carried out by recruiting patients attended by physicians from various medical centers and general hospitals all over Taiwan from July 1992 to June 1996. A total of 6,410 cases of traumatic spinal fracture were registered among which were 1,586 new cases of SCI. The results represented 70% of the scope of SCI in Taiwan. The observed average annual incidence of SCI in Taiwan was 18.8 per million population. The mean age was 46.1 years-old with a plateau distribution for over 20 years and older. Geriatric victims are a major group of SCI in Taiwan. The male to female ratio was 3 to 1. The leading causes of SCI were traffic accidents and accidental falls. Motorcycle collisions accounted for 62% of the traffic accidents, and as most of the motorcycle riders were not helmet users, head injury became the major associated injury of SCI in Taiwan. The effectiveness of the comprehensive care system for SCI patients in Taiwan is relatively good, as reflected by the low rates of complications of SCI, the low mortality rate (6.6%) and the high percentage (67.4%) of SCI patients achieving self-care ultimately at home after rehabilitation. The analysis of person days healthy life loss and quality adjusted survival time revealed that SCI patients in Taiwan required 4 years to cope with the morbidity, and on average, could return to the main stream of life for another 30 years.


Health Care Providers' Perceptions of Nutrition Support in Pediatric Oncology and Hematopoietic Stem Cell Transplant Patients.
One of the most common side effects of medical treatment for patients with an oncologic diagnosis is malnutrition. There is limited research that broadly assesses the perceptions of health care providers (HCPs) regarding nutrition support in the pediatric population. The purpose of this study was to describe the perceptions of nutrition support among pediatric oncology and hematopoietic stem cell transplant HCPs. The study used a cross-sectional descriptive design using a 31-item survey. Results of the survey indicated that nurses were more likely to initiate conversations about nutrition support during the first month of diagnosis, while midlevel providers and physicians initiated discussions in response to a change in nutritional status evidenced by decreased oral intake or weight loss. Participants reported resistance by patients and families more often for enteral nutrition compared with parenteral nutrition. Findings suggest a need to develop a more unified service line-based approach for initiating discussions related to nutrition support that incorporate patient and family perceptions.


Pic Protein From Enteroaggregative E. coli Induces Different Mechanisms for Its Dual Activity as a Mucus Secretagogue and a Mucinase.
A hallmark of enteroaggregative Escherichia coli (EAEC) infection is the formation of an intestinal biofilm, which comprises a mucus layer with immersed bacteria. Pic is an autotransporter secreted by EAEC, and other E. coli pathotypes, and has been involved in two apparently contradictory phenotypes, as a mucus secretagogue and as a mucinase. Here, we investigated this Pic dual activity, mucus secretagogue capability and mucinolytic activity, in human goblet cells that secrete MUC2 and MUC5AC. Pic induced mucus hypersecretion directly in the goblet cells, without other intestinal cell types involved. At the same time, Pic exhibited strong proteolytic activity on the secreted mucins. These activities were independent since a mutation in the serine protease motif (PicS258I) abolished mucin degradation while maintaining the mucus secretagogue activity intact. Furthermore, deoxycholic acid (DCA)-induced mucins were proteolytically degraded when goblet cells were co-incubated with DCA/Pic, while co-incubation with DCA/PicS258I induced a synergistic effect on mucus hypersecretion. Pic was more efficient degrading MUC5AC than MUC2, but no degradation was detected with Pic inactivated at the active site by mutation or pharmacological inhibition. Remarkably, Pic cleaved MUC2 and MUC5AC in the C-terminal domain, leaving N-terminal subproducts, impacting the feature of gel-forming mucins and allowing mucus layer penetration by EAEC. Astonishingly, Pic stimulated rapid mucin secretion in goblet-like cells by activating the intracellular calcium pathway resulting from the PLC signal transduction pathway, leading to the production of DAG and releasing IP3, a second messenger of calcium signaling. Therefore, the dual activity of Pic, as a mucus secretagogue and a mucinase, is relevant in the context of carbon source generation and mucus layer penetration, allowing EAEC to live within the layer of mucus but also access epithelial cells.


MicroRNA-148a-3p Directly Targets SERPINE1 to Suppress EMT-Mediated Colon Adenocarcinoma Progression.
This research aimed at clarifying the intracellular effect of SERPINE1 in the progression of colon adenocarcinoma (COAD) and the underlying mechanism. We obtained the expression profile of SERPINE1 in COAD via the Starbase database and verified it on COAD tissue samples through qRT-PCR and immunoblotting, respectively. Also, miRWalk, TargetScan and miRDB databases were adopted to generate the miRNA prediction that might target SERPINE1, and the gene target miR-148a-3p was confirmed using dual-luciferase assays. The effect of SERPINE1 and miR-148a-3p on COAD was further evaluated by cell experiments. MTT assay was used to detect the change of cell proliferation ability. The invasive and migratory capability of COAD cells was examined using transwell and would healing assays. Cell apoptosis was determined through flow cytometry. The expressions of genes and EMT-associated proteins were evaluated by qRT-PCR and immunoblotting. Further lucubration of the biological relevance of SERPINE1 and miR-148a-3p was conducted using rescue experiments. We found that the expression quantities of SERPINE1 in COAD tissues and cell lines were higher than those in corresponding non-cancerous tissues and normal cells. When SERPINE1 expression is reduced, EMT process is inhibited, invasion and proliferation ability of COAD cells are obviously reduced, and apoptosis level is increased. Moreover, SERPINE1 was identified as the target gene of miR-148a-3p. When the expression of miR-148a-3p was enhanced, it was found that the expression of SERPINE1 was reduced. miR-148a-3p played the similar effect of si-SERPINE1 that suppressed the COAD progression. Additionally, we found out that SERPINE1 is validated in hindering the tumor healing effect of miR148a-3p in COAD, including cell growth and invasion. Our study suggests that SERPINE1/miR-148a-3p axis has potential as prognostic markers of COAD and provides reference for the development of new therapies.


Improved specificity of newborn screening for congenital adrenal hyperplasia by second-tier steroid profiling using tandem mass spectrometry.
Newborn screening for congenital adrenal hyperplasia (CAH) involves measurement of 17alpha-hydroxyprogesterone (17-OHP), usually by immunoassay. Because this testing has been characterized by high false-positive rates, we developed a steroid profiling method that uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure 17-OHP, androstenedione, and cortisol simultaneously in blood spots. Whole blood was eluted from a 4.8-mm (3/16-inch) dried-blood spot by an aqueous solution containing the deuterium-labeled internal standard d(8)-17-OHP. 17-OHP, androstenedione, and cortisol were extracted into diethyl ether, which was subsequently evaporated and the residue dissolved in LC mobile phase. This extract was injected into a LC-MS/MS equipped with pneumatically assisted electrospray. The steroids were quantified in the selected-reaction monitoring mode by use of peak areas in reference to the stable-isotope-labeled internal standard. We analyzed 857 newborn blood spots, including 14 blood spots of confirmed CAH cases and 101 of false-positive cases by conventional screening. Intra- and interassay CVs for 17-OHP were 7.2-20% and 3.9-18%, respectively, at concentrations of 2, 30, and 50 microg/L. At a cutoff for 17-OHP of 12.5 microg/L and a cutoff of 3.75 for the sum of peak areas for 17-OHP and androstenedione divided by the peak area for cortisol, 86 of the 101 false-positive samples were within reference values by LC-MS/MS, whereas the 742 normal and 14 true-positive results obtained by conventional screening were correctly classified. Steroid profiling in blood spots can identify false-positive results obtained by conventional newborn screening for CAH.


Tumours of the heart. A study of 89 cases.
Eight-nine patients with heart tumours (76 metastatic and 13 primary) were studied. Metastatic tumours were usually clinically silent and most often came from lung carcinomas, lymphomas, malignant melanomas, and gastric and breast cancers. The primary tumours consisted of 11 myxomas, 1 fibrous histiocytoma and 1 haemangioma.


Nonalcoholic fatty liver disease in severely obese subjects.
Nonalcoholic fatty liver disease (NAFLD) has been consistently associated with obesity and insulin resistance. Nonalcoholic steatohepatitis (NASH) is a histological entity within NAFLD that can progress to cirrhosis. The exact prevalence of NASH in severe obesity is unknown. It is unclear whether differences in insulin sensitivity exist among subjects with NASH and simple fatty liver. To evaluate the prevalence and correlates of NASH and liver fibrosis in a racially diverse cohort of severely obese subjects. Ninety-seven subjects were enrolled. Liver biopsies, indirect markers of insulin resistance, metabolic parameters, and liver function tests were obtained. Thirty-six percent of subjects had NASH and 25% had fibrosis. No cirrhosis was diagnosed on histology. Markers of hyperglycemia, insulin resistance, and the metabolic syndrome but not body mass index were associated with the presence of NASH and fibrosis. Elevated transaminase levels correlated strongly with NASH and fibrosis but 46% subjects with NASH had normal transaminases. Subjects with NASH had more severe insulin resistance when compared to those with simple fatty liver. A signal detection model incorporating AST and the presence of diabetes predicted the presence of NASH while another incorporating ALT and HbA1C predicted the presence of fibrosis. NAFLD is associated with the metabolic syndrome rather than excess adipose tissue in severe obesity. Insulin resistance is higher in subjects with NASH versus those with simple fatty liver. Statistical models incorporating markers of liver injury and hyperglycemia may be useful in predicting the presence of liver pathology in this population.


Predictive Value of International Classification of Diseases Codes for Idiopathic Intracranial Hypertension in a University Health System.
Misclassification bias is introduced into medical claims-based research because of reliance on diagnostic coding rather than full medical record review. We sought to characterize this bias for idiopathic intracranial hypertension (IIH) and evaluate strategies to reduce it. A retrospective review of medical records was conducted using a clinical data warehouse containing medical records and administrative data from an academic medical center. Patients with 1 or more instances of International Classification of Diseases (ICD)-9 or -10 codes for IIH (348.2 or G93.2) between 1989 and 2017 and original results of neuroimaging (head CT or MRI), lumbar puncture, and optic nerve examination were included in the study. Diagnosis of IIH was classified as definite, probable, possible, or inaccurate based on review of medical records. The positive predictive value (PPV) for IIH ICD codes was calculated for all subjects, subjects with an IIH code after all testing was completed, subjects with high numbers of IIH ICD codes and codes spanning longer periods, subjects with IIH ICD codes associated with expert encounters (ophthalmology, neurology, or neurosurgery), and subjects with acetazolamide treatment. Of 1,005 patients with ICD codes for IIH, 103 patients had complete testing results and were included in the study. PPV of ICD-9/-10 codes for IIH was 0.63. PPV in restricted samples was 0.82 (code by an ophthalmologist n = 57), 0.70 (acetazolamide treatment n = 87), and 0.72 (code after all testing, n = 78). High numbers of code instances and longer duration between the first and last code instance also increased the PPV. An ICD-9 or -10 code for IIH had a PPV of 63% for probable or definite IIH in patients with necessary diagnostic testing performed at a single institution. Coding accuracy was improved in patients with an IIH ICD code assigned by an ophthalmologist. Use of coding algorithms considering treatment providers, number of codes, and treatment is a potential strategy to reduce misclassification bias in medical claims-based research on IIH. However, these are associated with a reduced sample size.


Exploration of characteristics and synthesis gas suitability for heat generation of coffee biomass pellets produced by single and co-pelletization.
Production of coffee beans generates various types of biomass that can be applied as bioenergy for drying and roasting the beans. Thus, the aims of this study were to explore the characteristics of coffee biomass pellets (CBPs) produced from coffee cherry pulp (CCP), coffee parchment (CPM), and expired green coffee beans (ECB) by single and co-pelletization. The CBPs were then used to produce the synthesis gas in a downdraft gasifier, and the syngas properties were investigated for further heat applications. The results showed that single and co-pelletization of CCP and CPM performed well. The CBPs had good physiochemical properties in shape, size, and atomic ratios. The higher heating value and energy density of CBPs were 19.25-24.29 MJ/kg and 12.09-14.87 GJ/m3. The ash from CBPs was rich in K2O, CaO and MgO oxides, and the CPM ash had the lowest initial deformation temperature at 1136 °C. The ash samples from CBPs also had different slagging and fouling indexes. The syngas from CBPs mainly contained H2 (6.85-9.30%), CO (12.15-18.85%), and CO2 (10.85-13.75%). The heating value and tar concentration of syngas from CBPs were 3.24-4.32 MJ/m3 and 21.75-30.92 g/m3. The main chemical compounds in tar were styrene, phenol, caffeine, and pyrrole according to GC-MS. These results indicate that CCP and CPM have potential for pelletization and gasification to generate heat needed for coffee bean processing.


Correlates of repeat lipid testing in patients with coronary heart disease.
Understanding the frequency and correlates of redundant lipid testing could identify areas for quality improvement initiatives aimed at improving the efficiency of cholesterol care in patients with coronary heart disease (CHD). To determine the frequency and correlates of repeat lipid testing in patients with CHD who attained low-density lipoprotein cholesterol (LDL-C) goals and received no treatment intensification. We assessed the proportion of patients with LDL-C levels of less than 100 mg/dL and no intensification of lipid-lowering therapy who underwent repeat lipid testing during an 11-month follow-up period. We performed logistic regression analyses to evaluate facility, provider, and patient characteristics associated with repeat testing. In total, we analyzed 35,191 patients with CHD in a Veterans Affairs network of 7 medical centers with associated community-based outpatient clinics. Frequency and correlates of repeat lipid testing in patients having CHD with LDL-C levels of less than 100 mg/dL and no further treatment intensification with lipid-lowering therapies. Of 27,947 patients with LDL-C levels of less than 100 mg/dL, 9200 (32.9%) had additional lipid assessments without treatment intensification during the following 11 months (12 ,686 total additional panels; mean, 1.38 additional panel per patient). Adjusting for facility-level clustering, patients with a history of diabetes mellitus (odds ratio [OR], 1.16; 95% CI, 1.10-1.22), a history of hypertension (OR, 1.21; 95% CI, 1.13-1.30), higher illness burden (OR, 1.39; 95% CI, 1.23-1.57), and more frequent primary care visits (OR, 1.32; 95% CI, 1.25-1.39) were more likely to undergo repeat testing, whereas patients receiving care at a teaching facility (OR, 0.74; 95% CI, 0.69-0.80) or from a physician provider (OR, 0.93; 95% CI, 0.88-0.98) and those with a medication possession ratio of 0.8 or higher (OR, 0.75; 95% CI, 0.71-0.80) were less likely to undergo repeat testing. Among 13,114 patients who met the optional LDL-C target level of less than 70 mg/dL, repeat lipid testing was performed in 8177 (62.4% of those with LDL-C levels of <70 mg/dL) during 11 follow-up months. One-third of patients having CHD with LDL-C levels at goal underwent repeat lipid panels. Our results highlight areas for quality improvement initiatives to reduce redundant lipid testing. These efforts would be more important if the forthcoming cholesterol guidelines adopt a medication dose-based approach in place of the current treat-to-target approach.


Relative efficacy of casein or soya protein combined with palm or safflower-seed oil on hyperuricaemia in rats.
Diets that ameliorate the adverse effects of uric acid (UA) on renal damage deserve attention. The effects of casein or soya protein combined with palm or safflower-seed oil on various serum parameters and renal histology were investigated on hyperuricaemic rats. Male Wistar rats administered with oxonic acid and UA to induce hyperuricaemia were fed with casein or soya protein plus palm- or safflower-seed oil-supplemented diets. Normal rats and hyperuricaemic rats with or without allopurinol treatment (150 mg/l in drinking water) were fed with casein plus maize oil-supplemented diets. After 8 weeks, allopurinol treatment and soya protein plus safflower-seed oil-supplemented diet significantly decreased serum UA in hyperuricaemic rats (one-way ANOVA; P < 0.05). In addition, soya protein and casein attenuated hyperuricaemia-induced decreases in serum albumin and insulin, respectively (two-way ANOVA; P < 0.05). Safflower-seed oil significantly decreased serum TAG and UA, whereas palm oil significantly increased serum cholesterol, TAG, blood urea N and creatinine. However, soya protein significantly decreased renal NO and nitrotyrosine and palm oil significantly decreased renal nitrotyrosine, TNF-alpha and interferon-gamma and increased renal transforming growth factor-beta. Casein with safflower-seed oil significantly attenuated renal tubulointerstitial nephritis, crystals and fibrosis. Comparing casein v. soya protein combined with palm or safflower-seed oil, the results support that casein with safflower-seed oil may be effective in attenuating hyperuricaemia-associated renal damage, while soya protein with safflower-seed oil may be beneficial in lowering serum UA and TAG.


Susceptibility and reactivity of sheep to Trichinella spiralis infection.
Susceptibility and reactive manifestation to Trichinella spiralis infection were studied in atypical hosts (sheep) for the period of 247 days. Sheep produced anti-Trichinella antibodies as early on Day 11 (low titer 1:200), with maximum reached at Day 35 (titer 1:800). From Day 42 the antibody level was declining with a negative result of examination on Day 70. Mice exhibited anti-Trichinella antibodies only on Day 32 (titer 1:200). This level was rising, reaching high titer (1:1600) on Day 56. This antibody level persisted until Day 156. In the following period, a rapid decrease in the titer was observed (Graph). On Day 32, T. spiralis larvae in sheep were present in all groups of the muscles examined. The highest larval counts during the entire experiment were detected in the masseter. The initially high counts in the diaphragm and tongue were reduced to only 1/4 or 1/10 at the end of the experiment. In mice, the larvae occurred evenly throughout the entire experiment (Tab. I). The first appearance of a capsule around the T. spiralis larva in muscles was observed on Day 32 p. i. No cell response was detected around the capsule (Fig. 1). Neither was any response observed around necrotizing larvae, even though the surrounding myofibrils were caused to die off (Fig. 2). Certain differences in the degree of myofibril degradation by larvae were evident as early as on Day 32. The least damaged myofibrils were those in the masseter, tongue and diaphragm. This finding correlates with the histological recovery of a different number of necrotized larvae from the individual muscle groups examined. Fresh blood extravassations around larvae were observed on Day 59 (Fig. 3). They could be caused by the migration of larvae to a parasitation site. Live uncapsulated larvae were also found on Day 115 p. i. (Fig. 4). An increased cellular presence around some larvae was observed on Day 84. The larvae surrounded by lymphocytes consequently died off, those without lymphocytic responses formed capsules and survived (Fig. 5). The necrotizing larvae were subject to a powerful phagocytic process, presented by histiocytes, forming multinuclear symplasms (Fig. 6). On Day 11 p. i., larvae inside a capsule were dying off as well. The initial stage of larval necrosis in a capsule is also accompanied by an increased lymphocytic response (Fig. 7). The condition of larvae in capsules and the cellular unresponsiveness as late as on Day 247 indicate the long-lasting viability of the larvae. The capsules surrounding T. spiralis larvae in mice were distinctly seen as early as on Day 32 p. i. Lymphocytic aggregations around the capsule were observed throughout the entire experiment (247 days)-Fig. 8.


Women's experience of post-traumatic growth following a traumatic birth: an interpretive phenomenological analysis.
Approximately 3-5% of women experience post-traumatic stress disorder following birth; positive experiences that can follow traumatic birth are under-researched. To explore how women experience post-traumatic growth following a traumatic birth. Interpretative Phenomenological Analysis was used to explore experiences of women who self-identified as having found positive benefits through coping with a traumatic birth. Eight women who had birthed in the past five years in the UK were recruited online and interviewed face-to-face. Three superordinate themes were developed: 'The total opposite to what I'd expected' (participants' struggle to understand and integrate their birth experience in the postnatal period); 'I see it a bit differently now' (processes experienced in coming to a place of growth); and 'A much better place' (experienced growth 'outcomes'). Faced with shattered assumptions whereby their birth experiences contrasted with their expectations, participants felt their identities as women and mothers were challenged. In overcoming these challenges, participants described actively striving to cope and make sense of their experience. Multiple factors facilitated this, notably partner support, telling their birth story, acknowledging the impact and developing a compassionate narrative. Although growth was experienced in ways commonly reported by survivors of challenging life events, some aspects appeared pertinent to birth trauma. Widening our understanding of the range of experiences following traumatic birth and making these narratives public may offer hope for some women and families and inform health professionals' education and practice. Further research would be needed before advocating interventions to foster post-traumatic growth.


Microsurgical treatment of neurovascular disease. Part I. Personnel, equipment, extracranial-intracranial anastomosis.
This study was prepared with the objective of directing the attention of the medical community to advances during the last decade in the operative treatment of cerebral ischemia, intracranial aneurysms, and cerebral and spinal vascular malformations. The requirements for specialized personnel, facilities, and instrumentation are detailed for those who are unfamiliar with the complexities of microneurosurgery. The importance of adequate training in the intricacies of extracranial-intracranial anastomotic procedures is stressed. Reported results of this operation are discussed. It is anticipated that a recently organized cooperative study will provide the data necessary for evaluation of long term benefits. Attention is given to grading the clinical status of patients with intracranial aneurysms in advance of surgery and the importance of operating upon good risk patients. How to manage the relatively large numbers of poor risk patients with aneurysms unsuitable for immediate operation is discussed. Sophisticated radiological procedures utilized in the diagnosis, management, and evaluation of therapy are described. The role and significance of properly administered anesthesia are discussed in relation to the performance of complicated and time-consuming microsurgery on the brain and spinal cord. The study group has tried to present this material objectively and to urge those individuals working in this difficult field to acquire the special skills and instrumentation of microneurosurgery, to implement the "team-of-experts" concept in the operating room, and to enlist the support of related disciplines.


Determination of Optimal Coronary Flow for the Preservation of "Donation after Circulatory Death" in Murine Heart Model.
Donation after circulatory death donors (DCD) have the potential to increase the number of heart transplants. The DCD hearts undergo an extended period of warm ischemia, which mandates the use of machine perfusion preservation if they are to be successfully recovered for transplantation. Because the minimum coronary artery flow needed to meet the basal oxygen demand (DCRIT) of a DCD heart during machine perfusion preservation is critical and yet unknown, we studied this in a DCD rat heart model. Adult male rats were anesthetized, intubated, heparinized, and paralyzed with vecuronium. The DCD hearts (n = 9) were recovered 30 minutes after circulatory death whereas non-DCD control hearts (n = 12) were recovered without circulatory death. Hearts were perfused through the aorta with an oxygenated Belzer Modified Machine Perfusion Solution (A3-Bridge to Life Ltd. Columbia, SC) at 15°C or 22°C starting at a flow index of 300 ml/100 g/min and decreasing by 40 ml/100 g/min every 10 minutes. Inflow (aortic) and outflow (inferior vena cava) perfusate samples were collected serially to assess the myocardial oxygen consumption index (MVO2) and O2 extraction ratio. The DCRIT is the minimum coronary flow below which the MVO2 becomes flow dependent. The MVO2, DCRIT, and oxygen extraction ratios were higher in DCD hearts compared with control hearts. The DCRIT for DCD hearts was achieved only at 15°C and was significantly higher (131.6 ± 7 ml/100 g/min) compared with control hearts (107.7 ± 8.4 ml/100 gm/min). The DCD hearts sustain warm ischemic damage and manifest higher metabolic needs during machine perfusion. Establishing adequate coronary perfusion is critical to preserving organ function for potential heart transplantation.


Delayed awakening after cardiac arrest: prevalence and risk factors in the Parisian registry.
Although prolonged unconsciousness after cardiac arrest (CA) is a sign of poor neurological outcome, limited evidence shows that a late recovery may occur in a minority of patients. We investigated the prevalence and the predictive factors of delayed awakening in comatose CA survivors treated with targeted temperature management (TTM). Retrospective analysis of the Parisian Region Out-of-Hospital CA Registry (2008-2013). In adult comatose CA survivors treated with TTM, sedated with midazolam and fentanyl, time to awakening was measured starting from discontinuation of sedation at the end of rewarming. Awakening was defined as delayed when it occurred after more than 48 h. A total of 326 patients (71 % male, mean age 59 ± 16 years) were included, among whom 194 awoke. Delayed awakening occurred in 56/194 (29 %) patients, at a median time of 93 h (IQR 70-117) from discontinuation of sedation. In 5/56 (9 %) late awakeners, pupillary reflex and motor response were both absent 48 h after sedation discontinuation. In multivariate analysis, age over 59 years (OR 2.1, 95 % CI 1.0-4.3), post-resuscitation shock (OR 2.6 [1.3-5.2]), and renal insufficiency at admission (OR 3.1 [1.4-6.8]) were associated with significantly higher rates of delayed awakening. Delayed awakening is common among patients recovering from coma after CA. Renal insufficiency, older age, and post-resuscitation shock were independent predictors of delayed awakening. Presence of unfavorable neurological signs at 48 h after rewarming from TTM and discontinuation of sedation did not rule out recovery of consciousness in late awakeners.


On the origin of gamma-ray bursts.
Gamma-ray bursts are the most energetic explosions in the Universe, occurring at cosmological distances. The initial phase of the emission from these bursts is predominantly of gamma rays and stems from a highly relativistic outflow. The nature of this emission is still under debate. Here, I present the interpretation that the peak in the photon spectrum can be attributed to the black-body emission of the photosphere of the outflow, having a temperature of approximately 109K. An additional non-thermal spectral component can be attributed to additional dissipation of the kinetic energy in the outflow. This two-component model can be well fitted to most instantaneous spectra. Interestingly, the thermal component exhibits a recurring behaviour over emission pulse structures. Both the temperature and the energy flux vary as broken power laws. During the pre-break phase, the temperature is approximately constant while the energy flux rises. Furthermore, the ratio of the observed thermal flux to the emergent flux increases as a power law over the whole pulse. It is argued that these observations hold the key to our understanding of the prompt emission and the properties of the site from which it emanates.


Alpha-Band Phase Modulates Bottom-up Feature Processing.
Recent studies reveal that attention operates in a rhythmic manner, that is, sampling each location or feature alternatively over time. However, most evidence derives from top-down tasks, and it remains elusive whether bottom-up processing also entails dynamic coordination. Here, we developed a novel feature processing paradigm and combined time-resolved behavioral measurements and electroencephalogram (EEG) recordings to address the question. Specifically, a salient color in a multicolor display serves as a noninformative cue to capture attention and presumably reset the oscillations of feature processing. We then measured the behavioral performance of a probe stimulus associated with either high- or low-salient color at varied temporal lags after the cue. First, the behavioral results (i.e., reaction time) display an alpha-band (~8 Hz) profile with a consistent phase lag between high- and low-salient conditions. Second, simultaneous EEG recordings show that behavioral performance is modulated by the phase of alpha-band neural oscillation at the onset of the probes. Finally, high- and low-salient probes are associated with distinct preferred phases of alpha-band neural oscillations. Taken together, our behavioral and neural results convergingly support a central function of alpha-band rhythms in feature processing, that is, features with varied saliency levels are processed at different phases of alpha neural oscillations.


Epicardial cell shape and maturation are regulated by Wt1 via transcriptional control of Bmp4.
The epicardium plays a crucial role in embryonic heart development and adult heart repair; however, the molecular events underlying its maturation remain unknown. Wt1, one of the main markers of the embryonic epicardium, is essential for epicardial development and function. Here, we analyse the transcriptomic profile of epicardial-enriched cells at different stages of development and from control and epicardial-specific Wt1 knockout (Wt1KO) mice. Transcriptomic and cell morphology analyses of epicardial cells from epicardial-specific Wt1KO mice revealed a defect in the maturation process of the mutant epicardium, including sustained upregulation of Bmp4 expression and the inability of mutant epicardial cells to transition into a mature squamous phenotype. We identified Bmp4 as a transcriptional target of Wt1, thus providing a molecular basis for the retention of the cuboidal cell shape observed in the Wt1KO epicardium. Accordingly, inhibition of the Bmp4 signalling pathway both ex vivo and in vivo rescued the cuboidal phenotype of the mutant epicardium. Our findings indicate the importance of the cuboidal-to-squamous transition in epicardial maturation, a process regulated by Wt1.


Efficacy and safety of intensive insulin therapy for critically ill neurologic patients: a meta-analysis.
Whether intensive insulin therapy (IIT) may improve clinical outcomes for patients admitted to intensive care units, especially critically ill neurologic patients, is still debated. In the present study, we performed a meta-analysis of literature comparing the efficacy and safety of IIT and conventional insulin therapy (CIT) for critically ill neurologic patients in terms of mortality, infection rate, neurologic outcome, and hypoglycemia. We searched for published reports of studies of randomized control trials (up to March 10, 2011) of patients admitted to neurologic intensive care units and investigated an IIT (target of blood glucose control <120 mg/dL) with a control of CIT. Data were abstracted by a standardized protocol. We retrieved reports of five studies involving 924 patients. The risk of mortality, infection rate, and neurologic outcome did not differ with IIT or CIT. However, the incidence of hypoglycemic episodes was significantly higher with IIT than CIT (78.8% vs. 48.9%), with a relative risk of 2.62 (95% confidence interval [CI]: 1.07-6.43; p < 0.04). As compared with CIT, IIT may not benefit critically ill neurologic patients in terms of mortality, infection rate, or neurologic outcome and in fact may be associated with increased hypoglycemic complications. Therefore, IIT cannot be recommended over conventional control for critical neurologic disease, but further study is warranted.


Denosumab in the treatment of glucocorticoid-induced osteoporosis: a systematic review and meta-analysis.
Glucocorticoid-induced osteoporosis (GIOP) is the most common form of secondary osteoporosis. In May 2018, denosumab was approved for the treatment of GIOP in men and women at high risk of fracture. We undertook a systematic review and meta-analysis to summarize the efficacy and safety of denosumab in the prevention and treatment of GIOP. We searched PubMed, CINAHL, American College of Rheumatology and American Society for Bone and Mineral Research meeting abstracts for relevant studies. We included studies in which subjects were taking systemic glucocorticoid therapy and were assigned to take denosumab or control therapy, and assessed the effect of treatment on areal bone mineral density (BMD), fractures and/or safety. Three eligible studies were included in the primary meta-analysis. Denosumab significantly increased lumbar spine BMD (2.32%, 95% CI 1.73%, 2.91%, P<0.0001) and hip BMD (1.52%, 95% CI 1.1%,1.94%, P<0.0001) compared to bisphosphonates. Adverse events, serious adverse events and fractures were similar between denosumab and bisphosphonate arms. Results suggest that denosumab is superior to bisphosphonates in its effects on lumbar spine and total hip BMD in patients with GIOP. There was no difference in the incidence of infections, adverse events or serious adverse events. Studies were underpowered to detect differences in the risk of fracture. Denosumab is a reasonable option for treatment of GIOP. However, further studies are needed to guide transitions off denosumab.


Effects of polarization dependent gain and dynamic birefringence of the SOA on the performance of the ultrafast nonlinear interferometer gate.
The model of ultrafast nonlinear interferometer gate accounting for the polarization dependent gain and dynamic birefringence has been developed. It is shown that these effects can lead to appearance of the satellite pulses and limit the achievable extinction ratio. The effect of dynamic birefringence can be completely eliminated by proper adjustment of SOA axes but it can strongly impair the performance of the gate when the axes are misaligned. The switching window should not be set to half of a bit-period to avoid interference of the satellites of neighbor pulses.


Nanoparticles as new tools for inhibition of cancer angiogenesis.
Angiogenesis is known as one of the hallmarks of cancer. Multiple lines evidence indicated that vascular endothelium growth factor (VEGF) is a key player in the progression of angiogenesis and exerts its functions via interaction with tyrosine kinase receptors (TKRs). These receptors could trigger a variety of cascades that lead to the supply of oxygen and nutrients to tumor cells and survival of these cells. With respect to pivotal role of angiogenesis in the tumor growth and survival, finding new therapeutic approaches via targeting angiogenesis could open a new horizon in cancer therapy. Among various types of therapeutic strategies, nanotechnology has emerged as new approach for the treatment of various cancers. Nanoparticles (NPs) could be used as effective tools for targeting a variety of therapeutic agents. According to in vitro and in vivo studies, NPs are efficient in depriving tumor cells from nutrients and oxygen by inhibiting angiogenesis. However, the utilization of NPs are associated with a variety of limitations. It seems that new approaches such as NPs conjugated with hydrogels could overcome to some limitations. In the present review, we summarize various mechanisms involved in angiogenesis, common anti-angiogenesis strategies, and application of NPs for targeting angiogenesis in various cancers.


[Interaction of the Cu(Lys)2 complex with the NADPH-dependent microsomal electron transport system and microsomal membrane].
The low molecular weight analog of superoxide dismutase, th Cu(Lys)2 complex inhibits the oxidation of type I (piperidinoanthraquinone) and type II(aniline) substrates catalyzed by cytochrome P-450. This fact is not associated with the conversion of cytochrome P-450 into the inactive form--cytochrome P-420. It was shown that the method of determining the activity of NADPH-cytochrome c-reductase by the cytochrome c reduction rate cannot be employed in the presence of the Cu(Lys)2 and Cu(Tyr)2 complexes. Potassium ferricyanide was used to demonstrate that Cu(Lys)2 does not affect the NADPH-cytochrome c-reductase activity. The inhibiting effect of Cu(Lys)2 on the microsomal oxidation of the substrates does not result from the interaction of the complex with the components of the NADPH-dependent microsomal electron transport system. The effect observed can be accounted for by the dismutase-like action of the complex on the superoxide radicals generated during the microsomal oxidation of the substrates. The interaction of Cu(Lys)2 with the membrane was studied by the ESR method using stable nitroxide rabicais. It was shown that the complex cannot freely diffuse through the lipid bilayer membrane. A model for Cu(Lys)2 incorporation into the membrane is proposed.


Morphological, morphometric, meristic and osteological evidence for two species of hake (Actinopterygii: Gadiformes: Merluccius) in Argentinean waters.
Morphologically, both classic and landmark-based morphometry and meristic analyses of 241 specimens of Merluccius, along with the re-examination of six paratype specimens of Merluccius hubbsi, the holotype and three paratypes of Merluccius patagonicus and the syntype of Merluccius australis revealed the presence of only two species of Merluccius in Argentinean waters. Internal structures (hyomandibula, urohyal and sagitta otolith) of M. hubbsi were compared to those reported for M. patagonicus and were shown to have identical morphology. Type specimens of M. patagonicus showed a complete overlap in morphometric and meristic characters with M. hubbsi, whereas M. australis had a greater number of second dorsal and anal-fin rays, and more lateral-line scales. In addition, M. australis had a smaller eye and longer snout than M. hubbsi and M. patagonicus. The results indicate that there is no evidence for a third hake species in Argentinean waters. Merluccius patagonicus is a synonym of M. hubbsi.


Activation of β2-adrenergic receptor by (R,R')-4'-methoxy-1-naphthylfenoterol inhibits proliferation and motility of melanoma cells.
(R,R')-4'-methoxy-1-naphthylfenoterol [(R,R')-MNF] is a highly-selective β2 adrenergic receptor (β2-AR) agonist. Incubation of a panel of human-derived melanoma cell lines with (R,R')-MNF resulted in a dose- and time-dependent inhibition of motility as assessed by in vitro wound healing and xCELLigence migration and invasion assays. Activity of (R,R')-MNF positively correlated with the β2-AR expression levels across tested cell lines. The anti-motility activity of (R,R')-MNF was inhibited by the β2-AR antagonist ICI-118,551 and the protein kinase A inhibitor H-89. The adenylyl cyclase activator forskolin and the phosphodiesterase 4 inhibitor Ro 20-1724 mimicked the ability of (R,R')-MNF to inhibit migration of melanoma cell lines in culture, highlighting the importance of cAMP for this phenomenon. (R,R')-MNF caused significant inhibition of cell growth in β2-AR-expressing cells as monitored by radiolabeled thymidine incorporation and xCELLigence system. The MEK/ERK cascade functions in cellular proliferation, and constitutive phosphorylation of MEK and ERK at their active sites was significantly reduced upon β2-AR activation with (R,R')-MNF. Protein synthesis was inhibited concomitantly both with increased eEF2 phosphorylation and lower expression of tumor cell regulators, EGF receptors, cyclin A and MMP-9. Taken together, these results identified β2-AR as a novel potential target for melanoma management, and (R,R')-MNF as an efficient trigger of anti-tumorigenic cAMP/PKA-dependent signaling in β2-AR-expressing lesions.


Hypotensive peptides from milk proteins.
Hypertension is the major controllable risk factor associated with cardiovascular disease (CVD) events such as myocardial infarction, stroke, heart failure, and end-stage diabetes. A 5 mm Hg decrease in blood pressure has been equated with approximately 16% decrease in CVD. In the U.S. alone current annual antihypertensive drug costs are approximately dollars 15 billion. The renin-angiotensin-aldosterone system is a target for blood pressure control. Cleavage of angiotensinogen by renin produces angiotensin I which is subsequently hydrolyzed by angiotensin-I-converting enzyme (ACE) to angiotensin II (a potent vasoconstrictor). Various side effects are associated with the use of ACE inhibitory drugs in the control of blood pressure including hypotension, increased potassium levels, reduced renal function, cough, angioedema, skin rashes, and fetal abnormalities. Milk proteins, both caseins and whey proteins, are a rich source of ACE inhibitory peptides. Several studies in spontaneously hypertensive rats show that these casokinins and lactokinins can significantly reduce blood pressure. Furthermore, a limited number of human studies have associated milk protein-derived peptides with statistically significant hypotensive effects (i.e., lower systolic and diastolic pressures). The advent of effective milk protein based functional food ingredients/nutraceuticals for the prevention/control of blood pressure therefore has the potential to significantly reduce global healthcare cost.


Evaluation of fertility outcome as a consequence of intravaginal inoculation with sperm-impairing micro-organisms in a mouse model.
The concept of infertility as a result of asymptomatic microbial colonization of the female reproductive tract has been neglected to date. However, increasing incidence of infertility and advanced research has drawn attention towards this idea. Many of these micro-organisms have been reported to bring about adverse changes in sperm parameters in vitro, but their in vivo potential to cause infertility is still a controversy. The present study was carried out to observe what effect the intravaginal inoculation of sperm-agglutinating Serratia marcescens and sperm-immobilizing Candida albicans had in the reproductive tract and consequently in fertility outcome. When these strains were intravaginally inoculated into female BALB/c mice at 10(4), 10(6) and 10(8) c.f.u. in 20 µl PBS for 10 consecutive days, with mating of mice on day 12, the results showed 100 % decrease in fertility in all groups as compared with control mice receiving PBS alone. Furthermore, no clinical or histopathological changes were observed in the reproductive organs (i.e. ovary, uterus and vagina), suggesting that colonization of the genital tract with sperm-impairing micro-organisms could be a feasible reason for female infertility.


Altered inhibition of Cx26 hemichannels by pH and Zn2+ in the A40V mutation associated with keratitis-ichthyosis-deafness syndrome.
Excessive opening of undocked Cx26 hemichannels in the plasma membrane is associated with disease pathogenesis in keratitis-ichthyosis-deafness (KID) syndrome. Thus far, excessive opening of KID mutant hemichannels has been attributed, almost solely, to aberrant inhibition by extracellular Ca(2+). This study presents two new possible contributing factors, pH and Zn(2+). Plasma pH levels and micromolar concentrations of Zn(2+) inhibit WT Cx26 hemichannels. However, A40V KID mutant hemichannels show substantially reduced inhibition by these factors. Using excised patches, acidification was shown to be effective from either side of the membrane, suggesting a protonation site accessible to H(+) flux through the pore. Sensitivity to pH was not dependent on extracellular aminosulfonate pH buffers. Single channel recordings showed that acidification did not affect unitary conductance or block the hemichannel but rather promoted gating to the closed state with transitions characteristic of the intrinsic loop gating mechanism. Examination of two nearby KID mutants in the E1 domain, G45E and D50N, showed no changes in modulation by pH or Zn(2+). N-bromo-succinimide, but not thiol-specific reagents, attenuated both pH and Zn(2+) responses. Individually mutating each of the five His residues in WT Cx26 did not reveal a key His residue that conferred sensitivity to pH or Zn(2+). From these data and the crystal structure of Cx26 that suggests that Ala-40 contributes to an intrasubunit hydrophobic core, the principal effect of the A40V mutation is probably a perturbation in structure that affects loop gating, thereby affecting multiple factors that act to close Cx26 hemichannels via this gating mechanism.


Pelleting and extrusion can ameliorate negative effects of toasting of rapeseed meal on protein digestibility in growing pigs.
Toasting time (TT) of rapeseed meal (RSM), the diet processing (DP) method and the interaction between both on the apparent CP digestion along the gastrointestinal tract and the apparent ileal digestibility (AID) of amino acids of growing pigs were investigated. The experiment consisted of a 3×3 factorial design of TT of RSM (0, 60 and 120 min) and DP method (mash, pelleting and extrusion). In total, 81 boars with a starting BW of 20 kg were euthanized 4 h after their last feeding. The gastrointestinal tract was dissected and the small intestine divided in three sections of similar length. Samples were collected from the stomach, 1.5 m from the ends of each of the three sections of the small intestine, and the rectum. The apparent digestibility (AD) of CP for each of the small intestine sections was used to calculate the rate of CP digestion. Increasing the TT of RSM resulted in lower protein solubility, lower lysine/reactive lysine contents and higher protein denaturation, indicative of the occurrence of protein aggregation and Maillard reactions. There were significant effects (P⩽0.01) of TT on the AD of CP in the different sections of the gastrointestinal tract. The rate of CP digestion of the 0 min toasted RSM diets was 23% and 35% higher than that of the 60 and 120 min toasted RSM diets, respectively. There was a significant interaction (P=0.04) between TT and DP for the AID of CP. Although pelleting of the 0 and 60 min toasted RSM diets did not change the AID of CP with respect to the mash diets, pelleting of the 120 min toasted RSM diet increased the AID of CP by 9.3% units. Extrusion increased the AID of CP of the 0 and 60 min toasted RSM diets by 3.4% and 4.3% units with respect to the mash diets, whereas extrusion of the 120 min toasted RSM diet increased the AID of CP by 6.9% units. Similar positive effects of pelleting and extrusion were obtained for the AID of lysine and reactive lysine, especially in the diets with higher TT. In conclusion, processing (pelleting and extrusion) of RSM containing diets can ameliorate the negative effects of RSM toasting on protein and amino acid digestibility; these effects were larger for the RSM toasted for longer times.


Effectiveness and safety of cabazitaxel chemotherapy for metastatic castration-resistant prostatic carcinoma on Turkish patients (The Anatolian Society of Medical Oncology).
Prostate cancer is among the most common cancers in males. Prostate cancer is androgen dependent in the beginning, but as time progresses, it becomes refractory to androgen deprivation treatment. At this stage, docetaxel has been used as standard treatment for years. Cabazitaxel has become the first chemotherapeutic agent which has been shown to increase survival for patients with metastatic Castrate Resistant Prostate Cancer (mCRPC) that progresses after docetaxel. Phase 3 TROPIC study demonstrated that cabazitaxel prolongs survival. In this study, we evaluated a total of 103 patients who took cabazitaxel chemotherapy for mCRPC diagnosis in 21 centers of Turkey, retrospectively. This study included patients who progressed despite docetaxel treatments, had ECOG performance score between 0-2, and used cabazitaxel treatment. Patients received cabazitaxel 25 mg/m2 at every 3 weeks, and prednisolone 5 mg twice a day. Median number of cabazitaxel cures was 5.03 (range: 1-17). Cabazitaxel response evaluation detected that 34% of the patients had a partial response, 22.3% had stable disease and 32% had a progressive disease. Grade 3-4 hematological toxicities were neutropenia (28.2%), neutropenic fever (14.5%), anemia (6.7%), and thrombocytopenia (3.8%). In our study, median progression-free survival (PFS) was 7.7 months and overall survival (OS) was 10.6 months. This study reflects toxicity profile of Turkish patients as a Caucasian race. We suggest that cabazitaxel is a safe and effective treatment option for mCRPC patients who progress after docetaxel. Moreover, ethnicity may play important roles both in treatment response and in toxicity profile.


Rationale for the Development of a Novel Clinical Grading Scale for Postoperative Facial Nerve Function: Results of a Multidisciplinary International Working Group.
The objective of the current study was to present the results of an international working group survey identifying perceived limitations of existing facial nerve grading scales to inform the development of a novel grading scale for assessing early postoperative facial paralysis that incorporates regional scoring and is anchored in recovery prognosis and risk of associated complications. Survey. A working group of 48 multidisciplinary clinicians with expertise in skull base, cerebellopontine angle, temporal bone, or parotid gland surgery. House-Brackmann grade is the most widely used system to assess facial nerve function among working group members (81%), although more than half (54%) agreed that the system they currently use does not adequately estimate the risk of associated complications, such as corneal injury, and confidence in interrater and intrarater reliability is generally low. Simplicity was ranked as the most important attribute of a novel postoperative facial nerve grading system to increase the likelihood of adoption, followed by reliability and accuracy. There was widespread consensus (91%) that the eye is the most critical facial region to focus on in the early postoperative setting. Members were invited to submit proposed grading systems in alignment with the objectives of the working group for subsequent validation. From these data, we plan to develop a simple, clinically anchored, and reproducible staging system with regional scoring for assessing early postoperative facial nerve function after surgery of the skull base, cerebellopontine angle, temporal bone, or parotid gland.


Quantitative in vivo microscopy: the return from the 'omics'.
The confluence of recent advances in microscopy instrumentation and image analysis, coupled with the widespread use of GFP-like proteins as reporters of gene expression, has opened the door to high-throughput in vivo studies that can provide the morphological and temporal context to the biochemical pathways regulating cell function. We are now able to quantify the concentration and three-dimensional distribution of multiple spectrally resolved GFP-tagged proteins. Using automatic segmentation and tracking we can then measure the dynamics of the processes in which these elements are involved. In this way, parallel studies are feasible where multiple cell colonies treated with drugs or gene expression repressors can be monitored and analyzed to study the dynamics of relevant biological processes.


Schizophrenia-An Overview.
Schizophrenia is a common, severe mental illness that most clinicians will encounter regularly during their practice. This report provides an overview of the clinical characteristics, epidemiology, genetics, neuroscience, and psychopharmacology of schizophrenia to provide a basis to understand the disorder and its treatment. This educational review is integrated with a clinical case to highlight how recent research findings can inform clinical understanding. The first theme considered is the role of early-life environmental and genetic risk factors in altering neurodevelopmental trajectories to predispose an individual to the disorder and leading to the development of prodromal symptoms. The second theme is the role of cortical excitatory-inhibitory imbalance in the development of the cognitive and negative symptoms of the disorder. The third theme considers the role of psychosocial stressors, psychological factors, and subcortical dopamine dysfunction in the onset of the positive symptoms of the disorder. The final theme considers the mechanisms underlying treatment for schizophrenia and common adverse effects of treatment. Schizophrenia has a complex presentation with a multifactorial cause. Nevertheless, advances in neuroscience have identified roles for key circuits, particularly involving frontal, temporal, and mesostriatal brain regions, in the development of positive, negative, and cognitive symptoms. Current pharmacological treatments operate using the same mechanism, blockade of dopamine D2 receptor, which contribute to their adverse effects. However, the circuit mechanisms discussed herein identify novel potential treatment targets that may be of particular benefit in symptom domains not well served by existing medications.


Safety profile of autologous macrophage therapy for liver cirrhosis.
Therapies to reduce liver fibrosis and stimulate organ regeneration are urgently needed. We conducted a first-in-human, phase 1 dose-escalation trial of autologous macrophage therapy in nine adults with cirrhosis and a Model for End-Stage Liver Disease (MELD) score of 10-16 (ISRCTN 10368050). Groups of three participants received a single peripheral infusion of 107, 108 or up to 109 cells. Leukapheresis and macrophage infusion were well tolerated with no transfusion reactions, dose-limiting toxicities or macrophage activation syndrome. All participants were alive and transplant-free at one year, with only one clinical event recorded, the occurrence of minimal ascites. The primary outcomes of safety and feasibility were met. This study informs and provides a rationale for efficacy studies in cirrhosis and other fibrotic diseases.


Site-Specific Profiling of N-Glycans in Drosophila melanogaster.
Drosophila melanogaster is a well-studied and highly tractable genetic model system for deciphering the molecular mechanisms underlying various biological processes. Although being one of the most critical post-translational modifications of proteins, the understanding of glycosylation in Drosophila is still lagging behind compared with that of other model organisms. In this study, we systematically investigated the site-specific N-glycan profile of Drosophila melanogaster using intact glycopeptide analysis technique. This approach identified the glycans, proteins, and their glycosites in Drosophila, as well as information on site-specific glycosylation, which allowed us to know which glycans are attached to which glycosylation sites. The results showed that the majority of N-glycans in Drosophila were high-mannose type (69.3%), consistent with reports in other insects. Meanwhile, fucosylated N-glycans were also highly abundant (22.7%), and the majority of them were mono-fucosylated. In addition, 24 different sialylated glycans attached with 16 glycoproteins were identified, and these proteins were mainly associated with developmental processes. Gene ontology analysis showed that N-glycosylated proteins in Drosophila were involved in multiple biological processes, such as axon guidance, N-linked glycosylation, cell migration, cell spreading, and tissue development. Interestingly, we found that seven glycosyltransferases and four glycosidases were N-glycosylated, which suggested that N-glycans may play a regulatory role in the synthesis and degradation of N-glycans and glycoproteins. To our knowledge, this work represents the first comprehensive analysis of site-specific N-glycosylation in Drosophila, thereby providing new perspectives for the understanding of biological functions of glycosylation in insects.


Leber's hereditary optic neuropathy is potentially associated with a novel m.5587T>C mutation in two pedigrees.
Mitochondrial (mt)DNA mutations have been revealed to be associated with Leber's hereditary optic neuropathy (LHON). The present study conducted clinical, genetic and molecular evaluations of two Han Chinese families. A total of 4 (3 men and 1 female) out of 14 matrilineal relatives in the families exhibited visual impairment with variable severity and age of onset. The average age of onset of visual loss was 20.5 years old. Molecular analysis of the complete mitochondrial genome in these pedigrees demonstrated that the three primary mutations associated with LHON were not detected; however, the homoplasmic m.5587T>C mutation was identified, which was localized at the end of the mitochondrially encoded transfer (t)RNA alanine gene and may alter the tertiary structure of this tRNA. Subsequently, this structural alteration may result in tRNA metabolism failure. In addition, distinct sets of mtDNA polymorphisms belonging to haplogroup F1 were detected in both families tested. The findings of the present study suggested that the m.5587T>C mutation may be involved in the pathogenesis of visual impairment. In addition, the mtDNA variant m.15024G>A(p.C93H) in the mitochondrially encoded cytochrome B gene was detected in both families, which exhibited evolutionary conservation, indicating it may serve a potential modifying role in the development of visual impairment associated with m.5587T>C mutation in these families. Furthermore, other modifying factors, including nuclear modifier genes, and environmental and personal factors may also contribute to the development of LHON in subjects carrying this mutation.


The characterization of RNA viruses in tropical seawater using targeted PCR and metagenomics.
Viruses have a profound influence on the ecology and evolution of plankton, but our understanding of the composition of the aquatic viral communities is still rudimentary. This is especially true of those viruses having RNA genomes. The limited data that have been published suggest that the RNA virioplankton is dominated by viruses with positive-sense, single-stranded (+ss) genomes that have features in common with those of eukaryote-infecting viruses in the order Picornavirales (picornavirads). In this study, we investigated the diversity of the RNA virus assemblages in tropical coastal seawater samples using targeted PCR and metagenomics. Amplification of RNA-dependent RNA polymerase (RdRp) genes from fractions of a buoyant density gradient suggested that the distribution of two major subclades of the marine picornavirads was largely congruent with the distribution of total virus-like RNA, a finding consistent with their proposed dominance. Analyses of the RdRp sequences in the library revealed the presence of many diverse phylotypes, most of which were related only distantly to those of cultivated viruses. Phylogenetic analysis suggests that there were hundreds of unique picornavirad-like phylotypes in one 35-liter sample that differed from one another by at least as much as the differences among currently recognized species. Assembly of the sequences in the metagenome resulted in the reconstruction of six essentially complete viral genomes that had features similar to viruses in the families Bacillarna-, Dicistro-, and Marnaviridae. Comparison of the tropical seawater metagenomes with those from other habitats suggests that +ssRNA viruses are generally the most common types of RNA viruses in aquatic environments, but biases in library preparation remain a possible explanation for this observation. Marine plankton account for much of the photosynthesis and respiration on our planet, and they influence the cycling of carbon and the distribution of nutrients on a global scale. Despite the fundamental importance of viruses to plankton ecology and evolution, most of the viruses in the sea, and the identities of their hosts, are unknown. This report is one of very few that delves into the genetic diversity within RNA-containing viruses in the ocean. The data expand the known range of viral diversity and shed new light on the physical properties and genetic composition of RNA viruses in the ocean.


Experimental Control for the Ovariectomized Rat Model: Use of Sham Versus Nonmanipulated Animal.
One of 2 models required by the U.S. Food and Drug Administration for registration of a treatment for osteoporosis, the ovariectomized (OVX) rat model, is widely used in scientific studies investigating sex hormone-deficient bone loss. The use of control nonhuman animals is critical because bone turnover may be affected by animal stress, use of anesthetic, and the mechanisms involved in wound healing. Historically, researchers have used sham-operated animals who undergo the same manipulations as the OVX rats, but ethical concerns require consideration of unmanipulated (unoperated) control animals to minimize animal distress and unnecessary procedures. Herein, we report the results of 3 studies including OVX, sham, and unmanipulated rats and the effects on bone mineral density and content (BMD/BMC) during 2 to 6 months postsurgery. Our data indicate that while OVX animals generally had lower BMD and BMC than animals in either of the control groups, no differences were observed between sham and unmanipulated animals at any of the time points assessed. However, because bone turnover is a long-term process, studies with longer duration and multiple endpoints are warranted to confirm these results.


An in situ XAS study of high surface-area IrO2 produced by the polymeric precursor synthesis.
Iridium oxide powders with a surface area of more than 1 m2 g-1 (4 m2 g-2 from the H-UPD charge) and iridium-oxide crystallites less than 10 nm across were synthesized by heat treating gels formed from citric acid, ethylene glycol and dihydrogen hexachloroiridate(iv) in air. The characteristics of the resulting material was found to be strongly dependent on the heat-treatment step in the synthesis. A single heat-treatment of the gel resulted in a material with a substantial fraction of elemental iridium metal, i.e. iridium in oxidation state zero (Ir0). Post-synthesis modification of the powder by potential cycling resulted in oxidation peaks consistent with the conversion of the metal phase to iridium oxide. Linear combination of the near-edge part of the X-ray absorption data (X-ray absorption near-edge spectroscopy, XANES) collected in situ during potential cycling and an analysis of the extended X-ray fine-structure (EXAFS) part of the spectrum showed that the overall metal fraction was not significantly affected by the cycling. The oxidation of the metal phase is therefore limited to a thin layer of oxide at the metal surface, and a significant part of the iridium is left inactive. A modification of the heat treatment procedure of the sample resulted in iridium oxide containing only insignificant amounts of elemental iridium metal.


Paradoxical allosteric effects of competitive inhibitors on neuronal alpha7 nicotinic receptor mutants.
Mutation of the conserved leucine residue, in the second transmembrane domain of the neuronal alpha7 acetylcholine receptor to a threonine (L247T) causes pleiotropic alterations of receptor properties. In this study we examined the effects of competitive inhibitors on the alpha7-L247T physiological responses. While the alpha7 competitive inhibitor dihydro-beta-erythroidine evoked a current comparable to that induced by ACh, other inhibitors such as methyllycaconitine (MLA) and alpha-bungarotoxin (alpha-Bgt) caused a blockade of alpha7-L247T to ACh activation. When applied in the absence of ACh, MLA or alpha-Bgt reduced the cell leakage current, showing that alpha7-L247T displays a significant fraction (10%) of spontaneously open channels. These data can be interpreted in terms of an allosteric model, assuming that the L247T mutant possesses a low isomerization constant L and that MLA and alpha-Bgt stabilize the closed, resting state.


A modulatory role for clathrin light chain phosphorylation in Golgi membrane protein localization during vegetative growth and during the mating response of Saccharomyces cerevisiae.
The role of clathrin light chain phosphorylation in regulating clathrin function has been examined in Saccharomyces cerevisiae. The phosphorylation state of yeast clathrin light chain (Clc1p) in vivo was monitored by [32P]phosphate labeling and immunoprecipitation. Clc1p was phosphorylated in growing cells and also hyperphosphorylated upon activation of the mating response signal transduction pathway. Mating pheromone-stimulated hyperphosphorylation of Clc1p was dependent on the mating response signal transduction pathway MAP kinase Fus3p. Both basal and stimulated phosphorylation occurred exclusively on serines. Mutagenesis of Clc1p was used to map major phosphorylation sites to serines 52 and 112, but conversion of all 14 serines in Clc1p to alanines [S(all)A] was necessary to eliminate phosphorylation. Cells expressing the S(all)A mutant Clc1p displayed no defects in Clc1p binding to clathrin heavy chain, clathrin trimer stability, sorting of a soluble vacuolar protein, or receptor-mediated endocytosis of mating pheromone. However, the trans-Golgi network membrane protein Kex2p was not optimally localized in mutant cells. Furthermore, pheromone treatment exacerbated the Kex2p localization defect and caused a corresponding defect in Kex2p-mediated maturation of the alpha-factor precursor. The results reveal a novel requirement for clathrin during the mating response and suggest that phosphorylation of the light chain subunit modulates the activity of clathrin at the trans-Golgi network.


SDF-1α/OPF/BP Composites Enhance the Migrating and Osteogenic Abilities of Mesenchymal Stem Cells.
In situ cell recruitment is a promising regenerative medicine strategy with the purpose of tissue regeneration without stem cell transplantation. This chemotaxis-based strategy is aimed at ensuring a restorative environment through the release of chemokines that promote site-specific migration of healing cell populations. Stromal cell-derived factor-1α (SDF-1α) is a critical chemokine that can regulate the migration of mesenchymal stem cells (MSCs). Accordingly, here, SDF-1α-loaded microporous oligo[poly(ethylene glycol) fumarate]/bis[2-(methacryloyloxy)ethyl] phosphate composites (SDF-1α/OPF/BP) were engineered and probed. SDF-1α/OPF/BP composites were loaded with escalating SDF-1α concentrations, namely, 0 ng/ml, 50 ng/ml, 100 ng/ml, and 200 ng/ml, and were cocultured with MSC. Scratching assay, Transwell assay, and three-dimensional migration model were utilized to assess the migration response of MSCs. Immunofluorescence staining of Runx2 and osteopontin (OPN), ELISA assay of osteocalcin (OCN) and alkaline phosphatase (ALP), and Alizarin Red S staining were conducted to assess the osteogenesis of MSCs. All SDF-1α/OPF/BP composites engendered a release of SDF-1α (>80%) during the first four days. SDF-1α released from the composites significantly promoted migration and osteogenic differentiation of MSCs documented by upregulated expression of osteogenic-related proteins, ALP, Runx2, OCN, and OPN. SDF-1α at 100 ng/ml was optimal for enhanced migration and osteogenic proficiency. Thus, designed SDF-1α/OPF/BP composites were competent in promoting the homing and osteogenesis of MSCs and thus offer a promising bioactive scaffold candidate for on-demand bone tissue regeneration.


A locally adaptive peano scanning algorithm.
This paper describes an algorithm which builds a ``Peano scanning,'' i.e., the reciprocal mapping, from [0, 1]n to [0, 1], of the well-known ``Peano curve.'' This Peano scanning is applied to a set of points in [0, 1]n and gives a one-dimensional image of it. Several applications of this technique have already been developed and are presented in this paper.


Genome-Wide Expression Profiling Unveils Autoimmune Response Signatures in the Perivascular Adipose Tissue of Abdominal Aortic Aneurysm.
Objective- Perivascular adipose tissue (PVAT) is thought to play a role in vascular homeostasis and in the pathogenesis of large vessel diseases, including abdominal aortic aneurysm (AAA). Herein, we tested the hypothesis that locally restricted transcriptional profiles characterize PVAT surrounding AAA, indicating specific dysfunctions associated with the disease. Approach and Results- Using a paired sample design to limit the effects of interindividual variation, we performed a microarray-based investigation of the PVAT transcriptome in 30 patients with AAA, comparing the adipose layer of the dilated abdominal aorta with that of the not-dilated aortic neck in each patient. Furthermore, we used a state-of-the-art data mining procedure to remove the effect of confounders produced by high-throughput gene expression techniques. We found substantial differences in PVAT gene expression clearly distinguishing the dilated from the not-dilated aorta, which increased in number and magnitude with increasing AAA diameter. Comparisons with other adipose depots (omental or subcutaneous fat) confirmed that gene expression changes are locally restricted. We dissected putative mechanisms associated with AAA PVAT dysfunction through a functional enrichment network analysis: both innate and adaptive immune-response genes along with genes related to cell-death pathways, metabolic processes of collagen, sphingolipids, aminoglycans, and extracellular matrix degradation were strongly overrepresented in PVAT of AAA compared with PVAT of the not-dilated aorta. Conclusions- Our results support a possible function of PVAT in AAA pathogenesis and suggest that AAA is an immunologic disease with an underlying autoimmune component. Interfering with these disease-specific pathways would clarify their precise role in AAA pathogenesis.


A randomized, placebo-controlled clinical trial of chiropractic and medical prophylactic treatment of adults with tension-type headache: results from a stopped trial.
Tension-type headache (TTH) is the most common headache experienced by adults in Western society. Only 2 clinical trials of spinal manipulation for adult tension-type headache have been reported, neither of which was fully controlled. In 1 trial, spinal manipulation was compared to amitriptyline. There is an urgent need for well-controlled studies of chiropractic spinal manipulation for TTH. This trial was stopped prematurely due to poor recruitment. The purposes of this report are (1) to describe the trial protocol, as it contained several novel features, (2) to report the limited data set obtained from our sample of completed subjects, and (3) to discuss the problems that were encountered in conducting this study. A randomized clinical trial was conducted with a factorial design in which adult TTH sufferers with more than 10 headaches per month were randomly assigned to four groups: real cervical manipulation + real amitriptyline, real cervical manipulation + placebo amitriptyline, sham cervical manipulation + real amitriptyline, and sham cervical manipulation + placebo amitriptyline. A baseline period of four weeks was followed by a treatment period of 14 weeks. The primary outcome was headache frequency obtained from a headache diary in the last 28 days of the treatment period. Nineteen subjects completed the trial. In the unadjusted analysis, a statistically significant main effect of chiropractic treatment was obtained (-2.2 [-10.2 to 5.8], P = .03) which was just below the 3-day reduction set for clinical importance. As well, a clinically important [corrected] effect of the combined therapies was obtained (-9 [-20.8 [corrected] to 2.9], P = .13), but this did not achieve statistical significance. In the adjusted analysis, neither the main effects of chiropractic nor amitriptyline were statistically significant or clinically important; however, the effect of the combined treatments was -8.4 (-15.8 to -1.1) which was statistically significant (P = .03) and reached our criterion for clinical importance. Although the sample size was smaller than initially required, a statistically significant and clinically important effect was obtained for the combined treatment group. There are considerable difficulties with recruitment of subjects in such a trial. This trial should be replicated with a larger sample.


Fecal microbiota transplantation for the treatment of Clostridium difficile infection: a systematic review.
By systematic review, we assessed the impact of fecal microbiota transplantation (FMT) for the treatment of Clostridium difficile (CD)-associated diarrhea. Fecal microbiota microbiota transplantation from a healthy donor into an individual with CD infection (CDI) can resolve symptoms. We conducted systematic searches in PubMed, SCOPUS, Web of Science, and Cochrane Library. The last search was run on February 8, 2013. The following Medical Subject Headings terms and keywords were used alone or in combination: Clostridium difficile; Clostridium infection; pseudomembranous colitis; feces; stools; fecal suspension; fecal transplantation; fecal transfer; fecal infusion; microbiota; bacteriotherapy; enema; nasogastric tube; colonoscopy; gastroscopy; fecal donation; donor. A critical appraisal of the clinical research evidence on the effectiveness and safety of FMT for the treatment of patients with CD-associated diarrhea was made. Twenty full-text case series, 15 case reports, and 1 randomized controlled study were included for the final analysis. Almost all patients treated with donors' fecal infusion experienced recurrent episodes of CD-associated diarrhea despite standard antibiotic treatment. Of a total of 536 patients treated, 467 (87%) experienced resolution of diarrhea. Diarrhea resolution rates varied according to the site of infusion: 81% in the stomach; 86% in the duodenum/jejunum; 93% in the cecum/ascending colon; and 84% in the distal colon. No severe adverse events were reported with the procedure. FMT seems efficacious and safe for the treatment of recurrent CDI. Hospitals should encourage the development of fecal transplantation programs to improve therapy of local patients.


A Survey on the Effect That Medical Cannabis Has on Prescription Opioid Medication Usage for the Treatment of Chronic Pain at Three Medical Cannabis Practice Sites.
The opioid epidemic continues to claim thousands of lives every year without an effective strategy useful in mitigating mortality. The use of medical cannabis has been proposed as a potential strategy to decrease opioid usage. The objective of this study was to determine how the use of medical cannabis affects prescribed opioid usage in chronic pain patients. We conducted an online convenience sample survey of patients from three medical cannabis practice sites who had reported using opioids. A total of 1181 patients responded, 656 were excluded for not using medical cannabis in combination with opioid use or not meeting the definition of chronic pain, leaving 525 patients who had used prescription opioid medications continuously for at least three months to treat chronic pain and were using medical cannabis in combination with their prescribed opioid use. Overall, 40.4% (n=204) reported that they stopped all opioids, 45.2% (n=228) reported some decrease in their opioid usage, 13.3% (n=67) reported no change in opioid usage, and 1.1% (n=6) reported an increase in opioid usage. The majority (65.3%, n=299) reported that they sustained the opioid change for over a year. Almost half (48.2%, n=241) reported a 40-100% decrease in pain while 8.6% (n=43) had no change in pain and 2.6% (n=13) had worsening pain. The majority reported improved ability to function (80.0%, n=420) and improved quality of life (87.0%, n=457) with medical cannabis. The majority (62.8%, n=323) did not want to take opioids in the future. While the change in pain level was not affected by age and gender, the younger age group had improved ability to function compared with the middle and older age groups. Patients in this study reported that cannabis was a useful adjunct and substitute for prescription opioids in treating their chronic pain and had the added benefit of improving the ability to function and quality of life.


Maternal Executive Functioning and Scaffolding in Families of Children with and without Parent-Reported ADHD.
Parental scaffolding robustly predicts child developmental outcomes, including improved self-regulation and peer relationships and fewer externalizing behaviors. However, few studies have examined parental characteristics associated with a parent's ability to scaffold. Executive functioning (EF) may be an important individual difference factor associated with maternal scaffolding that has yet to be examined empirically. Scaffolding may be particularly important for children with attention-deficit/hyperactivity disorder (ADHD) and disruptive behavior disorder (DBD) symptoms due to their core difficulties with inattention, disorganization, EF, and self-regulation, their need for greater parental structure, and higher-than-average rates of parental EF deficits. Yet, little research has examined child ADHD in relation to parental scaffolding. This cross-sectional study examined: (1) the association between maternal EF (as measured by the Hotel Test, Barkley's Deficits in Executive Functioning Scale, and Digit Span) and observed scaffolding, (2) the association between parent-reported child ADHD/DBD symptoms and scaffolding, and (3) the interaction between child ADHD/DBD symptoms and maternal EF in predicting scaffolding. In a sample of 84 mothers and their 5-10 year-old biological children (62% male) with and without parent-reported ADHD, we found that maternal EF, as measured by Digit Span and the Hotel Test, predicted observed maternal scaffolding. However, child ADHD/DBD symptoms did not significantly predict maternal scaffolding controlling for child age, maternal education, and maternal EF, nor did the interaction of maternal EF and parent-reported child ADHD/DBD symptoms. Working memory and task shifting may be key components of parental EF that could be targeted in interventions to improve parental scaffolding.


Improving general flexibility with a mind-body approach: a randomized, controlled trial using neuro emotional Technique®.
General flexibility is a key component of health, well-being, and general physical conditioning. Reduced flexibility has both physical and mental/emotional etiologies and can lead to musculoskeletal injuries and athletic underperformance. Few studies have tested the effectiveness of a mind-body therapy on general flexibility. The aim of this study was to investigate if Neuro Emotional Technique® (NET), a mind-body technique shown to be effective in reducing stress, can also improve general flexibility. The sit-and-reach test (SR) score was used as a measure of general flexibility. Forty-five healthy participants were recruited from the general population and assessed for their initial SR score before being randomly allocated to receive (a) two 20-minute sessions of NET (experimental group); (b) two 20-minute sessions of stretching instruction (active control group); or (c) no intervention or instruction (passive control group). After intervention, the participants were reassessed in a similar manner by the same blind assessor. The participants also answered questions about demographics, usual water and caffeine consumption, and activity level, and they completed an anxiety/mood psychometric preintervention and postintervention. The mean (SD) change in the SR score was +3.1 cm (2.5) in the NET group, +1.2 cm (2.3) in the active control group and +1.0 cm (2.6) in the passive control group. Although all the 3 groups showed some improvement, the improvement in the NET group was statistically significant when compared with that of either the passive controls (p = 0.015) or the active controls (p = 0.021). This study suggests that NET could provide an effective treatment in improving general flexibility. A larger study is required to confirm these findings and also to assess longer term effectiveness of this therapy on general flexibility.


Behavioural Effects of Using Sulfasalazine to Inhibit Glutamate Released by Cancer Cells: A Novel target for Cancer-Induced Depression.
Despite the lack of robust evidence of effectiveness, current treatment options for cancer-induced depression (CID) are limited to those developed for non-cancer related depression. Here, anhedonia-like and coping behaviours were assessed in female BALB/c mice inoculated with 4T1 mammary carcinoma cells. The behavioural effects of orally administered sulfasalazine (SSZ), a system xc- inhibitor, were compared with fluoxetine (FLX). FLX and SSZ prevented the development of anhedonia-like behaviour on the sucrose preference test (SPT) and passive coping behaviour on the forced swim test (FST). The SSZ metabolites 5-aminosalicylic acid (5-ASA) and sulfapyridine (SP) exerted an effect on the SPT but not on the FST. Although 5-ASA is a known anti-inflammatory agent, neither treatment with SSZ nor 5-ASA/SP prevented tumour-induced increases in serum levels of interleukin-1β (IL-1β) and IL-6, which are indicated in depressive disorders. Thus, the observed antidepressant-like effect of SSZ may primarily be attributable to the intact form of the drug, which inhibits system xc-. This study represents the first attempt at targeting cancer cells as a therapeutic strategy for CID, rather than targeting downstream effects of tumour burden on the central nervous system. In doing so, we have also begun to characterize the molecular pathways of CID.


Detection of human fetal fibrinogen in umbilical cord plasma by parallel assays of immunoreactive and thrombin-clottable protein.
Putative fetal fibrinogen was detected in normal human umbilical cord plasma by parallel assays of immunoreactive and thrombin-clottable protein. The 'excess' of immunoassayable fibrinogen was not due to products of proteolysis and the apparent 'insensitivity' to thrombin was reflected in delayed fibrin polymerization. Fetal fibrinogen was partially separable from the adult protein by two-dimensional immunoelectrophoresis.


Gene targeting to the ROSA26 locus directed by engineered zinc finger nucleases.
Targeted gene addition to mammalian genomes is central to biotechnology, basic research and gene therapy. For example, gene targeting to the ROSA26 locus by homologous recombination in embryonic stem cells is commonly used for mouse transgenesis to achieve ubiquitous and persistent transgene expression. However, conventional methods are not readily adaptable to gene targeting in other cell types. The emerging zinc finger nuclease (ZFN) technology facilitates gene targeting in diverse species and cell types, but an optimal strategy for engineering highly active ZFNs is still unclear. We used a modular assembly approach to build ZFNs that target the ROSA26 locus. ZFN activity was dependent on the number of modules in each zinc finger array. The ZFNs were active in a variety of cell types in a time- and dose-dependent manner. The ZFNs directed gene addition to the ROSA26 locus, which enhanced the level of sustained gene expression, the uniformity of gene expression within clonal cell populations and the reproducibility of gene expression between clones. These ZFNs are a promising resource for cell engineering, mouse transgenesis and pre-clinical gene therapy studies. Furthermore, this characterization of the modular assembly method provides general insights into the implementation of the ZFN technology.