Tumor-specific p53 sequences in blood and peritoneal fluid of women with epithelial ovarian cancer.
Free tumor DNA in body fluids may be an important biomarker. We tested whether tumor-specific mutated p53 DNA can be detected in blood and peritoneal fluid from women with epithelial ovarian cancer. Sequencing of tumor DNA identified somatic p53 mutations. Free DNA from matched blood or peritoneal fluid was evaluated for the tumor-specific p53 mutation using a ligase detection reaction. Sixty-nine of 137 tumors (50%) had p53 mutations. Plasma or serum from 21 (30%) of the 69 informative cases contained the tumor-specific p53 mutation. Circulating tumor was an independent predictor of decreased survival in multivariate analysis (P=.02). We detected tumor DNA in peritoneal fluid in 28 of 30 (93%) cases, including all 6 cases with negative cytology. One third of women with ovarian cancer have circulating tumor DNA and an associated reduced survival. Free tumor DNA can be detected in the majority of peritoneal fluid samples.


Drain Placement Does Not Increase Infectious Complications After Retromuscular Ventral Hernia Repair with Synthetic Mesh: an AHSQC Analysis.
The use of surgical drains after ventral hernia repair (VHR) remains controversial. Some have concerns of increased infectious complications; others advocate that drains reduce fluid accumulation and surgical site occurrences (SSO). The aim of our study was to investigate the impact of retromuscular drains on SSO following retromuscular VHR with synthetic mesh. Utilizing the Americas Hernia Society Quality Collaborative, patients between January 2013 and January 2016 undergoing retromuscular VHR with synthetic mesh were assessed for the presence of a drain. Propensity score matched patients (2 drains: 1 no drain) were evaluated for 30-day rates of SSO, surgical site infections (SSI) and SSO requiring procedural intervention (SSOPI). Five hundred eighty-one patients were identified as having undergone open, retromuscular VHR with synthetic mesh. Four hundred eighty-one patients with drains and 100 without drains. After matching, 300 patients were compared, 200 with drain placement and 100 without. Retromuscular drains were less likely to develop a noninfectious SSO (OR, 0.33). Drain placement was not associated with SSI (OR, 1.30) or SSOPI (OR, 0.94). Drain placement after retromuscular VHR with synthetic mesh is a common practice. Based on an analysis of early outcomes, surgical drains do not increase the risk of surgical infectious complications, and may be protective against some SSOs, such as seroma formation.


Is there a circadian variation in respiratory morbidity?
In a retrospective study of the time of presentation to an accident and emergency department patients with acute respiratory symptoms presented more commonly at night compared to a control group with abdominal pain. In a subsequent prospective study doctors from a GP deputizing service were called much more frequently at night by patients with asthma than by those with other symptoms. These findings refute the suggestion that the observed increased mortality from respiratory diseases at night results from reduced medical care as a consequence of a reluctance of patients to present during the night.


Prospects for the Use of Marine Sulfated Fucose-Rich Polysaccharides in Treatment and Prevention of COVID-19 and Post-COVID-19 Syndrome.
Symptoms of the new coronavirus infection that appeared in 2019 (COVID-19) range from low fever and fatigue to acute pneumonia and multiple organ failure. The clinical picture of COVID-19 is heterogeneous and involves most physiological systems; therefore, drugs with a wide spectrum of mechanism of action are required. The choice of the treatment strategy for post-COVID-19 syndrome is still a challenge to be resolved. Polysaccharides with a high fucose content derived from seaweed and marine animals can form the basis for the subsequent development of promising agents for the treatment of COVID-19 and post-COVID-19 syndrome. This class of biopolymers is characterized by a variety of biological activities, including antiviral, antithrombotic, anticoagulant, hemo-stimulating, anti-inflammatory and immune-regulatory. Low molecular weight derivatives of these polysaccharides, as well as synthetic oligosaccharides with a sufficient amount and sulfation type may be considered as the most promising compounds due to their better bioavailability, which undoubtedly increases their therapeutic potential.


Rapid detection of porcine DNA in processed food samples using a streamlined DNA extraction method combined with the SYBR Green real-time PCR assay.
A specialized DNA extraction method and a SYBR Green quantitative polymerase chain reaction (SyG-qPCR) assay were combined to generate a ready-to-use kit for rapid detection of porcine admixtures in processed meat products. Our qPCR assay utilized repetitive LINE-1 elements specific to the genome of Sus scrofa domesticus (pig) as a target and incorporated internal controls. We improved the genomic DNA extraction method, and reduced extraction times to the minimum. The method was validated for specificity, sensitivity (0.001% w/w) and robustness, and values were compared with those of a commercially available kit. We also tested our method using 121 processed food products and consistently detected amplification only in samples containing pork. Due to its efficiency and cost-effectiveness, our method represents a valuable new method for detecting food adulteration with pork that is superior to existing quality control approaches.


Secondary tricuspid valve regurgitation after isolated aortic valve replacement: outcomes and predictors.
We sought to assess the evolution of secondary tricuspid regurgitation (TR) after isolated aortic valve replacement (AVR) and its impact on mortality and to identify possible clinical and echocardiographic predictors of persisting and new-onset TR (TR de novo) after isolated AVR. Clinical and echocardiographic data of 441 patients, consecutively operated for isolated AVR between January 2017 and January 2020, were retrospectively collected. Four time points were included: preoperative, discharge, 3-6 months and last available follow-up. We followed patients with at least moderate TR (TR ≥2) over time and monitored the impact on survival. Logistic regression analysis was performed to identify possible predictors of persistent TR and TR de novo. Median follow-up was 33 months. Incidence of TR ≥2 changed over the time points. Twenty-three percent of patients with preoperative TR ≥2 had persistent TR at 3-6 months follow-up, and this phenomenon was predicted by age at regression analysis. Preoperative TR ≥2 was associated with a 3-fold higher risk to die. At 3- to 6-month follow-up, 12% of patients developed TR de novo. At least moderate preoperative mitral regurgitation (≥2) was predictive of TR de novo. Patients with TR ≥2 undergoing isolated AVR had worse long-term survival, and this was particularly evident in the elderly. Older patients were also more prone to have persistent TR after AVR. Some patients developed TR de novo after isolated AVR, but this did not affect survival.


Blood-brain barrier disruption in CCL2 transgenic mice during pertussis toxin-induced brain inflammation.
The chemokine CCL2 has an important role in the recruitment of inflammatory cells into the central nervous system (CNS). A transgenic mouse model that overexpresses CCL2 in the CNS shows an accumulation of leukocytes within the perivascular space surrounding vessels, and which infiltrate into the brain parenchyma following the administration of pertussis toxin (PTx). This study used contrast-enhanced magnetic resonance imaging (MRI) to quantify the extent of blood-brain barrier (BBB) disruption in this model pre- and post-PTx administration compared to wild-type mice. Contrast-enhanced MR images were obtained before and 1, 3, and 5 days after PTx injection in each animal. After the final imaging session fluorescent dextran tracers were administered intravenously to each mouse and brains were examined histologically for cellular infiltrates, BBB leakage and tight junction protein. BBB breakdown, defined as a disruption of both the endothelium and glia limitans, was found only in CCL2 transgenic mice following PTx administration and seen on MR images as focal areas of contrast enhancement and histologically as dextrans leaking from blood vessels. No evidence of disruption in endothelial tight junctions was observed. Genetic and environmental stimuli were needed to disrupt the integrity of the BBB in this model of neuroinflammation.


Perfusion-distention fixation of heart specimens: a key step in immortalizing heart specimens for wax infiltration and generating 3D imaging data sets for reconstruction and printed 3D models.
Perfusion-distention fixation by gravity is a fundamental cardiac predissection technique that may be used to preserve the 3D anatomy of a cardiac specimen. The equipment and steps are illustrated, and examples of cardiac specimens preserved in various methods including wet specimens, wax-infiltrated hearts, and 3D-printed models, created after performing this technique, are shown.


Juvenile polyposis coli: a facultative precancerosis with some similarities to ulcerative colitis?
We investigated the case of a 13-year-old male with juvenile polyposis (JP) to determine the extent of intraepithelial neoplasia and associated genetic changes, as well as cellular proliferation, within these polyps using immunohistochemistry with antibodies against p53, bcl-2, and Ki-67. Examination of the total proctocolectomy specimen revealed 70 polyps. The 18 largest polyps were investigated microscopically and disclosed the typical hamartomas with frequent erosions of the surface epithelium and reparative changes. Only one polyp showed focal low-grade intraepithelial neoplasia. The immunohistochemical studies revealed an expression of p53 and an abnormal Ki-67 pattern of the surface epithelium only within the neoplastic area. These findings may hint at a possible pathogenetic mechanism for the evolution of colorectal cancer in JP. As in ulcerative colitis, carcinomas in JP may develop along a dysplasia-carcinoma sequence resulting from permanent mechanical insults, inflammation, and repair rather than from an adenoma-carcinoma sequence as in familial adenomatous polyposis (FAP).


Norwood Operation with Right Ventricular-Pulmonary Artery Shunt Versus Comprehensive Stage II After Bilateral Pulmonary Artery Banding Palliation.
As a strategy for the primary Norwood operation, the right ventricular-pulmonary artery shunt is associated with satisfactory early outcome. However, use of this shunt after bilateral pulmonary artery banding remains controversial. This study compared the operative outcomes and late hemodynamics in patients who underwent the Norwood operation, preceded by bilateral pulmonary artery banding, with a right ventricular-pulmonary artery shunt or with bidirectional Glenn anastomosis (comprehensive stage II strategy). We retrospectively reviewed 38 patients who underwent the Norwood operation preceded by bilateral pulmonary artery banding between 2004 and 2017. Of these, 17 underwent the Norwood operation with a right ventricular-pulmonary artery shunt (Group S), whereas 21 underwent the comprehensive stage II strategy (Group G). 5 years after the Norwood operation, 10 (60%) and 17 (81%) patients in Group S and Group G, respectively, underwent the Fontan procedure. Group S showed significantly lower pressure in the superior vena cava after bidirectional Glenn anastomosis than Group G (13 ± 2 mmHg vs. 18 ± 3 mmHg; p < 0.01), but pressures were similar after the Fontan procedure. The right ventricular end-diastolic volume at 1 year post-Fontan procedure was significantly higher in Group S than in Group G (142 ± 41% vs. 91 ± 28%; p < 0.01). In terms of early outcomes, the Norwood operation with a right ventricular-pulmonary artery shunt enabled low pressure in the superior vena cava, but in the long term, this shunt adversely influenced the right ventricular volume.


Lymphocytic Esophagitis: Assessing Risk Factors and Clinical Outcomes.
Lymphocytic esophagitis is a rare esophageal condition. Our knowledge of potential risk factors and treatment outcomes of lymphocytic esophagitis is limited. To investigate potential risk factors associated with the development of lymphocytic esophagitis and compare clinical characteristics and treatment outcomes of patients diagnosed with lymphocytic esophagitis to patients diagnosed with eosinophilic esophagitis. This is a multicenter retrospective study. Lymphocytic esophagitis patients were identified based on pathology results between 1997 and 2019. Control groups consisted of patients with normal esophageal biopsies and patients diagnosed with eosinophilic esophagitis. Thirteen potential risk factors for lymphocytic esophagitis were analyzed using univariate and multivariate models including IBD, achalasia, hyperlipidemia, hypothyroidism, celiac sprue, CVID, H. pylori, thymoma, aspirin, opioids, ACE-I, metformin, and statin use. Comparative statistics were performed. Ninety-four adult patients with lymphocytic esophagitis, 344 with eosinophilic esophagitis, and 5202 control patients with normal esophageal biopsies were analyzed. Age older than 60 [adjusted odd ratio (AOR) 1.03, 95% CI 1.02-1.05, p = 0.001], aspirin use (2.7, 95% CI 1.4-4.9, p = 0.001), statin use (2.2, 95% CI 1.2-4.2, p = 0.01), or a diagnosis of achalasia (2.4, 95% 1.08-5.67, p = 0.03) were associated with lymphocytic esophagitis. Compared to eosinophilic esophagitis, lymphocytic esophagitis patients were more likely to respond to medical treatment (95% CI 2.54-12.8, p = 0.0001). Our data suggests that lymphocytic esophagitis is more likely to be found in older female patients and is significantly associated with achalasia, statin, and aspirin use. Compared to eosinophilic esophagitis, lymphocytic esophagitis is more likely to respond to treatment with medical therapy.


Multivariate calibration: applications to pharmaceutical analysis.
The principles of multivariate calibration (MC) are presented, with reference to the main objectives of this chellometrics technique: the reduction of the variance in the prediction of a response variable (generally, a chemical quantity) and the possibility of the determination of the response in complex matrices with no or limited sample preparation, as in the case of the determination of a drug in a medicament. In both cases MC uses the whole information in a spectrum (a series of predictors). The possibility of the improvement of the MC performances, eliminating some useless, noisy, predictors is shown. Variable selection has been performed using two original techniques: a stepwise elimination procedure, based on the normalised coefficients of the regression equation relating the response to the predictors and a technique based on iterative repetitions of the regression technique (partial least squares regression, PLS), each time by weighting the predictors by their normalised regression coefficient computed in the previous cycle. These strategies are illustrated by means of different data sets, a synthetic example and a real example where MC, applied to near infrared spectroscopy, is used in the analysis of a drug. In this case also the application of an original MC technique is shown, where a joint regression model is obtained for two different instruments.


Inhibition of Infected Red Blood Cell Binding to the Vascular Endothelium.
Plasmodium falciparum-infected red blood cells (IRBC) adhere to the endothelium via receptors expressed on the surface of vascular endothelial cells (EC) and sequester in the microvasculature of several organs. Sequestration is the primary step leading to complications related to the severity of malaria. In order to study this cytoadhesion phenomenon, IRBC in vitro binding assays have been developed using a monolayer of primary or transformed endothelial cells. Here we describe the methodology of an assay to inhibit the binding of IRBC on vascular endothelial cells under static adhesion conditions. Similar techniques could be used for conducting a binding inhibition assay under flow assay conditions using an appropriate device.


Effects of dalteparin on anti-Xa activities cannot be predicted in critically ill COVID-19 patients.
Critically ill COVID-19 patients are at high risk of thromboembolic events despite routine-dosed low-molecular-weight heparin thromboprophylaxis. However, in recent randomized trials increased-intensity thromboprophylaxis seemed futile and possibly even harmful. In this explorative pharmacokinetic (PK) study we measured anti-Xa activities on frequent timepoints in 15 critically ill COVID-19 patients receiving dalteparin and performed PK analysis by nonlinear mixed-effect modelling. A linear one-compartment model with first-order kinetics provided a good fit. However, wide interindividual variation in dalteparin absorption (variance 78%) and clearance (variance 34%) was observed, unexplained by routine clinical covariates. Using the final PK model for Monte Carlo simulations, we predicted increased-intensity dalteparin to result in anti-Xa activities well over prophylactic targets (0.2-0.4 IU/mL) in the majority of patients. Therapeutic-intensity dalteparin results in supratherapeutic anti-Xa levels (target 0.6-1.0 IU/mL) in 19% of patients and subtherapeutic levels in 22%. Therefore, anti-Xa measurements should guide high-intensity dalteparin in critically ill COVID-19 patients.


Two new cyclopentenones and a new furanone from Baeckea frutescens and their cytotoxicities.
Two new cyclopentenones, frutescencenones A (1) and B (2), and a new furanone derivative, frutescencenone C (3), together with two known cyclopentenones (4 and 5), were isolated from the leaves of Baeckea frutescens. Their structures were deduced by comprehensive spectroscopic analyses, including 1D and 2D NMR, and HREIMS data. Frutescencenone A (1) showed moderate growth inhibitory activity against human lung A549, pancreatic PSN-1, and breast MDA-MB-231 cancer cell lines, with IC50 values of 36.3μM, 38.2μM, and 29.3μM, respectively. In contrast, frutescencenone C (3) showed selective cytotoxic activity against PSN-1, with an IC50 value of 20.1μM.


Analysis of normal lung irradiation in radiosurgery treatments: a comparison of lung optimized treatment (LOT) on cyberknife, 4D target volume on helical tomotherapy, and DIBH on linear accelerator.
Quantitative retrospective analysis of the normal lung irradiation due to the variations of the ITV volume based on the techniques used for upper lobe (UL), mid lobe (ML), and lower lobe (LL) lung tumours when used with 2-view, 1-view, 0-view based LOT technique on Cyberknife, AveIP on Helical Tomotherapy, and DIBH on VMAT systems. In the treatment of lung tumours, patients medically inoperable or those who are unwilling to undergo surgery have the option to be treated using radiation therapy. There are many motion control techniques available for the treatment of the moving target, such as movement encompassment, respiratory gating, breath-hold, motion reduction, and tumour monitoring. ITV generation is dependent on technique and hence the volume of the PTVs will differ based on the technique used. This study aimed to determine the influence of these ITVs on the irradiated normal lung volume for UL, ML, and LL lung tumours for 23 patients. The mean difference in the PTV volumes generated with the 0-view technique was significant with that of 2-view and DIBH techniques (p-value < 0.04). The mean difference in the PTV volumes generated by 2-view and DIBH was small for UL, ML, and LL tumours. V5 of the combined lung with the 0-view method was 5% compared to the 2-view method for UL tumours (p-value = 0.04) and the same was 9.5%, and 16.8% for ML and LL tumours (p-value < 0.04). In contrast to all other techniques, lung volume parameters V5, V10, V20, and V30 for the 0-view technology were consistently higher irrespective of the tumour location in the lung. The observed maximum mean lung dose (MLD) was 6.2 Gy ± 2.7 Gy with the 0-view technique and the minimum was 3.85 Gy ± 1.75 Gy with the DIBH technique. The difference in MLD between DIBH and 2-view was negligible (p-value = 0.67). The MLD increased for LL tumours from 4 Gy to 6.5 Gy from the 2-view to 0-view technique (p-value = 0.009). There was a significant increase in MLD for LL tumours with the 0-view technique compared to AveIP (1.9 Gy, p-value = 0.04) and DIBH (2.0 Gy, p-value = 0.003) technique. For ML and UL tumours, except for 0-view and 1-view, the difference in the MLD between the rest of the methods was not significant (p-value > 0.11). In the treatment of lung tumour patients with SBRT, this study has demonstrated 2-view with Cyberknife and DIBH with VMAT treatment techniques have optimal normal lung tissue sparing. There was a significant increase in the average lung volume receiving 5%,10%, 20%, and 30% dose when comparing the 1-view, 0-view, AveIP, and DIBH techniques to the 2-view technique. However, DIBH with VMAT was dosimetrically advantageous for ML and LL tumours, while providing significantly shorter treatment times than any other technique studied.


Boxer's knuckle--dorsal capsular rupture of the metacarpophalangeal joint of a finger.
An injury to the dorsal aspect of the metacarpophalangeal joint of a finger usually damages the sagittal fibers of the extensor tendon mechanism. Rupture of the underlying joint capsule, a far more serious injury, has received scant attention in the medical literature. The problem is often unrecognized because the extensor tendon may remain in its normal midline position. Eight dorsal capsular ruptures are described in six patients who had closed direct trauma to either the knuckle of their index or long fingers. Most of the injuries occurred in professional boxers.


The relationship between maternal psychopathology and offspring incontinence and constipation at school age: A prospective cohort study.
There is evidence for an association between maternal psychopathology and offspring incontinence and constipation, but it is unclear if there is a critical/sensitive period of exposure to maternal depression and/or anxiety in the antenatal or postnatal period. Mothers from the Avon Longitudinal Study of Parents and Children provided data on their depression and anxiety (antenatal and postnatal) and their child's urinary and faecal incontinence and constipation at age 7 (6489 participants). We used multivariable logistic regression to examine evidence for independent effects of maternal depression/anxiety on offspring incontinence/constipation and to investigate if there was a critical/sensitive period of exposure. We examined evidence for causal intra-uterine effects using a negative control design. Postnatal maternal psychopathology was associated with an increased risk of offspring incontinence and constipation (e.g. postnatal anxiety and daytime wetting OR: 1.53; 95 % CI: 1.21-1.94). Data were consistent with a postnatal critical period model and there was evidence for an independent effect of maternal anxiety. Antenatal maternal psychopathology was associated with offspring constipation (e.g. antenatal anxiety OR: 1.57; 95 % CI: 1.25-1.98), but there was no evidence for a causal intra-uterine effect. Attrition and maternal reports without use of diagnostic criteria for incontinence/constipation are potential limitations. Children exposed to maternal postnatal psychopathology had a greater risk of incontinence/constipation, and maternal anxiety had stronger associations than depression. Health professionals should be vigilant to effects of maternal psychopathology on child development. Identification of mechanisms linking maternal psychopathology to child incontinence/constipation is required to inform evidence-based support.


Multivariate analysis of genome-wide data to identify potential pleiotropic genes for five major psychiatric disorders using MetaCCA.
Genome-wide association studies have been extensively applied in identifying SNP associated with major psychiatric disorders. However, the SNPs identified by the prevailing univariate approach only explain a small percentage of the genetic variance of traits, and the extensive data have shown the major psychiatric disorders have common biological mechanisms and the overlapping pathophysiological pathways. We applied the genetic pleiotropy-informed metaCCA method on summary statistics data from the Psychiatric Genomics Consortium Cross-Disorder Group to examine the overlapping genetic relations between the five major psychiatric disorders. Furthermore, to refine all genes, we performed gene-based association analyses for the five disorders respectively using VEGAS2. Gene enrichment analysis was applied to explore the potential functional significance of the identified genes. After metaCCA analysis, 1147 SNPs reached the Bonferroni corrected threshold (p < 1.06 × 10-6) in the univariate SNP-multivariate phenotype analysis, and 246 genes with a significance threshold (p < 3.85 × 10-6) were identified as potentially pleiotropic genes in the multivariate SNP-multivariate phenotype analysis. By screening the results of gene-based p-values, we identified 37 putative pleiotropic genes which achieved significance threshold in metaCCA analyses and were also associated with at least one disorder in the VEGAS2 analyses. Alternative approaches and experimental studies may be applied to check whether novel genes could still be identified/substantiated with these methods. The metaCCA method identified novel variants associated with psychiatric disorders by effectively incorporating information from different GWAS datasets. Our analyses may provide insights for some common therapeutic approaches of these five major psychiatric disorders based on the pleiotropic genes and common mechanisms identified.


Transcription of the prorenin gene in normal and diseased breast.
The angiotensin II type 1 (AT1) receptor is present in a wide variety of human and animal tissues, and is particularly abundant in epithelial cells. Because of this, and because it is known that tissue renin angiotensin systems (RASs) exist that have specific local functions, we investigated the expression and localisation of components of the RAS in normal and diseased breast tissue. Using a monoclonal antibody to the AT1 receptor, immunocytochemistry confirmed that the AT1 receptor was characteristically distributed in ductal epithelial cells in both normal and malignant tissue, and in most, although not all, cells in invasive tumours. Transcription of prorenin mRNA was studied by in situ hybridisation, using a DIG-ddUTP tail-labelled probe specific for the human prorenin gene. In normal tissue, and in cases of ductal carcinoma in situ, prorenin mRNA was distributed in myoepithelial cells and in a band of connective tissue cells completely surrounding the AT1-containing ductal epithelial cells. This prorenin transcribing tissue was disrupted and attenuated in invasive tumours, and in some of these, prorenin mRNA transcription could not be detected at all. Functions ascribed to the tissue RASs include regulation of mitosis and tissue modelling, as well as fluid and electrolyte transport. The results presented here strongly suggest the possibility that a tissue RAS may also be present in the breast, closely coupled to the provision of angiotensin II to the AT1 receptors in ductal epithelial cells. This mechanism is disrupted in cancer.


Double mitral valve. Report of a case and review of the literature.
A case of double mitral valve orifice associated with atrioventricular canal was found at operation. These two anomalies are frequently associated. Double mitral valve is usually subclinical and does not necessarily need to be corrected, depending on the associated anomalies, the extent of the defect, and the personal experience of the surgeon. The embryology of double mitral valve is complicated and still needs to be clarified.


Identification of the stimulated hemiretinae using a reduced number of PVEP trials.
Left and right hemifield pattern-reversal visual evoked potentials (PVEP) at P100 latency are characterised by typical field distributions of potential and the spectral parameter instantaneous frequency (IF). Both parameters can be utilised for the correct identification of the stimulated hemiretina in healthy volunteers (Hoffmann, K. et al. Electroenceph. clin. Neurophysiol., 100, 1996: 569-578). The aim of this study was the investigation of the robustness of instantaneous frequency for reduced numbers of averages. Hemifield PVEP of 15 volunteers (20 channel records) were analysed. The number of averages was reduced step-by-step (64-32-16-8-4-1). For each average, the time of P100 latency was determined by the Global field power maximum between 90 and 125 ms. The stimulated hemifield was identified using the potential or instantaneous frequency values at the occipital electrode positions O1 and O2: by maximal potential and instantaneous frequency on the stimulus-contralateral side. In summary, by reduced numbers of averages (as well as by single trials) the stimulated hemiretina was correctly identified more frequently on the basis of instantaneous frequency than of potential distribution. A number of 8 averages seems to be sufficient for the correct identification of the stimulus condition. Consequently, for the identification of left and right hemifield stimulations the recording time could be reduced immensely. Instantaneous frequency is suggested as an additional and robust parameter for the selective averaging of artefact-free trials during the recording of hemifield PVEP.


The relationship of addiction, tolerance, and dependence to alcohol and drugs: a neurochemical approach.
Alcohol and drug addiction are defined in behavioral terms as the preoccupation with, compulsive use of, and relapse to drugs that are descriptive and confirmatory. The basis of addiction may involve neurochemical changes in the brain that distort and redirect the drive states (instincts). Tolerance and dependence may only be incidentally associated with addiction as a result of a nonspecific adaptation by the body to the presence of a drug. The cellular adaptation may be the same in all organs. Addiction to alcohol and drugs may have no specific relationship to tolerance and dependence. Addiction occurs in the absence of observable tolerance and dependence to alcohol and drugs. Alcohol and drug addiction is probably more complex than tolerance and dependence. Addiction is difficult to study because of the variability of behavioral phenomena and the underlying intricacies of the neurosubstrates. Tolerance and dependence are still useful as they are indicators of drug use. It is a misconception that long term chronic use is necessary for tolerance and dependence to develop. Some studies have shown that tolerance can develop within hours and days to a single dose of alcohol or other drugs. Anxiety, depression and insomnia can occur after a single dose of ethanol in humans. These symptoms of withdrawal from the alcohol or drug constitute dependence. Redefining the criteria for addiction tolerance and dependence to alcohol and other drugs may be in order. A neurochemical model may provide a more definitive and uniform basis for considering addiction, tolerance, and dependence to alcohol and drugs.


[Systemic reactions in rats following the initiation of a local inflammatory process by subcutaneous administration of spirits of turpentine].
Local inflammation was induced in rats by single (1 x 4 ml/kg) or multiple (14 X 0.2 ml/animal) infections of turpentine. The induction of inflammatory processes in both groups resulted in anemia and granulocytosis following an initial leukopenia. Thrombopenia on the second day, followed by thrombocytosis, was also observed in both groups. Studies on blood chemistry parameters revealed a decline in serum albumin; elevation of alkaline phosphatase in serum was observed only after multiple injection of turpentine. In these animals an elevation in the weights of spleen and adrenals and a reduction in the weight of thymus were also found.


Nutrition and Preventative Medicine Across the Lifespan: Implementation of a Physician Assistant Team-Based Learning Curriculum on Pediatric Preventative Medicine.
Physician assistants (PAs) will increasingly fill an important primary care gap in pediatrics. This study implemented a pediatric preventative care curriculum using a team-based learning (TBL) strategy and then evaluated the effect on first-year PA students' knowledge and attitudes toward TBL as a teaching method. The curriculum was developed and implemented during a one-semester course. Students completed knowledge-based questions precourse and postcourse; the mean scores for individual students were then compared using a paired t test. Descripted statistics were used to analyze the postcourse survey that evaluated students' attitudes toward TBL. Of the 31 PA students who participated, 27 (87%) completed the surveys and were included in the analysis. The mean correct pretest and posttest scores were 17/39 and 31/39, respectively, resulting in a change of 14 points (p < .05). The majority of students (85%) rated TBL as effective, and most students (75%) reported that it led to retention of the presented content. A TBL-based curriculum in pediatric preventative care was effective in increasing PA student knowledge, and the students reported it as effective. This strategy could be considered for other PA courses.


Coupling endoplasmic reticulum stress to the cell-death program: a novel HSP90-independent role for the small chaperone protein p23.
The endoplasmic reticulum (ER) is the principal organelle for the biosynthesis of proteins, steroids and many lipids, and is highly sensitive to alterations in its environment. Perturbation of Ca(2+) homeostasis, elevated secretory protein synthesis, deprivation of glucose or other sugars, altered glycosylation and/or the accumulation of misfolded proteins may all result in ER stress, and prolonged ER stress triggers cell death. Studies from multiple laboratories have identified the roles of several ER stress-induced cell-death modulators and effectors through the use of biochemical, pharmacological and genetic tools. In the present work, we describe the role of p23, a small chaperone protein, in preventing ER stress-induced cell death. p23 is a highly conserved chaperone protein that modulates HSP90 activity and is also a component of the steroid receptors. p23 is cleaved during ER stress-induced cell death; this cleavage, which occurs close to the carboxy-terminus, requires caspase-3 and/or caspase-7, but not caspase-8. Blockage of the caspase cleavage site of p23 was associated with decreased cell death induced by ER stress. Immunodepletion of p23 or inhibition of p23 expression by siRNA resulted in enhancement of ER stress-induced cell death. While p23 co-immunoprecipitated with the BH3-only protein PUMA (p53-upregulated modulator of apoptosis) in untreated cells, prolonged ER stress disrupted this interaction. The results define a protective role for p23, and provide further support for a model in which ER stress is coupled to the mitochondrial intrinsic apoptotic pathway through the activities of BH3 family proteins.


Detector-based spectral CT with a novel dual-layer technology: principles and applications.
Detector-based spectral computed tomography is a novel dual-energy CT technology that employs two layers of detectors to simultaneously collect low- and high-energy data in all patients using standard CT protocols. In addition to the conventional polyenergetic images created for each patient, projection-space decomposition is used to generate spectral basis images (photoelectric and Compton scatter) for creating multiple spectral images, including material decomposition (iodine-only, virtual non-contrast, effective atomic number) and virtual monoenergetic images, on-demand according to clinical need. These images are useful in multiple clinical applications, including- improving vascular contrast, improving lesion conspicuity, decreasing artefacts, material characterisation and reducing radiation dose. In this article, we discuss the principles of this novel technology and also illustrate the common clinical applications. Teaching points • The top and bottom layers of dual-layer CT absorb low- and high-energy photons, respectively.• Multiple spectral images are generated by projection-space decomposition.• Spectral images can be generated in all patients scanned in this scanner.


The relationship between the ratio of interpedicular distance increase and the ratio of spinal canal compromise in thoracolumbar burst fractures.
The aim of the study was to investigate the relationship between the interpedicular distance increase ratio and the ratio of canal compromise in thoracolumbar burst fractures. Thirty-one patients (18 male and 13 female) with an average age of 30.8 (14-57) who had been treated for thoraco-lumbar burst fractures in the Department of Orthopaedics and Traumatology were included in the study. The initial anteroposterior radiographs of the patients were used to calculate the increase ratio of interpedicular distance (both from medial-to-medial and from center-to-center). The area measurements from the computerized tomography or magnetic resonans images were used to calculate the canal compromise. The relationship between the increase ratio of interpedicular distance and the ratio of canal compromise was investigated by correlation and linear regression analysis. There was a 'very good' correlation between the from medial-to-medial and from center-to-center measurements of interpedicular distance (Pearson correlation coefficient: 0.89, p<0.001). The correlation between the ratio of canal compromise and from medial-to-medial and from center-to-center measurements of interpedicular distance was 'good' with Pearson correlation coef-ficients of 0.60 and 0.63, respectively (p<0.001). No statistically significant relationships were found between the fracture levels, types, neurologic status of the patients, and the increase ratio of interpedicular distance or the ratio of canal compromise. Depending on the correlation coefficients which were obtained in this study: To predict the canal compromise from the ratio of interpedicular distance increase is not a reliable method for all of the patients.


After-Hours Versus Office-Hours Dental Injuries in Children: Does Timing Influence Outcome?
The aim of this study was to analyze the outcomes and factors associated with after-hours dental trauma. Study sample consisted of 1762 permanent teeth injuries in children, gender and age matched with office-hours injuries. Epidemiological and clinical data were collected from 4 university dental trauma centers. During median follow-up time of 4.3 years, complications have occurred in 14.5% of injured teeth. Age, type, and degree of tissue injury and after-hours time of injury were significantly associated with complications. Unfavorable outcomes were 34% more likely in the after-hours group compared with office-hours. Urgent treatment was significantly delayed in after-hours group with a delay of more than 3 hours in 90.5% versus 38.9% in the office-hours group. Multivariate regression model showed that after-hours time of injury was significant predictor of complications. Delayed urgent treatment was one of the main factors associated with unfavorable outcome of after-hours injuries.


Metastatic Spine Disease: Should Patients With Short Life Expectancy Be Denied Surgical Care? An International Retrospective Cohort Study.
Despite our inability to accurately predict survival in many cancer patients, a life expectancy of at least 3 mo is historically necessary to be considered for surgical treatment of spinal metastases. To compare health-related quality of life (HRQOL) in patients surviving <3 mo after surgical treatment to patients surviving >3 mo to assess the validity of this inclusion criteria. Patients who underwent surgery for spinal metastases between August 2013 and May 2017 were retrospectively identified from an international cohort study. HRQOL was evaluated using generic and disease-specific outcome tools at baseline and at 6 and 12 wk postsurgery. The primary outcome was the HRQOL at 6 wk post-treatment measured by the Spine Oncology Study Group Outcomes Questionnaire (SOSGOQ). A total of 253 patients were included: 40 patients died within the first 3 mo after surgery and 213 patients survived more than 3 mo. Patients surviving <3 mo after surgery presented with lower baseline performance status. Adjusted analyses for baseline performance status did not reveal a significant difference in HRQOL between both groups at 6 wk post-treatment. No significant difference in patient satisfaction at 6 wk with regard to their treatment could be detected between both groups. When controlled for baseline performance status, quality of life 6 wk after surgery for spinal metastasis is independent of survival. To optimize improvement in HRQOL for this patient population, baseline performance status should take priority over expected survival in the surgical decision-making process.


TLR2 signals via NF-κB to drive IL-15 production in salivary gland epithelial cells derived from patients with primary Sjögren's syndrome.
Toll-like receptors (TLRs) are pattern recognition receptors linking innate and adaptive immune responses, which resulted overexpressed in primary Sjögren's syndrome (pSS). Interleukin-15 (IL-15) is a pro-inflammatory cytokine which was recently demonstrated to be involved in pSS pathogenesis. The study was undertaken to clarify whether TLR2 is involved in the production of IL-15 in human salivary gland epithelial cells (SGEC) from pSS patients. SGEC primary cell cultures were established from pSS minor salivary gland tissues explanted from patients with a sure diagnosis of SS. After neutralization of TLR2 with a blocking monoclonal antibody, IL-15 production was assayed by immunoblotting and flow cytometry, IL-15 in the culture supernatants was measured by ELISA, and mRNA levels were assessed by RT-PCR and real-time PCR. The production of IL-15 by pSS SGEC decreased in culture supernatants and in protein lysates (p < 0.01) when TLR2 signaling was inhibited in pSS SGEC. In addition, a control at the transcriptional level was also detected; in fact, inhibition of nuclear factor (NF)-κB through the transfection of pSS SGEC with the dominant-negative inhibitory κBα proteins (IκBα) vector (IκBαDN) abrogated the stimulatory effect of TLR2 on IL-15 production. These data suggest that TLR2 activation is involved in the induction of IL-15 production by pSS SGEC and promotes inflammation through NF-κB activation. Therefore, therapeutic strategies that target TLR2/IL-15 pathway might be strong candidates for preventing or treating pSS.


Epilepsy is associated with an increased incidence of heart failure diagnoses.
Epilepsy is a complex disease with serious consequences for the quality of life and prognosis of those affected. The importance of comorbidities in disease progression and prognosis has gained increasing recognition in recent years. In the present study, we investigated the potential association between epilepsy and heart failure in an outpatient cohort in Germany. Using the IQVIA Disease Analyzer database, we identified a total of 9646 patients with late-onset epilepsy and a matched cohort of equal size without late-onset epilepsy who were followed up between 2005 and 2018. Cox regression models were used to evaluate the potential association between epilepsy and heart failure. Within 10 years of the index date, 28.6% of patients with epilepsy and 20.4% of patients without epilepsy had been diagnosed with HF (log-rank p < 0.001). The incidences were 36.3 cases per 1,000 patient years in the epilepsy cohort versus 23.1 cases in the non-epilepsy cohort. In regression analyses, epilepsy was significantly associated with the incidence of HF (Hazard Ratio (HR): 1.56, p < 0.001). The association was somewhat stronger in men (HR: 1.63, p < 0.001) than in women (HR: 1.49, p < 0.001). The HR in the epilepsy group decreased with increasing age. Our study provides strong evidence that epilepsy is associated with an increased incidence of heart failure. This finding should help raise awareness of this important comorbidity and could trigger specific cardiovascular screening programs in patients with epilepsy.


Vitamin A and systemic inflammation as protective factors in multiple sclerosis.
Vitamin A is important for the immune system, and might suppress inflammatory activity in multiple sclerosis (MS). We aimed to examine if vitamin A levels were associated with MS risk in samples collected prospectively and during gestation. We measured Retinol Binding Protein (RBP--a surrogate marker for vitamin A) and high-sensitivity C-reactive protein (hs-CRP) levels, in (1) prospectively collected biobank blood samples from MS cases and controls, and (2) gestational samples where the offspring had later developed MS, and gestational control samples. The risk of MS was calculated using matched multivariable logistic regression adjusted for confounders. In prospective samples, RBP levels within the second quintile (vs. the first) were associated with a lower MS risk (OR = 0.38, 95% CI 0.19-0.74). No effect on MS risk in the offspring by gestational RBP levels was found. In young subjects hs-CRP levels ≥10 mg/l in prospective samples were associated with a lower MS risk (OR = 0.36, 95% CI 0.14-0.95). Our results suggest that sub-optimal vitamin A levels may be associated with MS risk. The association between hs-CRP levels and MS risk in young subjects may support the role of the hygiene hypothesis in MS aetiology.


Deoxynivalenol contents in foodstuffs of organical and conventional production.
Samples of wheat flour, bread, noodles, rice, corn and corn based foods as well as oats and oats based foods of conventional and organical production were analysed for trichothecene toxins. In wheat flour, bread and noodels the median deoxynivalenol (DON) contents were lower in ecological than in conventional products with significant differences for what flour and bread. To estimate toxin uptake of the consumer a corrected mean of DON concentration was calculated, which was lower in wheat flour and bread for ecological than for conventional products. In noodles the corrected mean of organical products was higher than that of conventional ones.


Surgical resection of residual tumor after chemotherapy in non-seminomatous testicular cancer.
Fifteen patients with disseminated non-seminomatous testicular cancer, 13 of whom had advanced disease, underwent surgery for residual tumor after induction chemotherapy. Complete remissions were achieved in 7 of 9 patients with mediastinal or pulmonary metastases and in 2 of 6 patients with retroperitoneal metastases. Patients with alpha-fetoprotein (AFP) levels over 10(4) ng/ml at diagnosis and/or a positive AFP preoperatively failed to achieve complete remission. Complete remissions were obtained in all 8 patients who had resection of necrosis, mature teratoma, immature teratoma or mature teratoma with malignant foci, but in only 1 of 7 patients who had resection of embryonal carcinoma or yolk sac tumor with other components. Of 9 patients with complete remission, 8 have remained free of disease after a median follow-up time of 29 months (range 6-66 months) and one had a contralateral non-seminomatous testicular cancer removed after 60 months. In addition to being therapeutically successful, the combined use of chemotherapy followed by surgery for residual tumor may lead to a better understanding of the influence of chemotherapy on the biology of testicular carcinoma.


Nutrient turnover studies in alpine ecosystems : IV. Communities of the Central Alps and Comparative Survey.
The nutrient relations of five treeless plant communities on acid soils above siliceous rock of the Central Alps are investigated. Three of these communities, situated on Mt. Patscherkofel, are dominated by dwarf shrubs of the Ericaceae family: Loiseleurietum (P 1, 2175 m NN), Loiseleuria heath (P 2, 2000 m NN), and Vaccinium heath (P 3, 1980 m NN). The other two are bound to higher elevations (2500 m NN, at Timmelsjoch): Caricetum curvulae (T 1), forming the mats, and Salicetum herbaceae (T 2), covering the snow-beds.Phytomass productivity decreases with increasing altitude in the sequence P 3-P 2, P 1-T 1-T 2.Compared with the turf communities of the Northern Calcareous Alps, nitrogen reserves and experimental net-mineralization of the soils (0-15 cm) are extremely low in P 1, P 2, and P 3 (<0.5 g N/m2 mineralized per GS1). The fluctuation of N in the living above-ground phytomass during the GS is also low (about 1.6 g/m2 in P 1 and P 2; 2.2 g/m2 in P 3, although it exceeds the values of net mineralization. Additional uptake through mycorrhizal fungi or activation of mineralizing microbes in the rhizosphere by exudation is assumed.The P- and K-reserves are extremely small in the humic soils of P 2 and P 3, but somewhat higher in the more mineral soil of P 1. Mean lactatesoluble P of the three sites is low (0.3 g/m2 or less) whereas Klact (2.7-3.3 g/m2) is higher than the lowest level found in some turf communities, e.g. Caricetum firmae. The amounts of P in the phytomass are in the range of those of the turf communities and agree with the gradation in the mean Plact values (P 1 and P 3>P 2). There are, however, almost no discernible fluctuations of P in the phytomass, and the K-fluctuations are far below the mean Klact level.The Timmelsjoch communities generally have higher N/C-, P/C-, and K/C-ratios in the soils compared with those of Mt. Patscherkofel, although the N-reserves (g/m2) and the K-reserves (of T 1 only) are lower. The Plact values are higher than those from Mt. Patscherkofel and also exceed those of the calcareous turf communities. Klact is low in T 1 whereas in T 2 it is in the range of P 1, P 2, and P 3. Compared with T 1, T 2 has distinctly higher amounts of the three nutrients in the soils and a higher net mineralization of N, as well as higher values in the phytomass components and in the fluctuation of the latter.In conclusion, a general view is given (Fig. 9) of the most important nutrient parameters of the communities represented in this series, including some others of lower altitudes.


Transformation by murine sarcoma virus alters the sensitivity of clonal cells derived from NIH/3T3 mouse fibroblasts to interferon.
In contrast to normal clonal cells (A5) derived from NIH/3T3 mouse fibroblasts, another clone (A10) derived from the same source was found to be resistant to the anti-lytic-virus activity of IFN and to be deficient in the induction of (2'-5') oligoadenylate synthetase (2-5A synthetase) by IFN. Following infection of either A5 or A10 cells with Moloney murine sarcoma virus (MSV), a few transformed colonies were isolated, expanded, and tested for their sensitivity to IFN. It is clearly demonstrated that IFN exerts a specific anti-proliferative effect on both MSV-transformed A5 (MA5) and A10 (MA10) cells, as evident by a slower growth rate, a decreased rate of DNA synthesis, and a lower cloning efficiency in its presence. Furthermore, unlike the original A10 cells, the IFN-treated transformed counterpart (MA10 cells), as well as MA5 cells, were protected from the lytic effect of either mengovirus or vesicular stomatitis virus (VSV). In addition, IFN treatment inhibited the release of retroviral particles from the transformed cells. The level of 2-5A synthetase activity in the various transformed cell lines was then determined. Whereas in A10 cells an induction of less than twofold in the enzymatic activity was detected following IFN treatment, a four- to fivefold increase in this activity could be seen in MA10 cells.


Hemodynamic events in the peritoneal environment during pneumoperitoneum in dogs.
The purpose of this experimental study was to determine the hemodynamic conditions of intraperitoneal viscera during pneumoperitoneum by using either CO2 gas or helium (He) for insufflation. In 16 mongrel dogs (divided into a CO2 group and an He group) subjected to 14 mmHg pneumoperitoneum for 60 min, the following parameters were assessed at times before and 1, 2, 5, 15, 30, 45, and 60 min thereafter: (1) intestinal mucosal blood flow, by means of a laser-Doppler probe inplanted into a jejunal loop; (2) portal pressure and portal blood pCO2, through a catheter inserted via a mesenteric jejunal vein; (3) intramural jejunal pH (pHi), by means of a Tonometer, which expresses the degree of tissue ischemia; (4) inferior vena cava pressure and blood pCO2, through a catheter inserted via a femoral vein; and (5) from the systemic circulation pulse rate, arterial blood pressure, CO, CVP, PVP, SaO2, pCO2, and paO2 were measured through a catheter placed into a femoral artery and a Swan-Ganz thermodilution catheter inserted via external jugular vein: CI and SVR were then calculated. Jejunal mucosal blood flow was found decreased (P < 0.0001) and pHi revealed gut mucosal ischemia. Portal and inferior vena cava pressures were found to be elevated (P < 0.0001), as was blood pCO2 of these vessels (P < 0.001), in only the CO2 group. From the systemic circulation, arterial blood pressure, CO, CI, SaO2, and paO2 revealed a decrease (P < 0.001) while arterial pCO2 (only CO2 group), CVP, SVR, and PVP revealed an increase (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


CD43 is required for optimal growth inhibition of Mycobacterium tuberculosis in macrophages and in mice.
We explored the role of macrophage (Mphi) CD43, a transmembrane glycoprotein, in the pathogenesis of Mycobacterium tuberculosis. Using gene-deleted mice (CD43-/-), we assessed the association of the bacterium with distinct populations of Mphi and found that CD43-/- Mphi bound less M. tuberculosis than CD43+/+ Mphi. Increased infective doses did not abrogate this difference. However, reduced association due to the absence of CD43 could be overcome by serum components. Mphi from heterozygote mice, which express 50% of wild-type CD43, bound more bacteria than CD43-/- but less than CD43+/+, proving that the gene dose of CD43 correlates with binding of M. tuberculosis. Furthermore, the reduced ability of CD43-/- Mphi to bind bacteria was restricted to mycobacterial species. We also found that the survival and replication of M. tuberculosis within Mphi was enhanced significantly in the absence of CD43, making this the first demonstration that the mechanism of mycobacterial entry influences its subsequent growth. Most importantly, we show here that the absence of CD43 in mice aerogenically infected with M. tuberculosis results in an increased bacterial load during both the acute and chronic stages of infection and more rapid development of granulomas, with greater lung involvement and distinctive cellularity.


In vivo depletion of leukocytes and platelets following injection of T cell-specific antibodies into mice.
In vivo T cell depletion experiments are widely used to establish the role of these cells in a variety of immunological processes. Different clones of monoclonal antibody targeting the CD3 molecular complex (mainly 145-2C11 and 17A2) have been successfully used for T cell depletion. In the present work, we assessed the specificity of monoclonal antibody-mediated CD3 T cell depletion in mouse peripheral blood. We showed that treatment of BALB/C mice with monoclonal antibodies (clones 145-2C11 and 17A2) not only efficiently depletes T cells in vivo, but also leads to a substantial reduction in B cell, granulocyte and platelet counts. In contrast, T cell depletion using a combination of anti-CD4 and anti-CD8 antibodies was efficient and produced less deleterious effects on other blood cell populations. Therefore, the results obtained from T cell depletion experiments using anti-CD3 antibodies must be interpreted with caution prior to draw definitive conclusions on the role of T cells in a given immunological process.


[Effect of nociceptive stimulation on the mitotic activity of cutaneous epithelial cells surrounding a wound at different age periods].
Wounds, 1 cm in length, reaching the cutaneous muscle were inflicted to rats, aged 11 days, 2 and 2 1/2 months, under the right scapula. In 48 hours the animals were subjected to pain irritation with electric current. Mitotic activity was determined in the cells of the basal layer of cutaneous epithelium. Pain stimulation decreased the mitotic index of the cutaneous epithelium cells in animals of all the age groups under study considerably (several times). There was an increase of mitotic index from the early to later periods of postnatal ontogenesis.


Corrigendum: Association between cardiovascular risk factors and the severity of coronavirus disease 2019: Nationwide epidemiological study in Korea.
[This corrects the article DOI: 10.3389/fcvm.2021.732518.].


Status of Simulation-Based Training in Departments of Surgery in the United States.
Surgical simulation is particularly attractive because it allows training in a safe, controlled, and standardized environment. However, the status of surgical simulation among Departments of Surgery (DoS) in the United States is unknown. The objective of this study was to characterize the status of simulation-based training in DoS in the United States. A Qualtrics online survey was sent to 177 chairs of DoS in the United States in March 2018 regarding the utilization of surgical simulation in their department. Questions in the survey were focused on simulation capacities and activities as well as chairs' perception of the value and purpose of simulation. A total of 87 of 177 chairs responded to the survey (49% response rate). Most programs had either 20-50 trainees (42 of 87; 48%) or more than 50 trainees (37 of 87; 43%). Most chairs reported having a simulation center in their institution (85 of 87; 98%) or department (60 of 86; 70%) with a formal simulation curriculum for their trainees (83 of 87; 95%). Ninety percent (78 of 87) of DoS had protected time for simulation education for their residents, with most residents engaging in activities weekly or monthly (65 of 85; 76%). Although most chairs felt simulation improves patient safety (72 of 84; 86%) and is useful for practicing surgeons (68 of 84; 81%), only 40% reported that faculty use simulation to maintain technical skills and only 17% reported that faculty use simulation to address high complication rates. The vast majority of the DoS in the United States have established simulation activities for their trainees. However, engagement of faculty in simulation to maintain or improve their skills remains low.


A simplified measurement procedure and portable electronic photometer for disposable sensors based on ionophore-chromoionophore chemistry for potassium determination.
A field-portable photometer for potassium determination with disposable sensors has been developed. It can be applied to routine water and beverage analysis. The disposable sensor is based on ionophore-chromoionophore chemistry. A colour change in the sensing film is detected by measuring the transmitted intensity with a solid state photodetector. Optical excitation at 660 nm is emitted by a light-emitting diode (LED). Negative feedback for LED bias and thermal correction were included to improve system stability. Additionally, a measurement procedure is presented, characterized and validated for in situ photometer use and real-time results. This simplified procedure is based on prior preparation of the disposable sensor in its acidic form and on the use of an absorbance ratio as analytical parameter. The only requirement for analysis is prior equilibration with a buffered sample solution for 3 min and absorbance measurement before and after equilibration. Good sensitivity in the concentration range 5 muM to 100 mM and very good repetitively and stability were achieved that are comparable to those obtained with bulkier analytical instrumentation. Given the compact size, low weight, rapid response and low energy requirement of the electronic photometer developed here, this measurement system is suitable for potassium determination in the field.


Palivizumab in congenital heart disease: should international guidelines be revised?
Palivizumab has reduced the incidence of respiratory syncytial virus hospitalization in infants and children with congenital heart disease by 45%. Although the mortality rate of children with congenital heart disease hospitalized with respiratory syncytial virus infection has declined from 37% to approximately 3% over the past 3 decades, palivizumab has not been shown to improve mortality. There has been considerable controversy over the cost-effectiveness of administering palivizumab according to international guidelines, including children with congenital heart disease. In particular, the number of children that need to be treated with palivizumab to prevent one respiratory syncytial virus hospitalization increases dramatically in children > 12 months of age. As a result, the authors recommend that countries re-examine their recommendations for providing palivizumab up to age 24 months in children with congenital heart disease.


CRISPR/Cas9-induced transgene insertion and telomere-associated truncation of a single human chromosome for chromosome engineering in CHO and A9 cells.
Chromosome engineering techniques including gene insertion, telomere-associated truncation and microcell-mediated chromosome transfer (MMCT) are powerful tools for generation of humanised model animal, containing megabase-sized genomic fragments. However, these techniques require two cell lines: homologous recombination (HR)-proficient DT40 cells for chromosome modification, and CHO cells for transfer to recipient cells. Here we show an improved technique using a combination of CRISPR/Cas9-induced HR in CHO and mouse A9 cells without DT40 cells following MMCT to recipient cells. Transgene insertion was performed in CHO cells with the insertion of enhanced green fluorescence protein (EGFP) using CRISPR/Cas9 and a circular targeting vector containing two 3 kb HR arms. Telomere-associated truncation was performed in CHO cells using CRISPR/Cas9 and a linearised truncation vector containing a single 7 kb HR arm at the 5' end, a 1 kb artificial telomere at the 3' end. At least 11% and 6% of the targeting efficiency were achieved for transgene insertion and telomere-associated truncation, respectively. The transgene insertion was also confirmed in A9 cells (29%). The modified chromosomes were transferrable to other cells. Thus, this CHO and A9 cell-mediated chromosome engineering using the CRISPR/Cas9 for direct transfer of the modified chromosome is a rapid technique that will facilitate chromosome manipulation.


[A case of laryngeal carcinoma recurrence in the neck lymphatic system after total laryngectomy and combined method treatment].
A patient with inoperable recurrence of the larynx carcinoma in lymphatic neck system after total laryngectomy was described. Nonradical surgical treatment combined with radical radiotherapy allowed to achieve a five-year symptom-free survival in this case.


How Does Bridging Core Modification Alter the Photovoltaic Characteristics of Triphenylamine-Based Hole Transport Materials? Theoretical Understanding and Prediction.
Perovskite solar cells have gained immense interest from researchers owing to their good photophysical properties, low-cost production, and high power conversion efficiencies. Hole transport materials (HTMs) play a dominant role in enhancing the power conversion efficiencies (PCEs) and long diffusion length of holes and electrons in perovskite solar cells. In hole transport materials, modification of π-linkers has proved to be an efficient approach for enhancing the overall PCE of perovskite solar cells. In this work, π-linker modification of a recently synthesized H-Bi molecule (R) is achieved with novel π-linkers. After structural modifications, ten novel HTMs (HB1-HB10) with a D-π-D backbone are obtained. The structure-property relationship, and optoelectronic and photovoltaic characteristics of these newly designed hole transport materials are examined comprehensively and compared with reference molecules. In addition, different geometric parameters are also examined with the assistance of density functional theory (DFT) and time-dependent DFT. All the designed molecules exhibit narrow HOMO-LUMO energy gaps (Eg =2.82-2.99 eV) compared with the R molecule (Eg =3.05 eV). The designed molecules express redshifting in their absorption spectra with low values of excitation energy, which in return offer high power conversion efficiencies. Further, density of states and molecular electrostatic potential analysis is performed to locate the different charge sites in the molecules. The reorganizational energies of holes and electrons are found to have good values, suggesting that these novel designed molecules are efficient hole transport materials for perovskite solar cells. In addition, the low binding energy values of the designed molecules (compared with R) offer high current charge density. Finally, complex study of HB9:PC61 BM is also undertaken to understand the charge transfer between the molecules of the complex. The results of all analyses advocate that these novel designed HTMs are promising candidates for the construction of future high-performance perovskite solar cells.


Detecting shifts in ecosystem functioning: the decoupling of fundamental relationships with increased pollutant stress on sandflats.
In this study, we investigated the influence of low level contamination by copper, lead and zinc on the functioning of estuarine sandflat ecosystems by comparing the strength and variability of relationships between benthic macrofauna and fluxes (oxygen and nutrients) at three clean and three mildly contaminated sites. Specifically, as indicators of ecosystem functioning, we examined relationships between bivalve biomass, total benthic respiration and ammonium release, and ammonium uptake and benthic primary production. Furthermore, a small amount of organic matter was added to experimental plots at all sites (35 g/0.2 m²) to evaluate stress-on-stress responses relative to controls. Relationships were strongest at the clean sites (steepest slopes, highest r² values, lowest p-values) and weakest at the mildly contaminated sites and in organically enriched plots. Our results suggest that changes in ecosystem functioning may be occurring at mild (<ERL) levels of pollution that are unlikely to cause widespread mortality of macrofauna.


Correction: Cost comparison of nine-month treatment regimens with 20-month standardized care for the treatment of rifampicin-resistant/multi-drug resistant tuberculosis in Nigeria.
[This corrects the article DOI: 10.1371/journal.pone.0241065.].


Nanomedicines for back of the eye drug delivery, gene delivery, and imaging.
Treatment and management of diseases of the posterior segment of the eye such as diabetic retinopathy, retinoblastoma, retinitis pigmentosa, and choroidal neovascularization is a challenging task due to the anatomy and physiology of ocular barriers. For instance, traditional routes of drug delivery for therapeutic treatment are hindered by poor intraocular penetration and/or rapid ocular elimination. One possible approach to improve ocular therapy is to employ nanotechnology. Nanomedicines, products of nanotechnology, having at least one dimension in the nanoscale include nanoparticles, micelles, nanotubes, and dendrimers, with and without targeting ligands. Nanomedicines are making a significant impact in the fields of ocular drug delivery, gene delivery, and imaging, the focus of this review. Key applications of nanotechnology discussed in this review include a) bioadhesive nanomedicines; b) functionalized nanomedicines that enhance target recognition and/or cell entry; c) nanomedicines capable of controlled release of the payload; d) nanomedicines capable of enhancing gene transfection and duration of transfection; f) nanomedicines responsive to stimuli including light, heat, ultrasound, electrical signals, pH, and oxidative stress; g) diversely sized and colored nanoparticles for imaging, and h) nanowires for retinal prostheses. Additionally, nanofabricated delivery systems including implants, films, microparticles, and nanoparticles are described. Although the above nanomedicines may be administered by various routes including topical, intravitreal, intravenous, transscleral, suprachoroidal, and subretinal routes, each nanomedicine should be tailored for the disease, drug, and site of administration. In addition to the nature of materials used in nanomedicine design, depending on the site of nanomedicine administration, clearance and toxicity are expected to differ.


Giardia duodenalis: adhesion-deficient clones have reduced ability to establish infection in Mongolian gerbils.
The role of Giardia duodenalis surface molecules in the attachment of trophozoites to epithelial cells has been established through the dual strategies of characterizing G. duodenalis clones with deficient adhesion and blocking experiments with surface-specific monoclonal antibodies. Also, the infectivity of the analyzed clones was tested using Mongolian gerbils as experimental model. Two adhesion-deficient G. duodenalis clones, C6 and C7, were isolated from the wild type C5 clone which in turn was obtained from the WB strain. The adhesion efficiencies of C6 and C7 clones (48.2+/-4.9 and 32.6+/-2.4, respectively) were significantly lower as compared with WB strain or C5 clone (82.8+/-6.4 and 79.9+/-7.9). Analysis of radiolabel surface proteins by 1D and 2D SDS-PAGE revealed prominently labelled 28 and 88 kDa components in C6 and C7 clones and a major 200 kDa protein in the C5 clone and the WB strain. The 88 and 200 kDa components are acidic proteins by two-dimensional electrophoretic analyses. The most striking difference between wild-type and adhesion-deficient Giardia trophozoites was the reduced expression of a 200 kDa surface protein in the latter. Significantly, a mAb (IG3) specific for the 200 kDa protein that reacted with more than 99% of WB and C5 trophozoites and less than 1% of C6 and C7 trophozoites as determined by indirect immunofluorescence inhibited the adhesion of trophozoites from WB and C5 clone to Madin Darby Canine Kidney cells by 52% and 40.9%, respectively, suggesting a participation of this antigen in adherence. Finally, the functional relevance of trophozoite adhesion to epithelial cells was indicated by the reduced capacity of the adhesion-deficient clones to establish the infection in Mongolian gerbils.


Concomitant boost radiotherapy for muscle invasive bladder cancer.
To evaluate the feasibility and efficacy of a concomitant partial bladder boost schedule in radiotherapy for invasive bladder cancer, coupling a limited boost volume with shortening of the overall treatment time. Between 1994 and 1999, 50 patients with a T2-T4 N0M0 transitional cell carcinoma of the bladder received radiotherapy delivered in a short overall treatment time with a concomitant boost technique. With this technique a dose of 40 Gy in 2-Gy fractions was administered to the small pelvis with a concomitant boost limited to the bladder tumor area plus margin of 15 Gy in fractions of 0.75 Gy. The total tumor dose was 55 Gy in 20 fractions in 4 weeks. Toxicity was scored according to EORTC/RTOG toxicity criteria. The feasibility of the treatment was good. Severe acute toxicity >/=G3 was observed in seven patients (14%). Severe late toxicity >/=G3 was observed in six patients (13%). Thirty-seven patients (74%) showed a complete and five (10 %) a partial remission after treatment. The actuarial 3-year freedom of local progression was 55%. In external radiotherapy for muscle invasive bladder cancer a concomitant boost technique coupling a partial bladder boost with shortening of the overall treatment time provides a high probability of local control with acceptable toxicity.


Sampling errors in free energy simulations of small molecules in lipid bilayers.
Free energy simulations are a powerful tool for evaluating the interactions of molecular solutes with lipid bilayers as mimetics of cellular membranes. However, these simulations are frequently hindered by systematic sampling errors. This review highlights recent progress in computing free energy profiles for inserting molecular solutes into lipid bilayers. Particular emphasis is placed on a systematic analysis of the free energy profiles, identifying the sources of sampling errors that reduce computational efficiency, and highlighting methodological advances that may alleviate sampling deficiencies. This article is part of a Special Issue entitled: Biosimulations edited by Ilpo Vattulainen and Tomasz Róg.


Differential outcome of concurrent radiotherapy plus epidermal growth factor receptor inhibitors versus radiotherapy plus cisplatin in patients with human papillomavirus-related head and neck cancer.
Human papillomavirus (HPV)-related head and neck cancer has been associated with an improved prognosis in patients treated with radiotherapy (RT) +/- chemotherapy (CT); however, RT combined with epidermal growth factor receptor (EGFR) inhibitors has not been fully studied in this group of patients. Immunohistochemical expression of p16 and PCR of HPV16 DNA were retrospectively analyzed in tumor blocks from 108 stage III/IV head and neck cancer patients treated with RT+CT (56) or RT+EGFR inhibitors (52). Disease-free survival (DFS) and overall survival (OS) were analyzed by the Kaplan-Meier method. DNA of HPV16 was found in 12 of 108 tumors (11%) and p16 positivity in 18 tumors (17%), with similar rates in both arms of treatment. After a median follow-up time of 35 months (range 6-135), p16-positive patients treated with RT+EGFR inhibitors showed improved survival compared with those treated with RT+CT (2-year OS 88% vs. 60%, HR 0.18; 95% CI 0.04 to 0.88; p = 0.01; and 2-year DFS 75% vs. 47%, HR 0.17; 95% CI 0.03 to 0.8; p = 0.01). However, no differences were observed in p16-negative patients (2-year OS 56% vs. 53%, HR 0.97; 95% CI 0.55 to 1.7; p = 0.9; and 2-year DFS 43% vs. 45%, HR 0.99; 95% CI 0.57 to 1.7; p = 0.9). This is the first study to show that p16-positive patients may benefit more from RT+EGFR inhibitors than conventional RT+CT. These results are hypothesis-generating and should be confirmed in prospective trials.


Self-referral: social and demographic determinants of consulting behaviour.
This report, which is one of a series, is concerned with socio-demographic correlates of self-referral, and the extent to which the observed associations can be explained by variations in symptom prevalence and severity. Social class, marital status, employment status, and distance from health centre, all show small but significant associations with self-referral. For social class the effect appears to be mediated by symptoms. People in social classes IV and V, women who are widowed, divorced or separated, and people who live near the health centre are more likely to visit their general practitioner.


Non-redundancy of cytidine deaminases in class switch recombination.
Class switch recombination (CSR), somatic hypermutation, and gene conversion are immunoglobulin diversification mechanisms that are strictly dependent on the activity of the activation-induced cytidine deaminase (AID). The precise role and substrate(s) of AID in these processes remain to be well defined. The closest homologue of AID is APOBEC-1, a bona fide mRNA-editing enzyme, which shares with AID the ability to deaminate cytidines within single-stranded DNA in vitro and in prokaryotic cells. To determine whether APOBEC-1 can therefore substitute for AID in activated B cells, we expressed human AID, a catalytic mutant thereof, and rat APOBEC-1 in AID-deficient murine B cells. Whereas AID rescued CSR, neither the inactive mutant nor APOBEC-1 could complement AID deficiency. This indicates that cytidine deaminase activity is necessary but not sufficient to initiate CSR, and suggests that AID is specifically targeted to its cognate substrate, the immunoglobulin genes or a distinct mRNA, by an as-yet-unknown mechanism.


The challenges of PICU research: Lessons learned from a minimal-risk study with PICU parents.
Research conducted in pediatric intensive care units (PICUs) with families is essential to advancing evidenced-based practice and improving patient outcomes in this unique setting. However, several ethical, logistical, and methodological challenges have been cited in the literature as having a significant effect on the development of PICU research. Investigators at a large midwestern health care center encountered several challenges during the course of a minimal-risk, survey-based study with parents of PICU patients. This manuscript aims to highlight the challenges faced by the research team, which included challenges related to the environment of the PICU, the patients' length of stay, the health status of the patient, and the etiology of the patient's admission, as well as share the actions that the research team took to address these challenges.


Fluid absorption by skin tissue during intradermal injections through hollow microneedles.
Hollow microneedles are an emerging technology for delivering drugs and therapeutics, such as vaccines and insulin, into the skin. Although the benefits of intradermal drug delivery have been known for decades, our understanding of fluid absorption by skin tissue has been limited due to the difficulties in imaging a highly scattering biological material such as skin. Here, we report the first real-time imaging of skin tissue at the microscale during intradermal injections through hollow microneedles, using optical coherence tomography. We show that skin tissue behaves like a deformable porous medium and absorbs fluid by locally expanding rather than rupturing to form a single fluid filled cavity. We measure the strain distribution in a cross section of the tissue to quantify local tissue deformation, and find that the amount of volumetric expansion of the tissue corresponds closely to the volume of fluid injected. Mechanically restricting tissue expansion limits fluid absorption into the tissue. Our experimental findings can provide insights to optimize the delivery of drugs into skin for different therapeutic applications, and to better model fluid flow into biological tissue.


Characterization of calcium translocation across the plasma membrane of primary osteoblasts using a lipophilic calcium-sensitive fluorescent dye, calcium green C18.
The synthesis of Calcium Green C18, a lipophilic fluorescent calcium-sensitive dye, and its use as a monitor of Ca2+ efflux from cells is described. This indicator consists of a Calcium Green-1 molecule conjugated to a lipophilic 18-carbon alkyl chain which will intercalate into cell membranes. The Kd of the indicator for Ca2+ in aqueous solution (pH 7.2, 22 degrees C, ionic strength 0.1 M) is 0.23 +/- 0.04 microM and in the presence of liposomes is 0.062 +/- 0.007 microM. Due to its high negativity, the calcium chelating fluorophore faces the cell exterior, when loaded under a defined set of conditions. The dye was found largely on the surface of the cells when loaded at a concentration of 5 microM for 10 min at 37 degrees C. Five minutes after introduction of EGTA, 83-95% fluorescence disappeared, indicating that most of the fluorophore was on the cell surface. Photobleaching was minimal (3-13%). A confocal laser scanning microscope was used to detect and quantify fluorescence. Internalized dye was apparent in cells loaded for longer times (30-60 min) and in membrane-impaired cells, as shown by uptake of propidium iodide. Under defined confocal laser scanning microscope settings, a transient fluorescence at the periphery of approximately 30% of the cells was observed following 10(-8) M parathyroid hormone treatment, indicating the presence of outwardly directed calcium transport across the plasma membrane. Calcium efflux usually lasted 7-10 min, peaking at around 2-3 min. Changes in cell shape were also observed. Calcium efflux was shown to be sensitive to (a) 10 microM quercetin and 10 microM vanadate, partially specific inhibitors of plasma membrane Ca(2+)-ATPase, to (b) 0.1 mM trifluoperazine, an agent which renders calmodulin ineffective, and to (c) 10 mM neomycin sulfate, which blocks release of Ca2+ from intracellular stores. Thapsigargin (5 microM), an inhibitor of Ca(2+)-ATPase of the endoplasmic reticulum, prolonged fluorescence. These observations indicate that cell surface fluorescence was due to the capture of Ca2+ by Calcium Green C18 after Ca2+ had been translocated across osteoblast plasma membranes. Involvement of the plasma membrane Ca(2+)-ATPase, known to be present in osteoblasts in substantial amounts, is implicated.


Iso-Seq assembly and functional annotation of full-length transcriptome of turbot (Scophthalmus maximus) during bacterial infection.
Turbot (Scophthalmus maximus) is a flatfish, which is not only important in mariculture worldwide with the unique characteristic of body asymmetry, but also as an economically important species in aquaculture. Herein, we performed the first full-length transcriptome sequencing of turbot during the bacterial infection. A total of 307.1 Gb raw reads were obtained and processed with Iso-Seq, generating 187,509 high-quality redundant transcripts with an average length of 3005 base pairs. Benchmarking Universal Single-Copy Orthologs (BUSCO) analysis identified 81.5% complete BUSCOs and only 1.7% fragmented BUSCOs, suggesting a transcript structural completeness and functional diversity of this transcriptome. Moreover, the redundant transcripts were collapsed and compared to ENSEMBL reference with Cupcake and SQANTI3. Among 60,476 collapsed transcripts, we identified 12,059 annotated and 1684 novel genes. 42,956 (71.1%) transcripts provided new evidence for splice junctions identification. Furthermore, the untranslated region (UTR) identification was also benefited from the transcriptome. The open read frames prediction was conducted with PASApipeline. 42,118 transcripts were assigned with known function by aligning against Swiss-Prot or functional domain prediction. Taken together, the full-length transcriptome built in this study could provide important resources for immunologic research on turbot.


Frontal lobe role in simple arithmetic calculations: an fNIR study.
This study aimed to affirm the use of functional near-infrared spectroscopy (fNIR) in examining frontal lobe role during automatic (i.e., requires retrieval from long-term memory) and method-based (i.e., requires calculation) arithmetic processing. Adult university students (math difficulties [MD] and control) performed simple arithmetic calculations while monitored using an fNIR system designed to image regions within the frontal cortices. Addition and subtraction problems presented on a computer screen belonged to one of three categories: triples "under 10" (e.g., 2+3=?, 5-3=?), triples that "break 10" (e.g., 5+8=?, 13-5=?), or triples "including 10" (e.g., 10+7=?, 17-10=?). fNIR recordings indicated significant interactions between type of triple, operation, and group over left frontal lobe, and between type of triple and group over right frontal lobe. Within-group differences among controls were found in the "break 10" triples with higher DeOxyHb level recorded during subtraction processing. Between-group differences were found in the "break 10" and "including 10" triples for subtraction with higher levels of DeOxyHb recorded among controls. Results imply that among adults frontal lobe is still involved during simple mathematical processing and fNIR recordings can differentiate its role in adults of varying mathematical ability.


Bisalbumineamia in a bottlenosed dolphin (Tursiops truncatus).
Bisalbuminemia was found in a female bottlenosed dolphin (Turiops truncatus) on routine examination. There is no association with disease.


Paracetamol poisoning--impact of pack size restrictions.
Pack sizes of paracetamol available to the public were reduced by legislation in 1998. To study the effect of the legislation on paracetamol poisoning related admissions to an accident and emergency department and to a tertiary referral liver unit. Data were obtained from the University Hospitals, Birmingham, UK (UHB) Trust Information Service and the Queen Elizabeth Hospital liver unit records on (a) the number of patients admitted to hospital because of paracetamol overdose and (b) the number of patients admitted to the liver unit with parcetamol-induced hepatotoxicity. Prior to legislation, an average of 360 people a year were admitted to UHB. However, following the change in legislation the number of admissions has fallen to an average of 250 people per year. This represents a reduction of 31%. Prior to legislation, an average of 76 people per year were admitted to the liver unit. Post-legislation this number has dropped to an average of 38 per year representing a reduction of 50%. Legislation restricting paracetamol pack-size reduced the incidence and severity of poisoning.


When to recommend and to pay for first-line adjuvant breast cancer treatment? A structured review of the literature.
A structured review of studies on the health-economic evaluation of systemic adjuvant therapy for early-stage breast cancer was carried out. Of the eight articles that have been identified four were related to the cost-effectiveness of chemotherapy, three compared chemotherapy with combined chemotherapy and hormonal therapy and one compared tamoxifen (TAM) with third-generation aromatase inhibitors (ATIs). Results of the review indicate that the cost-utility of adjuvant breast cancer therapy is within the range of other oncological interventions. Adjuvant chemotherapy is most cost-effective in pre-menopausal women with node-positive breast cancer while cost-effectiveness decreases considerably with increasing age. Endocrine therapy with TAM is most cost-effective in ER-positive tumours with no significant age effect. The cost-utility of using the ATI anastrozole instead of TAM in adjuvant therapy cannot be conclusively assessed on the basis of the existing evidence.


The hydrogen sulphide-releasing derivative of diclofenac protects against ischaemia-reperfusion injury in the isolated rabbit heart.
Hydrogen sulphide (H(2)S) is an endogenous gaseous mediator active in the multilevel regulation of pathophysiological functions in mammalian cardiovascular tissues. This study investigated the pharmacological activity of a new H(2)S-releasing derivative of diclofenac, S-diclofenac (2-[(2,6-dichlorophenyl)amino]benzeneacetic acid 4-(3H-1,2-dithiole-3-thione-5-yl)-phenyl ester) in the isolated rabbit heart submitted to low-flow ischaemia-reperfusion damage. S-diclofenac (3, 10 and 30 microM), despite inhibiting prostacyclin generation by cardiac tissues, achieved dose-dependent normalization of coronary perfusion pressure, reducing left ventricular contracture during ischaemia and improving left ventricular developed pressure and +/-dP/dt(max) at reperfusion. Creatine kinase and lactate dehydrogenase activities in heart perfusates were significantly reduced during reperfusion. These effects were accompanied by substantial release of reduced glutathione (GSH), indicating that the H(2)S moiety may have up-regulated cysteine transport. The anti-ischaemic activities of S-diclofenac and the H(2)S-donor sodium hydro sulphide (NaHS) were partially prevented by the K(ATP) channel antagonist glibenclamide, suggesting a mechanism similar to H(2)S-induced cardioprotection in metabolic ischaemic preconditioning. Perfusion with the nitric oxide (NO) synthase inhibitor N(G)-monomethyl-L-arginine worsened the myocardial ischaemia-reperfusion damage, but this was dose-dependently prevented by S-diclofenac and NaHS, suggesting that the released H(2)S may have overcome NO deficiency. These data show that S-diclofenac had marked anti-ischaemic activity in ischaemic-reperfused rabbit hearts despite inhibition of prostaglandin generation. Increased GSH formation leading to activation of K(ATP) channels may have contributed to this beneficial effect. The pharmacological profile of S-diclofenac and its anti-inflammatory activity, with diminished gastrointestinal side effects, offer therapeutic applications in cardiovascular disease.


Analysis of human chromosome 11 by somatic cell genetics: reexamination of derivatives of human-hamster cell line J1.
Through the fusion of a CHO cell population to a human cell population, a hybrid cell line which has lost all human chromosomes except chromosome 11 was derived. This cell line, J1, does not appear to segregate human chromosome 11 during growth. A series of deletion segregants were isolated from J1 which had lost a portion of either the long, short, or both arms of chromosome 11. This panel of deletion segregants was used for mapping a number of genetic markers on the short arm of chromosome 11. Karyotypic analysis led to the interpretation that derivatives of J1 selected for the loss of cell surface antigens encoded by genes on the short arm of the chromosome had simple terminal deletions of this chromosome arm. More recently, we have applied recombinant DNA and in situ hybridization techniques to the analysis of the structure of chromosome 11. In the course of this analysis, we have obtained data that indicate that all J1 deletion segregants retain a small chromosomal segment containing the structural genes for insulin and HRAS1. Analysis of in situ hybridization data indicates that in cell lines in which a chromosome 11 fragment cannot be identified by karyotype analysis, human DNA has been translocated to a Chinese Hamster chromosome. These results suggest that the original interpretation of the karyotypes of deletion segregants derived from J1 as simple terminal deletions is not correct. A reanalysis of gene localization studies based on these deletion segregants suggests that some assignments of genes to specific bands on chromosome 11 should be reconsidered. In particular, data on additional deletion segregants are consistent with localization of the beta-globin gene complex to band 11p15. The data presented here suggest that in several hybrid derivatives of J1, a continuous DNA segment of approximately 10(7) base pairs in length which includes the insulin and HRAS1 (cellular homolog of retroviral oncogene Harvey ras) genes has been isolated from the remainder of the human genome. We propose that the stability of chromosome 11 in the original hybrid was due to complementation of a genetic defect in the original CHO cell parent by a gene located in close physical proximity to the insulin and HRAS1 genes on chromosome 11. Data are presented which test and support this hypothesis.


Neuroendocrine immune pathways in chronic arthritis.
The analysis and understanding of the complex effects of endocrine and nervous system alterations on the inflammatory process in human arthritis is far from complete. Such alterations are observed as decreased responsiveness of the hypothalamic-pituitary-adrenal axis, an inadequate production of cortisol in relation to inflammation, and - consequently - elevated sympathetic activity, alterations of sex hormone metabolism (loss of androgens), psychological alterations (with chronic fatigue and symptoms of depression due to elevated circulating cytokines), local reduction of synovial sympathetic innervation, altered metabolism of estrogens in the synovium, and high expression of estrogen receptors in synovial cells. An understanding of these alterations will help to identify the different neuroendocrine immune mechanisms involved in the pathophysiology of rheumatoid arthritis and could trigger research into novel therapeutic targets for the treatment of patients with rheumatoid arthritis.


Adrenal cortical carcinoma: preoperative demonstration of right atrial extension by sonography and computerized tomography.
We report a case of adrenal cortical carcinoma with tumor thrombus extending to the right atrium. Tumor extension was demonstrated preoperatively with sonography and computerized tomography. We recommend that both modalities be used when evaluating masses in the suprarenal space. The preoperative diagnosis of tumor extension via the adrenal vein into the inferior vena cava was confirmed at operation.


Community participation in primary care: what does it mean 'in practice'?
The New Zealand 2001 Primary Health Care Strategy requires primary health organisations (PHOs) to involve communities in their governance and be responsive to communities' needs. It is less clear what the implications of this policy are for general practices. This paper presents key findings from a national study undertaken in the wake of the 2001 primary care reforms on the purpose and process of involving communities in primary health care. A three-part qualitative process was employed. Initially, key informant interviews were undertaken with a diverse range of primary care policy makers, funders, practitioners and community stakeholders (n=42). A draft framework for involving communities in primary care was developed, and formed the basis of a consultation phase (n=26). The framework was further developed and then piloted in four PHOs around New Zealand. Views on community participation varied among different stakeholder groups in the sector. Most described it as a complex process of relationship-building over time, and one that is quite distinct from consumer feedback processes in general practice. For community representatives, it was a process of trust-building and information-sharing between communities and health professionals. These relationships enabled people to feel comfortable seeking care, and professionals to mould services to people's needs. This research highlights that community is much more than a setting of care. As citizens, members of disadvantaged communities are partners with general practices and PHOs, working with them to improve health equity by ensuring that services are responsive to their needs.


A multi-omic resource of wheat seed tissues for nutrient deposition and improvement for human health.
As a globally important staple crop, wheat seeds provide us with nutrients and proteins. The trend of healthy dietary has become popular recently, emphasizing the consumption of whole-grain wheat products and the dietary benefits. However, the dynamic changes in nutritional profiles of different wheat seed regions (i.e., the embryo, endosperm and outer layers) during developmental stages and the molecular regulation have not been well studied. Here, we provide this multi-omic resource of wheat seeds and describe the generation, technical assessment and preliminary analyses. This resource includes a time-series RNA-seq dataset of the embryo, endosperm and outer layers of wheat seeds and their corresponding metabolomic dataset, covering the middle and late stages of seed development. Our RNA-seq experiments profile the expression of 63,708 genes, while the metabolomic data includes the abundance of 984 metabolites. We believe that this was the first reported transcriptome and metabolome dataset of wheat seeds that helps understand the molecular regulation of the deposition of beneficial nutrients and hence improvements for nutritional and processing quality traits.


Effect of low-dose atropine on heart rate fluctuations during orthostatic load: a spectral analysis.
Spectral analysis was utilized in order to determine the influence of low (0.7 microgram/kg)-dose atropine sulfate injections on the isolated spectral power components of heart rate fluctuations in contrast to moderate (0.02 mg/dose)-dose atropine and unmedicated states in human subjects during orthostatic load. Low-dose atropine decreased mean heart rate and increased respiration-related spectral power compared with controls in both the supine and standing posture. In supine subjects total power (0.01-0.5 Hz) was unchanged compared with controls due to a decreasing trend of low-frequency (0.01-0.05 Hz) and mid-frequency (0.05-0.15 Hz) heart rate spectral power. Standing upright, there was a distinct increase in total power with significantly higher values than in the controls resulting from an enormous activation of mid-frequency heart rate fluctuations. The peak frequency of the mid-frequency component was increased with a strong tendency compared with controls. Moderate-dose atropine increased mean heart rate and decreased total power, mid-frequency and respiration-related heart rate spectral power and peak frequency compared with controls in both positions. Our results suggest that low-dose atropine affects the interaction between sympathetic and parasympathetic limbs in the autonomic control of cardiac function in a complex manner producing a differentiated pattern of heart rate fluctuations dependent on the body posture. We suggest that low-dose atropine augments and moderate-dose atropine attenuates the vagal cardiac efferent activity in each position.


Guanidinium-grafted polyethylenimine: an efficient transfecting agent for mammalian cells.
Polyethylenimine (PEI) is one of the most efficient polycationic non-viral gene delivery vectors. Its efficiency and cytotoxicity depends on molecular weight, with the 25-kDa PEI being most efficient but accompanied with cytotoxicity. In the present study, enhancement in gene delivery efficiency along with reduction in cytotoxicity by attachment of guanidinium side group was explored. The hypothesis was that the guanidination would lead to the delocalization of charge present on primary amines of the polymer thereby leading to enhancement in gene delivery efficiency along with reduction in cytotoxicity. The polymer was guanidinated using O-methylisourea hemisulfate and the chemical linkage characterized by FTIR spectroscopy. The hydrodynamic diameter of guanidinated PEI-DNA complexes was determined using DLS. Subsequently, these complexes were used for DNA binding assay and zeta-potential measurements, taking native PEI as reference. Further, guanidinated PEI-DNA complexes were investigated for their gene delivery efficacy on HEK 293 cells. The hydrodynamic diameter of guanidinated PEI-DNA complexes was found to be in the range of 176-548 nm. As expected, the zeta potential values increased, on increasing the N/P ratios. It was found that guanidinated PEI had higher transfection efficiency at the majority of the N/P ratios tested as compared to commercially available transfecting agent lipofectin and native PEI itself. The toxicity of guanidinated PEI-DNA complexes was also reduced considerably in comparison to PEI polymer, as determined by MTT colorimetric assay. Out of the various derivatives prepared, gPEI 56% was found to be the most efficient in in vitro transfection.