Embedding new technologies in practice - a normalization process theory study of point of care testing.
Many point of care diagnostic technologies are available which produce results within minutes, and offer the opportunity to deliver acute care out of hospital settings. Increasing access to diagnostics at the point of care could increase the volume and scope of acute ambulatory care. Yet these technologies are not routinely used in many settings. We aimed to explore how point of care testing is used in a setting where it has become 'normalized' (embedded in everyday practice), in order to inform future adoption and implementation in other settings. We used normalization process theory to guide our case study approach. We used a single case study design, choosing a community based ambulatory care unit where point of care testing is used routinely. A focused ethnographic approach was taken, including non-participant observation of all activities related to point of care testing, and semi-structured interviews, with all clinical staff involved in point of care testing at the unit. Data were analysed thematically, guided by normalization process theory. Fourteen days of observation and six interviews were completed. Staff had a shared understanding of the purpose, value and benefits of point of care testing, believing it to be integral to the running of the unit. They organised themselves as a team to ensure that point of care testing worked effectively; and one key individual led a change in practice to ensure more consistency and trust in procedures. Staff assessed point of care testing as worthwhile for the unit, their patients, and themselves in terms of job satisfaction and knowledge. Potential barriers to adoption of point of care testing were evident (including lack of trust in the accuracy of some results compared to laboratory testing; and lack of ease of use of some aspects of the equipment); but these did not prevent point of care testing from becoming embedded, because the importance and value attributed to it were so strong. This case study offers insights into successful adoption of new diagnostic technologies into every day practice. Such analyses may be critical to realising their potential to change processes of care.


Direct measurement of spin-lattice relaxation times of phosphorus metabolites in human myocardium.
T1 values of phosphorus metabolites visible in human cardiac 31P-MR spectra were determined in 12 volunteers at 1.5 T. Consecutive spectra were acquired with varying pulse repetition time (TR) from 1.6 to 24 s; volume selection was achieved with ISIS. T1's of creatine phosphate (CP), [gamma-P], [alpha-P], and [beta-P]ATP, 2-3 diphosphoglycerate, and phosphodiesters were 6.1 +/- 0.5, 5.4 +/- 0.5, 5.5 +/- 0.5, 5.8 +/- 1.0, 7.6 +/- 1.0, and 5.0 +/- 1.0 s, respectively. CP/ATP ratios showed little change with varying TR; linear regression of CP/ATP vs TR was of borderline significance (r = 0.28, P = 0.06). T1's for CP and ATP were also determined in standard solution (20 mM CP, 10 mM ATP) yielding T1CP of 8.7 +/- 0.2 and T1[gamma-P]-ATP of 9.9 +/- 0.7 s. Thus, T1's for CP and ATP were similar at 1.5 T in both human heart and standard solution. In human cardiac 31P-MR spectra, CP/ATP ratios may need little correction for partial saturation.


Treating Major Depression with Physical Activity: A Systematic Overview with Recommendations.
The purpose of this systematic overview was to determine the most effective mode and dose of physical activity (PA) for treating major depressive disorder (MDD), and to suggest guidelines and recommendations for clinicians. The selection process consisted of a comprehensive search that was conducted up until April 2014 in the following databases: PsycINFO, Medline, PubMed and Scopus. The inclusion criteria were: (1) a randomized controlled trial (RCT) design, (2) complete description of intensity, duration and frequency of the PA, (3) the participants had to be diagnosed with MDD according to Diagnostic Statistical Manual 4 th edition (DSM-IV) or International Classification of Disease tenth Revision (ICD-10) criteria (4) if the controls received any treatment, it had to be specified, (5) published after 1990, (6) consist of aerobic or anaerobic treatment PA, and (7) not be a pilotor preliminary study. A quality assessment of each study was conducted independently by two reviewers; this stringent selection process resulted in 12 reviewed studies. individually customized PA, for at least 30 minutes, preferably performed under supervision and with a frequency of at least three times per week is recommended when treating MDD. These recommendations must be viewed in light of the relatively few studies matching the inclusion criteria.


Inhibition of dopamine transporter activity by G protein βγ subunits.
Uptake through the Dopamine Transporter (DAT) is the primary mechanism of terminating dopamine signaling within the brain, thus playing an essential role in neuronal homeostasis. Deregulation of DAT function has been linked to several neurological and psychiatric disorders including ADHD, schizophrenia, Parkinson's disease, and drug addiction. Over the last 15 years, several studies have revealed a plethora of mechanisms influencing the activity and cellular distribution of DAT; suggesting that fine-tuning of dopamine homeostasis occurs via an elaborate interplay of multiple pathways. Here, we show for the first time that the βγ subunits of G proteins regulate DAT activity. In heterologous cells and brain tissue, a physical association between Gβγ subunits and DAT was demonstrated by co-immunoprecipitation. Furthermore, in vitro pull-down assays using purified proteins established that this association occurs via a direct interaction between the intracellular carboxy-terminus of DAT and Gβγ. Functional assays performed in the presence of the non-hydrolyzable GTP analog GTP-γ-S, Gβγ subunit overexpression, or the Gβγ activator mSIRK all resulted in rapid inhibition of DAT activity in heterologous systems. Gβγ activation by mSIRK also inhibited dopamine uptake in brain synaptosomes and dopamine clearance from mouse striatum as measured by high-speed chronoamperometry in vivo. Gβγ subunits are intracellular signaling molecules that regulate a multitude of physiological processes through interactions with enzymes and ion channels. Our findings add neurotransmitter transporters to the growing list of molecules regulated by G-proteins and suggest a novel role for Gβγ signaling in the control of dopamine homeostasis.


Imaging correlates of brain function in monkeys and rats isolates a hippocampal subregion differentially vulnerable to aging.
The hippocampal formation contains a distinct population of neurons organized into separate anatomical subregions. Each hippocampal subregion expresses a unique molecular profile accounting for their differential vulnerability to mechanisms of memory dysfunction. Nevertheless, it remains unclear which hippocampal subregion is most sensitive to the effects of advancing age. Here we investigate this question by using separate imaging techniques, each assessing different correlates of neuronal function. First, we used MRI to map cerebral blood volume, an established correlate of basal metabolism, in the hippocampal subregions of young and old rhesus monkeys. Second, we used in situ hybridization to map Arc expression in the hippocampal subregions of young and old rats. Arc is an immediate early gene that is activated in a behavior-dependent manner and is correlated with spike activity. Results show that the dentate gyrus is the hippocampal subregion most sensitive to the effects of advancing age, which together with prior studies establishes a cross-species consensus. This pattern isolates the locus of age-related hippocampal dysfunction and differentiates normal aging from Alzheimer's disease.


A Systematic Review and Meta- Analysis of Green-Light Laser Vaporization for Superficial Bladder.
The related research of green-light laser vaporization in the treatment of non-muscle invasive bladder cancer (NMIBC) is limited. This study focused on analyzing the effectiveness and safety of it from the perspective of an extensive literature review. A comprehensive search of CNKI, WanFang, VIP, PubMed, Embase, and CENTRAL databases for photoselective vaporization of bladder tumor and transurethral resection of bladder tumor treatment of non-muscle invasive bladder cancer (NMIBC). The search included studies from January 1996 to December 2019. Two reviewers independently screened literature, extracted data, assessed the risk of bias of included studies. RevMan 5.3 software was used for Meta-analysis. A total of 18 RCTs involving 1648 patients met the predefined criteria. Meta-analysis data demonstrated that the PVBT group exhibited a significant advantage over the TURBT group in intraoperative obturator nerve reflex (RR = 0.09, 95% CI [0.04, 0.18], P< 0.001)and bladder perforation (RR = 0.14, 95% CI [0.07, 0.28], P< 0.001) and postoperative 1-year recurrence (RR = 0.52, 95% CI [0.40, 0.67], P< 0.001). The PVBT procedure has advantages over TURBT in the amount of surgical bleeding (MD = -17.27, 95% CI [-24.73, -9.81], P< 0.001) and the length of hospital stay (MD = -2.80, 95% CI [-3.82, -1.87], P< 0.001), bladder irrigation time (MD = -0.95, 95% CI [-1.49, -0.42], P< 0.001), and catheter indwelling time (MD = -2.60, 95% CI [-3.30, -1.90], P< 0.001). There was no difference between the two types of surgery in the incidence of postoperative urethral stricture (RR = 0.53, 95% CI [0.15, 1.83], P = 0.32) and the length of surgery (MD = -2.46, 95% CI [-5.37, 0.46], P = 0.10). Our systematic review and meta-analysis suggests that PVBT is better than TURBT as an alternative treatment for patients with NMIBC in safe aspect. However, whether it is equally effective in terms of oncological control remains to be elucidated, and additional high quality RCTs are needed to confirm our findings.


Parameters influencing the accuracy and practical applicability of UV indicator cards.
In order to evaluate the potential hazard for the skin and the eye presented by solar ultraviolet radiation, and to take appropriate precautions, it is necessary to characterise the biological effective irradiance or dose. Recently, inexpensive UV indicator cards became available that in principle would provide an attractive means to roughly indicate the local level of erythemal solar UV irradiance. We have characterised the properties of a number of different types of UV indicator cards. Several parameters which may influence the colour of the cards were examined with both outdoor trials under solar UV as well as indoor trials using a filtered xenon arc lamp. Our findings show that the tested cards do not give an appropriate estimation of the effective irradiance due to their spectral sensitivity and their temperature dependence. The application area of the tested UV indicator cards is therefore limited to certain temperature ranges and to seasons where a certain ratio between solar UV-A and solar UV-B occurs.


Vascularization of cytochrome oxidase-rich blobs in the primary visual cortex of squirrel and macaque monkeys.
The close correlation between energy supply by blood vessels and energy consumption by cellular processes in the brain is the basis of blood flow-related functional imaging techniques. Regional differences in vascular density can be detected using high-resolution functional magnetic resonance imaging. Therefore, inhomogeneities in vascularization might help to identify anatomically distinct areas noninvasively in vivo. It was reported previously that cytochrome oxidase-rich blobs in the striate cortex of squirrel monkeys are characterized by a notably higher vascular density (42% higher than interblob regions). However, blobs have so far never been identified in vivo on the basis of their vascular density. Here, we analyzed blobs of the primary visual cortex of squirrel monkeys and macaques with respect to the relationship between vascularization and cytochrome oxidase activity. By double staining with cytochrome oxidase enzyme histochemistry to define the blobs and collagen type IV immunohistochemistry to quantify the blood vessels, a close correlation between oxidative metabolism and vascularization was confirmed and quantified in detail. The vascular length density in cytochrome oxidase blobs was on average 4.5% higher than in the interblob regions, a difference almost one order of magnitude smaller than previously reported. Thus, the vascular density that is closely associated with local average metabolic activity is a structural equivalent of cerebral metabolism and blood flow. However, the quantitative differences in vascularization between blob and interblob regions are small and below the detectability threshold of the noninvasive hemodynamic imaging methods of today.


Performance comparison of second- and third-generation sequencers using a bacterial genome with two chromosomes.
The availability of diverse second- and third-generation sequencing technologies enables the rapid determination of the sequences of bacterial genomes. However, identifying the sequencing technology most suitable for producing a finished genome with multiple chromosomes remains a challenge. We evaluated the abilities of the following three second-generation sequencers: Roche 454 GS Junior (GS Jr), Life Technologies Ion PGM (Ion PGM), and Illumina MiSeq (MiSeq) and a third-generation sequencer, the Pacific Biosciences RS sequencer (PacBio), by sequencing and assembling the genome of Vibrio parahaemolyticus, which consists of a 5-Mb genome comprising two circular chromosomes. We sequenced the genome of V. parahaemolyticus with GS Jr, Ion PGM, MiSeq, and PacBio and performed de novo assembly with several genome assemblers. Although GS Jr generated the longest mean read length of 418 bp among the second-generation sequencers, the maximum contig length of the best assembly from GS Jr was 165 kbp, and the number of contigs was 309. Single runs of Ion PGM and MiSeq produced data of considerably greater sequencing coverage, 279× and 1,927×, respectively. The optimized result for Ion PGM contained 61 contigs assembled from reads of 77× coverage, and the longest contig was 895 kbp in size. Those for MiSeq were 34 contigs, 58× coverage, and 733 kbp, respectively. These results suggest that higher coverage depth is unnecessary for a better assembly result. We observed that multiple rRNA coding regions were fragmented in the assemblies from the second-generation sequencers, whereas PacBio generated two exceptionally long contigs of 3,288,561 and 1,875,537 bps, each of which was from a single chromosome, with 73× coverage and mean read length 3,119 bp, allowing us to determine the absolute positions of all rRNA operons. PacBio outperformed the other sequencers in terms of the length of contigs and reconstructed the greatest portion of the genome, achieving a genome assembly of "finished grade" because of its long reads. It showed the potential to assemble more complex genomes with multiple chromosomes containing more repetitive sequences.


Color Doppler-US guidance in transjugular placement of intrahepatic portosystemic shunts.
Percutaneous placement of portosystemic shunts requires access to the portal system from a transjugular approach. Color Doppler sonography was used to direct the transjugular puncture in intrahepatic portosystemic shunt procedures in four patients. In each case, the technique allowed quick, safe transjugular puncture of the portal vein and close real-time monitoring of the procedure.


Adeno-associated viral vector serotypes 1 and 5 targeted to the neonatal rat and pig striatum induce widespread transgene expression in the forebrain.
Viral vector-mediated gene transfer has emerged as a powerful means to target transgene expression in the central nervous system. Here we characterized the efficacy of serotypes 1 and 5 recombinant adeno-associated virus (rAAV) vectors encoding green fluorescent protein (GFP) after stereotaxic delivery to the neonatal rat and minipig striatum. The efficiency of GFP expression and the phenotype of GFP-positive cells were assessed within the forebrain at different time points up to 12 months after surgery. Both rAAV1-GFP and rAAV5-GFP delivery resulted in transduction of the striatum as well as striatal input and output areas, including large parts of the cortex. In both species, rAAV5 resulted in a more widespread transgene expression compared to rAAV1. In neonatal rats, rAAV5 also transduced several other areas such as the olfactory bulbs, hippocampus, and septum. Phenotypic analysis of the GFP-positive cells, performed using immunohistochemistry and confocal microscopy, showed that most of the GFP-positive cells by either serotype were NeuN-positive neuronal profiles. The rAAV5 vector further displayed the ability to transduce non-neuronal cell types in both rats and pigs, albeit at a low frequency. Our results show that striatal delivery of rAAV5 vectors in the neonatal brain represents a useful tool to express genes of interest both in the basal ganglia and the neocortex. Furthermore, we apply, for the first time, viral vector-mediated gene transfer to the pig brain providing the opportunity to study effects of genetic manipulation in this non-primate large animal species. Finally, we generated an atlas of the Göttingen minipig brain for guiding future studies in this large animal species.


Comparative morphology of the hepatic and coronary artery walls. Part I. Differences in the distribution and intensity of non-atherosclerotic intimal thickening and atherosclerosis.
We compared the morphology of the hepatic artery intima which is free of atherosclerosis, with that of the typically atherosclerosis-prone coronary artery. The study group comprised material collected from 116 autopsies. Subjects were aged between 1 month and 90 years. Specimens were collected from grossly normal arterial segments. An already known fact was confirmed: the presence and remarkable intensity of non-atherosclerotic intimal thickening in the coronary arteries since the beginning of life. On the contrary, hepatic artery intimal thickening was incidental during the first 4 decades of life and regular later on. During all the decades intimal thickness was decreased in the hepatic artery in comparison to the coronary artery. Initial atherosclerosis was noted during the 4th decade of life in the coronary artery and during the 8th and 9th decades in the hepatic artery. Intimal thickness was always lower in women compared to men. These findings reveal a positive correlation between the intensity of intimal hyperplasia and the incidence of atherosclerosis. The intimal pattern is of limited value in the determination of an unidentified subject age who underwent an autopsy. The thickness of the hepatic artery intima, which surpasses 200 microns indicates that the subject is at least 40 years of age, while initial atherosclerosis--the 8th or 9th decades of life.


Pleural plaque profiles on the chest radiographs and CT scans of asbestos-exposed Japanese construction workers.
Pleural plaques are asymptomatic focal thickenings of the pleura and considered the hallmark of asbestos exposure. However, it is often difficult to detect pleural plaques on chest x-rays (CXR). In a retrospective study, using chest CT scans of 140 Japanese asbestos-exposed construction workers who have probable or definite findings of pleural plaque on CXR; firstly, we proposed plaque morphology-based classification for CXR findings, and then we examined if those classified findings could be confirmed as pleural plaques on CT scans. Our morphology-based classification of pleural plaque findings included nine types. The percentages of confirmed pleural plaques on CT scans by type (number of confirmed pleural plaque on CT/number of observed on CXR) were 93% (40/43) for straight, 89% (56/63) for diamond, 88% (7/8) for double, 83% (19/23) for tapered medially, 80% (20/25) for parallel, 77% (23/30) for crescent, 79% (11/14) for tenting, 72% (18/25) for tapered-laterally (long type), and 0% (0/9) for tapered-laterally (short type). When added to the ILO classification, morphology-based classification of CXR pleural plaque findings makes its detection easier and hence chest radiograph continues to be a suitable tool for screening asbestos-related pleural plaques based on its simplicity, low radiation exposure, wide availability and cost-effectiveness.


Tumour Treating Fields in combination with pemetrexed and cisplatin or carboplatin as first-line treatment for unresectable malignant pleural mesothelioma (STELLAR): a multicentre, single-arm phase 2 trial.
Tumour Treating Fields (TTFields) are a regional, antimitotic treatment for solid tumours, which is based on the delivery of low-intensity alternating electric fields. The aim of the STELLAR study was to test the activity of TTFields delivered to the thorax in combination with systemic chemotherapy for the front-line treatment of patients with unresectable malignant pleural mesothelioma. STELLAR was a prospective, single-arm, phase 2 trial done at 12 European academic and non-academic sites (five in Italy, three in Poland, one in France, one in Belgium, one in Spain, and one in the Netherlands) for treatment-naive patients with histologically confirmed unresectable malignant pleural mesothelioma. Patients were aged at least 18 years, had an Eastern Cooperative Oncology Group performance status of 0-1, and at least one measurable or evaluable lesion according to modified Response Evaluation Criteria in Solid Tumors for mesothelioma. Patients received continuous TTFields at a frequency of 150 kHz to the thorax and concomitant chemotherapy with intravenous pemetrexed (500 mg/m2 on day 1) plus intravenous platinum (either cisplatin 75 mg/m2 on day 1 or carboplatin area under the curve 5 on day 1) every 21 days for up to six cycles. Patients not progressing after completion of chemotherapy received TTFields as maintenance treatment until progression, patient or physician decision, or unacceptable toxic effects. The primary endpoint of the trial was overall survival. Survival analyses were done in the intention-to-treat population, and safety analyses were done in all patients who received at least 1 day of TTFields treatment. This trial is registered with ClinicalTrials.gov, NCT02397928. Between Feb 9, 2015 and March 21, 2017, 80 patients were enrolled in the study. Median follow-up was 12·5 months (IQR 7·4-16·6). Median overall survival was 18·2 months (95% CI 12·1-25·8). The most common grade 3 or worse adverse events were anaemia (nine [11%] patients), neutropenia (seven [9%]), and thrombocytopenia (four [5%]). Skin reaction was the only adverse event associated with TTFields and was reported as grade 1-2 in 53 (66%) patients, and as grade 3 in four (5%) patients. No treatment-related deaths were observed. The trial showed encouraging overall survival results, with no increase in systemic toxicity. TTFields (150 kHz) delivered to the thorax concomitant with pemetrexed and platinum was an active and safe combination for front-line treatment of unresectable malignant pleural mesothelioma. Further investigation in a randomised trial is warranted. Novocure.


Critical value reporting: a survey of 36 clinical laboratories in South Africa.
Critical value policies are used by clinical laboratories to decide when to notify caregivers of life-threatening results. Despite their widespread use, critical value policies have not been published locally. A survey was designed to determine critical value policies for haematology tests in South Africa. A survey was carried out on 136 identified laboratories across South Africa in January 2013. Of these, 36 responded. Data collected included critical value policies, critical values for haematology parameters, and critical value reporting. Of the 36 laboratories surveyed, 11.1% (n=4) were private, 33.3% (n=12) were affiliated to academic institutions and 55.6% (n=20) were peripheral or regional National Health Laboratory Service laboratories. All the laboratories confirmed that they had a critical value policy, and 83.3% of such policies were derived from local clinical opinion. Mean low and high critical limits for the most frequently listed tests were as follows: haemoglobin <6 and >20 g/dl, platelet count <41 and >1 000 ×10(9)/l, white cell count <2 and >46 ×10(9)/l, activated partial thromboplastin time >101 seconds, and international normalised ratio >6. In almost all cases critical value reporting was performed by the technologist on duty (97.2%). The majority of laboratories required that the person notified of the critical value be the doctor who ordered the test or the caregiver directly involved in the patient's care (83.3%); 73.3% of laboratories indicated that they followed an algorithm if the doctor/caregiver could not be reached. Each laboratory is responsible for establishing clinically relevant critical limits. Clinicians should be involved in developing the laboratory's critical value policy. The findings of this survey may be of value to local laboratories that are in the process of establishing or reviewing critical value policies.


Comparison of ranibizumab alone versus ranibizumab with targeted retinal laser for branch retinal vein occlusion with macular edema.
To determine the effect of ultra-widefield fluorescein angiography (UWFFA)-guided targeted retinal photocoagulation (TRP) in branch retinal vein occlusion (BRVO) with macular edema after intravitreal Ranibizumab (RBZ). 33 eyes of 32 treatment naïve patients diagnosed as BRVO with macular edema were prospectively randomized to 0.5 mg Ranibizumab only (RBZ group) (n = 17) or Ranibizumab with UWFFA-guided laser (RBZ + TRP group) (n = 16). Both groups received three injections at monthly intervals and PRN henceforth. RBZ + TRP group additionally underwent UWFFA-guided TRP of peripheral capillary nonperfusion areas 1 week post injection. Outcome measures included improvement in visual acuity, central subfoveal thickness (CST), and the number of injections required with a minimum follow-up of 9 months. Both groups showed significant improvement in mean BCVA (25.7 ± 8.19 letters, P < 0.001 vs. 23.38 ± 7.56 letters, P < 0.001; in RBZ and RBZ + TRP group, respectively) and reduction in mean central subfoveal thickness (379.12 ± 242.7 μm, P < 0.001 vs. 253.75 ± 137.9 μm, P < 0.001 in RBZ and RBZ + TRP group, respectively) at 9 months. The number of injections in the RBZ group (5.76 ± 1.3) was significantly greater than RBZ + TRP (4.06 ± 0.99) (P < 0.001). Both groups had significant improvement in contrast sensitivity and mean deviation on visual fields; however, the difference between the groups was not significant (P = 0.62 and P = 0.79, respectively). UWFFA-guided TRP reduced the number of injections of Ranibizumab in patients having BRVO with macular edema, while maintaining similar benefits in the improvement of BCVA, central subfoveal thickness without deleterious effect on the visual field, and contrast sensitivity.


The interplay between tissue growth and scaffold degradation in engineered tissue constructs.
In vitro tissue engineering is emerging as a potential tool to meet the high demand for replacement tissue, caused by the increased incidence of tissue degeneration and damage. A key challenge in this field is ensuring that the mechanical properties of the engineered tissue are appropriate for the in vivo environment. Achieving this goal will require detailed understanding of the interplay between cell proliferation, extracellular matrix (ECM) deposition and scaffold degradation. In this paper, we use a mathematical model (based upon a multiphase continuum framework) to investigate the interplay between tissue growth and scaffold degradation during tissue construct evolution in vitro. Our model accommodates a cell population and culture medium, modelled as viscous fluids, together with a porous scaffold and ECM deposited by the cells, represented as rigid porous materials. We focus on tissue growth within a perfusion bioreactor system, and investigate how the predicted tissue composition is altered under the influence of (1) differential interactions between cells and the supporting scaffold and their associated ECM, (2) scaffold degradation, and (3) mechanotransduction-regulated cell proliferation and ECM deposition. Numerical simulation of the model equations reveals that scaffold heterogeneity typical of that obtained from [Formula: see text]CT scans of tissue engineering scaffolds can lead to significant variation in the flow-induced mechanical stimuli experienced by cells seeded in the scaffold. This leads to strong heterogeneity in the deposition of ECM. Furthermore, preferential adherence of cells to the ECM in favour of the artificial scaffold appears to have no significant influence on the eventual construct composition; adherence of cells to these supporting structures does, however, lead to cell and ECM distributions which mimic and exaggerate the heterogeneity of the underlying scaffold. Such phenomena have important ramifications for the mechanical integrity of engineered tissue constructs and their suitability for implantation in vivo.


Selective cyclooxygenase inhibitors increase paclitaxel sensitivity in taxane-resistant ovarian cancer by suppressing P-glycoprotein expression.
The purpose of this study was to investigate whether selective cyclooxygenase (COX) inhibitors promote paclitaxel-induced apoptosis in taxane-resistant ovarian cancer cells by suppressing MDR1/P-glycoprotein (P-gp) expression. Taxane-resistant ovarian cancer cells were cultured with paclitaxel alone or combined with a selective COX inhibitors. The expression patterns of MDR1/P-gp and the ability of COX inhibitors to inhibit growth of taxane-resistant ovarian cancer cells were measured. The efficacy of prostaglandin E2 (PGE2) supplementation was measured to evaluate the mechanisms involved in suppressing MDR1 gene expression. P-gp was upregulated in taxane-resistant ovarian cancer cells compared to paired paclitaxel-sensitive ovarian cancer cells. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that selective COX inhibitors significantly enhanced the cytotoxic effects of paclitaxel in taxane-resistant ovarian cancer cells via a prostaglandin-independent mechanism. These increased apoptotic effects were further verified by measuring an increased percentage of cells in sub-G1 stage using flow cytometry. Selective COX inhibitors suppressed MDR1 and P-gp expression. Moreover, combined treatment with paclitaxel and selective COX inhibitors increased poly (ADP-ribose) polymerase (PARP) cleavage in taxane-resistant ovarian cancer cells. Selective COX inhibitors significantly promote paclitaxel-induced cell death in taxane-resistant ovarian cancer cells in a prostaglandin-independent manner. COX inhibitors could be potent therapeutic tools to promote paclitaxel sensitization of taxane-resistant ovarian cancers by suppressing MDR1/P-gp, which is responsible for the efflux of chemotherapeutic agents.


Lipid Nanoparticles: A Novel Approach for Brain Targeting.
Brain is a delicate organ, separated from general circulation and is characterized by the presence of relatively impermeable Blood Brain Barrier (BBB). The BBB maintains homeostasis in the brain thus restricting the entrance of foreign bodies and several molecules from reaching the brain. As a result several promising molecules do not reach the target site and fail to produce in vivo response. Nevertheless, lipid nanoparticles are taken up readily by the brain because of their lipophilic nature. The bioacceptable and biodegradable nature of lipid nanoparticles makes them less toxic and suited for brain targeting. In the present review the BBB, mechanism of transport across the BBB, strategies to bypass the blood-brain barrier have been presented. The aptness of lipid nanoparticles for brain targeting has been highlighted. The proposed mechanism of uptake of the lipid nanoparticles, methods of prolonging the plasma retention and various methods of preparation for formulation of effective delivery systems for brain targeting have been included and dealt in this review. Lipid based formulations can be designated as the current and future generation of drug delivery systems as these possess tremendous potential to bypass BBB and reach the target site due to their small size and ability to dodge the reticular endothelial system. However, these nanostructures need to be investigated intensively to successfully reach the clinical trials stage.


Individuals with high bone mass have increased progression of radiographic and clinical features of knee osteoarthritis.
High bone mass (HBM) is associated with an increased prevalence of radiographic knee OA (kOA), characterized by osteophytosis. We aimed to determine if progression of radiographic kOA, and its sub-phenotypes, is increased in HBM and whether observed changes are clinically relevant. A cohort with and without HBM (L1 and/or total hip bone mineral density Z-score≥+3.2) had knee radiographs collected at baseline and 8-year follow-up. Sub-phenotypes were graded using the OARSI atlas. Medial/lateral tibial/femoral osteophyte and medial/lateral joint space narrowing (JSN) grades were summed and Δosteophytes, ΔJSN derived. Pain, function and stiffness were quantified using the WOMAC questionnaire. Associations between HBM status and sub-phenotype progression were determined using multivariable linear/poisson regression, adjusting for age, sex, height, baseline sub-phenotype grade, menopause, education and total body fat mass (TBFM). Generalized estimating equations accounted for individual-level clustering. 169 individuals had repeated radiographs, providing 330 knee images; 63% had HBM, 73% were female, mean (SD) age was 58 (12) years. Whilst HBM was not clearly associated with overall Kellgren-Lawrence measured progression (RR = 1.55 [0.56.4.32]), HBM was positively associated with both Δosteophytes and ΔJSN individually (adjusted mean differences between individuals with and without HBM 0.45 [0.01.0.89] and 0.15 [0.01.0.29], respectively). HBM individuals had higher WOMAC knee pain scores (β = 7.42 [126.96.36.199]), largely explained by adjustment for osteophyte score (58% attenuated) rather than JSN (30% attenuated) or TBFM (16% attenuated). The same pattern was observed for symptomatic stiffness and functional limitation. HBM is associated with osteophyte progression, which appears to contribute to increased reported pain, stiffness and functional loss.


First report of Hepatozoon sp. in the Oregon spotted frog, Rana pretiosa.
From 2005 through 2008, we screened 650 Oregon spotted frogs (Rana pretiosa) from three populations in central Oregon, USA, for hemoparasites. A Hepatozoon sp., not previously reported in R. pretiosa, was found in one population of frogs, mostly as intracellular gamonts at a prevalence of 10.5% with parasitemias ranging from 0.02% to greater than 42% of erythrocytes within individual frogs. Intra-and extracellular merozoites were present, but rare. A potential vector, the mosquito Culex boharti, was common throughout the habitat of the population carrying Hepatozoon sp.


Identification of the aryl hydrocarbon receptor target gene TiPARP as a mediator of suppression of hepatic gluconeogenesis by 2,3,7,8-tetrachlorodibenzo-p-dioxin and of nicotinamide as a corrective agent for this effect.
The environmental toxin TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin, dioxin) produces diverse toxic effects including a lethal wasting syndrome whose hallmark is suppressed hepatic gluconeogenesis. All TCDD toxicities require activation of the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor. Whereas the mechanism for AHR induction of target genes is well understood, it is not known how AHR activation produces any TCDD toxicity. This report identifies for the first time an AHR target gene, TiPARP (TCDD-inducible poly(ADP-ribose) polymerase, PARP7) that can mediate a TCDD toxicity, i.e. suppression of hepatic gluconeogenesis. TCDD suppressed hepatic glucose production, expression of key gluconeogenic genes, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pase), and NAD(+) levels, and increased PARP activity and TiPARP expression. TCDD also increased acetylation and ubiquitin-dependent proteosomal degradation of the peroxisome proliferator-activated receptor γ coactivator 1 α (PGC1α), a coactivator of PEPCK and G6Pase transcription. TiPARP overexpression reproduced TCDD effects on glucose output and NAD(+) levels whereas TiPARP silencing diminished them. TiPARP overexpression also increased PGC1α acetylation and decreased PGC1α levels. In contrast, silencing of cytochromes P450 (CYP) 1A, main AHR-induced genes, did not alter TCDD suppression of gluconeogenesis. The vitamin B3 constituent, nicotinamide (NAM), prevented TCDD suppression of glucose output, NAD(+), and gluconeogenic genes and stabilized PGC1α. The corrective effects of NAM could be attributed to increased NAD(+) levels and suppression of AHR target gene induction. The results reveal that TiPARP can mediate a TCDD effect, that the AHR is linked to PGC1α function and stability and that NAM has novel AHR antagonist activity.


Optical and high-frequency electrical properties of the lunar sample.
Reflectivity and polarization laws for the powder sample and its spectrum are close to the mean for the lunar maria. Solid samples show a marked absorption feature at 1 micron. The low albedo appears to be due to a surface coating on dust grains rather than to volume absorption. The high-frequency electrical properties resemble those of a fine powder made from typical dense terrestrial rocks and are consistent with previous estimates from ground-based radar observations. The differential mass spectrum is almost constant from 100 micron particles down to 0.1 micron particles; most particles are smaller than 0.3 micron. Their shapes disclose a variety of processes of generation.


Gianotti-Crosti syndrome. A review of ten cases not associated with hepatitis B.
Studies of data from ten cases of infantile acrodermatitis and from eight cases reported in the North American literature disclose distinctive papular dermatosis of the face and extremities, often related to virus infection. None of our eight patients who were tested had evidence of hepatitis B infections, although transaminase values were elevated in two. All five patients who were tested had lymphocytosis. Six patients had antecedent upper respiratory tract symptoms. Data from our cases and from the other previously reported cases indicate that the eruption is a virus-related response. Although the hepatitis virus has been the most frequently encountered causative agent to date, other viruses, including Epstein-Barr virus, coxsackievirus, and parainfluenza virus, may produce a similar cutaneous response.


Backside wear of modular ultra-high molecular weight polyethylene tibial inserts.
The capture mechanisms of modular tibial total knee components may allow relative micromotion between the insert and the base-plate, leading to wear at the nonarticulating (backside) surface. Although retrieved components often display laxity in the capture mechanism in the unloaded condition, the magnitude of the relative motion that actually occurs under physiologic conditions has not been determined. This study was performed to assess the impact of different modes of knee-loading on the relative micromotion between the insert and the base-plate and the relationship between the duration that the implant had been in situ and the severity of backside wear. Twenty-one posterior-stabilized total knee replacements of one common design (Insall-Burstein II) were retrieved at one to 100 months after implantation. The extent and severity of backside wear was graded with use of stereomicroscopy. All components were soaked in a bath (of physiologic saline solution at 37 degrees C for four days prior to reassembly. The relative micromotion between the insert and the base-plate of each specimen was measured in vitro in two different conditions: with no axial load and with a combination of loads and torques simulating the stance phase of gait. The capture mechanism laxity between the insert and the tibial base-plate in the unloaded condition was approximately eight times larger than the micromotion measured during simulated gait. The capture mechanism laxity allowed a mean (and standard deviation) of 618 +/- 226 micro m of total relative micromotion compared with 103 +/- 54 micro m of relative micromotion during the gait cycle. Under both loading conditions, the predominant direction of interface motion was medial-lateral. No correlation was found between the magnitude of capture mechanism laxity and the relative micromotion measured during simulated gait (p = 0.11). Larger polyethylene protrusions on the backside surface did not correlate with less micromotion (p = 0.48) or with capture mechanism laxity (p = 0.06). For the implant design that was studied, capture mechanism laxity between the modular insert and the base-plate in the unloaded condition was an order of magnitude larger than and not indicative of the micromotion that occurred during simulated physiologic loading. In addition, polyethylene protrusions into the screw-holes of tibial base-plates did not seat or lock the insert in place and reduce relative motion. While some clearance between the insert and the base-plate is required to allow assembly of modular tibial components at the time of surgery, the amount of relative interface motion during a functional activity such as normal gait, which can produce potentially damaging wear debris, is unknown. However, the compressive forces applied to the articular surface during a functional activity may substantially reduce micromotion between the insert and the base-plate relative to the unloaded condition.


The effects of unsupervised exercise training on physical activity and physiological factors after supervised cardiac rehabilitation.
Exercise maintenance after supervised cardiac rehabilitation is important in maintaining both physical activity and physiological factors, such as peak and muscle strength (MS), associated with reduced mortality. However, there is no evidence of the effects of unsupervised exercise training and MS training on physical activity and physiological factors after supervised cardiac rehabilitation of Japanese cardiac patients. We conducted a randomized, controlled trial to evaluate the effect of unsupervised exercise training on physical activity and selected physiological factors after supervised cardiac rehabilitation. Eighteen myocardial infarction (MI) patients (16 men, 2 women; mean age 66.3 years) were recruited following completion of a supervised recovery-phase cardiac rehabilitation program. Patients were randomly assigned to a MS training (n=10) or control group (n=8). Baseline measurements of physical activity, peak , and MS were performed at the end of supervised recovery-phase cardiac rehabilitation (6 months after the onset of MI: T1). Six months later, after going through an unsupervised exercise program (12 months after the onset of MI: T2) exercise maintenance, peak , MS, and physical activity were remeasured. The MS training group performed low-intensity MS training and walking over the second 6-month period; the control group performed walking exercise only. All patients maintained their exercise training. At T2, there were no significant differences in peak values between the MS training and control groups. There was also no significant difference in physical activity (mean number of steps per week) between the MS training and control groups. However, MS was significantly higher in the MS training group than in the control group. We concluded that unsupervised exercise training and low-level MS training performed after supervised cardiac rehabilitation may effectively maintain not only physical activity and peak but increase MS.


In vitro comparison of two centrally threaded, positive-profile transfixation pin designs for use in third metacarpal bones in horses.
To compare heat generation during insertion, pullout strength, and associated microdamage between a self-tapping positive profile transfixation pin (STTP) and nontapping positive profile transfixation pin (NTTP). 30 pairs of third metacarpal bones (MC3s) from adult equine cadavers. One MC3 of each pair was assigned to the STTP group; the other was assigned to the NTTP group. The assigned pin was inserted into the diaphysis in a lateral to medial direction. Bone temperature increase during pilot-hole drilling and pin insertion was recorded at 1 mm from the final thread position with wire thermocouples at cis and trans cortices. Resistance to axial extraction before and after cyclic loading was measured in a material testing device, and microstructural damage caused by transfixation pin insertion was assessed with scanning electron microscopy. The STTP group developed a significant increase in bone temperature, compared with the NTTP group. No significant difference was found between the mean maximal pullout strength of the STTP and the NTTP in both non-cyclic-loaded and cyclic-loaded groups. Microdamage to the bone-pin interface was lower when the STTP versus the NTTP was used, but more bone debris was apparent after inserting the STTP. Because of the significant increase in temperature generation and debris accumulation despite similar pullout strengths and lesser microfracture formation, the STTP likely poses a higher risk of bone necrosis and potential loosening than the NTTP. This might be corrected by redesign of the tapping aspect of the STTP.


Estimated volumetric bone mineral density in a rural Thai men and women: Khon Kaen Osteoporosis Study (KKOS).
The authors examined the areal bone mineral density (aFNBMD) and estimated volumetric bone mineral density at the femoral neck (vFNBMD) in rural Thai men and women. A total of 181 men and 255 women, between 20 and 84 years of age, living in rural areas of Khon Kaen province, were randomly selected. Areal FNBMD and estimated v FNBMD were determined using dual energy X-ray absorptiometry (DPX-IQ, GE Lunar Corp, Madison, WI). Men had a significantly higher aFNBMD than women, whereas the estimated vFNBMD was similar regardless of sex. The peak for the aFNBMD vs. vFNBMD was observed between 20 and 29 vs. 30 and 39 years of age in men and women, respectively. The prevalence of osteoporosis in men and women using estimated vFNBMD vs. aFNBMD cut-offs was 19 and 14.2 vs 11.8 and 26 percent, respectively. Prevalence increased with age. Estimated vFNBMD shows only small sex-correlated differences in bone density. Estimated vFNBMD was more sensitive than aFNBMD, when used to define the osteoporotic cut-offs in men, while it was less sensitive than aFNBMD in women.


Interleukin 10 gene transfer prevents experimental colitis in rats.
The development of colitis in interleukin 10 (IL-10) deficient mice, together with the known anti-inflammatory and immunomodulatory properties of this cytokine have prompted consideration of IL-10 as a treatment for inflammatory bowel disease (IBD). However, studies using hrIL-10 in IBD models have yielded inconsistent results. To examine the therapeutic potential of overexpressing the IL-10 gene before and after the induction of experimental colitis in rats. Gene transfer was achieved by intraperitoneal injection of non-replicating human type 5 adenovirus bearing the IL-10 gene, either 24 hours before or one hour after intrarectal administration of dinitrobenzene sulphonic acid in rats. Colonic damage and inflammation was assessed macroscopically and by measuring myeloperoxidase activity and leukotriene B4 concentrations. Gene transfer increased IL-10 protein in serum for up to six days. IL-10 gene transfer prior to colitis improved colitis macroscopically and histologically, and significantly reduced colonic myeloperoxidase activity and leukotriene B4 concentrations. In contrast, IL-10 gene transfer after the onset of colitis had no beneficial effect. Gene therapy using an adenovirus-IL-10 construct was successful in preventing but not in reversing experimental colitis in the rat.


Family treatment of child anxiety: outcomes, limitations and future directions.
Anxiety of childhood is a common and serious condition. The past decade has seen an increase in treatment-focussed research, with recent trials tending to give greater attention to parents in the treatment process. This review examines the efficacy of family-based cognitive behaviour therapy and attempts to delineate some of the factors that might have an impact on its efficacy. The choice and timing of outcome measure, age and gender of the child, level of parental anxiety, severity and type of child anxiety and treatment format and content are scrutinised. The main conclusions are necessarily tentative, but it seems likely that Family Cognitive Behaviour Therapy (FCBT) is superior to no treatment, and, for some outcome measures, also superior to Child Cognitive Behaviour Therapy (CCBT). Where FCBT is successful, the results are consistently maintained at follow-up. It appears that where a parent is anxious, and this is not addressed, outcomes are less good. However, for children of anxious parents, FCBT is probably more effective than CCBT. What is most clear is that large, well-designed studies, examining these factors alone and in combination, are now needed.


A role of oligodendrocytes in information processing.
Myelinating oligodendrocytes enable fast propagation of action potentials along the ensheathed axons. In addition, oligodendrocytes play diverse non-canonical roles including axonal metabolic support and activity-dependent myelination. An open question remains whether myelination also contributes to information processing in addition to speeding up conduction velocity. Here, we analyze the role of myelin in auditory information processing using paradigms that are also good predictors of speech understanding in humans. We compare mice with different degrees of dysmyelination using acute multiunit recordings in the auditory cortex, in combination with behavioral readouts. We find complex alterations of neuronal responses that reflect fatigue and temporal acuity deficits. We observe partially discriminable but similar deficits in well myelinated mice in which glial cells cannot fully support axons metabolically. We suggest a model in which myelination contributes to sustained stimulus perception in temporally complex paradigms, with a role of metabolically active oligodendrocytes in cortical information processing.


The effects of opposition and gender on knee kinematics and ground reaction force during landing from volleyball block jumps.
The aim of this study was to examine the effect of opposition and gender on knee kinematics and ground reaction force during landing from a volleyball block jump. Six female and six male university volleyball players performed two landing tasks: (a) an unopposed and (b) an opposed volleyball block jump and landing. A 12-camera motion analysis system (120 Hz) was used to record knee kinematics, and a force platform (600 Hz) was used to record ground reaction force during landing. The results showed a significant effect for level of opposition in peak normalized ground reaction force (p = .04), knee flexion at ground contact (p = .003), maximum knee flexion (p = .001), and knee flexion range of motion (p = .003). There was a significant effect for gender in maximum knee flexion (p = .01), knee flexion range of motion (p = .001), maximum knee valgus angle (p = .001), and knee valgus range of motion (p = .001). The changes in landing biomechanics as a result of opposition suggest future research on landing mechanics should examine opposed exercises, because opposition may significantly alter neuromuscular responses.


Long head biceps tenodesis with a knotless cinch suture anchor: a biomechanical analysis.
The purpose of this study was to evaluate the initial fixation strength of 3 techniques of arthroscopic tenodesis of the long head of the biceps (LHB). Eighteen human cadaveric shoulders were randomly assigned to one of 3 simulated arthroscopic biceps tenodesis techniques-simple suture (SS), Krakow stitch (KS), or lasso loop (LL)-combined with a knotless fixation implant (3.5-mm Piton Anchor; Tornier, Minneapolis, MN). Biomechanical parameters were evaluated by cyclic loading and load to failure. The mean failure load (P = .007) was 158.3 ± 32.2 N, 109.8 ± 41.1 N, and 46.6 ± 3.8 N for the KS, SS, and LL techniques, respectively. Mean stiffness was greater (statistically significant) in the KS (21.4 ± 3.0 N/mm) and SS (20.7 ± 7.9 N/mm) treatment groups compared with the LL group (4.5 ± 1.5 N/mm) (P = .011). Biceps tenodesis performed with a more secure tendon suturing technique, such as the Krakow technique, provides superior ultimate and fatigue strength and thus may be more secure in clinical application and yield better clinical results. The mechanical properties of the LL technique were especially poor in comparison. Although more complex suturing techniques for arthroscopic biceps tenodesis can be technically challenging, more secure tendon fixation may improve clinical outcomes.


Prediction of parturition in bitches utilizing continuous vaginal temperature measurement.
The objective of this study was to determine sensitivity and specificity of a body temperature decline in bitches to predict parturition. Temperature loggers were placed into the vaginal cavity of 16 pregnant bitches on day 56-61 after estimated ovulation or first mating. This measurement technique has been validated previously and enabled continuous sampling of body temperature. The temperature loggers were expelled from the vagina before delivery of the first pup. The computed values for specificity (77-92%) were higher than sensitivity (53-69%), indicating a more precise prognosis of parturition not occurring. In conclusion, our findings may assist interpreting vaginal temperature measurements in order to predict parturition in bitches.


Critical Factors Driving the High Volumetric Uptake of Methane in Cu₃(btc)₂.
A thorough experimental and computational study has been carried out to elucidate the mechanistic reasons for the high volumetric uptake of methane in the metal-organic framework Cu3(btc)2 (btc(3-) = 1,3,5-benzenetricarboxylate; HKUST-1). Methane adsorption data measured at several temperatures for Cu3(btc)2, and its isostructural analogue Cr3(btc)2, show that there is little difference in volumetric adsorption capacity when the metal center is changed. In situ neutron powder diffraction data obtained for both materials were used to locate four CD4 adsorption sites that fill sequentially. This data unequivocally shows that primary adsorption sites around, and within, the small octahedral cage in the structure are favored over the exposed Cu(2+) or Cr(2+) cations. These results are supported by an exhaustive parallel computational study, and contradict results recently reported using a time-resolved diffraction structure envelope (TRDSE) method. Moreover, the computational study reveals that strong methane binding at the open metal sites is largely due to methane-methane interactions with adjacent molecules adsorbed at the primary sites instead of an electronic interaction with the metal center. Simulated methane adsorption isotherms for Cu3(btc)2 are shown to exhibit excellent agreement with experimental isotherms, allowing for additional simulations that show that modifications to the metal center, ligand, or even tuning the overall binding enthalpy would not improve the working capacity for methane storage over that measured for Cu3(btc)2 itself.


Field-scale Variation in Colloid Dispersibility and Transport: Multiple Linear Regressions to Soil Physico-Chemical and Structural Properties.
Water-dispersible soil colloids (WDC) act as carriers for sorbing chemicals in macroporous soils and hence constitute a significant risk for the aquatic environment. The prediction of WDC readily available for facilitated chemical transport is an unsolved challenge. This study identifies key parameters and predictive indicators for assessing field-scale variation of WDC. Samples representing three measurement scales (1- to 2-mm aggregates, intact 100-cm rings, and intact 6283 cm columns) were retrieved from the topsoil of a 1.69-ha agricultural field in a 15-m by 15-m grid to determine colloid dispersibility, mobilization, and transport. The amount of WDC was determined using (i) a laser diffraction method on 1- to 2-mm aggregates and (ii) an end-over-end shaking method on 100-cm intact rings. The accumulated amount of colloids leached from 20-cm by 20-cm intact columns was determined as a measure of the integrated colloid mobilization and transport. The WDC and the accumulated colloid transport were higher in samples from the northern part of the field. Using multiple linear regression (MLR) analyses, WDC or amount of colloids transported were predicted at the three measurement scales from 24 measured, geo-referenced parameters to identify parameters that could serve as indicator parameters for screening for colloid dispersibility, mobilization, and transport. The MLR analyses were performed at each sample scale using all, only northern, and only southern field locations. Generally, the predictive power of the regression models was best on the smallest 1- to 2-mm aggregate scale. Overall, our results suggest that different drivers controlled colloid dispersibility and transport at the three measurement scales and in the two subareas of the field.


Anti-ganglioside antibody induction by swine (A/NJ/1976/H1N1) and other influenza vaccines: insights into vaccine-associated Guillain-Barré syndrome.
Receipt of an A/NJ/1976/H1N1 "swine flu" vaccine in 1976, unlike receipt of influenza vaccines used in subsequent years, was strongly associated with the development of the neurologic disorder Guillain-Barré syndrome (GBS). Anti-ganglioside antibodies (e.g., anti-GM(1)) are associated with the development of GBS, and we hypothesized that the swine flu vaccine contained contaminating moieties (such as Campylobacter jejuni antigens that mimic human gangliosides or other vaccine components) that elicited an anti-GM(1) antibody response in susceptible recipients. Surviving samples of monovalent and bivalent 1976 vaccine, comprising those from 3 manufacturers and 11 lot numbers, along with several contemporary vaccines were tested for hemagglutinin (HA) activity, the presence of Campylobacter DNA, and the ability to induce anti-Campylobacter and anti-GM(1) antibodies after inoculation into C3H/HeN mice. We found that, although C. jejuni was not detected in 1976 swine flu vaccines, these vaccines induced anti-GM(1) antibodies in mice, as did vaccines from 1991-1992 and 2004-2005. Preliminary studies suggest that the influenza HA induces anti-GM(1) antibodies. Influenza vaccines contain structures that can induce anti-GM(1) antibodies after inoculation into mice. Further research into influenza vaccine components that elicit anti-ganglioside responses and the role played by these antibodies (if any) in vaccine-associated GBS is warranted.


Electrophysiological correlates of the retrieval of emotional and non-emotional context.
In two experiments, words were presented in negatively or neutrally valenced sentences. At test, subjects made old/new recognition judgments to these words. In Experiment 2 only, for words judged old, subjects also indicated whether the words had been studied in a neutral or a negative context. In Experiment 1, left parietal old/new event-related brain potential (ERP) effects were larger and more sustained when elicited by words that had been studied in negative sentences, and a right frontal old/new effect was elicited by these words exclusively. In Experiment 2, the left parietal and right frontal effects elicited by old words correctly assigned to their study context were equivalent in size regardless of the nature of the context; a third ERP old/new effect, maximal over posterior scalp regions, was seen only for words from negative contexts. The findings indicate that incidental retrieval of emotional context gives rise to greater activation in neural systems supporting conscious recollection than does retrieval of nonemotional context. When contextual retrieval is intentional, recollection of emotional and non-emotional information are associated with equivalent engagement of these systems. The findings from Experiment 2 suggest that additional neural circuitry may be activated selectively by emotionally valenced episodic information.


[Inhibitory effects of puerarin on invasion and metastasis of oophoroma cells HO-8910].
To investigate the inhibitory effect of puerarin on invasive and metastatic abilities of tumor cells, and its possible mechanism through observing its impacts on the migratory, adhesive and invasive capacities of human oophoroma cells HO-8910 to the artificial recombined basement membrane. Expression of estrogen receptor (ER) in HO-8910 cells was detected using PCR assay. Effects of puerarin on HO-8910 proliferation was detected with MTT assay; on its adhesion potential was tested with cell-matrigel adhesion assay, and on invasive and migratory capacities were measured with Transwell matrigel invasion assay and Transwell motility assay respectively. ER was positively expressed in HO-8910 cells. After being treated with 20 micromol/L puerarin for 12 h, the adhesive test showed that OD value in the tested group was significantly lower than that in the control (P < 0.01), the inhibiting rate reached 50.63%; and the Transwell assay showed a significant lowering of penetrated cells (P < 0.01), the inhibition rate for invasion was 38.59% and that for motility migration 40.63%. The number of penetrated cells was lower in the group intervened with combination of Puerarin and estrogen than in the group intervened with estrogen alone, 33.40 +/- 3.30 vs 48.05 +/- 3. 56 for invasion and 35.35 +/- 3.03 vs 52.45 +/- 1.04 for motility (all P < 0.01). Puerarin can inhibit the adhesion, invasion and migration of HO-8910 cells, plays an antagonist effect against the stimulation of estrogen on the malignant behavior of tumor cells.


Essential role of Tip60-dependent recruitment of ribonucleotide reductase at DNA damage sites in DNA repair during G1 phase.
A balanced deoxyribonucleotide (dNTP) supply is essential for DNA repair. Here, we found that ribonucleotide reductase (RNR) subunits RRM1 and RRM2 accumulated very rapidly at damage sites. RRM1 bound physically to Tip60. Chromatin immunoprecipitation analyses of cells with an I-SceI cassette revealed that RRM1 bound to a damage site in a Tip60-dependent manner. Active RRM1 mutants lacking Tip60 binding failed to rescue an impaired DNA repair in RRM1-depleted G1-phase cells. Inhibition of RNR recruitment by an RRM1 C-terminal fragment sensitized cells to DNA damage. We propose that Tip60-dependent recruitment of RNR plays an essential role in dNTP supply for DNA repair.


Peripheral neuropathy: pathogenic mechanisms and alternative therapies.
Peripheral neuropathy (PN), associated with diabetes, neurotoxic chemotherapy, human immunodeficiency virus (HIV)/antiretroviral drugs, alcoholism, nutrient deficiencies, heavy metal toxicity, and other etiologies, results in significant morbidity. Conventional pain medications primarily mask symptoms and have significant side effects and addiction profiles. However, a widening body of research indicates alternative medicine may offer significant benefit to this patient population. Alpha-lipoic acid, acetyl-L-carnitine, benfotiamine, methylcobalamin, and topical capsaicin are among the most well-researched alternative options for the treatment of PN. Other potential nutrient or botanical therapies include vitamin E, glutathione, folate, pyridoxine, biotin, myo-inositol, omega-3 and -6 fatty acids, L-arginine, L-glutamine, taurine, N-acetylcysteine, zinc, magnesium, chromium, and St. John's wort. In the realm of physical medicine, acupuncture, magnetic therapy, and yoga have been found to provide benefit. New cutting-edge conventional therapies, including dual-action peptides, may also hold promise.


Clinical history for diagnosis of dementia in men: Caerphilly Prospective Study.
Diagnosis of dementia often requires specialist referral and detailed, time-consuming assessments. To investigate the utility of simple clinical items that non-specialist clinicians could use, in addition to routine practice, to diagnose all-cause dementia syndrome. Cross-sectional diagnostic test accuracy study. Participants were identified from the electoral roll and general practice lists in Caerphilly and adjoining villages in South Wales, UK. Participants (1225 men aged 45-59 years) were screened for cognitive impairment using the Cambridge Cognitive Examination, CAMCOG, at phase 5 of the Caerphilly Prospective Study (CaPS). Index tests were a standardised clinical evaluation, neurological examination, and individual items on the Informant Questionnaire for Cognitive Disorders in the Elderly (IQCODE). Two-hundred and five men who screened positive (68%) and 45 (4.8%) who screened negative were seen, with 59 diagnosed with dementia. The model comprising problems with personal finance and planning had an area under the curve (AUC) of 0.92 (95% confidence interval [CI] = 0.86 to 0.97), positive likelihood ratio (LR+) of 23.7 (95% CI = 5.88 to 95.6), negative likelihood ratio (LR-) of 0.41 (95% CI = 0.27 to 0.62). The best single item for ruling out was no problems learning to use new gadgets (LR- of 0.22, 95% CI = 0.11 to 0.43). This study found that three simple questions have high utility for diagnosing dementia in men who are cognitively screened. If confirmed, this could lead to less burdensome assessment where clinical assessment suggests possible dementia.


Molecular basis of oocyte-paracrine signalling that promotes granulosa cell proliferation.
Oocytes regulate follicle growth by secreting paracrine growth factors that act on neighbouring granulosa cells (GCs). Those factors identified to date are mainly members of the transforming growth factor-beta (TGFbeta) superfamily, but little is known about which specific receptor/signalling system(s) they employ. This study was conducted to determine the requisite pathways utilised by oocytes to promote GC proliferation. We used an established oocyte-secreted mitogen bioassay, where denuded mouse oocytes are co-cultured with mural GCs. Oocytes, growth differentiation factor-9 (GDF9), TGFbeta1 and activin-A all promoted GC DNA synthesis, but bone-morphogenetic protein 6 (BMP6) did not. Subsequently, we tested the capacity of various TGFbeta superfamily receptor ectodomains (ECD) to neutralise oocyte- or specific growth factor-stimulated GC proliferation. The BMP type-II receptor (BMPR-II) ECD antagonised oocyte and GDF9 bioactivity dose-dependently, but had no or minimal effect on TGFbeta1 and activin-A bioactivity, demonstrating its specificity. The TGFbetaR-II, activinR-IIA and activinR-IIB ECDs all failed to neutralise oocyte- or GDF9-stimulated GC DNA synthesis, whereas they did antagonise the activity of their respective native ligands. An activin receptor-like kinase (ALK) 4/5/7 inhibitor, SB431542, also antagonised both oocyte and GDF9 bioactivity in a dose-dependent manner. Consistent with these findings, oocytes, GDF9 and TGFbeta1 all activated SMAD2/3 reporter constructs in transfected GC, and led to phosphorylation of SMAD2 proteins in treated cells. Surprisingly, oocytes did not activate the SMAD1/5/8 pathway in transfected GCs although exogenous BMP6 did. This study indicates that oocyte paracrine factors primarily utilise a similar signalling pathway first identified for GDF9 that employs an unusual combination of TGFbeta superfamily receptors, the BMPR-II and a SMAD2/3 stimulatory ALK (4, 5 or 7), for transmitting their mitogenic actions in GC. This cell-signalling pathway may also have relevance in the hypothalamic-pituitary axis and in germ-somatic cell interactions in the testis.


Subcortical differences among youths with attention-deficit/hyperactivity disorder compared to those with bipolar disorder with and without attention-deficit/hyperactivity disorder.
A significant number of children with bipolar disorder (BP) have co-morbid attention-deficit/hyperactivity disorder (ADHD). It is unknown if these children have neuroimaging findings unique to their co-morbid presentation, or if their brain findings are similar to children diagnosed with BP alone. Fifty three children with Diagnostic and Statistical Manual of Mental Disorders, 4(th) edition (DSM-IV) BP (23 with ADHD, 30 without), 29 healthy controls (HC), and 23 children with ADHD, similar in sex and age, had magnetic resonance imaging (MRI) scans on a 1.5T GE scanner. Volumetric assessments were performed for basal ganglia and limbic subcortical structures. Youths with ADHD had smaller caudate and putamen volumes compared to both BP groups and they had moderately smaller total amygdala volumes compared to the other three groups. Youths with BP + ADHD had moderately larger nucleus accumbens volumes than HC, and females in both BP groups had smaller hippocampal volumes compared to ADHD and HC. No differences were found between the BP and BP + ADHD groups. These data suggest that morphometric subcortical volumes in youths with BP + ADHD are more similar to those in youths with BP. They do not share subcortical neuroanatomic correlates with the ADHD group. These findings suggest that BP + ADHD is a subtype of pediatric BP rather than severe ADHD.


Neutron scattering in the proximate quantum spin liquid α-RuCl3.
The Kitaev quantum spin liquid (KQSL) is an exotic emergent state of matter exhibiting Majorana fermion and gauge flux excitations. The magnetic insulator α-RuCl3 is thought to realize a proximate KQSL. We used neutron scattering on single crystals of α-RuCl3 to reconstruct dynamical correlations in energy-momentum space. We discovered highly unusual signals, including a column of scattering over a large energy interval around the Brillouin zone center, which is very stable with temperature. This finding is consistent with scattering from the Majorana excitations of a KQSL. Other, more delicate experimental features can be transparently associated with perturbations to an ideal model. Our results encourage further study of this prototypical material and may open a window into investigating emergent magnetic Majorana fermions in correlated materials.


Cardiac responses associated with affective processing of unpleasant film stimuli.
The autonomic basis of cardiac reactions to unpleasant film stimuli was investigated. Film clips depicting major surgery, threats of violence, and neutral material were presented to 46 subjects. Self-report measures of emotion were obtained, as well as heart rate, respiration rate, respiratory sinus arrhythmia, T-wave amplitude and skin conductance level. Resting vagal tone was estimated in a paced breathing task prior to film viewing. Spontaneous blink rate was also taken as a measure of visual engagement during film viewing. Coherent increases in sympathetic activation accompanied the film containing violent threats, whereas the surgery film yielded greater electrodermal activation, as well as heart rate deceleration and T-wave increase. These data support the hypothesis of differential autonomic response patterns to specifically unpleasant material. As compared with threat and neutral films, greater blink rate inhibition was observed during the surgery film. Individual differences in parasympathetic cardiac control measured at rest were able to discriminate cardiac response patterns during film viewing.


Pattern of gynaecological cancers in University of Nigeria Teaching Hospital, Enugu, south eastern Nigeria.
Gynaecological cancers are among the leading causes of cancer related deaths worldwide. The objective of this study was to determine the pattern and relative frequencies of gynecological cancers as seen at the University of Nigeria Teaching Hospital (UNTH), Ituku/Ozalla, Enugu, Southeastern Nigeria. An analysis of retrospective clinical data from the ward admissions and discharge books and the operating theatre record books. During the 10-year period under study (2000-2009), 407 cases of gynaecological cancers were admitted into the hospital. The age range of these women was 21-80years with mean age of 54.6 +/- 14.2 years. The commonest gynaecological cancers in women less than 30 years of age were choriocarcinoma (36.1%) and ovarian cancer (33.3%). After 30 years, cervical cancer became the commonest cancer. Overall, cervical cancer was by far the commonest gynecological cancer constituting 78% of all the cases, followed by ovarian cancer (8.9%), choriocarcinoma (4.3%), endometrial cancer (4.1%), vulva cancer (4.0%) and leiomyosarcoma (0.5%). There was no case of vaginal cancer during the study period. Anaemia was the commonest sequelae, complicating 55% of the cases, followed by genital fistulae (12%), infertility (3%), renal failure (3%), and pulmonary complications (2%). Cervical cancer is by far the commonest gynaecological cancer in Enugu and there is high incidence of this cancer even in young women below the age of 30 years. The commonest gynaecological cancer in women less than 30 years is choriocarcinoma, and anaemia is the commonest sequelae of all gynaecological cancers.


argos Is required for projection of photoreceptor axons during optic lobe development in Drosophila.
The Drosophila argos gene encodes a secreted protein with an epidermal growth factor (EGF) motif, which acts as an inhibitor of cell recruitment in the developing eye and wing. Here, we have analyzed the role of argos during optic lobe development. argos expression was observed in the optic lobes throughout the developmental stages. In argos mutants, neuropiles failed to develop normally during embryonic and larval stages, and photoreceptor axons did not project properly into the lamina. Ubiquitous expression of argos, under control of the hsp70 promoter, rescued the defects in optic lobes. We have found that glial cells failed to differentiate in the larval optic lobes of argos mutants. Correspondingly, in loss-of-function repo mutants, whose glial cells also fail to differentiate, photoreceptor axons showed the impaired projection pattern similar to the argos phenotype. These results suggest that glial cells play a role for guidance of photoreceptor axons. The loss-of-function Star mutation (StarX155) dominantly suppressed the defects in the argos optic lobes, suggesting that these two genes act in an antagonistic fashion during optic lobe development.


Pyloric ceca of fish: a "new" absorptive organ.
The functions of the blind appendages attached to the proximal intestine of many fish, the pyloric ceca, have been disputed. Hence we recorded morphological parameters and nutrient uptake rates in the ceca and intestine of four fish species (rainbow trout, cod, largemouth bass, and striped bass) with various degrees of cecal development (the ceca contribute 70, 69, 42, and 16% of the total postgastric surface area, respectively). Proline and glucose uptake, measured in vitro, is similar in the ceca and proximal intestine. For these two solutes in these four species, and for 10 other solutes (9 amino acids and 1 dipeptide) in trout, the ceca contribute about the same percentage to uptake as to total gut area. Trout ceca and intestine have similar membrane-bound disaccharidase activity. Separate experiments with trout fed either graded glass beads or a radiopaque marker and then X-rayed show that the ceca fill and empty with particles less than 150 microns and over the same time course as the proximal intestine. Thus ceca are an adaptation for increasing intestinal surface area without increasing the length or thickness of the intestine itself. Fish ceca are entirely different from the distally located ceca of birds and mammals, which have fermentation functions.


In Vivo Cytochrome P450 3A Isoenzyme Activity and Pharmacokinetics of Imatinib in Relation to Therapeutic Outcome in Patients With Chronic Myeloid Leukemia.
Cytochrome P450 3A (CYP3A) isoenzyme metabolic activity varies between individuals and is therefore a possible candidate of influence on the therapeutic outcome of the tyrosine kinase inhibitor imatinib in patients with chronic myeloid leukemia (CML). The aim of this study was to investigate the influence of CYP3A metabolic activity on the plasma concentration and outcome of imatinib in patients with CML. Forty-three patients with CML were phenotyped for CYP3A activity using quinine as a probe drug and evaluated for clinical response parameters. Plasma concentrations of imatinib and its main metabolite, CGP74588, were determined using liquid chromatography-mass spectrometry. Patients with optimal response to imatinib after 12 months of therapy did not differ in CYP3A activity compared to nonoptimal responders (quinine metabolic ratio of 14.69 and 14.70, respectively; P = 0.966). Neither the imatinib plasma concentration nor the CGP74588/imatinib ratio was significantly associated with CYP3A activity. The CYP3A activity does not influence imatinib plasma concentrations or the therapeutic outcome. These results indicate that although imatinib is metabolized by CYP3A enzymes, this activity is not the rate-limiting step in imatinib metabolism and excretion. Future studies should focus on other pharmacokinetic processes so as to identify the major contributor to patient variability in imatinib plasma concentrations.


NHS Health Check comorbidity and management: an observational matched study in primary care.
The NHS Health Check programme completed its first 5 years in 2014, identifying those at highest risk of cardiovascular disease and new comorbidities, and offering behavioural change support and treatment. To describe the coverage and impact of this programme on cardiovascular risk management and identification of new comorbidities. Observational 5-year study from April 2009 to March 2014, in 139 of 143 general practices in three clinical commissioning groups (CCGs) in east London. A matched analysis compared comorbidity in NHS Health Check attendees and non-attendees. A total of 252 259 adults aged 40-74 years were eligible for an NHS Health Check and, of these, 85 122 attended in 5 years. Attendance increased from 7.3% (10 900/149 867) in 2009 to 17.0% (18 459/108 525) in 2013 to 2014, representing increasing coverage from 36.4% to 85.0%. Attendance was higher in the more deprived quintiles and among South Asians. Statins were prescribed to 11.5% of attendees and 8.2% of non-attendees. In a matched analysis, newly-diagnosed comorbidity was more likely in attendees than non-attendees, with odds ratios for new diabetes 1.30 (95% confidence interval [CI] = 1.21 to 1.39), hypertension 1.50 (95% CI = 1.43 to 1.57), and chronic kidney disease 1.83 (95% CI = 1.52 to 2.21). The NHS Health Check programme provision in these CCGs was equitable, with recent coverage of 85%. Statins were 40% more likely to be prescribed to attendees than non-attendees, providing estimated absolute benefits of public health importance. More new cases of diabetes, hypertension, and chronic kidney disease were identified among attendees than a matched group of non-attendees.


Treatment of peritoneal carcinomatosis with Pressurized IntraPeritoneal Aerosol Chemotherapy - PIPAC-OPC2.
Peritoneal carcinomatosis (PC) is a common endpoint in both gastrointestinal and non-gastrointestinal cancers, and PC is treated as other systemic metastases - unfortunately with disappointing results and considerable side-effects. Pressurized IntraPeritoneal Aerosol Chemotherapy (PIPAC) is a new method of applying traditional chemotherapy, and preliminary data indicate that PIPAC is safe, able to stabilize or improve quality of life, and can induce an objectively measurable reduction in disease burden in PC. PIPAC-OPC2 is a prospectively controlled Phase II, single center, one-arm, open-label clinical trial investigating the treatment effect of PIPAC in patients with histological or cytological proven PC from gastrointestinal, ovarian or primary peritoneal cancer. Eligible patients will receive PIPAC in series of three using a combination of doxorubicin (1.5 mg/m2) and cisplatin (7.5 mg/m2) for non-colorectal cancer patients (PIPAC C/D), and oxaliplatin (92 mg/m2) in patients with PC of colorectal origin (PIPAC OX). Patients are monitored by: (1) repeated measurements of the Peritoneal Regression Grading Score (PRGS) in biopsies obtained from metal clips marked areas, (2) Quality-of-Life (QoL) questionnaires, (3) Magnetic Resonance Imaging (MRI) and (4) Prognostic Nutritional Index (PNI). Adverse events and surgical complications will be recorded according to the 30 days definition. The primary outcome of PIPAC-OPC2 is to evaluate if PIPAC can induce major or complete response (PRGS 1 or 2) within a series of three PIPAC procedures. Secondarily this study investigates changes in QoL and MRI as a staging and response evaluation tool. The secondary outcomes will be used to create a model that may predict which of the patients will benefit from PIPAC treatment. It is expected that PIPAC directed therapy can induce major or complete response in 50 % of patients with PC of colorectal origin and in 30 % of patients with PC of non-colorectal origin - and at the same time stabilize or even improve quality of life. This trial may provide data regarding the utility of MRI as a staging and response evaluation tool in patients with PC. The study is registered with ClinicalTrials.gov Identifier NCT03287375 and the European Clinical Trials Database (EudraCT) number 2016-003394-18.


Growth of Francisella tularensis LVS in macrophages: the acidic intracellular compartment provides essential iron required for growth.
Murine macrophages supported exponential intracellular growth of Francisella tularensis LVS in vitro with a doubling time of 4 to 6 h. LVS was internalized and remained in a vacuolar compartment throughout its growth cycle. The importance of endosome acidification to intracellular growth of this bacterium was assessed by treatment of LVS-infected macrophages with several different lysosomotropic agents (chloroquine, NH4Cl, and ouabain). Regardless of the agent used or its mechanism of action, macrophages treated with agents that blocked endosome acidification no longer supported replication of LVS. Over several experiments for each lysosomotropic agent, the number of CFU of LVS recovered from treated macrophage cultures was equivalent to the input inoculum (approximately 10(4) CFU) at 72 h. In contrast, over 10(8) CFU was consistently recovered from untreated cultures. Pretreatment of macrophages with these endosome acidification inhibitors did not alter their ingestion of bacteria. Further, the effects of the inhibitors were completely reversible: inhibitor-pretreated LVS-infected macrophages washed free of the agent and cultured in medium fully supported LVS growth over 72 h. Endosome acidification is an important cellular event essential for release of iron from transferrin. The growth-inhibitory effects of both chloroquine and NH4Cl were completely reversed by addition of ferric PPi, a transferrin-independent iron source, at a neutral pH but not by addition of excess holotransferrin. Thus, intracellular localization in an acidic vesicle which facilitates the availability of iron essential for Francisella growth is a survival tactic of this bacterium, and iron depletion is one mechanism that macrophages use to inhibit its growth.


Isolation of Murine Primary Aortic Smooth Muscle Cells.
Vascular smooth muscle cells (VSMCs) have been cultured for decades to study the role of these cells in cardiovascular disorders. The most common source of VSMCs is the rat aorta. Here we show the adaptation of this method to isolate and culture mouse aortic VSMCs. The advantage of this method is that there are many more transgenic mouse lines available compared to rats. The protocol consists of the isolation of the aorta, the liberation of vascular cells by the action of collagenase, culturing of VSCMs, and analyzing filamentous actin and alpha smooth muscle actin by fluorescence microscopy. VSCMs can be further used to study mechanisms underlying cardiovascular diseases. Graphic abstract: Figure 1.Working steps.


Silkworm fungal infection model for identification of virulence genes in pathogenic fungus and screening of novel antifungal drugs.
The silkworm infection model has the potential to replace conventional animal models for evaluation of the efficacy and toxicity of investigational antifungal agents. Silkworms are relatively inexpensive, can be simply grown in large numbers and can be easily infected with pathogenic fungi, including mutant strains. Antifungal agents can then be injected into the silkworm either via the hemolymph to mimic intravenous administration or directly into the gut for oral administration, and their antifungal effect can be evaluated. Common features regarding the mechanisms of pharmacokinetics between the silkworm and mammals result in consistent therapeutic effectiveness of antifungal agents. ASP2397, a promising new antifungal agent, was discovered using the silkworm model. The conclusion is that silkworms can be a more ethical and less expensive alternative to standard animal models, particularly for the identification and testing of new antifungal agents.


Bicarbonate increases tumor pH and inhibits spontaneous metastases.
The external pH of solid tumors is acidic as a consequence of increased metabolism of glucose and poor perfusion. Acid pH has been shown to stimulate tumor cell invasion and metastasis in vitro and in cells before tail vein injection in vivo. The present study investigates whether inhibition of this tumor acidity will reduce the incidence of in vivo metastases. Here, we show that oral NaHCO(3) selectively increased the pH of tumors and reduced the formation of spontaneous metastases in mouse models of metastatic breast cancer. This treatment regimen was shown to significantly increase the extracellular pH, but not the intracellular pH, of tumors by (31)P magnetic resonance spectroscopy and the export of acid from growing tumors by fluorescence microscopy of tumors grown in window chambers. NaHCO(3) therapy also reduced the rate of lymph node involvement, yet did not affect the levels of circulating tumor cells, suggesting that reduced organ metastases were not due to increased intravasation. In contrast, NaHCO(3) therapy significantly reduced the formation of hepatic metastases following intrasplenic injection, suggesting that it did inhibit extravasation and colonization. In tail vein injections of alternative cancer models, bicarbonate had mixed results, inhibiting the formation of metastases from PC3M prostate cancer cells, but not those of B16 melanoma. Although the mechanism of this therapy is not known with certainty, low pH was shown to increase the release of active cathepsin B, an important matrix remodeling protease.


Simple and adaptable R implementation of WHO/ISH cardiovascular risk charts for all epidemiological subregions of the world.
The World Health Organisation and International Society of Hypertension (WHO/ISH) cardiovascular disease (CVD) risk assessment charts have been implemented in many low- and middle-income countries as part of the WHO Package of Essential Non-Communicable Disease (PEN) Interventions for Primary Health Care in Low-Resource settings. Evaluation of the WHO/ISH cardiovascular risk charts and their use is a key priority and since they only exist in paper or PDF formats, we developed a simple R implementation of the charts for all epidemiological subregions of the world. The main strengths of this implementation are that it is built in a free, open-source, coding language with simple syntax, can be modified by the user, and can be used with a standard computer.


Oncornavirus expression in human x mouse hybrid cells segregating mouse chromosomes.
Human x mouse hybrid clones obtained by fusing transformed human (VA2) cells with embryonic mouse brain cells were tested for their ability to spontaneously express type C virus particles. It had been previously shown that these hybrid cells preferentially retained human chromosomes while mouse chromosomes were lost. The culture fluid from one cell line was found to contain type C particle markers in abundance, and typical budding C particles were observed in the cells by electron microscopy. In contrast, no particle markers were detected in the culture fluid from parental cells and several other hybrid cell lines. Subclones of the virus-positive cell line continued to lose mouse chromosomes and were found to vary more than 100-fold in their culture fluid DNA polymerase activity. The hybrid cell viruses, termed HMV1, banded in a sucrose gradient between 1.14 and 1.16 g/ml, possessed viral group-specific antigens, and exhibited B-tropic host range for replication in mouse embryo cells, but did not replicate in human cells when directly applied. The virus did not transform mouse cells but was able to rescue the defective murine sarcoma virus from sarcoma-positive, helper-virus-negative cells. Activity of the DNA polymerase associated with HMV1 was similar to the activity of Rauscher murine leukemia virus (MuLV) DNA polymerase in its preference for poly(rA) over poly(dA) as a template, use of endogenous template, detergent requirement, and inhibition by antiserum directed against MuLV.DNA polymerase. The results suggest that human x mouse hybrid cells segregating mouse chromosomes can spontaneously express endogenous type C viruses and that such hybrid cell lines may be used for the isolation of latent mammalian oncornaviruses and analysis of viral gene regulation.


Protection from noise-induced hearing loss by Kv2.2 potassium currents in the central medial olivocochlear system.
The central auditory brainstem provides an efferent projection known as the medial olivocochlear (MOC) system, which regulates the cochlear amplifier and mediates protection on exposure to loud sound. It arises from neurons of the ventral nucleus of the trapezoid body (VNTB), so control of neuronal excitability in this pathway has profound effects on hearing. The VNTB and the medial nucleus of the trapezoid body are the only sites of expression for the Kv2.2 voltage-gated potassium channel in the auditory brainstem, consistent with a specialized function of these channels. In the absence of unambiguous antagonists, we used recombinant and transgenic methods to examine how Kv2.2 contributes to MOC efferent function. Viral gene transfer of dominant-negative Kv2.2 in wild-type mice suppressed outward K(+) currents, increasing action potential (AP) half-width and reducing repetitive firing. Similarly, VNTB neurons from Kv2.2 knock-out mice (Kv2.2KO) also showed increased AP duration. Control experiments established that Kv2.2 was not expressed in the cochlea, so any changes in auditory function in the Kv2.2KO mouse must be of central origin. Further, in vivo recordings of auditory brainstem responses revealed that these Kv2.2KO mice were more susceptible to noise-induced hearing loss. We conclude that Kv2.2 regulates neuronal excitability in these brainstem nuclei by maintaining short APs and enhancing high-frequency firing. This safeguards efferent MOC firing during high-intensity sounds and is crucial in the mediation of protection after auditory overexposure.


Assessing motor imagery ability in younger and older adults by combining measures of vividness, controllability and timing of motor imagery.
With the population aging, a large number of patients undergoing rehabilitation are older than 60 years. Also, since the use of motor imagery (MI) training in rehabilitation is becoming more popular, it is important to gain a better knowledge about the age-related changes in MI ability. The main goal of this study was to compare MI ability in younger and older adults as well as to propose a new procedure for testing this ability. Thirty healthy young subjects (mean age: 22.9±2.7 years) and 28 healthy elderly subjects (mean age: 72.4±5.5 years) participated in the experiment. They were administered three tests aimed at assessing three dimensions of MI: (1) the kinesthetic and visual imagery questionnaire (KVIQ) to assess MI vividness; (2) a finger-thumb opposition task to assess MI controllability; and (3) a chronometric task to assess the timing of MI. On average, the younger and older groups showed similar results on the KVIQ and the chronometric task, but the younger group was more accurate at the finger-thumb opposition task. Interestingly, there was a large variability in the performance within both groups, emphasizing the importance of considering each person individually regarding MI ability, whatever his age. Finally, we propose two indexes of MI ability to identify the potential of persons to engage in MI training programs. Future studies are needed to confirm the predictive value of these MI indexes and define inclusion/exclusion thresholds for their use as a screening tool in both younger and older adults.


Renal Medullary Interstitial COX-2 (Cyclooxygenase-2) Is Essential in Preventing Salt-Sensitive Hypertension and Maintaining Renal Inner Medulla/Papilla Structural Integrity.
COX (cyclooxygenase)-derived prostaglandins regulate renal hemodynamics and salt and water homeostasis. Inhibition of COX activity causes blood pressure elevation. In addition, chronic analgesic abuse can induce renal injury, including papillary necrosis. COX-2 is highly expressed in the kidney papilla in renal medullary interstitial cells (RMICs). However, its role in blood pressure and papillary integrity in vivo has not been definitively studied. In mice with selective, inducible RMIC COX-2 deletion, a high-salt diet led to an increase in blood pressure that peaked at 4 to 5 weeks and was associated with increased papillary expression of AQP2 (aquaporin 2) and ENac (epithelial sodium channel) and decreased expression of cystic fibrosis transmembrane conductance regulator. With continued high-salt feeding, the mice with RMIC COX-2 deletion had progressive decreases in blood pressure from its peak. After return to a normal-salt diet for 3 weeks, blood pressure remained low and was associated with a persistent urinary concentrating defect. Within 2 weeks of institution of a high-salt diet, increased apoptotic RMICs and collecting duct cells could be detected in papillae with RMIC deletion of COX-2, and by 9 weeks of high salt, there was a striking loss of the papillae. Therefore, RMIC COX-2 expression plays a crucial role in renal handling water and sodium homeostasis, preventing salt-sensitive hypertension and maintaining structural integrity of papilla.


Breast Cancer Litigation in the 21st Century.
Approximately 15% of general surgeons practicing in the United States face a medical malpractice lawsuit each year. This study aimed to determine the reasons for litigation for breast cancer care during the past 17 years by reviewing a public legal database. The LexisNexis legal database was queried using a comprehensive list of terms related to breast cancer, identifying all cases from 2000 to 2017. Data were abstracted, and descriptive analyses were performed. The study identified 264 cases of litigation pertaining to breast cancer care. Delay in breast cancer diagnosis was the most common reason for litigation (n = 156, 59.1%), followed by improperly performed procedures (n = 26, 9.8%). The medical specialties most frequently named in lawsuits as primary defendants were radiology (n = 76, 28.8%), general surgery (n = 74, 28%), and primary care (n = 52, 19.7%). The verdict favored the defendant in 145 cases (54.9%) and the plantiff in 60 cases (22.7%). In 59 cases (22.3%), a settlement was reached out of court. The median plaintiff verdict payouts ($1,485,000) were greater than the settlement payouts ($862,500) (p = 0.04). Failure to diagnose breast cancer in a timely manner was the most common reason for litigation related to breast cancer care in the United States. General surgery was the second most common specialty named in the malpractice cases studied. Most cases were decided in favor of the defendant, but when the plaintiff received a payout, the amount often was substantial. Identifying the most common reasons for litigation may help decrease this rate and improve the patient experience.


Long-term recovery of diaphragm strength in neuralgic amyotrophy.
Diaphragm paralysis is a recognized complication of neuralgic amyotrophy that causes severe dyspnoea. Although recovery of strength in the arm muscles, when affected, is common, there are little data on recovery of diaphragm function. This study, therefore, re-assessed diaphragm strength in cases of bilateral diaphragm paralysis due to neuralgic amyotrophy that had previously been diagnosed at the authors institutions. Fourteen patients were recalled between 2 and 11 yrs after the original diagnosis. Respiratory muscle and diaphragm strength were measured by volitional manoeuvres as maximal inspiratory pressure and sniff transdiaphragmatic pressure. Cervical magnetic phrenic nerve stimulation was used to give a nonvolitional measure of diaphragm strength: twitch transdiaphragmatic pressure. Only two patients remained severely breathless. Ten of the 14 patients had evidence of some recovery of diaphragm strength, in seven cases to within 50% of the lower limit of normal. The rate of recovery was variable: one patient had some recovery after 2 yrs, and the rest took 3 yrs or more. In conclusion, in most patients with diaphragm paralysis due to neuralgic amyotrophy, some recovery of the diaphragm strength occurs, but the rate of recovery may be slow.