Head and neck cancer, a rehabilitation approach.
Survival from head and neck cancer is not uncommon. However, treatment is often radical and may compromise the patient's appearance, function, and quality of life. The most common modalities of treatment, employed either singly or in combination, are chemotherapy, radiation, and surgery. Each treatment modality involves unique complications and compromises, many of which are amenable to rehabilitation techniques. A role of the occupational therapist on a cancer rehabilitation team is described and an overview of the therapeutic procedures for the evaluation and treatment of common problems recognized in these patients is presented. The material is derived both from clinical experience of the rehabilitation team and from a literature review employed in developing the rehabilitation program.


Fictive locomotion in the fourth thoracic ganglion of the crayfish, Procambarus clarkii.
Bath application of muscarinic agonists induced rhythmic motor activity in an in vitro preparation of the thoracic nervous system of the crayfish, Procambarus clarkii. In 70% of the cases, the rhythm was organized into 1 of the 2 normal patterns: "backward" walking or "forward" walking. In the rest (30%), the ganglion produced either a series of bursts of impulses or no rhythm at all, just an increase in the tonic activity. When it was isolated from all ascending and descending afferents, the fourth thoracic ganglion was still able to generate rhythmic motor output during bath application of muscarinic agonists. In certain motor neurons, muscarinic agonists induced plateau potentials. Under these conditions, some of these motor neurons were able to change the period of the motor pattern, which might suggest that these motor neurons were part of the central pattern generator (CPG) for locomotion. In the presence of 5 x 10(-6)M TTX, the membrane potential of these motor neurons continued to oscillate with organized rhythmic membrane potential oscillations into 1 of the 2 patterns. Under these conditions, current injection into certain motor neurons demonstrated that they continued to affect the CPG. Two classes of walking leg interneurons have been found. First, there are those with a sustained membrane potential: injection of a steady depolarizing current into some of these interneurons induced rhythmic activity in all thoracic motor nerves, even in the absence of any pharmacological activation. Second, there are those with an oscillating membrane potential: these seemed to enable silent motor neurons to be involved in an ongoing rhythm.


PCNA interacts with hHus1/hRad9 in response to DNA damage and replication inhibition.
The hHus1 and several hRad proteins are involved in the control of DNA integrity checkpoints, although the mechanisms underlying these processes are unknown. Using a yeast two-hybrid system to detect protein-protein interactions, we found that human proliferating cell nuclear antigen (PCNA), a protein known to function in both DNA replication and repair, interacts with the human checkpoint-related protein Hus1 (hHus1). In human skin fibroblast cells, exposure to ionizing radiation of hydroxyurea triggers translocation of hHus1 from the cytosol to the nucleus, where it associates with PCNA as well as another checkpoint protein, hRad9. This nuclear translocation and the complex formation or hHus1 with PCNA and hRad9 correlate closely with changes in cell cycle distribution in response to radiation exposure. These results suggest that this multi-protein complex may be important for coordinating cell-cycle progression, DNA replication and repair of damaged DNA.


The relationship of left ventricular mass and geometry to incident cardiovascular events: the MESA (Multi-Ethnic Study of Atherosclerosis) study.
The purpose of this study was to evaluate the relationship of left ventricular (LV) mass and geometry measured with cardiac magnetic resonance imaging (MRI) to incident cardiovascular events in the MESA (Multi-Ethnic Study of Atherosclerosis) study. MRI is highly accurate for evaluation of heart size and structure and has not previously been used in a large epidemiologic study to predict cardiovascular events. A total of 5,098 participants in the MESA study underwent cardiac MRI at the baseline examination and were followed up for a median of 4 years. Cox proportional hazard models were constructed to predict the end points of coronary heart disease (CHD), stroke, and heart failure (HF) after adjustment for cardiovascular risk factors. A total of 216 incident events were observed during the follow-up period. In adjusted models, the end points of incident CHD and stroke were positively associated with increased LV mass-to-volume ratio (CHD, hazard ratio [HR]: 2.1 per g/ml, p = 0.02; stroke, HR: 4.2 per g/ml, p = 0.005). In contrast, LV mass showed the strongest association with incident HF events (HR: 1.4 per 10% increment, p < 0.0001). The HF events occurred primarily in participants with LV hypertrophy, that is, >or=95th percentile of LV mass (HR: 8.6, 95% confidence interval: 3.7 to 19.9, reference group <50th percentile of LV mass). The LV size was related to incident HF, stroke, and CHD in this multiethnic cohort. Whereas body size-adjusted LV mass alone predicted incident HF, concentric ventricular remodeling predicted incident stroke and CHD.


Utilizing small nutrient compounds as enhancers of exercise-induced mitochondrial biogenesis.
Endurance exercise, when performed regularly as part of a training program, leads to increases in whole-body and skeletal muscle-specific oxidative capacity. At the cellular level, this adaptive response is manifested by an increased number of oxidative fibers (Type I and IIA myosin heavy chain), an increase in capillarity and an increase in mitochondrial biogenesis. The increase in mitochondrial biogenesis (increased volume and functional capacity) is fundamentally important as it leads to greater rates of oxidative phosphorylation and an improved capacity to utilize fatty acids during sub-maximal exercise. Given the importance of mitochondrial biogenesis for skeletal muscle performance, considerable attention has been given to understanding the molecular cues stimulated by endurance exercise that culminate in this adaptive response. In turn, this research has led to the identification of pharmaceutical compounds and small nutritional bioactive ingredients that appear able to amplify exercise-responsive signaling pathways in skeletal muscle. The aim of this review is to discuss these purported exercise mimetics and bioactive ingredients in the context of mitochondrial biogenesis in skeletal muscle. We will examine proposed modes of action, discuss evidence of application in skeletal muscle in vivo and finally comment on the feasibility of such approaches to support endurance-training applications in humans.


Phase II trials of fosquidone, (GR63178A), in colorectal, renal and non-small cell lung cancer. CRC Phase II Clinical Trials Committee.
A total of 61 eligible patients with metastatic cancer have been treated in a series of Phase II trials of the novel pentacyclic pyrroloquinone, fosquidone. Tumour types were colorectal (23), renal (21), and non small cell lung (17). No patient had received prior chemotherapy. The drug was given intravenously as a 20 min infusion at the dose of 120 mg-2 on days 1 to 5 every 3 weeks. Treatment was well tolerated; the only significant side effects being mild nausea and generalised musculo-skeletal pains. Response was assessed after two cycles of therapy. No patient achieved an objective partial response. A total of nine patients demonstrated stable disease for a median duration of 11 weeks. Using this schedule of administration, fosquidone has no significant antitumour activity in this group of tumours.


High-sensitivity detection of BCR-ABL kinase domain mutations in imatinib-naive patients: correlation with clonal cytogenetic evolution but not response to therapy.
Mutations in the kinase domain (KD) of BCR-ABL are the leading cause of acquired imatinib resistance. In some cases, identical mutations were detected at relapse and in pretherapeutic specimens, consistent with selection of resistant clones in the presence of drug. However, the incidence of KD mutations in imatinibnaive patients, irrespective of response to therapy, is unknown. We studied mutation frequency in 66 patients with chronic myelogenous leukemia (CML), using cDNA sequencing and allele-specific oligonucleotide-polymerase chain reaction (ASO-PCR) assays for 8 common mutations. Thirteen patients were positive by ASO-PCR only, 1 by ASO-PCR and sequencing, and 1 by sequencing only (overall frequency, 22.7%). T315I was most frequent (12% of patients). Eleven of the 14 patients with positive ASO-PCR had follow-up samples available for sequencing. Wild-type sequence was detected in 6 of 11, 2 different mutations in 1 of 11, and identical mutations in 4 of 11 patients, 2 of whom had achieved major cytogenetic response. In multivariate analysis mutation detection was associated with clonal cytogenetic evolution, exposure to 6-Thioguanine, and a low platelet count, but not with response to imatinib, event-free survival, and overall survival. KD mutants present at low levels do not invariably lead to relapse, and additional factors are required to induce a fully drug-resistant phenotype.


A method to increase the power of multiple testing procedures through sample splitting.
Consider the standard multiple testing problem where many hypotheses are to be tested, each hypothesis is associated with a test statistic, and large test statistics provide evidence against the null hypotheses. One proposal to provide probabilistic control of Type-I errors is the use of procedures ensuring that the expected number of false positives does not exceed a user-supplied threshold. Among such multiple testing procedures, we derive the most powerful method, meaning the test statistic cutoffs that maximize the expected number of true positives. Unfortunately, these optimal cutoffs depend on the true unknown data generating distribution, so could never be used in a practical setting. We instead consider splitting the sample so that the optimal cutoffs are estimated from a portion of the data, and then testing on the remaining data using these estimated cutoffs. When the null distributions for all test statistics are the same, the obvious way to control the expected number of false positives would be to use a common cutoff for all tests. In this work, we consider the common cutoff method as a benchmark multiple testing procedure. We show that in certain circumstances the use of estimated optimal cutoffs via sample splitting can dramatically outperform this benchmark method, resulting in increased true discoveries, while retaining Type-I error control. This paper is an updated version of the work presented in Rubin et al. (2005), later expanded upon by Wasserman and Roeder (2006).


Towards practical soft X-ray spectromicroscopy of biomaterials.
Scanning transmission X-ray microscopy (STXM) is being developed as a new tool to study the surface chemical morphology and biointeractions of candidate biomaterials with emphasis on blood compatible polymers. STXM is a synchrotron based technique which provides quantitative chemical mapping at a spatial resolution of 50 nm. Chemical speciation is provided by the near edge X-ray absorption spectral (NEXAFS) signal. We show that STXM can detect proteins on soft X-ray transparent polymer thin films with monolayer sensitivity. Of great significance is the fact that measurements can be made in situ, i.e. in the presence of an overlayer of the protein solution. The strengths, limitations and future potential of STXM for studies of biomaterials are discussed.


Correlates of HIV infection among incarcerated women: implications for improving detection of HIV infection.
The prevalence of HIV infection in correctional settings is several-fold higher than found in community settings. New approaches to identifying HIV infection among prisoners are urgently needed. In order to determine the HIV seroprevalence and to identify the correlates of HIV infection among female prisoners, an anonymous, but linked HIV serosurvey was conducted at Connecticut's sole correctional facility for women (census=1,100). After removing all individual identifiers for inmates' standardized clinical and risk behavior information, data are linked by a third source to blinded HIV-testing information by a third party. This three-step sequential process allows for anonymous HIV testing that can still be linked with deidentified clinical and behavioral data. Of the 3,315 subjects with complete information, 250 (7.5%) were HIV+. Of these, 157 (63%) self-reported being HIV+. Using multiple logistic regression analysis, having sex with a known HIV+ person [adjusted odds ratio (AOR)=9.1] and injection drug use (AOR=6.1) were the most highly correlated risk factors for HIV, whereas leukopenia (AOR=9.4) and hypoalbuminemia (AOR=7.2) were the most significant laboratory markers. Other independent correlates of HIV included self-report of syphilis (AOR=1.9) or genital herpes infection (AOR=2.7) and being Black (AOR=2.1) or Hispanic (AOR=2.2). The prevalence of HIV and HIV-risk behaviors is high among incarcerated women. Existing voluntary HIV counseling and testing programs do not completely target high-risk groups who remain part of the evolving epidemic. Defined demographic, behavioral, and clinical assessments may provide useful information for encouraging targeted counseling and testing. Newer targeted approaches merit further study to determine the effectiveness of this approach. Alternative methods of facilitating more widespread HIV testing, such as saliva tests, rapid serologic tests, and more routine testing in high HIV-prevalence areas should be considered both for clinical and for public health benefits.


Reconstitution of biological molecular generators of electric current. Transhydrogenase.
Direct measurement of the electrogenic activity of purified mitochondrial transhydrogenase has been carried out. To this end, beef-heart transhydrogenase was isolated and reconstituted with phospholipids to form proteoliposomes. The transhydrogenase proteoliposomes were incorporated into a membrane filter impregnated with a decane solution of phospholipids. It is shown that addition of substrates of either the forward (NADPH and NAD+) or the reverse (NADH and NADP+) transhydrogenase reaction gives rise to an electric potential difference across the proteoliposome-treated membrane filter. The electric vector depends upon the direction of the reaction. The proteoliposome-supplemented compartment charges negatively in the case of the forward reaction and positively in the case of the reverse one. Addition of the reaction products after substrates equalizes the potentials. The transhydrogenase-treated membrane filter retains the ability to perform transhydrogenase-linked electrogenesis after removal of excess non-incorporated proteoliposomes. The electric potential difference reaching 20 mV immediately after the transhydrogenase substrate addition, slowly decreases due to accumulation of the reaction products. Such decay is prevented when the mixture is supplemented with the substrate-regenerating and product-utilizing enzymic systems. Under these conditions, a steady continuous electric current of about 10 pA can be observed.


Living Donor Kidney Transplantation: Improving Education Outside of Transplant Centers about Live Donor Transplantation--Recommendations from a Consensus Conference.
Living donor kidney transplantation (LDKT) offers better quality of life and clinical outcomes, including patient survival, compared with remaining on dialysis or receiving a deceased donor kidney transplant. Although LDKT education within transplant centers for both potential recipients and living donors is very important, outreach and education to kidney patients in settings other than transplant centers and to the general public is also critical to increase access to this highly beneficial treatment. In June 2014, the American Society of Transplantation's Live Donor Community of Practice, with the support of 10 additional sponsors, convened a consensus conference to determine best practices in LDKT, including a workgroup focused on developing a set of recommendations for optimizing outreach and LDKT education outside of transplant centers. Members of this workgroup performed a structured literature review, conducted teleconference meetings, and met in person at the 2-day conference. Their efforts resulted in consensus around the following recommendations. First, preemptive transplantation should be promoted through increased LDKT education by primary care physicians and community nephrologists. Second, dialysis providers should be trained to educate their own patients about LDKT and deceased donor kidney transplantation. Third, partnerships between community organizations, organ procurement organizations, religious organizations, and transplant centers should be fostered to support transplantation. Fourth, use of technology should be improved or expanded to better educate kidney patients and their support networks. Fifth, LDKT education and outreach should be improved for kidney patients in rural areas. Finally, a consensus-driven, evidence-based public message about LDKT should be developed. Discussion of the effect and potential for implementation around each recommendation is featured, particularly regarding reducing racial and socioeconomic disparities in access to LDKT. To accomplish these recommendations, the entire community of professionals and organizations serving kidney patients must work collaboratively toward ensuring accurate, comprehensive, and up-to-date LDKT education for all patients, thereby reducing barriers to LDKT access and increasing LDKT rates.


Exploring peptide-membrane interactions with coarse-grained MD simulations.
The interaction of α-helical peptides with lipid bilayers is central to our understanding of the physicochemical principles of biological membrane organization and stability. Mutations that alter the position or orientation of an α-helix within a membrane, or that change the probability that the α-helix will insert into the membrane, can alter a range of membrane protein functions. We describe a comparative coarse-grained molecular dynamics simulation methodology, based on self-assembly of a lipid bilayer in the presence of an α-helical peptide, which allows us to model membrane transmembrane helix insertion. We validate this methodology against available experimental data for synthetic model peptides (WALP23 and LS3). Simulation-based estimates of apparent free energies of insertion into a bilayer of cystic fibrosis transmembrane regulator-derived helices correlate well with published data for translocon-mediated insertion. Comparison of values of the apparent free energy of insertion from self-assembly simulations with those from coarse-grained molecular dynamics potentials of mean force for model peptides, and with translocon-mediated insertion of cystic fibrosis transmembrane regulator-derived peptides suggests a nonequilibrium model of helix insertion into bilayers.


Valeriana officinalis Root Extract Modulates Cortical Excitatory Circuits in Humans.
Valeriana officinalis extract (VE) is a popular herbal medicine used for the treatment of anxiety and sleep disorders. Although the anxiolytic and sedative effects are mainly attributed to the modulation of GABA-ergic transmission, the mechanism of action has not been fully investigated in humans. Noninvasive brain stimulation protocols can be used to elucidate the mechanisms of action of psychoactive substances at the cortical level in humans. In this study, we investigated the effects of a single dose of VE on cortical excitability as assessed with transcranial magnetic stimulation (TMS). Fifteen healthy volunteers participated in a double-blind, randomized, cross-over, placebo-controlled study. Subjects were required to take either 900 mg of VE (valerenic acid 0.8%) or placebo (an equal dose of vitamin E). Motor cortex excitability was studied by single and paired TMS before and at 1 h and 6 h after the oral administration. Cortical excitability was assessed using different TMS parameters: resting motor threshold, motor-evoked potential amplitude, cortical silent period, short-interval intracortical inhibition, and intracortical facilitation. Furthermore, we assessed sensorimotor integration by short-latency and long-latency afferent inhibition. We found a significant reduction in ICF, without any significant changes in other TMS measures of motor cortex excitability. The amount of ICF returned to baseline value 6 h after the intake of the VE. A single oral dose of VE modulates intracortical facilitatory circuits. Our results in healthy subjects could be predictive markers of treatment response in patients and further support the use of pharmaco-TMS to investigate the neuropsychiatric effects of herbal therapies in humans.


Biallelic C1QBP Mutations Cause Severe Neonatal-, Childhood-, or Later-Onset Cardiomyopathy Associated with Combined Respiratory-Chain Deficiencies.
Complement component 1 Q subcomponent-binding protein (C1QBP; also known as p32) is a multi-compartmental protein whose precise function remains unknown. It is an evolutionary conserved multifunctional protein localized primarily in the mitochondrial matrix and has roles in inflammation and infection processes, mitochondrial ribosome biogenesis, and regulation of apoptosis and nuclear transcription. It has an N-terminal mitochondrial targeting peptide that is proteolytically processed after import into the mitochondrial matrix, where it forms a homotrimeric complex organized in a doughnut-shaped structure. Although C1QBP has been reported to exert pleiotropic effects on many cellular processes, we report here four individuals from unrelated families where biallelic mutations in C1QBP cause a defect in mitochondrial energy metabolism. Infants presented with cardiomyopathy accompanied by multisystemic involvement (liver, kidney, and brain), and children and adults presented with myopathy and progressive external ophthalmoplegia. Multiple mitochondrial respiratory-chain defects, associated with the accumulation of multiple deletions of mitochondrial DNA in the later-onset myopathic cases, were identified in all affected individuals. Steady-state C1QBP levels were decreased in all individuals' samples, leading to combined respiratory-chain enzyme deficiency of complexes I, III, and IV. C1qbp-/- mouse embryonic fibroblasts (MEFs) resembled the human disease phenotype by showing multiple defects in oxidative phosphorylation (OXPHOS). Complementation with wild-type, but not mutagenized, C1qbp restored OXPHOS protein levels and mitochondrial enzyme activities in C1qbp-/- MEFs. C1QBP deficiency represents an important mitochondrial disorder associated with a clinical spectrum ranging from infantile lactic acidosis to childhood (cardio)myopathy and late-onset progressive external ophthalmoplegia.


Association of Macular Thickness With Age and Age-Related Macular Degeneration in the Carotenoids in Age-Related Eye Disease Study 2 (CAREDS2), An Ancillary Study of the Women's Health Initiative.
To evaluate the relationship of retinal layer thickness with age and age-related macular degeneration (AMD) in the Carotenoids in Age-Related Eye Disease Study 2. Total retinal thickness within the macular area, and individual layer thickness was determined for CAREDS2 participants (n = 906 eyes, 473 women) from the Women's Health Initiative using Heidelberg optical coherence tomography (OCT). Mean measurements within the OCT grid were compared across age tertiles (69-78, 78-83, and 83-101 years) and AMD outcomes. Mean retinal thickness in the central circle, inner ring, and outer ring were 277 ± 34 µm, 326 ± 20 µm, and 282 ± 15 µm, respectively. Thickness did not vary by age in the central circle, but decreased with age in the inner and outer circles (P ≤ 0.004). Specifically, ganglion cell (GCL), inner plexiform, and outer nuclear (ONL) layer thickness decreased with age (P ≤ 0.003). Age-adjusted retinal thickness in all three circles did not vary by AMD outcomes (486 without AMD and 413 with AMD). However, individual layers showed changes with GCL and photoreceptor thinning and retinal pigment epithelial thicknening in eyes with late AMD. After controlling for age and AMD, higher ONL thickness was associated with better visual acuity. In this cohort of older women, a decrease in perifoveal thickness was associated with increasing age, particularly in the inner retinal layers. Variabilty in thickness in AMD eyes was primarily due to outer retinal layers. Among all retinal layers, the ONL plays an important role in preserving visual acuity. The study provides a deeper understanding of age related changes to the retinal layers and their effect on visual loss.


Response of dairy cows to high doses of a sustained-release bovine somatotropin administered during two lactations. 2. Health and reproduction.
Eighty-two lactating Holstein cows in their first, second, or third lactation received either one, three, or five concurrent i.m. injections of a unit dose (.6 g) of zinc methionyl bST (sometribove) or five doses of the vehicle. Injections were given at 14-d intervals from 60 +/- 3 d postpartum until the end of lactation or necropsy. Thirty-eight animals were continued on treatment for a 2nd yr. Sometribove did not affect the incidence of ketosis, milk fever, tetany, or pneumonia. Digestive disorders, primarily cows going off feed, were increased by bST during yr 1 only. The incidence of lameness was increased by bST in some time frames because of an increase in the 3.0-g bST group. Lameness was not associated with treatment-specific histopathologic changes or with abnormalities in cartilage or bone. Reproductive health generally was unaffected by treatment, but delayed conception and increased incidence of abortion were noted. Incidence of cystic ovaries was unaffected by bST. Pregnancy rates were decreased during the 100-d breeding interval of yr 1 but not during the 215-d interval of yr 2. The incidence of clinical mastitis was increased by bST, primarily at the 3.0-g dose. During the 2-yr study, 0, 3, 3, and 2 cows died or became moribund on 0, .6, 1.8, and 3.0 g of bST, respectively. Health issues identified for further evaluation included lameness and clinical mastitis for the 3.0-g group and early removal from the herd and decreased reproductive performance for all bST groups. Bovine somatotropin caused no treatment-specific toxic effects in dairy cows even at 3.0 g every 14 d.


The use of a novel drinkometer system for assessing pharmacological treatment effects on ethanol consumption in rats.
There are numerous studies in the preclinical alcohol research field showing that pharmacological interventions and many other manipulations can influence ethanol (EtOH) consumption in a free-choice paradigm in rats. Most of these studies are based on 24-hour measurements. These studies provide a measure of the total amount of EtOH consumed per day, but do not provide information on the drinking patterns within this period of measurement. Here, we used a novel drinkometer system in combination with Fourier analysis to provide detailed information on drinking patterns. Our automated drinkometer system measures fluid consumption by means of high-precision sensors attached to the drinking bottles in the home cage of the rat and thereby ameliorates several limitations of a classical lickometer-based drinkometer system. As an example of its application, we used the alcohol deprivation effect (ADE) model for relapse-like drinking and tested as a reference compound lamotrigine, which has a robust effect on the ADE. Fourier analysis was chosen as the main strategy for 24-hour drinking pattern recognition during water/EtOH drinking. Under baseline conditions, voluntary EtOH consumption in rats can be expressed as characteristic oscillations that follow diurnal activity and differ in their amplitude, depending on the EtOH concentration. This diurnal drinking rhythmicity was altered during a relapse condition. Furthermore, lamotrigine given during the ADE did not significantly affect the drinking frequency or the number of approaches to the EtOH bottles when compared to vehicle-treated animals. However, EtOH intake during a drinking approach was dramatically reduced. The use of the drinkometer system and mathematical modeling allows the characterization of treatment effects on relapse-like drinking with a great level of detail. One use of such detailed information may lie in its translational predictability. For instance, owing to lamotrigine treatment's lack of effect on EtOH drinking frequency or the number of approaches to the EtOH bottles, this compound might not be effective in relapse prevention per se but may reduce hedonic EtOH effects and could therefore be used in alcohol-dependent patients if harm reduction is the primary goal of treatment.


Cerebral ischemia during hemodialysis-finding the signal in the noise.
Hemodialysis patients have multiple risk factors for small vessel cerebrovascular disease and cognitive dysfunction. Hemodialysis itself may cause clinically significant neurological injury through repetitive cerebral ischemia. However, supporting evidence to date consists of epidemiological associations, expert opinion, and small, single-centre studies of variable methodological quality. Isolating the impact of intra-dialytic hemodynamic instability from underlying renal and vascular disease on clinically relevant functional outcomes would require very large, controlled studies, given the heterogeneity and confounding comorbidities of the population, and the complex relationship between blood pressure and cerebral oxygen delivery. There has been an increase in complementary physiological studies looking directly at intra-dialytic cerebral oxygen balance, which have provided supporting evidence for the occurrence of cerebral ischemia, often independently of hemodynamics. Data suggesting a relationship between these measures of oxygen balance and functional outcomes is only hypothesis-generating at this stage. We advocate the testing of interventions that aim to reduce intra-dialytic cerebral hypoxia (rather than hypotension) in sufficiently powered studies, followed by correlation with validated, longitudinal assessment of clinically relevant neurological damage.


Prediction of development of fatigue during a simulated ambulance work task from physical performance tests.
The aims of this study were (1) to identify which physical performance tests could best explain the development of fatigue during a simulated ambulance work task, (2) to investigate the effect of height and weight and (3) to investigate in what respects these findings differ between female and male ambulance personnel. Forty-eight male and 17 female ambulance personnel completed a test battery assessing cardio-respiratory capacity, muscular strength and endurance, and co-ordination. The subjects also completed a simulated ambulance work task -- carrying a loaded stretcher. The work task was evaluated by development of fatigue. Univariate and multiple regression analyses were used to investigate to what extent the tests in the test battery were able to explain the variance of developed fatigue. The explained variance was higher for female than for male ambulance personnel (time > 70% of HRpeak: R2 = 0.75 vs 0.10, accumulated lactate: R2 = 0.62 vs 0.42, perceived exertion: R2 = 0.75 vs 0.10). Significant predictors in the models were VO2max, isometric back endurance, one-leg rising, isokinetic knee flexion and shoulder extension strength. Height, but not weight, could further explain the variance. The high physical strain during carrying the loaded stretcher implies the importance of investigating whether improved performance, matching the occupational demands, could decrease the development of fatigue during strenuous tasks.


Advanced phase chronic myeloid leukaemia (CML) in the tyrosine kinase inhibitor era - a report from the Swedish CML register.
The primary goal in management of chronic phase (CP) chronic myeloid leukaemia (CML) is to prevent disease progression to accelerated phase (AP) or blast crisis (BC). We have evaluated progression rates in a decentralised healthcare setting and characterised patients progressing to AP/BC on TKI treatment. Using data from the Swedish CML register, we identified CP-CML patients diagnosed 2007-2011 who progressed to AP/BC within 2 yrs from diagnosis (n = 18) as well as patients diagnosed in advanced phase during 2007-2012 (n = 36) from a total of 544 newly diagnosed CML cases. We evaluated baseline characteristics, progression rates, outcome and adherence to guidelines for monitoring and treatment. The cumulative progression rate at 2 yrs was 4.3%. All 18 progression cases had been treated with imatinib, and six progressed within 6 months. High-risk EUTOS score was associated to a higher risk of progression. Insufficient cytogenetic and/or molecular monitoring was found in 33%. Median survival after transformation during TKI treatment was 1.4 yrs. In those presenting with BC and AP, median survival was 1.6 yrs and not reached, respectively. In this population-based setting, progression rates appear comparable to that reported from clinical trials, with similar dismal patient outcome. Improved adherence to CML guidelines may minimise the risk of disease progression.


Surface enhanced resonance Raman spectroscopy of iron Hangman complexes on electrodes during electrocatalytic oxygen reduction: advantages and problems of common drycast methods.
Drycast methods have been used frequently in recent decades to adsorb a range of synthetic catalysts on electrodes. The uncoordinated multilayers that are formed via this immobilization method can however have a strong impact on the electrocatalytic reaction pathway as slow electron transfer and intermolecular interactions can alter the chemistry of the catalysts on the surface. To gain insight into the structure of Fe porphyrin Hangman catalysts during electrocatalytic oxygen reduction a combination of electrochemistry and surface enhanced resonance Raman spectroscopy (SERRS) was applied. The Hangman complexes were attached to the electrodes via different methods and the influence of the immobilisation technique on oxygen chemistry was studied. In multilayer systems, new intermediates could be identified via potential dependent SERRS that were not present in solution or in monolayer systems under catalytic conditions. A comparison of Raman spectra obtained either via Soret or Q-band excitation showed that the porphyrin symmetry is strongly distorted under reducing conditions, which was interpreted by the transient formation of dimer complexes during catalysis.


A Specific Mapping Study Using Fluorescence Sentinel Lymph Node Detection in Patients with Intermediate- and High-risk Prostate Cancer Undergoing Extended Pelvic Lymph Node Dissection.
Sentinel lymph node (SLN) detection techniques have the potential to change the standard of surgical care for patients with prostate cancer. We performed a lymphatic mapping study and determined the value of fluorescence SLN detection with indocyanine green (ICG) for the detection of lymph node metastases in intermediate- and high-risk patients undergoing radical prostatectomy and extended pelvic lymph node dissection. A total of 42 patients received systematic or specific ICG injections into the prostate base, the midportion, the apex, the left lobe, or the right lobe. We found (1) that external and internal iliac regions encompass the majority of SLNs, (2) that common iliac regions contain up to 22% of all SLNs, (3) that a prostatic lobe can drain into the contralateral group of pelvic lymph nodes, and (4) that the fossa of Marcille also receives significant drainage. Among the 12 patients who received systematic ICG injections, 5 (42%) had a total of 29 lymph node metastases. Of these, 16 nodes were ICG positive, yielding 55% sensitivity. The complex drainage pattern of the prostate and the low sensitivity of ICG for the detection of lymph node metastases reported in our study highlight the difficulties related to the implementation of SNL techniques in prostate cancer. There is controversy about how extensive lymph node dissection (LND) should be during prostatectomy. We investigated the lymphatic drainage of the prostate and whether sentinel node fluorescence techniques would be useful to detect node metastases. We found that the drainage pattern is complex and that the sentinel node technique is not able to replace extended pelvic LND.


Secret to friendship: Children make inferences about friendship based on secret sharing.
Secrets carry valuable social information. Because the content of secrets can be damaging to the secret-keeper's reputation, people should only disclose their secrets to people whom they trust. Therefore, tracking which people know each other's secrets can be used as cue of social relationships: If one person tells another person a secret, those people are likely friends. Here, in 5 studies with 3- to 12-year-old children (total N = 452), we examined the developmental trajectory of reasoning about secret sharing as an indication of third-party friendship. By age 6, but not before, children expected that a person would be friends with someone that she told a secret. We replicated this main finding across four studies by comparing secret sharing to other cues of affiliation. Children treated sharing a secret as a stronger cue to friendship than sharing a physical object (Study 1), sharing a fact (Studies 2-4), or sharing membership on the same sports team (Study 3). Although younger children did not understand that secret sharing indicated friendship, they did expect people to be more likely to disclose their secrets to friends than to nonfriends (Study 5). Taken together, our results indicate that children understand the social significance of sharing secrets and use secret sharing to make important predictions about the social world. Specifically, children infer social relationships based on which people know each other's secrets and expect others to share secrets selectivity with friends. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


The effect of a cardiovascular educational intervention on healthcare utilization and costs.
To evaluate healthcare utilization and costs following a cardiovascular disease (CVD) screening and educational special intervention (SI) compared with a control intervention (CIN) at 1 year in the Family-Based Intervention Trial for Heart Health. Participants randomized to SI for screening and periodic lifestyle counseling were compared with participants randomized to CIN for resource utilization and associated costs at 1 year. A total of 421 participants (67% women and 37% minorities) were healthy family members of hospitalized patients with CVD who had 1-year follow-up resource utilization data. Resource utilization was systematically measured using a standardized questionnaire in both study groups and was validated by medical records in a subsample. Outcomes included provider visits, diagnostic studies, laboratory assessment, medication use, behavioral program enrollment, emergency department (ED) visits, hospital admissions, and healthcare costs. At 1 year, there were significantly fewer overall provider visits (P = .04) and psychiatrist visits (P = .03) in SI versus CIN. There was a nonsignificant trend toward fewer ED visits, decreased hospital admissions, and shorter inpatient length of stay in SI versus CIN. Estimated healthcare expenditures for CIN exceeded those for SI by $590 per participant. The cost of the 1-year intervention was $95 per participant. A 1-year standardized low-cost screening and educational intervention was associated with significantly fewer provider visits and with a nonsignificant trend toward reduced healthcare utilization for several parameters. The long-term effect on outcomes and costs deserves further study.


Correlation between microsatellite discrepancy scores and transplant outcome after haemopoietic SCT for pediatric ALL.
Microsatellite analyses show that self-reported ethnicity often correlates poorly with true genetic ancestry. As unknown ancestral differences could potentially have an impact on transplant outcome, we developed an average allele length discrepancy (AALD) score to assess allele length discrepancy between donor/recipient (D/R) using microsatellites analysed routinely in post-transplant chimeric assessment. This was then compared with outcome in a homogeneously treated cohort of pediatric patients undergoing high-resolution sibling or matched unrelated donor transplantation for acute lymphoblastic leukemia (ALL). AALD scores formed a numeric continuum ranging from 0 to 1.4 (median 0.76) for sibling pairs and 0.8-2.17 (median 1.6) for high-resolution matched unrelated donor (HR-MUD) pairs. There was a trend for worse OS with increasing AALD score, which reached statistical significance above a threshold of 1.7 for OS. Patients whose transplants had an AALD score of ⩾1.8 had a risk of non-relapse mortality 4.9 times greater (P=0.025) and relapse risk three times greater (P=0.058) than those scoring <1.8. This approach will now be explored in a Centre International for Blood and Marrow Transplantation Research (CIBMTR) study of 750 D/R pairs across all disease groups; if confirmed, it has the potential to improve donor selection for patients with multiple prospective donors.


Photoparoxysmal responses in the electroencephalograms of some ethnic groups of the Cape Peninsula.
A comparative investigation of the frequency of photoparoxysmal responses (PPR) in the white, mixed and African populations of the Cape Peninsula is presented, based on 9697 consecutive referrals for EEG. PPR were observed in 2.5% of the white, in 1.3% of the mixed and in 0.9% of the African population groups. In the present material there were more patients with epilepsy in the mixed than in the white group, the frequencies changing with age in the two groups. Minor interrelationships of PPR exist with sex, age and race considered separately. Another minor relationship could be demonstrated between PPR and epilepsy. Three-factor interaction exists among epilepsy, age and race, and a further three-way interaction could be shown among sex, age and epilepsy. These relations and interactions were all significant below the 1% level. Standard EEG records (including response to hyperventilation) sensitivity limits to PPR, and the types of seizures themselves, did not differ significantly between the two main ethnic groups.


Taking Respect Seriously: Clinical Research and the Demands of Informed Consent.
There is broad agreement among research ethicists that investigators have a duty to obtain the informed consent of all subjects who participate in their research trials. On a common view, the duty to obtain this informed consent follows from the need to respect persons and their autonomous decisions. However, the nature of informed consent and the demands it places on investigators are open to dispute and recently have been challenged. Respect for persons, it has been claimed, does not require investigators to guarantee that the subjects enrolled in their trials comprehend the risk/benefit information disclosed to them or even that they appreciate the difference between research and therapy. According to this critique, the significance of defects in informed consent, like therapeutic misconception or unrealistic optimism, has been greatly exaggerated. This article reevaluates informed consent in clinical research in light of this critique. It not only rebuts the main points the critics raise, but also shows that other points they raise can be accepted by a doctrine of informed consent that resembles the common view in maintaining that autonomous authorization is central to informed consent.


Modern subsurface bacteria in pristine 2.7 Ga-old fossil stromatolite drillcore samples from the Fortescue Group, Western Australia.
Several abiotic processes leading to the formation of life-like signatures or later contamination with actual biogenic traces can blur the interpretation of the earliest fossil record. In recent years, a large body of evidence showing the occurrence of diverse and active microbial communities in the terrestrial subsurface has accumulated. Considering the time elapsed since Archaean sedimentation, the contribution of subsurface microbial communities postdating the rock formation to the fossil biomarker pool and other biogenic remains in Archaean rocks may be far from negligible. In order to evaluate the degree of potential contamination of Archean rocks by modern microorganisms, we looked for the presence of living indigenous bacteria in fresh diamond drillcores through 2,724 Myr-old stromatolites (Tumbiana Formation, Fortescue Group, Western Australia) using molecular methods based on the amplification of small subunit ribosomal RNA genes (SSU rDNAs). We analyzed drillcore samples from 4.3 m and 66.2 m depth, showing signs of meteoritic alteration, and also from deeper "fresh" samples showing no apparent evidence for late stage alteration (68 m, 78.8 m, and 99.3 m). We also analyzed control samples from drilling and sawing fluids and a series of laboratory controls to establish a list of potential contaminants introduced during sample manipulation and PCR experiments. We identified in this way the presence of indigenous bacteria belonging to Firmicutes, Actinobacteria, and Alpha-, Beta-, and Gammaproteobacteria in aseptically-sawed inner parts of drillcores down to at least 78.8 m depth. The presence of modern bacterial communities in subsurface fossil stromatolite layers opens the possibility that a continuous microbial colonization had existed in the past and contributed to the accumulation of biogenic traces over geological timescales. This finding casts shadow on bulk analyses of early life remains and makes claims for morphological, chemical, isotopic, and biomarker traces syngenetic with the rock unreliable in the absence of detailed contextual analyses at microscale.


Pharmacological approaches to the treatment of obstructive sleep apnoea.
Currently the treatment of choice for symptomatic obstructive sleep apnoea (OSA) is continuous positive airway pressure (CPAP). Some patients with OSA do not tolerate CPAP or have insufficiently severe symptoms to justify its use; for these patients, drug therapy would be a desirable potential therapeutic alternative. To summarize the current evidence on the effectiveness of drug therapy in patients with OSA. A systematic review of randomized controlled trials was performed to investigate the effects of drug therapy on OSA. Searches of bibliographical databases revealed 33 trials investigating the effects of 27 different drugs on OSA severity and/or symptoms. The mechanisms by which these drugs are supposed to improve OSA include, amongst others, an increase in tone of the upper airways, an increase in ventilatory drive, a reduction in airway resistance, and alterations in surface tension forces in the upper airway. In most of these studies there was no significant effect on OSA observed. However, there is evidence from a few small trials that some drugs, especially those thought to increase upper airway muscle tone, have the potential to reduce OSA severity; but further data from larger studies of adequate duration are needed.


Cross-sectional associations between central and general adiposity with albuminuria: observations from 400,000 people in UK Biobank.
Whether measures of central adiposity are more or less strongly associated with risk of albuminuria than body mass index (BMI), and by how much diabetes/levels of glycosylated haemoglobin (HbA1c) explain or modify these associations, is uncertain. Ordinal logistic regression was used to estimate associations between values of central adiposity (waist-to-hip ratio) and, separately, general adiposity (BMI) with categories of urinary albumin-to-creatinine ratio (uACR) in 408,527 UK Biobank participants. Separate central and general adiposity-based models were initially adjusted for potential confounders and measurement error, then sequentially, models were mutually adjusted (e.g. waist-to-hip ratio adjusted for BMI, and vice versa), and finally they were adjusted for potential mediators. Levels of albuminuria were generally low: 20,425 (5%) had a uACR ≥3 mg/mmol. After adjustment for confounders and measurement error, each 0.06 higher waist-to-hip ratio was associated with a 55% (95%CI 53-57%) increase in the odds of being in a higher uACR category. After adjustment for baseline BMI, this association was reduced to 32% (30-34%). Each 5 kg/m2 higher BMI was associated with a 47% (46-49%) increase in the odds of being in a higher uACR category. Adjustment for baseline waist-to-hip ratio reduced this association to 35% (33-37%). Those with higher HbA1c were at progressively higher odds of albuminuria, but positive associations between both waist-to-hip ratio and BMI were apparent irrespective of HbA1c. Altogether, about 40% of central adiposity associations appeared to be mediated by diabetes, vascular disease and blood pressure. Conventional epidemiological approaches suggest that higher waist-to-hip ratio and BMI are independently positively associated with albuminuria. Adiposity-albuminuria associations appear strong among people with normal HbA1c, as well as people with pre-diabetes or diabetes.


Cohesins and condensins orchestrate the 4D dynamics of yeast chromosomes during the cell cycle.
Duplication and segregation of chromosomes involves dynamic reorganization of their internal structure by conserved architectural proteins, including the structural maintenance of chromosomes (SMC) complexes cohesin and condensin. Despite active investigation of the roles of these factors, a genome-wide view of dynamic chromosome architecture at both small and large scale during cell division is still missing. Here, we report the first comprehensive 4D analysis of the higher-order organization of the Saccharomyces cerevisiae genome throughout the cell cycle and investigate the roles of SMC complexes in controlling structural transitions. During replication, cohesion establishment promotes numerous long-range intra-chromosomal contacts and correlates with the individualization of chromosomes, which culminates at metaphase. In anaphase, mitotic chromosomes are abruptly reorganized depending on mechanical forces exerted by the mitotic spindle. Formation of a condensin-dependent loop bridging the centromere cluster with the rDNA loci suggests that condensin-mediated forces may also directly facilitate segregation. This work therefore comprehensively recapitulates cell cycle-dependent chromosome dynamics in a unicellular eukaryote, but also unveils new features of chromosome structural reorganization during highly conserved stages of cell division.


Identification of {beta}-catenin binding regions in colon cancer cells using ChIP-Seq.
Deregulation of the Wnt/β-catenin signaling pathway is a hallmark of colon cancer. Mutations in the adenomatous polyposis coli (APC) gene occur in the vast majority of colorectal cancers and are an initiating event in cellular transformation. Cells harboring mutant APC contain elevated levels of the β-catenin transcription coactivator in the nucleus which leads to abnormal expression of genes controlled by β-catenin/T-cell factor 4 (TCF4) complexes. Here, we use chromatin immunoprecipitation coupled with massively parallel sequencing (ChIP-Seq) to identify β-catenin binding regions in HCT116 human colon cancer cells. We localized 2168 β-catenin enriched regions using a concordance approach for integrating the output from multiple peak alignment algorithms. Motif discovery algorithms found a core TCF4 motif (T/A-T/A-C-A-A-A-G), an extended TCF4 motif (A/T/G-C/G-T/A-T/A-C-A-A-A-G) and an AP-1 motif (T-G-A-C/T-T-C-A) to be significantly represented in β-catenin enriched regions. Furthermore, 417 regions contained both TCF4 and AP-1 motifs. Genes associated with TCF4 and AP-1 motifs bound β-catenin, TCF4 and c-Jun in vivo and were activated by Wnt signaling and serum growth factors. Our work provides evidence that Wnt/β-catenin and mitogen signaling pathways intersect directly to regulate a defined set of target genes.


[National survey on the use of induced labor by obstetricians. Study Group on Induced Labor].
A strong rise in the use of induced labor has been observed in France. The aim of this work was to analyze the different methods used for achieving induction of labor and their implications. One out of four French obstetricians were randomly selected to answer a questionnaire on their practice for achieving induction of labor. Four hundred of the 997 obstetricians answered the questionnaire. Univariate and multivariate analysis was applied. A high rate of induced labor was correlated with some areas of the country and with private practice. Certain methods were used in spite of opposing advice by experts in the field: elective induction of labor with unfavorable cervix, use of prostaglandins in elective induction of labor, induction of labor in cases of scarred uterus or breech presentation, use of misoprostol. Some methods were still used in spite of their poor efficacy: intravenous oxytocin used with unfavorable cervix, use of intravensou PGE2. This study would show that theory and practice are often distinctly different. Induction of labor is currently used on a far wider scale than ever before. We obviously need studies for careful assessment of the circumstances in which induction of labor is used in order to improve methods and indications of such a clinical practice.


Life cycle emission distributions within the economy: implications for life cycle impact assessment.
Refinements of methods for life cycle impact assessment (LCIA) are directed at removing unjustified simplifications and quantifying and reducing uncertainties in results. The amount of uncertainty reduction that is actually achieved through LCIA method refinement depends on the structure of the life cycle inventory model. We investigate the general structure of inventory models using an economic input/output (I/O) life cycle assessment model of the U.S. economy. In particular, we study the results of applying a streamlining algorithm to the I/O LCA model. The streamlining algorithm retains only those "branches" of the process tree that are jointly required to account for a specified fraction of the total impacts upstream of each point in the tree. We examine the implications of these "tree pruning" results for site-informed LCIA. Percentiles are presented for U.S. commodities and several important pollutants, for the share of total upstream emissions contributed by the set of processes in each supply tier, that is, each set of processes that directly supply inputs to another set of processes Capturing at least 90% of the total direct plus upstream emissions for criteria air pollutants and toxic releases for at least 75% of the commodities in the U.S. economy requires full modeling of direct emissions plus the first five supply tiers. The requirements for capturing a high percentage (e.g., >80%) of total emissions vary widely across products or commodities. To capture more than 60% of total emissions for more than half of all commodities requires models with more than 4,000 process instances. To well characterize the total impacts of products, life cycle impact assessment methods must characterize foreground process impacts in a site-informed way and mean impacts of far-removed processes in an unbiased way.


Focusing method for nondestructive measurement of optical fiber index profiles.
A new method to determine the refractive-index distributions of the cores of optical fibers is presented. The fiber is immersed in index matching oil to eliminate light refraction at the outer core boundary. Collimated light is passed transversely through the fiber core and is detected with a microscope focused just above the plane of the core boundary. The intensity distribution in the image plane is recorded with the aid of a computer-controlled video-analysis system. The refractive-index profile of the fiber core is then computed from this intensity distribution. In this paper we present error estimates and show how well the focusing method agrees with the interferometric slab method. The repeatability of the index measurement is approximately 2%, an error which is easily attributable to incident light level fluctuations.


Pixelated high-index ring Bragg fibers.
A new type of Anti Resonant Reflecting Optical Waveguide (ARROW) fiber with a low refractive index contrast is reported. This waveguide is similar to a Bragg fiber for which the high index rings are replaced by discontinuous rings made of circular High Index Inclusions (HII). As compared to conventional Bragg fibers, such a new structure enables true Photonic BandGap (PBG) guidance and limits the number of cladding modes located within the high index regions, thus enhancing the guiding properties. A Mode Field Diameter (MFD) of 26 μm is reported at a wavelength of 1400 nm. Single Mode (SM) behavior is also observed beyond 1400 nm for a 1 m-long fiber.


Complex formation of platelet thrombospondin with plasminogen. Modulation of activation by tissue activator.
Thrombospondin (TSP), a multifunctional alpha-granule glycoprotein of platelets, binds fibrinogen, fibronectin, heparin, and histidine-rich glycoprotein and thus may play an important role in regulating thrombotic influences at vessel surfaces. In this study we have demonstrated that purified human platelet TSP formed a complex with purified human plasminogen (Plg). Complex formation was detected by rocket immunoelectrophoresis of mixtures of the purified radiolabeled proteins. Significant complex formation of fluid-phase Plg with adsorbed TSP was also demonstrated by enzyme-linked immunosorbent assay (ELISA). The complex formation was specific, saturable, and inhibited by excess fluid-phase TSP, with an apparent KD of approximately 35 nM. In both ELISA and rocket immunoelectrophoresis systems, complex formation was inhibited by 10 mM epsilon-amino-n-caproic acid, implying that there is a role for the lysine binding sites of Plg in mediating the interaction. TSP also formed a complex with plasmin as detected by ELISA but did not directly inhibit plasmin activity measured with a synthetic fluorometric substrate or with a 125I-fibrin plate assay. TSP, when incubated with Plg before addition to 125I-fibrin plates significantly inhibited the generation of plasmin activity by tissue plasminogen activator (TPA) in a manner that was calcium dependent. A kinetic study of Plg activation by TPA in the presence of TSP demonstrated that Michaelis-Menten kinetics were followed and that TSP acted as a noncompetitive inhibitor. These studies support the hypothesis that TSP, acting as a multifunctional regulator in focal areas of active hemostasis, could serve as a prothrombotic influence, leading to increased deposition of fibrin.


Effect of atropine on alveolar gas mixing in man.
1. Atropine is known to diminish bronchomotor tone. In order to investigate the acute effect of atropine on respiration and alveolar gas mixing, a dose of 2.4 mg was given intravenously. 2. Ten normal male volunteers were each studied three times with a nitrogen washout method, once before administration of atropine and then 20 min and 60 min thereafter. 3. After the administration of atropine there was a reduction in tidal volume, a slight increase in frequency of respiration and an increase in series dead space. The tidal mixing volume showed a fall of 25%. In spite of the reduced alveolar dead space the effective mixing volume fell by 29%. Multi-breath alveolar mixing efficiency fell by 3.5%. 4. Multi-breath alveolar mixing efficiency was found to be less with smaller tidal mixing volumes, a fall of 518 ml in the latter causing a reduction of 17.2% in mixing efficiency. 5. A reduction of 100 ml in tidal volume in normal subjects was associated with a decrease of 6.9% in alveolar mixing efficiency. In the subjects receiving atropine tidal volume reduced by 96 ml, but the observed fall in alveolar mixing efficiency was only 3.5%, This suggests an improvement in alveolar mixing of 3.4% due to the administration of atropine. Despite this small improvement, the mixing efficiency is still only 66%. The residual inefficiency of 34% cannot therefore be explained on the basis of bronchomotor tone.


Nanopore sequencing for rapid diagnostics of salmonid RNA viruses.
Analysis of pathogen genome variation is essential for informing disease management and control measures in farmed animals. For farmed fish, the standard approach is to use PCR and Sanger sequencing to study partial regions of pathogen genomes, with second and third-generation sequencing tools yet to be widely applied. Here we demonstrate rapid and accurate sequencing of two disease-causing viruses affecting global salmonid aquaculture, salmonid alphavirus (SAV) and infectious salmon anaemia virus (ISAV), using third-generation nanopore sequencing on the MinION platform (Oxford Nanopore Technologies). Our approach complements PCR from infected material with MinION sequencing to recover genomic information that matches near perfectly to Sanger-verified references. We use this method to present the first SAV subtype-6 genome, which branches as the sister to all other SAV lineages in a genome-wide phylogenetic reconstruction. MinION sequencing offers an effective strategy for fast, genome-wide analysis of fish viruses, with major potential applications for diagnostics and robust investigations into the origins and spread of disease outbreaks.


Inhibitors of TLR-9 act on multiple cell subsets in mouse and man in vitro and prevent death in vivo from systemic inflammation.
In parallel with the discovery of the immunostimulatory activities of CpG-containing oligodeoxynucleotides, several groups have reported specific DNA sequences that could inhibit activation by CpG-containing oligodeoxynucleotides in mouse models. We show that these inhibitory sequences, termed IRS, inhibit TLR-9-mediated activation in human as well as mouse cells. This inhibitory activity includes proliferation and IL-6 production by B cells, and IFN-alpha and IL-12 production by plasmacytoid dendritic cells. Our studies of multiple cell types in both mice and humans show the optimal IRS to contain a GGGG motif within the sequence, and the activity to require a phosphorothioate backbone. Although the GGGG motif readily itself leads to formation of a tetrameric oligodeoxynucleotide structure, inhibitory activity resides exclusively in the single-stranded form. When coinjected with a CpG oligodeoxynucleotide in vivo, IRS were shown to inhibit inflammation through a reduction in serum cytokine responses. IRS do not need to be injected at the same site to inhibit, demonstrating that rapid, systemic inhibition of TLR-9 can be readily achieved. IRS can also inhibit a complex pathological response to ISS, as shown by protection from death after massive systemic inflammation induced by a CpG-containing oligodeoxynucleotides.


Effect of staphylococcal alpha-toxin on phagocytosis of staphylococci by human polymorphonuclear leukocytes.
Evidence is presented which shows that alpha-toxin elaborated by Staphylococcus aureus can affect certain of the biological functions of human polymorphonuclear leukocytes (PMN). The pretreatment of PMNs with low doses (less than 10 hemolytic units) of purified toxin enhanced their ability to phagocytose and to kill serum-opsonized staphylococci. At higher doses (greater than or equal to 10 hemolytic units), a certain amount of cell damage was caused (detectable by trypan blue uptake), which resulted in a reduction of their phagocytic capacity. These effects were not seen either with heat-inactivated (60 degrees C, 30 min) or with antibody-neutralized toxin. It is thought that the toxin can bind onto the membrane of the PMN, either exposing additional receptor sites for opsonized bacteria or speeding up the transport of staphylococci across the membrane. Once ingested, the bacteria were more readily killed by the PMNs.


[Role of compression bandage with elevated level of pressure in decreasing incidence of venous thrombosis in high-risk patients].
The study was aimed at assessing efficacy and safety of using graduated compression bandage with the level of pressure in the area of the middle ankle amounting to 20-40 mm Hg as compared to the conventional profile (10-20 mm Hg in the ankle area) in the composition of comprehensive prevention of postoperative venous thromboembolic complications (VTEC) in patients of a high-risk group. For this purpose we carried out a prospective randomized clinical study including a total of 100 surgical patients (general surgery, neurosurgery) from a group of high risk for the development of VTEC with the presence of 3 and more conditions predisposing to venous thrombosis. All patients were subdivided into two subgroups 50 patients each. Patients in both groups received anticoagulants in standard preventive doses from the first day after surgery or in the remote period. The Study group patients were treated by applying on the ankle a bandage of medium distensibility with the target pressure in the ankle area of 20-40 mm Hg measured by means of a portable manometer. The control group patients were treated by a similar bandage applied onto the ankle and femur with a pressure of 10-20 mm Hg. In all cases a special lining material was placed under the bandage. With the purpose of active screening of postoperative VTECs, prior to the beginning of the study and then each 3-5 days we performed ultrasound angioscanning and in case thrombosis was revealed we performed static perfusion scintigraphy of the lungs and/or echocardiography for verification of pulmonary embolism. The patients who died were subjected to post-mortem examination. We also assessed the frequency of correcting the bandage on the femur and crus. The incidence rate of postoperative venous thrombosis in the Study Group amounted to 16.0% (95% CI: 5.8-26.2%) and in the Control Group 36.0% (95% CI: 22.7-49.3%, p=0.039), with no significant differences in the incidence of proximal thrombosis or pulmonary embolism revealed. Maximal reduction of the development of venous thrombosis was revealed in patients with lower-limb paralysis amounting to 69.5%. The frequency of the development of cutaneous lesions under the bandage did not differ significantly between the subgroups, being 8% and 12%, respectively. The total number of corrections of the compression in the Control Group was significantly higher, amounting to 0.64±0.23 time a day versus 0.32±0.05 t/d (p<0.0001). The use of an elevated compression profile makes it possible to decrease the incidence of postoperative VTECs, especially in patients with palsy, not increasing the rate of cutaneous lesions under the bandages. Femoral compression provides no additional reduction of the incidence of thrombosis but creates an additional necessity of correcting the bandages.


Acinetobacter: an old friend, but a new enemy.
Acinetobacter emerged as a significant nosocomial pathogen during the late 1970s, probably as a consequence, at least in part, of increasing use of broad-spectrum antibiotics in hospitals. Most clinically significant isolates belong to the species Acinetobacter baumannii or its close relatives, with many infections concentrated in intensive care, burns or high dependency units treating severely ill or debilitated patients. Large outbreaks can occur in such units, involving the infection or colonisation of numerous patients by specific epidemic strains of A. baumannii. Recently, a particular problem has concerned cross-infection of injured military patients repatriated from combat regions of the world (e.g. Iraq and Afghanistan). Carbapenems have previously been the treatment of choice for infected patients, but increasing reports worldwide now describe A. baumannii isolates resistant to all conventional antimicrobial regimens. Data to support therapeutic use of the limited number of new antimicrobial agents (e.g. tigecycline) with in-vitro activity against these pathogens are still very limited. Detailed advice concerning prevention and control of outbreaks caused by multidrug-resistant strains of acinetobacter is available from the UK Health Protection Agency. In addition to antibiotic prescribing policies and audit, these measures focus on reinforcing standard infection control procedures and precautions, with particular attention to thorough cleaning of patient areas to take account of the long-term survival of acinetobacter after drying and inadequate disinfection. Despite these measures, the problem continues to escalate, with many hospitals worldwide now reporting outbreaks caused by multidrug-resistant strains of acinetobacter.


Hyperemesis gravidarum: is an ultrasound scan necessary?
Traditionally, in cases of hyperemesis gravidarum (HG), an ultrasound evaluation is recommended to confirm viability and to exclude multiple pregnancies and gestational trophoblastic disease (GTD). Our aim was to perform a case-control study to evaluate the incidence of these findings. Each case of HG was matched for gestational age with the next ultrasound examination performed in an asymptomatic pregnancy. The findings were compared between the two groups. Two hundred and eighty-six cases of HG were matched with 286 asymptomatic women. The total number of viable pregnancies was higher in the HG group (280/286, 97.9%) than that in the control group (265/286, 92.6%; P = 0.006). The incidence of twins was 3.1% in each group (P > 0.999). The incidence of early pregnancy failure was 0.7% in women with HG compared to 7.0% in asymptomatic women (odds ratio 0.09, 95% CI 0.01-0.04, P < 0.0001). The one case of GTD was in the HG group; however, this case also presented with vaginal bleeding. Pregnancies complicated by HG had a similar risk of twin pregnancy, and a lower risk of early pregnancy failure compared to controls. In the absence of vaginal bleeding, there was no increase in GTD in women with HG. We conclude that an ultrasound scan is not clinically necessary in women presenting with HG, other than for maternal reassurance.


Effects of antithyroid drugs on radioiodine treatment: systematic review and meta-analysis of randomised controlled trials.
To determine the effect of adjunctive antithyroid drugs on the risk of treatment failure, hypothyroidism, and adverse events after radioiodine treatment. Meta-analysis. Electronic databases (Cochrane central register of controlled trials, Medline, Embase) searched to August 2006 and contact with experts. Review methods Three reviewers independently assessed trial eligibility and quality. Pooled relative risks for treatment failure and hypothyroidism after radioiodine treatment with and without adjunctive antithyroid drugs were calculated with a random effects model. We identified 14 relevant randomised controlled trials with a total of 1306 participants. Adjunctive antithyroid medication was associated with an increased risk of treatment failure (relative risk 1.28, 95% confidence interval 1.07 to 1.52; P=0.006) and a reduced risk for hypothyroidism (0.68, 0.53 to 0.87; P=0.006) after radioiodine treatment. We found no difference in summary estimates for the different antithyroid drugs or for whether antithyroid drugs were given before or after radioiodine treatment. Antithyroid drugs potentially increase rates of failure and reduce rates of hypothyroidism if they are given in the week before or after radioiodine treatment, respectively.


Back to the forest: exploring forest transitions in Candelaria Loxicha, Mexico.
Declining profitability of agriculture and/or higher prices of forest products and services typically drive an increase in forest cover. This article examines changes in forest cover in Candelaria Loxicha, Mexico. Forest cover increased in the area as a result of coffee cultivation in coffee forest-garden systems. Dependence on forest products and services, and not prices of forest products, drive the process in our study site. Low international coffee prices and high labor demand outside the community might pull farmers out of agriculture, but they do not completely abandon the lands. A diversification in income sources prevents land abandonment and contributes to maintaining rural populations and coffee forest gardens.


Effects of moderate-volume, high-load lower-body resistance training on strength and function in persons with Parkinson's disease: a pilot study.
Background. Resistance training research has demonstrated positive effects for persons with Parkinson's disease (PD), but the number of acute training variables that can be manipulated makes it difficult to determine the optimal resistance training program. Objective. The purpose of this investigation was to examine the effects of an 8-week resistance training intervention on strength and function in persons with PD. Methods. Eighteen men and women were randomized to training or standard care for the 8-week intervention. The training group performed 3 sets of 5-8 repetitions of the leg press, leg curl, and calf press twice weekly. Tests included leg press strength relative to body mass, timed up-and-go, six-minute walk, and Activities-specific Balance Confidence questionnaire. Results. There was a significant group-by-time effect for maximum leg press strength relative to body mass, with the training group significantly increasing their maximum relative strength (P < .05). No other significant interactions were noted (P > .05). Conclusions. Moderate volume, high-load weight training is effective for increasing lower-body strength in persons with PD.


Different roles of BDNF in nucleus accumbens core versus shell during the incubation of cue-induced cocaine craving and its long-term maintenance.
Brain-derived neurotrophic factor (BDNF) contributes to diverse types of plasticity, including cocaine addiction. We investigated the role of BDNF in the rat nucleus accumbens (NAc) in the incubation of cocaine craving over 3 months of withdrawal from extended access cocaine self-administration. First, we confirmed by immunoblotting that BDNF levels are elevated after this cocaine regimen on withdrawal day 45 (WD45) and showed that BDNF mRNA levels are not altered. Next, we explored the time course of elevated BDNF expression using immunohistochemistry. Elevation of BDNF in the NAc core was detected on WD45 and further increased on WD90, whereas elevation in shell was not detected until WD90. Surface expression of activated tropomyosin receptor kinase B (TrkB) was also enhanced on WD90. Next, we used viral vectors to attenuate BDNF-TrkB signaling. Virus injection into the NAc core enhanced cue-induced cocaine seeking on WD1 compared with controls, whereas no effect was observed on WD30 or WD90. Attenuating BDNF-TrkB signaling in shell did not affect cocaine seeking on WD1 or WD45 but significantly decreased cocaine seeking on WD90. These results suggest that basal levels of BDNF transmission in the NAc core exert a suppressive effect on cocaine seeking in early withdrawal (WD1), whereas the late elevation of BDNF protein in NAc shell contributes to incubation in late withdrawal (WD90). Finally, BDNF protein levels in the NAc were significantly increased after ampakine treatment, supporting the novel hypothesis that the gradual increase of BDNF levels in NAc accompanying incubation could be caused by increased AMPAR transmission during withdrawal.


Saturated Fats and Health: A Reassessment and Proposal for Food-Based Recommendations: JACC State-of-the-Art Review.
The recommendation to limit dietary saturated fatty acid (SFA) intake has persisted despite mounting evidence to the contrary. Most recent meta-analyses of randomized trials and observational studies found no beneficial effects of reducing SFA intake on cardiovascular disease (CVD) and total mortality, and instead found protective effects against stroke. Although SFAs increase low-density lipoprotein (LDL) cholesterol, in most individuals, this is not due to increasing levels of small, dense LDL particles, but rather larger LDL particles, which are much less strongly related to CVD risk. It is also apparent that the health effects of foods cannot be predicted by their content in any nutrient group without considering the overall macronutrient distribution. Whole-fat dairy, unprocessed meat, and dark chocolate are SFA-rich foods with a complex matrix that are not associated with increased risk of CVD. The totality of available evidence does not support further limiting the intake of such foods.


[P53 gene mutation in gallbladder cancer].
Gallbladder cancer frequency and mortality renders it one of the most important neoplastic diseases in Chile. P53 tumor suppressor gene has been studied in most types of cancer, but there is scarce information about it in gallbladder cancer. To study the frequency of P53 gene mutation in gallbladder cancer in the ninth region of Chile. In 25 pathological samples of gallbladder cancer, the direct amplification and sequencing of p53 gene exons 5,6,7,8-8 was possible. Seventeen punctual mutations were observed in 13 cases (52%). There were 10 transitions, 5 transversions, one insertion (codon 194) and one deletion (codon 186). Eight cases had mutations in exon 5, six had mutations in exon 6, two had mutations in exon 7 and one had mutations in exons 8-9. In 14 of 25 cases, gene p53 protein was positive. When immunohistochemical expression of gene p53 protein was positive in more than 20% of cells, there was a high correlation between genetic alterations and immunohistochemical expression of the protein, with a specificity, sensitivity, positive and negative predictive values over 80%. P53 gene mutation is observed in a high proportion of gallbladder cancers at it can be accurately detected with conventional immunohistochemical techniques. The importance of this gene in the genesis of this carcinoma should be determined studying preneoplastic lesions and early carcinomas.


Heterogeneous pattern of selective pressure for PRRT2 in human populations, but no association with autism spectrum disorders.
Inherited and de novo genomic imbalances at chromosome 16p11.2 are associated with autism spectrum disorders (ASD), but the causative genes remain unknown. Among the genes located in this region, PRRT2 codes for a member of the synaptic SNARE complex that allows the release of synaptic vesicles. PRRT2 is a candidate gene for ASD since homozygote mutations are associated with intellectual disability and heterozygote mutations cause benign infantile seizures, paroxysmal dyskinesia, or hemiplegic migraine. Here, we explored the contribution of PRRT2 mutations in ASD by screening its coding part in a large sample of 1578 individuals including 431 individuals with ASD, 186 controls and 961 individuals from the human genome Diversity Panel. We detected 24 nonsynonymous variants, 1 frameshift (A217PfsX8) and 1 in-frame deletion of 6 bp (p.A361_P362del). The frameshift mutation was observed in a control with no history of neurological or psychiatric disorders. The p.A361_P362del was observed in two individuals with autism from sub-Saharan African origin. Overall, the frequency of PRRT2 deleterious variants was not different between individuals with ASD and controls. Remarkably, PRRT2 displays a highly significant excess of nonsynonymous (pN) vs synonymous (pS) mutations in Asia (pN/pS = 4.85) and Europe (pN/pS = 1.62) compared with Africa (pN/pS = 0.26; Asia vs Africa: P = 0.000087; Europe vs Africa P = 0.00035; Europe vs Asia P = P = 0.084). We also showed that whole genome amplification performed through rolling cycle amplification could artificially introduce the A217PfsX8 mutation indicating that this technology should not be performed prior to PRRT2 mutation screening. In summary, our results do not support a role for PRRT2 coding sequence variants in ASD, but provide an ascertainment of its genetic variability in worldwide populations that should help researchers and clinicians to better investigate the role of PRRT2 in human diseases.


Epistasis and natural selection shape the mutational architecture of complex traits.
The evolutionary trajectories of complex traits are constrained by levels of genetic variation as well as genetic correlations among traits. As the ultimate source of all genetic variation is mutation, the distribution of mutations entering populations profoundly affects standing variation and genetic correlations. Here we use an individual-based simulation model to investigate how natural selection and gene interactions (that is, epistasis) shape the evolution of mutational processes affecting complex traits. We find that the presence of epistasis allows natural selection to mould the distribution of mutations, such that mutational effects align with the selection surface. Consequently, novel mutations tend to be more compatible with the current forces of selection acting on the population. These results suggest that in many cases mutational effects should be seen as an outcome of natural selection rather than as an unbiased source of genetic variation that is independent of other evolutionary processes.


Regular effective hologram regions for computer generated holography.
An effective hologram region (EHR) based approach is presented to speed up the computation of computer generated holograms (CGHs). The object space is predivided into subspaces, and an EHR for each subspace is predefined according to the maximum spatial frequency of interference fringes, light diffraction efficiency, and CGH binarization effect. To compute the hologram of an object, the object points are first categorized according to which subspace they are located in, and then their holograms are calculated using the corresponding EHRs. As each EHR usually takes up only a portion of the hologram plate, the CGH computational load is thus reduced. This new approach is highly suitable for large hologram display systems. In addition, when compared to the reconstructed image using the conventional approach, our experimental results show that more noise can be blocked off and the reconstructed image appears sharper without noticeable brightness reduction.


Reorganization of the centrosome and associated microtubules during the morphogenesis of a mouse cochlear epithelial cell.
Reorganization of centrosomal microtubule-organizing centres and the minus ends of microtubules occurs as the centrosomal ends of large microtubule bundles are repositioned and anchored to cell junctions in certain epithelial cells called inner pillar cells in the mouse organ of Corti. The microtubule bundle that assembles in each cell consists of two distinct microtubule arrays that run closely alongside each other. Both arrays are attached to the cell surface at their upper and lower ends. One of the arrays spans the entire length of a cell but the other is confined to its lower portion. Initially, about 3,000 microtubules elongate downwards from an apically situated centrosome in each cell. Subsequently, the minus ends of these microtubules, and the centrosome and its two centrioles, migrate for about 12 microns to the tip of a laterally directed projection. Then, a meshwork of dense material accumulates to link microtubule minus ends and the centrosome to cell junctions at the tip of the projection. Pericentriolar satellite bodies, which form after the initial burst of microtubule nucleation, may represent a condensed and inactive concentration of microtubule-nucleating elements. Surprisingly, as a cell matures, about 2,000 microtubules are eliminated from the centrosomal end of the microtubule bundle. However, about 2,000 microtubules are added to the basal portion of each bundle at levels that are remote with respect to the location of the centrosome. Possibly, these microtubules have escaped from the centrosome. If this is the case, then both the plus and minus ends of most of the errant microtubules are captured by sites at the cell surface where the ends are finally anchored. Alternatively, each cell possesses at least one other major microtubule-nucleating site (which does not possess centrioles) in addition to its centrosome.


Multiple sclerosis symptoms and spasticity management: new data.
Spasticity, perceived by patients as muscle rigidity and spasms, is a common symptom in multiple sclerosis (MS). It is associated with functional impairment that can exacerbate other MS symptoms and reduce quality of life. Pharmacological treatment options are limited and frequently ineffective. Treatment adherence is a key issue to address in these patients. The efficacy and safety of 9-delta-tetrahydrocannabinol:cannabidiol (THC:CBD) oromucosal spray for treatment of MS spasticity were demonstrated in four Phase III trials. Observational studies and registry data subsequently confirmed the effectiveness and tolerability of THC:CBD oromucosal spray under everyday practice conditions. Among patients who respond to treatment, THC:CBD oromucosal spray has been shown to produce positive improvements in gait parameters and to normalize muscle fibers.


Sensitivity of sea urchin fertilization to pH varies across a natural pH mosaic.
In the coastal ocean, temporal fluctuations in pH vary dramatically across biogeographic ranges. How such spatial differences in pH variability regimes might shape ocean acidification resistance in marine species remains unknown. We assessed the pH sensitivity of the sea urchin Strongylocentrotus purpuratus in the context of ocean pH variability. Using unique male-female pairs, originating from three sites with similar mean pH but different variability and frequency of low pH (pHT ≤ 7.8) exposures, fertilization was tested across a range of pH (pHT 7.61-8.03) and sperm concentrations. High fertilization success was maintained at low pH via a slight right shift in the fertilization function across sperm concentration. This pH effect differed by site. Urchins from the site with the narrowest pH variability regime exhibited the greatest pH sensitivity. At this site, mechanistic fertilization dynamics models support a decrease in sperm-egg interaction rate with decreasing pH. The site differences in pH sensitivity build upon recent evidence of local pH adaptation in S. purpuratus and highlight the need to incorporate environmental variability in the study of global change biology.


Triamterene inhibits the delayed rectifier potassium current (IK) in guinea pig ventricular myocytes.
In humans, proarrhythmia during therapy with action potential-prolonging drugs can be associated with hypokalemia often provoked by concomitant administration of diuretic agents. Consequently, therapy with class III antiarrhythmics and K(+)-sparing diuretics, such as triamterene, may be indicated. Triamterene, along with its K(+)-sparing properties, exhibits other pharmacological effects. In the heart, it can increase action potential duration (guinea pig atria and papillary muscles), protect against reperfusion-induced arrhythmias (rat), and increase the QT interval (humans). Therefore, studies were undertaken to assess effects of triamterene on cardiac K+ repolarizing currents. Guinea pig ventricular myocytes were superfused at 30 degrees C with Cd(2+)-containing solution to block Isi and held at -40 mV to inactivate INa. Currents were measured in the whole-cell configuration of the patch-clamp technique. The delayed rectifier outward current (IK) was elicited by short (250-millisecond) and long (5000-millisecond) depolarizing pulses, and time-independent currents were assessed by a rapid ramp test protocol. After high-voltage long pulses (+50 mV; 5000 milliseconds), tail current amplitude of the slow component of IK (IKs) was decreased 36 +/- 6% (n = 6) and 51 +/- 8% (n = 6) by triamterene 10(-5) and 10(-4) mol/L, respectively. After low-voltage short pulses (-20 mV; 250 milliseconds), tail current amplitude corresponding essentially to the rapid component of IK (IKr) was decreased only 14 +/- 11% (n = 9) and 19 +/- 10% (n = 10) by triamterene 10(-5) and 10(-4) mol/L, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Objective burden, resources, and other stressors among informal cancer caregivers: a hidden quality issue?
A great deal of clinical cancer care is delivered in the home by informal caregivers (e.g. family, friends), who are often untrained. Caregivers' context varies widely, with many providing care despite low levels of resources and high levels of additional demands. Changes in health care have shifted much cancer care to the home, with limited data to inform this transition. We studied the characteristics, care tasks, and needs of informal caregivers of cancer patients. Caregivers of seven geographically and institutionally defined cohorts of newly diagnosed colorectal and lung cancer patients completed self-administered questionnaires (n = 677). We combined this information with patient survey and chart abstraction data and focused on caregivers who reported providing, unpaid, at least 50% of the patient's informal cancer care. Over half of caregivers (55%) cared for a patient with metastatic disease, severe comorbidity, or undergoing current treatment. Besides assisting with activities of daily living, caregivers provided cancer-specific care such as watching for treatment side effects (68%), helping manage pain, nausea or fatigue (47%), administering medicine (34%), deciding whether to call a doctor (30%), deciding whether medicine was needed (29%), and changing bandages (19%). However, half of caregivers reported not getting training perceived as necessary. In addition, 49% of caregivers worked for pay, 21% reported poor or fair health, and 21% provided unpaid care for other individuals. One in four reported low confidence in the quality of the care they provided. Much assistance for cancer patients is delivered in the home by informal caregivers, often without desired training, with a significant minority having limited resources and high additional demands. Future research should explore the potentially high yield of addressing caregiver needs in improving quality of cancer care and both survivors' and caregivers' outcomes.


Pelvic lymph node dissection alone versus adjuvant radiotherapy in node positive penile cancer: A systematic review.
The management options for regional lymph nodes (LNs) in men with penile cancer include surveillance, surgery, and chemotherapy. The use of radiotherapy (RT) for nodal disease follows tradition and single-institution policies. We aimed to analyse the existing evidence regarding the management of penile cancer patients with suspected or known metastatic pelvic LNs using pelvic LN dissection (PLND) with RT versus PLND or RT alone. A systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, with no filters for language or time. The search was conducted in EMBASE, MEDLINE/PubMed, and Cochrane Library. Inclusion criteria were adult men with penile cancer and suspected metastatic pelvic LNs, undergoing PLND with or without RT or RT alone. Primary outcomes included disease-specific survival and locoregional recurrence. Secondary outcomes included overall survival and complications of therapy. A total of 552 articles were identified. Only eight retrospective studies were eligible for inclusion (including 406 patients). All studies had a high risk of bias. None of the studies reported the use of neoadjuvant RT. Indications for PLND varied but were usually two or more clinically positive inguinal nodes with or without extracapsular extension. Adjuvant RT was mainly used in positive pelvic LNs or pN2/pN3 stages. The rate of locoregional recurrence following adjuvant RT was 70%. Complications of treatment were reported in two studies only. There is insufficient evidence to recommend the use of adjuvant RT following PLND in penile cancer patients. The quality of evidence is low due to the retrospective design and high risk of bias. Randomized clinical trials are required to assess the efficacy and safety of adjuvant RT and PLND.


Lipid-Mediated Regulation of Embedded Receptor Kinases via Parallel Allosteric Relays.
Membrane-anchored receptors are essential cellular signaling elements for stimulus sensing, propagation, and transmission inside cells. However, the contributions of lipid interactions to the function and dynamics of embedded receptor kinases have not been described in detail. In this study, we used amide hydrogen/deuterium exchange mass spectrometry, a sensitive biophysical approach, to probe the dynamics of a membrane-embedded receptor kinase, EnvZ, together with functional assays to describe the role of lipids in receptor kinase function. Our results reveal that lipids play an important role in regulating receptor function through interactions with transmembrane segments, as well as through peripheral interactions with nonembedded domains. Specifically, the lipid membrane allosterically modulates the activity of the embedded kinase by altering the dynamics of a glycine-rich motif that is critical for phosphotransfer from ATP. This allostery in EnvZ is independent of membrane composition and involves direct interactions with transmembrane and periplasmic segments, as well as peripheral interactions with nonembedded domains of the protein. In the absence of the membrane-spanning regions, lipid allostery is propagated entirely through peripheral interactions. Whereas lipid allostery impacts the phosphotransferase function of the kinase, extracellular stimulus recognition is mediated via a four-helix bundle subdomain located in the cytoplasm, which functions as the osmosensing core through osmolality-dependent helical stabilization. Our findings emphasize the functional modularity in a membrane-embedded kinase, separated into membrane association, phosphotransferase function, and stimulus recognition. These components are integrated through long-range communication relays, with lipids playing an essential role in regulation.


Total shoulder arthroplasty with an all-polyethylene pegged bone-ingrowth glenoid component: a clinical and radiographic outcome study.
Loosening of the glenoid component continues to be the foremost cause of medium and long-term failure of shoulder replacements. The purpose of this study was to evaluate the clinical and radiographic results of a minimally cemented all-polyethylene pegged glenoid component designed for biologic fixation. Forty-four shoulders in forty-one patients with a mean age of sixty-six years underwent total shoulder arthroplasty with a pegged bone-ingrowth glenoid component. Outcome data included the American Shoulder and Elbow Surgeons questionnaire, the Simple Shoulder Test, and visual analog scales. A detailed radiographic analysis was performed by two board-certified musculoskeletal radiologists who were blinded to clinical and patient-reported outcomes. The radiographs were evaluated with regard to the presence of radiolucent lines at the bone-cement interface, implant seating, and the radiodensity between the flanges of the central peg. The mean duration of clinical follow-up was four years and the mean duration of radiographic follow-up was three years. Twenty shoulders had perfect seating and radiolucency grades, thirty had increased radiodensity between the flanges of the central peg, and three demonstrated osteolysis. Radiodensity about the uncemented central peg at the time of the latest follow-up was positively associated with perfect seating and radiolucency grades on the initial postoperative radiographs (p = 0.03, Fisher exact test). The Simple Shoulder Test score, the American Shoulder and Elbow Surgeons score, and all visual analog scale scores had improved significantly (p < 0.01) at the time of the latest follow-up. Total shoulder arthroplasty with a minimally cemented, all-polyethylene, pegged glenoid implant can yield stable and durable fixation at short to medium-term follow-up (mean, four years).


The spatial distribution and relative abundance of gap-junctional connexin40 and connexin43 correlate to functional properties of components of the cardiac atrioventricular conduction system.
Electrical coupling between heart muscle cells is mediated by specialised regions of sarcolemmal interaction termed gap junctions. In previous work, we have demonstrated that connexin42, a recently identified gap-junctional protein, is present in the specialised conduction tissues of the avian heart. In the present study, the spatial distribution of the mammalian homologue of this protein, connexin40, was examined using immunofluorescence, confocal scanning laser microscopy and quantitative digital image analysis in order to determine whether a parallel distribution occurs in rat. Connexin40 was detected by immunofluorescence in all main components of the atrioventricular conduction system including the atrioventricular node, atrioventricular bundle, and Purkinje fibres. Quantitation revealed that levels of connexin40 immunofluorescence increased along the axis of atrioventricular conduction, rising over 10-fold between atrioventricular node and atrioventricular bundle and a further 10-fold between atrioventricular bundle and Purkinje fibres. Connexin40 and connexin43, the principal gap-junctional protein of the mammalian heart, were co-localised within atrioventricular nodal tissues and Purkinje fibres. By applying a novel photobleach/double-labelling protocol, it was demonstrated that connexin40 and connexin43 are co-localised in precisely the same Purkinje fibre myocytes. A model, integrating data on the spatial distribution and relative abundance of connexin40 and connexin43 in the heart, proposes how myocyte-type-specific patterns of connexin isform expression account for the electrical continuity of cardiac atrioventricular conduction.