Phase-derivative-based estimation of a digital reference wave from a single off-axis digital hologram.
This paper describes a method to obtain an estimated digital reference wave from a single off-axis digital hologram that matches the actual experimental reference wave as closely as possible. The proposed method is independent of a reference flat plate and speckles. The digital reference wave parameters are estimated directly from the recorded phase information. The parameters include both the off-axis tilt angle and the curvature of the reference wave. Phase derivatives are used to extract the digital reference wave parameters without the need for a phase unwrapping process. Thus, problems associated with phase wrapping are avoided. Experimental results for the proposed method are provided. The simulated effect of the digital reference wave parameters on the reconstructed image phase distribution is shown. The pseudo phase gradient originating from incorrect estimation of the digital reference wave parameters and its effect on object reconstruction are discussed.


Imprinted Nesp55 influences behavioral reactivity to novel environments.
Genomic imprinting results in parent-of-origin-dependent monoallelic expression of selected genes. Although their importance in development and physiology is recognized, few imprinted genes have been investigated for their effects on brain function. Gnas is a complex imprinted locus whose gene products are involved in early postnatal adaptations and neuroendocrine functions. Gnas encodes the stimulatory G-protein subunit Gsalpha and two other imprinted protein-coding transcripts. Of these, the Nesp transcript, expressed exclusively from the maternal allele, codes for neuroendocrine secretory protein 55 (Nesp55), a chromogranin-like polypeptide associated with the constitutive secretory pathway but with an unknown function. Nesp is expressed in restricted brain nuclei, suggesting an involvement in specific behaviors. We have generated a knockout of Nesp55 in mice. Nesp55-deficient mice develop normally, excluding a role of this protein in the severe postnatal effects associated with imprinting of the Gnas cluster. Behavioral analysis of adult Nesp55 mutants revealed, in three separate tasks, abnormal reactivity to novel environments independent of general locomotor activity and anxiety. This phenotype may be related to prominent Nesp55 expression in the noradrenergic locus coeruleus. These results indicate a role of maternally expressed Nesp55 in controlling exploratory behavior and are the first demonstration that imprinted genes affect such a fundamental behavior.


Binaural characteristics of units in the owl's brainstem auditory pathway: precursors of restricted spatial receptive fields.
The barn owl uses binaural phase and intensity differences for sound localization. These two cues also determine the receptive fields of specialized neurons in the inferior colliculus. The main aim of this study was to investigate where neuronal sensitivity to the binaural cues emerges in the brainstem auditory nuclei, and how this sensitivity reaches the neurons in the inferior colliculus. The owl's phase-sensitive neurons are selective to microsecond phase differences of high frequency signals, unlike mammalian phase-sensitive neurons which are restricted to low frequency signals. In certain nuclei virtually all of the neurons are sensitive to either phase differences or intensity differences, but not to both. These nuclei form two distinctly separate pathways that converge at the inferior colliculus where neurons selective to both phase and intensity differences occur. In contrast to the mammalian auditory system, the owl's phase- and intensity difference-sensitive pathways are not segregated into low frequency and high frequency channels.


Collectin-11 Is an Important Modulator of Retinal Pigment Epithelial Cell Phagocytosis and Cytokine Production.
In this paper, we report previously unknown roles for collectin-11 (CL-11, a soluble C-type lectin) in modulating the retinal pigment epithelial (RPE) cell functions of phagocytosis and cytokine production. We found that CL-11 and its carbohydrate ligand are expressed in both the murine and human neural retina; these resemble each other in terms of RPE and photoreceptor cells. Functional analysis of murine RPE cells showed that CL-11 facilitates the opsonophagocytosis of photoreceptor outer segments and apoptotic cells, and also upregulates IL-10 production. Mechanistic analysis revealed that calreticulin on the RPE cells is required for CL-11-mediated opsonophagocytosis whereas signal-regulatory protein α and mannosyl residues on the cells are involved in the CL-11-mediated upregulation of IL-10 production. This study is the first to demonstrate the role of CL-11 and the molecular mechanisms involved in modulating RPE cell phagocytosis and cytokine production. It provides a new insight into retinal health and disease and has implications for other phagocytic cells.


Synergistic effect of fasting-mimicking diet and vitamin C against KRAS mutated cancers.
Fasting-mimicking diets delay tumor progression and sensitize a wide range of tumors to chemotherapy, but their therapeutic potential in combination with non-cytotoxic compounds is poorly understood. Here we show that vitamin C anticancer activity is limited by the up-regulation of the stress-inducible protein heme-oxygenase-1. The fasting-mimicking diet selectivity reverses vitamin C-induced up-regulation of heme-oxygenase-1 and ferritin in KRAS-mutant cancer cells, consequently increasing reactive iron, oxygen species, and cell death; an effect further potentiated by chemotherapy. In support of a potential role of ferritin in colorectal cancer progression, an analysis of The Cancer Genome Atlas Database indicates that KRAS mutated colorectal cancer patients with low intratumor ferritin mRNA levels display longer 3- and 5-year overall survival. Collectively, our data indicate that the combination of a fasting-mimicking diet and vitamin C represents a promising low toxicity intervention to be tested in randomized clinical trials against colorectal cancer and possibly other KRAS mutated tumors.


The effect of weight controllability beliefs on prejudice and self-efficacy.
An experiment was conducted to test for the presence of prejudice towards obesity and whether weight controllability beliefs information reduces this prejudice and impacts on a person's own healthy eating self-efficacy. The experiment randomly allocated 346 participants (49 males) into one of three conditions: controllable contributors toward obesity condition (e.g., information about personal control about diet and exercise); uncontrollable contributors toward obesity condition (e.g., information about genes, factors in society); and a control condition with no information given. Prejudice was present in 81% of the sample. High prejudice was predicted by low self-efficacy for exercise and weight. Weight controllability beliefs information had no significant effect on prejudice levels or exercise or healthy eating self-efficacy levels. Future research directions are discussed.


Imatinib-induced bone edema: case report and review of literature.
Imatinib mesylate represents a revolution in the management of patients with metastatic gastrointestinal stromal tumors (GISTs). More recently, postoperative imatinib has been shown to improve both disease-free and overall survivals in patients with a high risk of recurrence. This article presents a well-documented case of a patient with painful and reversible bone edema related to imatinib.


A reliability study of the rapid emergency triage and treatment system for children.
To evaluate inter- and intrarater reliability of a new Scandinavian triage system for children, the Rapid Emergency Triage and Treatment System-pediatric (RETTS-p). Two observational studies were conducted at the Pediatric Emergency Department (PED), St. Olav's University Hospital, Trondheim, Norway. Using RETTS-p, nurses assign one of five triage priority levels to each patient on the basis of clinical signs and symptoms evaluations and vital parameter measurements. Study 1: Prior to the introduction of RETTS-p in 2012, all nurses in the PED completed a theoretical and practical training. Four months later, 19 nurses triaged 20 fictive but realistic pediatric cases two times 9 months apart (Waves A and B). Study 2: Nurse pairs consisting of a regular nurse and a research nurse simultaneously and independently triaged 200 pediatric patients who were referred with various common medical and surgical complaints. Study 1: Kendall's W for Waves A and B were 0.822 and 0.844, respectively. Using a mixed linear model, we found no difference in triage priority levels between Waves A and B. Compared to a consensus level made by the research group, the nurses rated 85.1 % fictive cases correctly, and 99 % were rated correctly or within one adjacent priority score. Study 2: The interrater correlation coefficient in a linear mixed model was 0.762, confirming a high interrater reliability in real-life triaging. We found a very high degree of agreement between nurses who used RETTS-p to prioritize children, both in a theoretical case scenarios study, but also in real-life triaging. RETTS-p may be a credible and robust triage system, but it has not been validated yet.


Evidence basis for integrated management of mineral metabolism in patients with end-stage renal disease.
Treatment of mineral metabolism is a mainstay of dialysis care including some of its most widely used and costly pharmaceuticals. Although many mineral metabolites are associated with increased risk of mortality, cardiovascular disease, and other morbidities, few clinical trials are available to guide therapy and most focus on single drug approaches. In practice, providers manage many aspects of mineral metabolism simultaneously in integrated treatment approaches that incorporate multiple agents and changes in the dialysis prescription. The present review discusses the rationale and existing evidence for evaluating integrated, as opposed to single drug, approaches in mineral metabolism. Drugs used to treat mineral metabolism have numerous, and sometimes, opposing effects on biochemical risk factors, such as fibroblast growth factor 23 (FGF23), calcium, and phosphorus. Although vitamin D sterols raise these risk markers when lowering parathyroid hormone (PTH), calcimimetics lower them. Trials demonstrate that combined approaches best 'normalize' the mineral metabolism axis in end-stage renal disease (ESRD). Observations embedded within major trials of calcimimetics reveal that adjustment of calcium-based binders and dialysate calcium is a common approach to adverse effects of these drugs with some initial, but inconclusive, evidence that these co-interventions may impact outcomes. The multiple, and often opposing, biochemical effects of many mineral metabolism drugs provides a strong rationale for studying integrated management strategies that consider combinations of drugs and co-interventions as a whole. This remains a current gap in the field with opportunities for clinical trials.


Sampling strategies and variability in fruit pulp micronutrient contents of west and central african bananas and plantains (Musa species).
The variability in fruit micronutrient contents in a selection of Central and West African Musa varieties cultivated under standardized field conditions was studied. Analysis of the within-fruit, within-hand, and within-plant as well as the between-plant variations demonstrated that both provitamin A carotenoids (pVACs) and mineral micronutrient (Fe, Zn) contents vary significantly across all sample groups. The variations in pVACs contents appear to be at least partly related to differences in the developmental status of the fruit, but the observed trends were genotype-specific. The mean pVACs concentrations per genotype indicated that there is substantial genetic variation in the fruit pVACs contents between Musa cultivars, with orange-fleshed plantain varieties (AAB) having generally higher fruit pVACs contents than dessert bananas (AAA). It was not possible to identify consistent trends between the sampling position and fruit Fe/Zn contents. Once the within-bunch micronutrient variability has been accounted for, the mean variations in fruit micronutrient contents between individual plants of a variety generally fell to within acceptable limits. Results are discussed within the framework of standardizing sampling and developing strategies to screen for the nutritional values of new and existing Musa varieties.


In situ hybridization and immunohistochemical analysis of cytomegalovirus-associated ileal perforation.
A patient with the acquired immunodeficiency syndrome (AIDS) with a cytomegalovirus (CMV) infection localized in the area of an ileal Kaposi sarcoma resulting in perforation is described. Because only one case of such an association of Kaposi sarcoma with CMV leading to perforation has been reported, the presence and distribution of CMV-related nucleic acids and proteins in the affected segment of intestine were evaluated. By using in situ hybridization and immunocytochemical analyses the presence of CMV was shown within epithelial, endothelial, smooth muscle, and inflammatory cells at the site of perforation. This study not only confirmed that CMV can be detected in virtually all components of the intestinal wall despite the absence of distinctive cytomegalic changes, but also generated critical information that illustrates the usefulness of immunohistochemistry and in situ hybridization in elucidating the pathogenesis of CMV-associated lesions. These findings lend further support to the concept that CMV plays a pivotal role in the pathogenesis of intestinal perforation and emphasize the critical importance of in situ hybridization in gaining insight into the mechanisms of CMV-induced injury.


Eye movement patterns during the recognition of three-dimensional objects: preferential fixation of concave surface curvature minima.
This study used eye movement patterns to examine how high-level shape information is used during 3D object recognition. Eye movements were recorded while observers either actively memorized or passively viewed sets of novel objects, and then during a subsequent recognition memory task. Fixation data were contrasted against different algorithmically generated models of shape analysis based on: (1) regions of internal concave or (2) convex surface curvature discontinuity or (3) external bounding contour. The results showed a preference for fixation at regions of internal local features during both active memorization and passive viewing but also for regions of concave surface curvature during the recognition task. These findings provide new evidence supporting the special functional status of local concave discontinuities in recognition and show how studies of eye movement patterns can elucidate shape information processing in human vision.


Myalgia in Patients with Dermatomyositis and Polymyositis Is Attributable to Fasciitis Rather Than Myositis: A Retrospective Study of 32 Patients who Underwent Histopathological Examinations.
To determine the association between fasciitis and the clinical variables in patients with dermatomyositis (DM) and polymyositis (PM). We retrospectively reviewed the medical records of 32 patients (24 DM, 8 PM) with newly diagnosed DM and PM and in whom fascia and muscle specimens were histopathologically examined. The relationship between fasciitis and the clinical variables was statistically analyzed. These included age, sex, myalgia, muscle weakness, creatine kinase (CK) and aldolase activities, anti-Jo1 antibody, interstitial lung disease, and malignancy. Twenty (62.5%) of the 32 patients who underwent the histopathological examination of a fascia specimen had fasciitis, including 18 (75%) of 24 patients with DM and 2 (25%) of 8 patients with PM. The frequency of fasciitis was significantly higher among the patients with DM than among the patients with PM (p < 0.05). Histopathologically, fasciitis in PM was very mild in comparison to that in DM. The frequency of myalgia in patients with fasciitis was significantly higher than that in patients without fasciitis (p < 0.05). However, myalgia was not associated with myositis. There were no significant differences in the patients with and without fasciitis in age, sex, manual muscle test 8 scores, CK or aldolase activities, or the presence of anti-Jo1 antibodies and malignancy. The frequency of fasciitis was significantly higher among patients with DM than among those with PM. Fasciitis, rather than myositis, was associated with myalgia.


Helicobacter pylori and autoimmune disease: cause or bystander.
Helicobacter pylori (H. pylori) is the main cause of chronic gastritis and a major risk factor for gastric cancer. This pathogen has also been considered a potential trigger of gastric autoimmunity, and in particular of autoimmune gastritis. However, a considerable number of reports have attempted to link H. pylori infection with the development of extra-gastrointestinal autoimmune disorders, affecting organs not immediately relevant to the stomach. This review discusses the current evidence in support or against the role of H. pylori as a potential trigger of autoimmune rheumatic and skin diseases, as well as organ specific autoimmune diseases. We discuss epidemiological, serological, immunological and experimental evidence associating this pathogen with autoimmune diseases. Although over one hundred autoimmune diseases have been investigated in relation to H. pylori, we discuss a select number of papers with a larger literature base, and include Sjögrens syndrome, rheumatoid arthritis, systemic lupus erythematosus, vasculitides, autoimmune skin conditions, idiopathic thrombocytopenic purpura, autoimmune thyroid disease, multiple sclerosis, neuromyelitis optica and autoimmune liver diseases. Specific mention is given to those studies reporting an association of anti-H. pylori antibodies with the presence of autoimmune disease-specific clinical parameters, as well as those failing to find such associations. We also provide helpful hints for future research.


Safety potential of crash avoidance features, improved headlights, and V2V-enhanced technologies for older drivers.
Age-related changes and frailty are among the reasons that older drivers are overrepresented in certain crash types. Vehicle safety features that address these crash types may therefore deliver greater safety benefits for older drivers than for other age groups even though they are designed for the general population. U.S. crash data from 2016-2019 were used to estimate the proportion of crash involvements and fatal and nonfatal driver injuries for older (70 years old and above) and middle-aged (35-54 years old) drivers from crash scenarios to which current crash avoidance features, improved headlights, and forthcoming vehicle-to-vehicle (V2V)-connected intersection-assistance features could be relevant. Risk ratios were then calculated to determine the relative benefits of each technology for older drivers compared with middle-aged drivers. Combined, these technologies were potentially relevant to 65 % of older driver and 72 % of middle-aged driver fatalities during the study period. Intersection assistance features showed the most promise for older drivers. Such features were potentially relevant to 32 % of older driver crash involvements, 38 % of older driver injuries, and 31 % of older driver fatalities. Intersection assistance features were significantly more likely to be relevant to older driver deaths than middle-aged driver fatalities (RR, 3.52; 95 % CI, 3.33-3.71). Vehicle technologies have the potential to substantially reduce or mitigate crashes and the injuries that they cause for everyone, but the potential safety impact of each technology varies by driver age because different age groups are over- or underrepresented in specific crash scenarios. With the older driver population growing, these findings underscore the need to bring intersection assistance technologies to the consumer market. At the same time, everyone stands to benefit from currently available crash avoidance features and improved headlights, so their use should be promoted among all drivers.


The anticonvulsant, lamotrigine decreases spontaneous glutamate release but increases spontaneous GABA release in the rat entorhinal cortex in vitro.
It has been suggested that the anticonvulsant effect of lamotrigine resides with it's ability to block voltage gated Na-channels at presynaptic sites, thus stabilizing the presynapse, and, consequently, reducing the release of synaptic transmitters. Neurochemical studies have shown that it can inhibit the veratrine-stimulated release of the excitatory transmitter, glutamate from cortical tissue, but that at slightly higher concentrations it also reduces the release of the inhibitory transmitter, GABA. In the present study we examined the effect of the drug on the release of these transmitters at synapses in the rat entorhinal cortex, using the whole-cell patch clamp technique to record spontaneous excitatory (EPSCs) and inhibitory postsynaptic currents (IPSCs). Lamotrigine reduced the frequency, but not the amplitude of spontaneous EPSCs. This clearly indicated a presynaptic effect to reduce the release of glutamate. However, the same effect was observed when we tested the drug on miniature EPSCs, recorded in the presence of TTX and Cd, showing that blockade of Na-channels or Ca-channels was not a prerequisite for inhibition of glutamate release. In contrast to it's effects on EPSCs, lamotrigine increased both the frequency and amplitude of spontaneous IPSCs, suggesting that the drug was acting presynaptically to enhance GABA release. Again, similar effects were seen with miniature IPSCs recorded in TTX. These opposite effects of lamotrigine on glutamate and GABA release are similar to those we have reported previously with phenytoin, and suggest that reciprocal modulation of the background release of the major excitatory and inhibitory transmitters may be a significant factor in dampening excitability in pathologically hyperexcitable cortical networks.


Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA.
Fiber mRNA of adenovirus 2 has been used as a template for RNA-dependent DNA polymerase. The resulting cDNA/RNA hybrids have been inserted at the Pst I site of the plasmid vector pBR322 after A:T tailing. One recombinant plasmid, pJAW 43, has been characterized in detail and shown to contain sequences from the main body of fiber mRNA, the three leaders common to most late adenoviral mRNAs and a fourth leader found in some species of fiber mRNA. The complete DNA sequence of the leader region has been determined and does not contain the initiation codon AUG, although this codon does occur immediately downstream from the junction between the fourth leader and the main body of the fiber mRNA. The first leader (map coordinate 16.6) is 41 nucleotides long, the second (from 19.6) is 71 nucleotides, the third (from 26.6) is 88 nucleotides and the fourth (from 78.5) is 181 nucleotides. The location of junctions between viral leaders and intervening sequences has been determined by reference, where possible, to sequences of the adenovirus 2 genome. Although the presence of short repeated sequences at the boundaries of intervening sequences and leaders makes it impossible to locate the splice point unambiguously, all of the leader-intervening sequence junctions can be arranged to stress a common feature--the presence of the dinucleotides GT and AG at the 5' and 3' ends, respectively, of the intervening sequences. This prototype sequence, which has also been recognized at or near the splice points in other eucaryotic systems, is possibly part of a larger unit which serves as a recognition site for specific excision-ligation events that ultimately lead to the production of mature mRNAs.


Nitrogen deposition changes ectomycorrhizal communities in Swiss beech forests.
Atmospheric pollution has implications for the health and diversity of temperate forests covering large parts of central Europe. Long-term elevated anthropogenic deposition of nitrogen (N) is driving forest ecosystems from the limitation by N to other nutrients and is found to affect tree health and ectomycorrhizal fungi (EMF), which most trees depend on for nutrient uptake. However, the consequence of EMF community changes for trees remains unclear. Therefore, we investigated changes in EMF communities on root tips and in soil of beech forests along a N deposition gradient ranging between 16 and 33kgNha-1a-1, where high N deposition was found to negatively affect tree growth and nutrient levels. The most important factors significantly explaining variation in root tip and mycelium EMF community composition in both root tips and mesh bags were increased N deposition, base saturation, growing season temperature and precipitation. With increasing N deposition, fine root length, EMF root colonization, EMF diversity on root tips and in soil, and production of extramatrical mycelium decreased significantly. Foliar P and potassium (K) were positively associated with increasing EMF diversity and we found EMF community composition to be associated with foliar P and N:P ratio. The decrease in root colonization, mesh bag ingrowth and abundance of the important species Cenococcum geophilum as well as high biomass species with increasing N availability clearly indicate repercussions for belowground carbon allocation, although some indicator species for high N deposition and low foliar P have long mycelia and may reflect a potential optimization of host P uptake. Our study supports the hypothesis that the decrease in nutrient uptake in beech forests across Europe is related to changes in EMF communities and suggests that continued high N deposition changes soil carbon and nutrient cycles, thereby affecting forest ecosystem health.


Maternal cell-mediated immunity to the fetus in human pregnancy.
Cell-mediated sensitisation of the mother to her fetus is not a regular event in human pregnancy, and the role of suppressor cells and blocking antibodies in preventing this sensitisation is unresolved. There is as yet little evidence to support the hypothesis that maternal immune recognition of the fetus is essential for the success of the pregnancy. Further progress in this area will depend on systematic investigations of the development of specific maternal humoral and cell-mediated responses to the fetus throughout gestation, and whether a deficiency of these responses results in the failure of the pregnancy.


Phase I trial of cyclosporine-induced autologous graft-versus-host disease in patients with multiple myeloma undergoing high-dose chemotherapy with autologous stem-cell rescue.
To determine the feasibility and toxicity of inducing autologous graft-versus-host disease (GVHD) with cyclosporine in patients with multiple myeloma undergoing autologous stem-cell transplantation. Fourteen multiple myeloma patients with a median age of 50 years (range, 41 to 63) were enrolled. The median time from diagnosis to transplant was 651 days (range, 229 to 3,353). Ten patients had primary refractory disease, two were in first remission, and two were responsive to salvage therapy. The preparative regimen consisted of thiotepa, busulfan, and cyclophosphamide. Cyclosporine was administered daily for 28 days after the stem-cell infusion, and the dose was adjusted to maintain whole-blood cyclosporine levels between 50 and 150 ng/dL in the first seven patients (low-level group) and between 150 and 300 ng/dL in the other seven patients (high-level group). All patients achieved neutrophil engraftment a median of 11 days after transplant. Four patients developed > or = grade 2 hepatic toxicity, six developed > or = grade 2 nephrotoxicity, and four developed reversible cardiac toxicity. Only one treatment-related death occurred. Cyclosporine was withheld in seven patients for a median of 6 days because of renal and/or liver dysfunction. One patient developed clinical skin GVHD, which responded to corticosteroid therapy. Six patients developed histologic evidence of GVHD without clinical signs of GVHD (subclinical GVHD). The incidence of clinical and subclinical GVHD was similar in both cyclosporine groups. Three of 11 patients assessable for response achieved remissions. Three patients experienced disease progression 80, 160, and 354 days after transplant. Ten patients are alive without progression between 56 and 444 days after transplant. Induction of autologous GVHD by posttransplant cyclosporine is feasible and well tolerated in patients with multiple myeloma.


Consumption of cashew nuts does not influence blood lipids or other markers of cardiovascular disease in humans: a randomized controlled trial.
The US Food and Drug Administration (FDA) approved a qualified health claim for tree nuts and reduction of cardiovascular disease. However, cashews are excluded from that claim due to their content of saturated fats, which is predominantly stearic acid. Because stearic acid is neutral with respect to blood lipids, several studies have been conducted to test the effect of cashew nuts on blood lipids, and these studies have produced conflicting results. The aim of this study was to conduct a highly controlled intervention to determine the effect of cashews fed at the amount specified in the health claim on risk factors for cardiovascular disease. A total of 42 adults participated in a controlled-feeding study conducted as a randomized crossover trial with 2 treatment phases. The volunteers were provided the same base diet in both treatment phases, with no additions during the control phase and with the addition of 1.5 servings (42 g) of cashews/d for the cashew nut phase. During the cashew nut phase, the amount of all foods was decreased proportionally to achieve isocaloric overall diets in the 2 phases. After 4 wk of intervention, assessments included blood lipids, blood pressure, central (aortic) pressure, augmentation index, blood glucose, endothelin, proprotein convertase subtilisin/kexin type 9 (PCSK9), adhesion molecules, and clotting and inflammatory factors. There were no significant differences in blood lipids, blood pressure, augmentation index, blood glucose, endothelin, adhesion molecules, or clotting factors in this weight-stable cohort. PCSK9 was significantly decreased after cashew consumption, although there was no change in LDL cholesterol. Consumption of 1.5 servings of cashew nuts/d, the amount associated with the FDA qualified health claim for tree nuts and cardiovascular disease, did not positively or adversely affect any of the primary risk factors for cardiovascular disease. This trial was registered at clinicaltrials.gov as NCT02628171.


Inadvertently Developed Ventricular Fibrillation during Electrophysiologic Study and Catheter Ablation: Incidence, Cause, and Prognosis.
Ventricular fibrillation (VF) can inadvertently occur during electrophysiologic study (EPS) or catheter ablation. We investigated the incidence, cause, and progress of inadvertently developed VF during EPS and catheter ablation. We reviewed patients who had developed inadvertent VF during EPS or catheter ablation. Patients who developed VF during programmed ventricular stimulation to induce ventricular tachycardia or VF were excluded. Inadvertent VF developed in 11 patients (46.7±9.3 years old) among 2624 patients (0.42%); during catheter ablation for atrial fibrillation (AF) in nine patients, frequent ventricular premature beats (VPBs) in one, and Wolff-Parkinson-White (WPW) syndrome were observed in one. VF was induced after internal cardioversion in six AF patients due to incorrect R-wave synchronization of a direct current shock. Two AF patients showed spontaneous VF induction during isoproterenol infusion while looking for AF triggering foci. The remaining AF patient developed VF after rapid atrial pacing to induce AF, but the catheter was accidentally moved to the right ventricular (RV) apex. A patient with VPB ablation spontaneously developed VF during isoproterenol infusion. The focus of VPB was in the RV outflow tract and successfully ablated. A patient with WPW syndrome developed VF after rapid RV pacing with a cycle length of 240 ms. Single high energy (biphasic 150-200 J) external defibrillation was successful in all patients, except in two, who spontaneously terminated VF. The procedure was uneventfully completed in all patients. At a mean follow-up period of 17.4±15.5 months, no patient presented with ventricular arrhythmia. Although rare, inadvertent VF can develop during EPS or catheter ablation. Special caution is required to avoid incidental VF during internal cardioversion, especially under isoproterenol infusion.


Systematic Review of Intraoperative Anesthesia Handoffs and Handoff Tools.
Intraoperative handoffs between anesthesia clinicians are critical for care continuity. However, such handoffs pose a significant threat to patient safety. This systematic review synthesizes the empirical evidence on the (a) effect of intraoperative handoffs on outcomes and (b) effect of intraoperative handoff tools on outcomes. All studies on intraoperative handoffs and handoff tools published until September 2019, in any study setting and population, and with no prespecified criteria on the type of comparison and outcome were included. Data extracted from the included studies were aggregated to identify common patterns related to the type of surgery, clinician(s) involved, patient population, handoff tool, the tool design approach (where relevant), tool implementation strategies, and finally, all reported clinical and process outcomes. Quality of studies was assessed using the Newcastle-Ottawa Scale (NOS). Fourteen studies met the inclusion criteria. All included studies used adult patients. Eight studies were retrospective cohort studies that used administrative or electronic health record (EHR)-based databases to investigate the effects of intraoperative handoffs on morbidity and mortality. These studies included a total of 680,855 surgeries, with 139,426 of these surgeries having at least 1 handoff (20.47%). Seven of the studies found a positive association between intraoperative handoffs and considered outcomes. However, a pooled meta-analysis across these studies was not feasible across the retrospective studies due to differing surgical populations and varying definitions of the considered outcomes. Six studies used a nonrandomized prospective design to evaluate the effects of handoff tools on process-based outcomes such as clinician satisfaction, information transfer, handoff duration, and adherence. Five of the 6 handoff tools were checklist based. All prospective tool-based studies relied on small samples and reported a significant improvement on the considered process-based outcomes. The median quality score among retrospective (median [interquartile range {IQR}] = 9 [1]) was significantly higher than that of prospective (median [IQR] = 5 [1.5]) studies (U = 21, P = .0017). This systematic review provides a unique appraisal of the current state of intraoperative handoff research. To improve the quality and outcomes of handoffs, future efforts should focus on design and implementation of standardized handoff tools integrated within EHR systems, consider the use of similar metrics for evaluating handoff process and clinical outcomes, and improve the execution and reporting of studies using standard protocols and guidelines.


Logistic regression analysis of multiple interosseous hand-muscle activities using surface electromyography during finger-oriented tasks.
Intrinsic hand muscles are densely located in the hand, and the myoelectric observation from the surface is sometimes unreliable because of some outside influences that may interfere with the signals. In the present study, we evaluated the activities of multiple interosseous hand-muscles which densely located in the hand, through analyzing the surface electromyographic signals during finger-oriented tasks using univariate and multivariate logistic regression models. Ten healthy subjects participated in our experiment, and isometrically exercised each finger one by one in flexed form. The result of a univariate analysis with the power and amplitude domain predictor variables of the surface electromyographic signals showed significant consistency between the activated finger and the inserted finger of the dorsal interosseous muscles to the proximal phalanx (P < 0.001). Meanwhile, the results of a multivariate analysis showed a higher correlation of the regression model of the fourth dorsal interosseous muscle during the action of the ring finger using frequency-domain variables (the Nagelkerke R2 = 0.716 when the median frequency was used), compared to the model without the frequency-domain variables (the Nagelkerke R2 = 0.583). Our result showed that the logistic regression models have a particular possibility for the analysis of the surface electromyographic signals of densely located hand-muscle activities related to the finger-oriented tasks.


Epidemiology of epilepsy in Pakistan: review of literature.
To review literature pertinent to the epidemiology of epilepsy in developing countries with special reference to Pakistan. All the studies published in medical journals related to epilepsy in Pakistan were systematically reviewed. Important findings from various studies are summarized. Overall prevalence of epilepsy in Pakistan is estimated to be 9.99 per 1000 population. Highest prevalence is seen in people younger than 30 years of age. A slight decrease in prevalence is noted between the ages of 40 and 59. Higher prevalence is observed in rural population. Etiology of epilepsy is more commonly identified in pediatric population. Epilepsy was considered idiopathic in 21 to 76% cases. Only 27.5% epileptic persons in urban areas and 1.9% in the rural areas were treated with AEDs. The burden of epilepsy is not fully evaluated and understood. Generalized seizures were the most common seizure type noted. Knowledge about epilepsy and its care is extremely low. Epilepsy is a common medical problem in Paksitan, more prevalent is rural population. The majority of people with epilepsy are treated inadequately or inappropriately.


Asian-Pacific consensus statement on the management of chronic hepatitis B: a 2012 update.
Large volume of new data on the natural history and treatment of chronic hepatitis B virus (HBV) infection have become available since 2008. These include further studies in asymptomatic subjects with chronic HBV infection and community-based cohorts, the role of HBV genotype/naturally occurring HBV mutations, the application of non-invasive assessment of hepatic fibrosis and quantitation of HBV surface antigen and new drug or new strategies towards more effective therapy. To update HBV management guidelines, relevant new data were reviewed and assessed by experts from the region, and the significance of the reported findings was discussed and debated. The earlier "Asian-Pacific consensus statement on the management of chronic hepatitis B" was revised accordingly. The key terms used in the statement were also defined. The new guidelines include general management, indications for fibrosis assessment, time to start or stop drug therapy, choice of drug to initiate therapy, when and how to monitor the patients during and after stopping drug therapy. Recommendations on the therapy of patients in special circumstances, including women in childbearing age, patients with antiviral drug resistance, concurrent viral infection, hepatic decompensation, patients receiving immune suppression or chemotherapy and patients in the setting of liver transplantation and hepatocellular carcinoma, are also included.


Preinjury beta blockers are associated with increased mortality in geriatric trauma patients.
Beta-blockade decreases mortality and morbidity in selected older patient populations undergoing noncardiac general surgery. We hypothesized that preinjury beta blockade would increase mortality in geriatric trauma patients, given beta-blockers inhibit patient's physiologic responses to hypovolemic shock. Patients older than 65 years admitted to a level I trauma center were identified by the trauma registry. Medical records were reviewed for demographic and injury information. Preinjury beta blockade was determined by review of nurse and pharmacy admission histories. Logistic regression was used to determine whether there was any correlation between mortality and the use of preinjury beta blockers. Separate models were developed based on the presence or the absence of head injury. Of the 1,598 patients older than 65 years admitted between 1996 and 2006, 1,479 met inclusion criteria. Primary reason for exclusion was lack of documentation. Two hundred seventy-three patients were taking beta blockers before their trauma, and 14.7% died before discharge. Mortality in patients not taking beta blockers was 13.4%. Mortality in patients with head injury was 25.9%, significantly associated with warfarin use (OR 2.5, 95% CI 1.3-4.8). In patients without head injury, preinjury beta blockade had a significant association with mortality (OR 2.1, 95% CI 1.1-4.3). Many factors associated with mortality in elderly trauma patients are similar to the younger patient population. Unique to this population are increased comorbidities and use of prescription medications. Beta blockers, one of these common medications, are associated with increased mortality in the elderly.


Isolation of cytomegalovirus-specific cytotoxic T-lymphocytes from gut-associated lymphoid tissue (GALT) of HIV type 1-infected subjects.
Cytomegalovirus (CMV) can be an important opportunistic infection in HIV-1-infected patients, particularly when the CD4+ T-cell count drops below 50 lymphocytes/mm3. CMV-associated disease, including retinitis, pneumonitis, gastroenteritis, and encephalitis, is estimated to affect up to 40% of AIDS patients. We have studied the cellular immune response to CMV in gut-associated lymphoid tissue (GALT) of HIV-1-infected patients. Two patients with chronic diarrhea of unknown etiology were examined by flexible sigmoidoscopy and upper endoscopy. Biopsy specimens were obtained from lymphoid-associated tissue sites in rectum and duodenum. Both patients were seropositive for CMV IgG, but had not been treated with ganciclovir, and neither had clinical signs of CMV disease. Mononuclear cell cultures were established from GALT and blood and assayed for the presence of CMV-specific CD8+ T cells. CD8+ T-cell phenotype and function were assessed by MHC Class I tetramer staining, using an HLA-A*0201 tetramer complex specific for peptide 495-503 (NLVPMVATV) of CMV lower matrix protein pp65, and by a standard 51Cr release assay. CMV pp65-specific cytotoxic lymphocytes (CTL) were detected in GALT and blood MNC from both patients. These results demonstrate that HIV-1-infected subjects seropositive for CMV, but without active CMV gastrointestinal disease, harbor CMV-specific CTL in intestinal lymphoid tissue. This is the first report of isolation of CMV-specific CTL in GALT and will lead to greater understanding of the pathogenesis of CMV disease in human mucosal tissue.


Learning curves, acquisition, and retention of skills trained with the endoscopic sinus surgery simulator.
As an initial step in evaluating the effectiveness of training otolaryngology residents on an endoscopic sinus surgery simulator (ES3), we have assessed the ability of the ES3 to train persons inexperienced in sinus surgery (medical students) to perform certain simulated procedural tasks needed in endoscopic sinus surgery (ESS). A total of 26 medical students were enrolled and trained on the ES3 following a preset protocol in the three levels of difficulty and complexity (novice, intermediate, and advanced modes). In the novice mode (three-dimensional abstract images are used to teach the use of endoscopic surgical equipment), medical students displayed a steep learning curve within three to five trials on the simulator and after an additional four to five trials, they reached a plateau in their learning curves to within 90% of that of experienced sinus surgeons. In the intermediate mode (ESS is performed on a simulated patient with teaching aids), medical students were able to reach a plateau in their learning curves to within 80% of that of experienced surgeons. This performance was sustained in the advanced mode (simulated sinus surgery without teaching aids). We observed that medical students, who had novice or intermediate mode training interrupted with an interval of 11-60 days, were able to resume their training without deviation from their prior learning curves. Intensive, proctored training on the ES3 can train inexperienced persons to perform simulated ESS within a reasonable approximation of the performance of experienced sinus surgeons on the ES3 and the training that an inexperienced person receives on the simulator is not short term but is retained over a period of at least 2 months.


Statins May Reduce Breast Cancer Risk, Particularly Hormone Receptor-Negative Disease.
Estrogen and progesterone receptor-negative breast cancer disproportionately affects young women and African Americans, has a poor prognosis, and lacks an effective chemoprevention agent. 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, known as "statins," are appealing candidate agents for breast cancer chemoprevention because of their demonstrated safety after decades of widespread use. In preclinical studies, statins inhibit multiple cancer-associated pathways in both hormone receptor (HR)-negative and HR-positive cell lines. Epidemiologic studies of statins and breast cancer show inconsistent results, with some suggesting a reduction in HR-negative breast cancer incidence in lipophilic statin users. However, large meta-analyses show no association between statin use and overall risk of breast cancer, although most did not evaluate tumor HR status. Multiple phase 1 and 2 prevention studies of statins for breast cancer risk reduction are ongoing. If results are promising, they may justify a randomized trial of statins for breast cancer chemoprevention, with a focus on HR-negative disease.


Contemporary use and outcome of Cabrol shunt in type A aortic dissection surgery: insight from China 5A study.
Cabrol shunt has been introduced for surgical repair of type A aortic dissection (TAAD) without robust evidence supporting its routine preventive use. Adult patients with TAAD from China 5A study were included if surgically repaired between 2016 and 2022. Primary outcome was operative mortality according to Society of Thoracic Surgeons criterion. Overall, we compared clinical outcomes in patients with and without Cabrol shunt, and subgroup analysis were further examined between Cabrol shunt and outcome among patients with or without root replacement. 3283 patients were finally identified for analysis, with median age of 51 (IQR 41-59) years, 2389 men, and 2201 treated with Cabrol shunt technique. Cabrol shunt-treated patients were more severely ill before surgery than those without Cabrol shunt. Overall, the rate of operative mortality was 6.6% (146/2201 in Cabrol shunt group and 71/1082 in non-Cabrol shunt group), with no association between Cabrol shunt and operative mortality (OR 1.012 (95% CI 0.754 to 1.357); p=0.938). Stratified by root replacement, Cabrol shunt was associated with similar risk of operative mortality either in patients without root replacement (OR 1.054 (0.747 to 1.487); p=0.764) or in patients with root replacement (OR 1.194 (0.563 to 2.536); p=0.644) (P interaction=0.765). Results were similar in multiple sensitivity analysis. Cabrol shunt was not associated with either a greatly lowered or an increased risk of operative mortality, regardless of aortic root replacement. Our study did not support the use of Cabrol shunt as a routine preventive strategy in the treatment of TAAD. NCT04398992.


Minimally invasive sacroiliac joint fusion: one-year outcomes in 40 patients.
Background. SI joint pain is difficult to diagnose due to overlapping symptoms of the lumbar spine, and until recently, treatment options have been limited. The purpose of this retrospective study is to report on the safety and effectiveness of MIS SI joint arthrodesis using a series of triangular, porous plasma coated implants in patients refractory to conservative care. Methods. We report on the first 40 consecutive patients with one-year follow-up data that underwent MIS SI joint fusion with the iFuse Implant System (SI-BONE, Inc., San Jose, CA) by a single surgeon. Medical charts were reviewed for demographics, perioperative metrics, complications, pain scores, and satisfaction. Results. Mean age was 58 years (range 30-81) and 75% of patients were female. Postoperative complications were minimal and included transient trochanteric bursitis (5%), facet joint pain (20%), and new low back pain (2.5%). There were no reoperations at one year. Mean pain score improved from 8.7 (1.5 SD) at baseline to 0.9 (1.6) at 12 months, a 7.8-point improvement (P < .001). Patient satisfaction was very high. Conclusions. The results of this case series reveal that MIS SI joint fusion using the iFuse Implant System is a safe and effective treatment option in carefully selected patients.


Muscle-tendon interaction and elastic energy usage in human walking.
The present study was designed to explore how the interaction between the fascicles and tendinous tissues is involved in storage and utilization of elastic energy during human walking. Eight male subjects walked with a natural cadence (1.4 +/- 0.1 m/s) on a 10-m-long force plate system. In vivo techniques were employed to record the Achilles tendon force and to scan real-time fascicle lengths for two muscles (medial gastrocnemius and soleus). The results showed that tendinous tissues of both medial gastrocnemius and soleus muscles lengthened slowly throughout the single-stance phase and then recoiled rapidly close to the end of the ground contact. However, the fascicle length changes demonstrated different patterns and amplitudes between two muscles. The medial gastrocnemius fascicles were stretched during the early single-stance phase and then remained isometrically during the late-stance phase. In contrast, the soleus fascicles were lengthened until the end of the single-stance phase. These findings suggest that the elastic recoil takes place not as a spring-like bouncing but as a catapult action in natural human walking. The interaction between the muscle fascicles and tendinous tissues plays an important role in the process of release of elastic energy, although the leg muscles, which are commonly accepted as synergists, do not have similar mechanical behavior of fascicles in this catapult action.


Genomic rearrangements of the PRPF31 gene account for 2.5% of autosomal dominant retinitis pigmentosa.
To determine whether genomic rearrangements in the PRPF31 (RP11) gene are a frequent cause of autosomal dominant retinitis pigmentosa (adRP) in a cohort of patients with adRP. In a cohort of 200 families with adRP, disease-causing mutations have previously been identified in 107 families. To determine the cause of disease in the remaining families, linkage testing was performed with markers for 13 known adRP loci. In a large American family, evidence was found of linkage to the PRPF31 gene, although DNA sequencing revealed no mutations. SNP testing throughout the genomic region was used to determine whether any part of the gene was deleted. Aberrant segregation of a SNP near exon 1 was observed, leading to the testing of additional SNPs in the region. After identifying an insertion-deletion mutation, the remaining 92 families were screened for genomic rearrangements in PRPF31 with multiplex ligation-dependent probe amplification (MLPA). Five unique rearrangements were identified in the 93 families tested. In the large family used for linkage exclusion testing, an insertion-deletion was found that disrupts exon 1. The other four mutations identified in the cohort were deletions, ranging from 5 kb to greater than 45 kb. Two of the large deletions encompass all PRPF31 as well as several adjacent genes. The two smaller deletions involve either 5 or 10 completely deleted exons. In an earlier long-term study of 200 families with adRP, disease-causing mutations were identified in 53% of the families. Mutation-testing by sequencing missed large-scale genomic rearrangements such as insertions or deletions. MLPA was used to identify genomic rearrangements in PRPF31 in five families, suggesting a frequency of approximately 2.5%. Mutations in PRPF31 now account for 8% of this adRP cohort.


Single-limb landing biomechanics are altered and patellar tendinopathy related pain is reduced with acute infrapatellar strap application.
Patellar tendinopathy, a common condition of the knee, is often treated with patellar tendon straps to control pain during dynamic activity. Little is known regarding their effect on pain, landing kinematics and kinetics with their application. The purpose of this study was to determine if patellar tendon straps influenced pain, kinematics at landing and ground reaction forces in individuals with patellar tendinopathy versus healthy controls. Thirty participants with patellar tendinopathy and 30 controls participated. They completed single-limb landings with and without patellar tendon straps while pain, three-dimensional kinematics and vertical ground reaction forces were measured. A multivariate analysis of variance was completed to determine the differences in strapping condition and group for the dependent variables. Individuals with patellar tendinopathy demonstrated a significant decrease in pain (no strap=37.1±22.1mm (mean±SD), strap=28.0±18.5mm (mean±SD)). With the strap at landing all participants displayed less hip rotation (F=7.16, p=.01), knee adduction (F=10.20, p=.002), ankle inversion (F=4.60, p=.04), and peak vertical ground reaction force (F=7.30, p=.009). Patellar tendon straps reduced pain in those with patellar tendinopathy. Additionally, with the strap, individuals landed in a more neutral alignment and decreased landing forces which could provide a benefit to those with patellar tendinopathy.


Near-Stasis in the Long-Term Diversification of Mesozoic Tetrapods.
How did evolution generate the extraordinary diversity of vertebrates on land? Zero species are known prior to ~380 million years ago, and more than 30,000 are present today. An expansionist model suggests this was achieved by large and unbounded increases, leading to substantially greater diversity in the present than at any time in the geological past. This model contrasts starkly with empirical support for constrained diversification in marine animals, suggesting different macroevolutionary processes on land and in the sea. We quantify patterns of vertebrate standing diversity on land during the Mesozoic-early Paleogene interval, applying sample-standardization to a global fossil dataset containing 27,260 occurrences of 4,898 non-marine tetrapod species. Our results show a highly stable pattern of Mesozoic tetrapod diversity at regional and local levels, underpinned by a weakly positive, but near-zero, long-term net diversification rate over 190 million years. Species diversity of non-flying terrestrial tetrapods less than doubled over this interval, despite the origins of exceptionally diverse extant groups within mammals, squamates, amphibians, and dinosaurs. Therefore, although speciose groups of modern tetrapods have Mesozoic origins, rates of Mesozoic diversification inferred from the fossil record are slow compared to those inferred from molecular phylogenies. If high speciation rates did occur in the Mesozoic, then they seem to have been balanced by extinctions among older clades. An apparent 4-fold expansion of species richness after the Cretaceous/Paleogene (K/Pg) boundary deserves further examination in light of potential taxonomic biases, but is consistent with the hypothesis that global environmental disturbances such as mass extinction events can rapidly adjust limits to diversity by restructuring ecosystems, and suggests that the gradualistic evolutionary diversification of tetrapods was punctuated by brief but dramatic episodes of radiation.


Predictors of decline in CD4 lymphocytes in a cohort of homosexual men infected with human immunodeficiency virus.
Longitudinal data on four visits scheduled at 6 month intervals were available on a cohort of 1,827 homosexual men who were human immunodeficiency virus (HIV) seropositive at entry. To identify predictors of the rate of decline of CD4 T-lymphocytes, we used an autoregressive model that relates CD4 counts to predictor variables, while adjusting for previous CD4 counts. Significant predictors of steeper decline of CD4 counts were high CD8 count, low hemoglobin, low platelets, high serum IgA, high cytomegalovirus (CMV) antibody, and low HIV antibody. Using the fitted model, a subject with an initial deficit of 314 CD4 cells (median value of study sample) with respect to seronegative subjects and with average values in all other predictors is estimated to lose approximately 53 cells in a 6 month period (95% C.I. = 45-61 cells). Contrasting this estimate to the one obtained with similar methods in intravenous drug users, it is suggested that a faster rate of decline is present among i.v. drug users. This analysis provides evidence that several covariates in addition to previous number of CD4 counts have significant predictive power for estimating the decline in CD4 counts in HIV seropositive subjects.


In pursuit of formaldehyde: causally explanatory models and falsification.
Falsification no longer is the cornerstone of philosophy of science; but it still looms widely that scientists ought to drop an explanatory hypothesis in view of negative results. We shall argue that, to the contrary, negative empirical results are unable to disqualify causally explanatory hypotheses-not because of the shielding effect of auxiliary assumptions but because of the fact that the causal irrelevance of a factor cannot empirically be established. This perspective is elaborated at a case study taken from the history of plant physiology: the formaldehyde model of photosynthesis, which for about sixty years (1870s to 1930s) dominated the field-despite the fact that in these sixty years all the attempts to conclusively demonstrate even the presence of formaldehyde in plants failed.


The 1994 annual lecture of the Society for the Study of Addiction. Commissions (Royal and other) on drug misuse: who needs them?
In the context of current calls in the United Kingdom for a Royal Commission on drugs, the political antecedents and consequences of nine major commissions of enquiry into drugs established by governments in the English-speaking world during the last 100 years are described, together with available information on their modus operandi and costs. The different interest groups calling for reform of UK drug laws are described and the broad basis on which a putative Royal Commission might proceed is suggested. Finally, analysis of previous commissions is used to identify factors likely to lead to a new Commission having a substantial influence on government policy on drugs.


Response of the lower respiratory tract to injury. Mechanisms of repair of the parenchymal cells of the alveolar wall.
Although the lower respiratory tract is frequently exposed to injurious agents, the lung does possess some ability to effect repair and thus restore the damaged alveolar wall to normal; however, in some circumstances, normal repair is not possible. The result is often a markedly deranged alveolus, with improper proportions of epithelial cells (eg, relatively more cuboidal type-2-like cells), a loss of endothelial cells or migration of endothelial cells into improper locations, and a proliferation of interstitial fibroblasts with an accompanying deposition of a collagenous extracellular matrix (ie, fibrosis). Although the development of "fibrosis" is frequently thought to be a form of attempted "repair" of an injured alveolar wall, this concept is not clearly established; it is possible that the expansion of fibroblastic numbers in the alveolar wall is part of the disease process itself, resulting from alveolar macrophagic activation, rather than an attempt by the macrophage to "repair" an injured alveolar wall. Thus, it is not known if the development of fibrosis represents "healing" and thus is beneficial (as a localized scar "heals" a localized incision in the skin) or whether it represents part of the disease process itself. The distinction is important, as it is unclear whether therapy should be directed against the development of fibrosis per se. If fibroblastic expansion and deposition of the connective tissue products of these fibroblasts are a useful form of repair, prevention of this process may cause future loss of pulmonary function. Alternatively, if "fibrosis" compromises pulmonary function (particularly decreased compliance), prevention of fibrosis might be beneficial. It is apparent, therefore, that what is needed is an understanding of the processes that lead to alveolar parenchymal cellular repair and how such processes might be manipulated for the benefit of the patient.


A fully organic retinal prosthesis restores vision in a rat model of degenerative blindness.
The degeneration of photoreceptors in the retina is one of the major causes of adult blindness in humans. Unfortunately, no effective clinical treatments exist for the majority of retinal degenerative disorders. Here we report on the fabrication and functional validation of a fully organic prosthesis for long-term in vivo subretinal implantation in the eye of Royal College of Surgeons rats, a widely recognized model of retinitis pigmentosa. Electrophysiological and behavioural analyses reveal a prosthesis-dependent recovery of light sensitivity and visual acuity that persists up to 6-10 months after surgery. The rescue of the visual function is accompanied by an increase in the basal metabolic activity of the primary visual cortex, as demonstrated by positron emission tomography imaging. Our results highlight the possibility of developing a new generation of fully organic, highly biocompatible and functionally autonomous photovoltaic prostheses for subretinal implants to treat degenerative blindness.


Faster Adaptation in Smaller Populations: Counterintuitive Evolution of HIV during Childhood Infection.
Analysis of HIV-1 gene sequences sampled longitudinally from infected individuals can reveal the evolutionary dynamics that underlie associations between disease outcome and viral genetic diversity and divergence. Here we extend a statistical framework to estimate rates of viral molecular adaptation by considering sampling error when computing nucleotide site-frequencies. This is particularly beneficial when analyzing viral sequences from within-host viral infections if the number of sequences per time point is limited. To demonstrate the utility of this approach, we apply our method to a cohort of 24 patients infected with HIV-1 at birth. Our approach finds that viral adaptation arising from recurrent positive natural selection is associated with the rate of HIV-1 disease progression, in contrast to previous analyses of these data that found no significant association. Most surprisingly, we discover a strong negative correlation between viral population size and the rate of viral adaptation, the opposite of that predicted by standard molecular evolutionary theory. We argue that this observation is most likely due to the existence of a confounding third variable, namely variation in selective pressure among hosts. A conceptual non-linear model of virus adaptation that incorporates the two opposing effects of host immunity on the virus population can explain this counterintuitive result.


CutL: an alternative to Kulldorff's scan statistics for cluster detection with a specified cut-off level.
When searching for epidemiological clusters, an important tool can be to carry out one's own research with the incidence rate from the literature as the reference level. Values exceeding this level may indicate the presence of a cluster in that location. This paper presents a method of searching for clusters that have significantly higher incidence rates than those specified by the investigator. The proposed method uses the classic binomial exact test for one proportion and an algorithm that joins areas with potential clusters while reducing the number of multiple comparisons needed. The sensitivity and specificity are preserved by this new method, while avoiding the Monte Carlo approach and still delivering results comparable to the commonly used Kulldorff's scan statistics and other similar methods of localising clusters. A strong contributing factor afforded by the statistical software that makes this possible is that it allows analysis and presentation of the results cartographically.


Theaflavin derivatives in black tea and catechin derivatives in green tea inhibit HIV-1 entry by targeting gp41.
Theaflavin derivatives and catechin derivatives are the major polyphenols in black tea and green tea, respectively. Several tea polyphenols, especially those with galloyl moiety, can inhibit HIV-1 replication with multiple mechanisms of action. Here we showed that the theaflavin derivatives had more potent anti-HIV-1 activity than catechin derivatives. These tea polyphenols could inhibit HIV-1 entry into target cells by blocking HIV-1 envelope glycoprotein-mediated membrane fusion. The fusion inhibitory activity of the tea polyphenols was correlated with their ability to block the formation of the gp41 six-helix bundle, a fusion-active core conformation. Computer-aided molecular docking analyses indicate that these tea polyphenols, theaflavin-3,3'-digallate (TF3) as an example, may bind to the highly conserved hydrophobic pocket on the surface of the central trimeric coiled coil formed by the N-terminal heptad repeats of gp41. These results indicate that tea, especially black tea, may be used as a source of anti-HIV agents and theaflavin derivatives may be applied as lead compounds for developing HIV-1 entry inhibitors targeting gp41.


Fear Avoidance Beliefs and Risk of Long-Term Sickness Absence: Prospective Cohort Study among Workers with Musculoskeletal Pain.
Musculoskeletal pain is common in the population. Negative beliefs about musculoskeletal pain and physical activity may lead to avoidance behavior resulting in absence from work. The present study investigates the influence of fear avoidance beliefs on long-term sickness absence. Workers of the general working population with musculoskeletal pain (low back, neck/shoulder, and/or arm/hand pain; n = 8319) from the Danish Work Environment Cohort Study were included. Long-term sickness absence data were obtained from the Danish Register for Evaluation and Marginalization (DREAM). Time-to-event analyses (cox regression) controlled for various confounders estimated the association between fear avoidance beliefs (very low, low, moderate [reference category], high, and very high) at baseline and long-term sickness absence (LTSA; ≥6 consecutive weeks) during a 2-year follow-up. During the 2-year follow-up, 10.2% of the workers experienced long-term sickness absence. In the fully adjusted model, very high-level fear avoidance increased the risk of LTSA with hazard ratio (HR) of 1.48 (95% CI 1.15-1.90). Similar results were seen analyses stratified for occupational physical activity, i.e., sedentary workers (HR 1.72 (95% CI 1.04-2.83)) and physically active workers (HR 1.48 (95% CI 1.10-2.01)). A very high level of fear avoidance is a risk factor for long-term sickness absence among workers with musculoskeletal pain regardless of the level of occupational physical activity. Future interventions should target fear avoidance beliefs through information and campaigns about the benefits of staying active when having musculoskeletal pain.


[PCR and ELISA--in vitro alternatives to the mouse-bioassay for assessing the botulinum-neurotoxin-C1 production potential in environmental samples?].
Botulism is one of the most important bird diseases world-wide and is caused by the intoxication with Botulinum-Neurotoxin-C1 (BoNt-C1), which is produced by toxigenic clostridia under appropriate conditions. Avian botulism leads regularly to large losses among the migrating bird populations breeding and resting at the saltwater pools of the Austrian national park Neusiedler See-Seewinkel. Despite of its ethical dubiousness and its high technical expense the mouse-bioassay is still used as the routine standard method for the detection of BoNt-C1. According to the 3R-concept, in vitro alternative methods for the qualitative detection of BoNt-C1 (immunostick-ELISA) and a corresponding BoNt-C1 gene fragment (nested-PCR) were established. In order to estimate the BoNt-C1 production potential the methods were tested with sediment samples from different saltwater pools subjected to cultivation conditions appropriate for in vitro BoNt-C1-production. With the mouse-bioassay, 52 out of 77 samples were found to have a positive toxin production potential. The immunostick-ELISA showed a similar sensitivity as the mouse-bioassay and exhibited a highly significant positive correlation (r=0.94; p<0.001) with the mouse-bioassay in detecting BoNt-C1. The nested-PCR approach revealed higher numbers of positive BoNt-C1 gene fragment detections as compared to the direct toxin analysis approaches. A weak correlation (r=0.21; p=0.07) with the mouse-bioassay was discernible, no correlation was found with the immunostick-ELISA (r=0.09; p=0.46). Obviously, the PCR approach detected the BoNt-C1 gene fragment in some of the samples where no toxin expression has occurred. Thus it is suggested that the qualitative immunostick-ELISA represents a potential in-vitro alternative to the mouse-bioassay for assessing the BoNt-C1 production potential in environmental samples. In contrast, qualitative BoNt-C1 gene fragment detection via PCR led to an overestimation of the actual toxin production potential.


Gore Iliac Branch Endoprosthesis for treatment of bilateral common iliac artery aneurysms.
The Gore Iliac Branch Endoprosthesis (IBE; W. L. Gore & Associates, Flagstaff, Ariz) has recently been approved by the Food and Drug Administration for treatment of common iliac artery (CIA) aneurysms. Despite early excellent results in clinical trial, none of 63 patients were treated for bilateral iliac aneurysms. The goal of this study was to examine real-world experience using the Gore IBE for bilateral CIA aneurysms. A retrospective review of an international multicenter (16 U.S., 8 European) experience using the Gore IBE to treat bilateral CIA aneurysms was performed. Cases were limited to those occurring after Food and Drug Administration approval (February 2016) in the United States and after CE mark approval (November 2013) in Europe. Demographics of the patients, presentation, anatomic characteristics, and procedural details were captured. There were 47 patients (45 men; mean age, 68 years; range, 41-84 years) treated with bilateral Gore IBEs (27 U.S., 20 European). Six patients (12.7%) were symptomatic and 12 (25.5%) patients were treated primarily for CIA aneurysm (aorta <5.0 cm). Mean CIA diameter was 40.3 mm. Four patients had aneurysmal internal iliac arteries (IIAs). Two of these were sealed proximally at the IIA aneurysm neck and two required coil embolization of IIA branches to achieve seal in the largest first-order branches. Technical success was achieved in 46 patients (97.9%). No type I or type III endoleaks were noted. There was no significant perioperative morbidity or mortality. IIA branch adjunctive stenting was required in four patients (one IIA distal dissection, three kinks). On follow-up imaging available for 40 patients (85.1%; mean, 6.5 months; range, 1-36 months), 12 type II endoleaks (30%) and no type I or type III endoleaks were detected. Two of 80 (2.5%) IIA branches imaged were occluded; one was intentionally sacrificed perioperatively. Preservation of bilateral IIAs in repair of bilateral CIA aneurysms can be performed safely with excellent technical success and short-term patency rates using the Gore IBE device. Limb and branch occlusions are rare, usually are due to kinking, and can almost always be treated successfully with stenting.


Engineering Graphene Grain Boundaries for Plasmonic Multi-Excitation and Hotspots.
Surface plasmons, merging photonics and electronics in nanoscale dimensions, have been the cornerstones in integrated informatics, precision detection, high-resolution imaging, and energy conversion. Arising from the exceptional Fermi-Dirac tunability, ultrafast carrier mobility, and high-field confinement, graphene offers excellent advantages for plasmon technologies and enables a variety of state-of-the-art optoelectronic applications ranging from tight-field-enhanced light sources, modulators, and photodetectors to biochemical sensors. However, it is challenging to co-excite multiple graphene plasmons on one single graphene sheet with high density, a key step toward plasmonic wavelength-division multiplexing and next-generation dynamical optoelectronics. Here, we report the heteroepitaxial growth of a polycrystalline graphene monolayer with patterned gradient grain boundary density, which is synthesized by creating diverse nanosized local growth environments on a centimeter-scale substrate with a polycrystalline graphene ring seed in chemical vapor deposition. Such geometry enables plasmonic co-excitation with varied wavelength diversification in the nanoscale. Via using high-resolution scanning near-field optical microscopy, we demonstrate rich plasmon standing waves, even bright plasmonic hotspots with a size up to 3 μm. Moreover, by changing the grain boundary density and annealing, we find the local plasmonic wavelengths are widely tunable, from 70 to 300 nm. Theoretical modeling supports that such plasmonic versatility is due to the grain boundary-induced plasmon-phonon interactions through random phase approximation. The seed-induced heteroepitaxial growth provides a promising way for the grain boundary engineering of two-dimensional materials, and the controllable grain boundary-based plasmon co-generation and manipulation in one single graphene monolayer will facilitate the applications of graphene for plasmonics and nanophotonics.


Hyperglucagonemia and insulin-mediated glucose metabolism.
The effect of chronic physiologic hyperglucagonemia on basal and insulin-mediated glucose metabolism was evaluated in normal subjects, using the euglycemic insulin clamp technique (+50, +100, and +500 microU/ml). After glucagon infusion fasting glucose increased from 76 +/- 4 to 93 +/- 2 mg/dl and hepatic glucose production (HGP) rose from 1.96 +/- 0.08 to 2.25 +/- 0.08 mg/kg X min (P less than 0.001). Basal glucose oxidation after glucagon increased (P less than 0.05) and correlated inversely with decreased free fatty acid concentrations (r = -0.94; P less than 0.01) and decreased lipid oxidation (r = -0.75; P less than 0.01). Suppression of HGP and stimulation of total glucose disposal were impaired at each insulin step after glucagon (P less than 0.05-0.01). The reduction in insulin-mediated glucose uptake was entirely due to diminished non-oxidative glucose utilization. Glucagon infusion also caused a decrease in basal lipid oxidation and an enhanced ability of insulin to inhibit lipid oxidation and augment lipid synthesis. These results suggest that hyperglucagonemia may contribute to the disturbances in glucose and lipid metabolism in some diabetic patients.


Rapid onset of neuronal death induced by blockade of either axoplasmic transport or action potentials in afferent fibers during brain development.
We have investigated how neurons in the optic tecta of embryonic day 16 chick embryos depend for survival on their afferents from the retina. To distinguish between activity-mediated effects and other, "trophic," ones, we compared the effects on the tectal neurons of blocking intraocular axoplasmic transport (with colchicine) or action potentials (by means of TTX). Both interventions rapidly induced the appearance of dying (pyknotic) neurons in the tectum, with major increases in their number occurring within 13 hr post-colchicine and within 9 hr post-TTX. Following both drugs, the dying neurons were morphologically similar, and in both cases the cell death depended on protein synthesis. However, the effects of colchicine and of TTX could be dissociated, since the most superficial tectal neurons became pyknotic only in response to colchicine, and, with a sufficiently short survival time (9 hr), the deep cells of the stratum griseum centrale became pyknotic only in response to TTX. We hence argue that the survival of the tectal neurons depends on their ongoing maintenance by substances released from retinotectal axon terminals, the release being activity dependent in the case of the deep neurons but independent of activity in the case of the superficial ones.


Guidelines for the diagnosis of leukemia or lymphoma in children.
Our progress in understanding and treating pediatric acute leukemia and lymphoma is one of the success stories of cancer biology and management. Event-free survival for children with acute leukemia or lymphoma has improved progressively during the past four decades. The advances in the cell and molecular biology of acute leukemia and lymphoma that have been made can help determine both prognosis and therapy; however, the pathologic evaluation of bone marrow and lymph node specimens must be directed appropriately to benefit from these advances. We present guidelines for the pathologic evaluation of bone marrow and lymph node specimens to maximize the chance that each patient will benefit from our increased understanding of these diseases.


Development of six PROMIS pediatrics proxy-report item banks.
Pediatric self-report should be considered the standard for measuring patient reported outcomes (PRO) among children. However, circumstances exist when the child is too young, cognitively impaired, or too ill to complete a PRO instrument and a proxy-report is needed. This paper describes the development process including the proxy cognitive interviews and large-field-test survey methods and sample characteristics employed to produce item parameters for the Patient Reported Outcomes Measurement Information System (PROMIS) pediatric proxy-report item banks. The PROMIS pediatric self-report items were converted into proxy-report items before undergoing cognitive interviews. These items covered six domains (physical function, emotional distress, social peer relationships, fatigue, pain interference, and asthma impact). Caregivers (n = 25) of children ages of 5 and 17 years provided qualitative feedback on proxy-report items to assess any major issues with these items. From May 2008 to March 2009, the large-scale survey enrolled children ages 8-17 years to complete the self-report version and caregivers to complete the proxy-report version of the survey (n = 1548 dyads). Caregivers of children ages 5 to 7 years completed the proxy report survey (n = 432). In addition, caregivers completed other proxy instruments, PedsQL™ 4.0 Generic Core Scales Parent Proxy-Report version, PedsQL™ Asthma Module Parent Proxy-Report version, and KIDSCREEN Parent-Proxy-52. Item content was well understood by proxies and did not require item revisions but some proxies clearly noted that determining an answer on behalf of their child was difficult for some items. Dyads and caregivers of children ages 5-17 years old were enrolled in the large-scale testing. The majority were female (85%), married (70%), Caucasian (64%) and had at least a high school education (94%). Approximately 50% had children with a chronic health condition, primarily asthma, which was diagnosed or treated within 6 months prior to theinterview. The PROMIS proxy sample scored similar or better on the other proxy instruments compared to normative samples. The initial calibration data was provided by a diverse set of caregivers of children with a variety of common chronic illnesses and racial/ethnic backgrounds. The PROMIS pediatric proxy-report item banks include physical function (mobility n = 23; upper extremity n = 29), emotional distress (anxiety n = 15; depressive symptoms n = 14; anger n = 5), social peer relationships (n = 15), fatigue (n = 34), pain interference (n = 13), and asthma impact (n = 17).


Interposition of a connective tissue graft or a collagen matrix to enhance wound stability - an experimental study in dogs.
The aim of this study was to evaluate the role of a connective tissue graft (CTG) or a collagen matrix (CM) interpositioned between flaps and non-shedding hard surfaces on wound stability. Sixty bone dehiscence defects were prepared in five Beagle dogs. Three treatments were performed in 12 sites per dog: (1) repositioned flaps were sutured onto instrumented dentin surfaces (control), (2) repositioning of flaps with an interpositioned CTG and (3) repositioning of flaps with the application of a CM. To allow postoperative healing with n = 5 for 1, 3, 7 and 14 days before evaluation, the sutures were removed, incision lines retraced and tensile forces applied to the flaps. The minimum magnitude of forces required to detach the flaps from the wound bed was recorded. After 1 week of healing, 6 N had to be applied to disrupt flaps from their wound bed in the CTG group. In the control group, a similar magnitude of resistance was achieved after 2 weeks (6.1 N). Flap resistance to tearing was highest in the CTG group (maximum 9.1 N) 2 weeks postoperatively. On the third postoperative day, the mean tearing forces of all groups differed significantly, displaying a 50% lower resistance to tearing in the CM compared to the CTG group. In comparison, flap resistance to tearing forces established earlier and in higher magnitude in sites with an interpositioned CTG than in flaps repositioned on dentin or CM. Application of a CTG, sutured to a non-shedding hard surface, significantly increased flap resistance to tearing when applying disrupting forces compared to controls. A less pronounced effect was achieved by interpositioning of a CM.


Dynamic Glycosylation Governs the Vertebrate COPII Protein Trafficking Pathway.
The COPII coat complex, which mediates secretory cargo trafficking from the endoplasmic reticulum, is a key control point for subcellular protein targeting. Because misdirected proteins cannot function, protein sorting by COPII is critical for establishing and maintaining normal cell and tissue homeostasis. Indeed, mutations in COPII genes cause a range of human pathologies, including cranio-lenticulo-sutural dysplasia (CLSD), which is characterized by collagen trafficking defects, craniofacial abnormalities, and skeletal dysmorphology. Detailed knowledge of the COPII pathway is required to understand its role in normal cell physiology and to devise new treatments for disorders in which it is disrupted. However, little is known about how vertebrates dynamically regulate COPII activity in response to developmental, metabolic, or pathological cues. Several COPII proteins are modified by O-linked β-N-acetylglucosamine (O-GlcNAc), a dynamic form of intracellular protein glycosylation, but the biochemical and functional effects of these modifications remain unclear. Here, we use a combination of chemical, biochemical, cellular, and genetic approaches to demonstrate that site-specific O-GlcNAcylation of COPII proteins mediates their protein-protein interactions and modulates cargo secretion. In particular, we show that individual O-GlcNAcylation sites of SEC23A, an essential COPII component, are required for its function in human cells and vertebrate development, because mutation of these sites impairs SEC23A-dependent in vivo collagen trafficking and skeletogenesis in a zebrafish model of CLSD. Our results indicate that O-GlcNAc is a conserved and critical regulatory modification in the vertebrate COPII-dependent trafficking pathway.


Bovine serum albumin adsorption to iron-oxide coated sands can change microsphere deposition mechanisms.
Particulate colloids often occur together with proteins in sewage-impacted water. Using Bovine Serum Albumin (BSA) as a surrogate for protein in sewage, column experiments investigating the capacity of iron-oxide coated sands to remove latex microspheres from water revealed that microsphere attenuation mechanisms depended on antecedent BSA coverage. Dual pulse experiment (DPE) results suggested that where all BSA was adsorbed, subsequent multiple pore volume microsphere breakthrough curves reflected progressively reduced colloid deposition rates with increasing adsorbed BSA content. Modeling colloid responses suggested adsorption of 1 μg BSA generated the same response as blockage by between 7.1 × 10(8) and 2.3 × 10(9) deposited microspheres. By contrast, microsphere responses in DPEs where BSA coverage of the deposition sites approached/reached saturation revealed the coated sand maintained a finite capacity to attenuate microspheres, even when incapable of further BSA adsorption. Subsequent microsphere breakthrough curves demonstrated the matrix's colloid attenuation capacity progressively increased with continued microsphere deposition. Experimental findings suggested BSA adsorption on the sand surface approaching/reaching saturation generated attractive deposition sites for colloids, which became progressively more attractive with further colloid deposition (filter ripening). Results demonstrate that adsorption of a single type of protein may either enhance or inhibit colloid mobility in saturated porous media.


Universal Surface Engineering of Transition Metals for Superior Electrocatalytic Hydrogen Evolution in Neutral Water.
The development of low-cost hybrid water splitting-biosynthetic systems that mimic natural photosynthesis to achieve solar-to-chemical conversion is of great promise for future energy demands, but often limited by the kinetically sluggish hydrogen evolution reaction (HER) on the surface of nonprecious transition metal catalysts in neutral media. It is thus highly desirable to rationally tailor the reaction interface to boost the neutral HER catalytic kinetics. Herein, we report a general surface nitrogen modification of diverse transition metals (e.g., iron, cobalt, nickel, copper, and nickel-cobalt alloy), accomplished by a facile low-temperature ammonium carbonate treatment, for significantly improved hydrogen generation from neutral water. Various physicochemical characterization techniques including synchrotron X-ray absorption spectroscopy (XAS) and theory modeling demonstrate that the surface nitrogen modification does not change the chemical composition of the underlying transition metals. Notably, the resulting nitrogen-modified nickel framework (N-Ni) exhibits an extremely low overpotential of 64 mV at 10 mA cm-2, which is, to our knowledge, the best among those nonprecious electrocatalysts reported for hydrogen evolution at pH 7. Our combined experimental results and density functional theory (DFT) calculations reveal that the surface electron-rich nitrogen simultaneously facilitates the initial adsorption of water via the electron-deficient H atom and the subsequent dissociation of the electron-rich HO-H bond via H transfer to N on the nickel surface, beneficial to the overall hydrogen evolution process.


In Situ One-Step Fluorescence Labeling Strategy of Exosomes via Bioorthogonal Click Chemistry for Real-Time Exosome Tracking In Vitro and In Vivo.
Exosomes are cellular components with promising uses in cancer diagnostics and therapeutics, and their imaging and tracking are essential to study their biological properties. Herein, we report on an in situ one-step fluorescence labeling strategy for exosomes via bioorthogonal click chemistry. First, exosome donor cancer cells were treated with tetraacetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) to generate unnatural azide groups (-N3) on their surface via metabolic glycoengineering. Then, the azide groups were labeled with near-infrared fluorescent dye-conjugated dibenzylcyclooctyne (DBCO-Cy5) via bioorthogonal click chemistry. After 2 days of incubation, the DBCO-Cy5-labeled exosomes (Cy5-Exo) were successfully secreted from the donor cancer cells and were isolated via classical ultracentrifugation, providing a high-yield of fluorescent dye-labeled exosomes. This in situ one-step bioorthogonal click chemistry offers improved labeling efficiency, biocompatibility, and imaging sensitivy compared to standard exosomes (ST-Exo), purified with classical ultracentrifugation or carbocyanine lipophilic dye (DiD)-labeled exosomes (DiD-Exo) in vitro. In particular, the Cy5-Exo were successfully taken up by A549 cells in a time-dependent manner, and they could escape from lysosome confinement, showing their possible use as a delivery carrier of therapeutic drugs or imaging agents. Finally, intraveneously injected Cy5-Exo were noninvasively tracked and imaged via near-infrared fluorescence (NIRF) imaging in tumor-bearing mice. This new fluorescence labeling strategy for natural exosomes may be useful to provide better understanding of their theranostic effects in many biomedical applications.


PRC1-labeled microtubule bundles and kinetochore pairs show one-to-one association in metaphase.
In the mitotic spindle, kinetochore microtubules form k-fibers, whereas overlap or interpolar microtubules form antiparallel arrays containing the cross-linker protein regulator of cytokinesis 1 (PRC1). We have recently shown that an overlap bundle, termed bridging fiber, links outermost sister k-fibers. However, the relationship between overlap bundles and k-fibers throughout the spindle remained unknown. Here, we show that in a metaphase spindle more than 90% of overlap bundles act as a bridge between sister k-fibers. We found that the number of PRC1-GFP-labeled bundles per spindle is nearly the same as the number of kinetochore pairs. Live-cell imaging revealed that kinetochore movement in the equatorial plane of the spindle is highly correlated with the movement of the coupled PRC1-GFP-labeled fiber, whereas the correlation with other fibers decreases with increasing distance. Analysis of endogenous PRC1 localization confirmed the results obtained with PRC1-GFP PRC1 knockdown reduced the bridging fiber thickness and interkinetochore distance throughout the spindle, suggesting a function of PRC1 in bridging microtubule organization and force balance in the metaphase spindle.


Molecular phylogenetics reveal multiple tertiary vicariance origins of the African rain forest trees.
Tropical rain forests are the most diverse terrestrial ecosystems on the planet. How this diversity evolved remains largely unexplained. In Africa, rain forests are situated in two geographically isolated regions: the West-Central Guineo-Congolian region and the coastal and montane regions of East Africa. These regions have strong floristic affinities with each other, suggesting a former connection via an Eocene pan-African rain forest. High levels of endemism observed in both regions have been hypothesized to be the result of either 1) a single break-up followed by a long isolation or 2) multiple fragmentation and reconnection since the Oligocene. To test these hypotheses the evolutionary history of endemic taxa within a rain forest restricted African lineage of the plant family Annonaceae was studied. Molecular phylogenies and divergence dates were estimated using a Bayesian relaxed uncorrelated molecular clock assumption accounting for both calibration and phylogenetic uncertainties. Our results provide strong evidence that East African endemic lineages of Annonaceae have multiple origins dated to significantly different times spanning the Oligocene and Miocene epochs. Moreover, these successive origins (c. 33, 16 and 8 million years--Myr) coincide with known periods of aridification and geological activity in Africa that would have recurrently isolated the Guineo-Congolian rain forest from the East African one. All East African taxa were found to have diversified prior to Pleistocene times. Molecular phylogenetic dating analyses of this large pan-African clade of Annonaceae unravels an interesting pattern of diversification for rain forest restricted trees co-occurring in West/Central and East African rain forests. Our results suggest that repeated reconnections between the West/Central and East African rain forest blocks allowed for biotic exchange while the break-ups induced speciation via vicariance, enhancing the levels of endemicity. These results provide an explanation for present day distribution patterns and origins of endemicity for African rain forest trees. Moreover, given the pre-Pleistocene origins of all the studied endemic East African genera and species, these results also offer important insights for setting conservation priorities in these highly diversified but threatened ecosystems.


Systematic studies of all PIH proteins in zebrafish reveal their distinct roles in axonemal dynein assembly.
Construction of motile cilia/flagella requires cytoplasmic preassembly of axonemal dyneins before transport into cilia. Axonemal dyneins have various subtypes, but the roles of each dynein subtype and their assembly processes remain elusive in vertebrates. The PIH protein family, consisting of four members, has been implicated in the assembly of different dynein subtypes, although evidence for this idea is sparse. Here, we established zebrafish mutants of all four PIH-protein genes: pih1d1, pih1d2, ktu, and twister, and analyzed the structures of axonemal dyneins in mutant spermatozoa by cryo-electron tomography. Mutations caused the loss of specific dynein subtypes, which was correlated with abnormal sperm motility. We also found organ-specific compositions of dynein subtypes, which could explain the severe motility defects of mutant Kupffer's vesicle cilia. Our data demonstrate that all vertebrate PIH proteins are differently required for cilia/flagella motions and the assembly of axonemal dyneins, assigning specific dynein subtypes to each PIH protein.


Accuracy of the actuator disc-RANS approach for predicting the performance and wake of tidal turbines.
The actuator disc-RANS model has widely been used in wind and tidal energy to predict the wake of a horizontal axis turbine. The model is appropriate where large-scale effects of the turbine on a flow are of interest, for example, when considering environmental impacts, or arrays of devices. The accuracy of the model for modelling the wake of tidal stream turbines has not been demonstrated, and flow predictions presented in the literature for similar modelled scenarios vary significantly. This paper compares the results of the actuator disc-RANS model, where the turbine forces have been derived using a blade-element approach, to experimental data measured in the wake of a scaled turbine. It also compares the results with those of a simpler uniform actuator disc model. The comparisons show that the model is accurate and can predict up to 94 per cent of the variation in the experimental velocity data measured on the centreline of the wake, therefore demonstrating that the actuator disc-RANS model is an accurate approach for modelling a turbine wake, and a conservative approach to predict performance and loads. It can therefore be applied to similar scenarios with confidence.


An open trial of OKT3 in patients with multiple sclerosis.
We report our experience with treatment with Muromonab-CD3 (Orthoclone OKT3) of 16 patients with multiple sclerosis (MS) who were in a progressive phase of their disease (n = 13) or in an acute severe attack lasting longer than 1 month without recovery (n = 3). We induced acute severe T-cytopenia with OKT3. Fifteen patients completed treatment for 10 days. Side effects were common and severe and included hypotension, nausea and vomiting, diarrhea, fever, and myalgia. In two of two patients tested, there was a transient though major rise in the levels of interferon gamma and tumor necrosis factor in the first 12 hours of treatment. Nonetheless, we did not detect new clinical or MRI activity of MS during the period of treatment, although many patients deteriorated transiently in disability scores. At the conclusion of follow-up, only four patients had deteriorated by 1.0 or more points on the Expanded Disability Status Scale of Kurtzke (EDSS) (73% stabilization rate). Of those patients who deteriorated, two died of complications of MS (EDSS 10). Only two patients had clinical improvement at 1 year follow-up. The attendant toxicity of OKT3 makes it unlikely that it will play a major role in the treatment of MS.


Immune-like phagocyte activity in the social amoeba.
Social amoebae feed on bacteria in the soil but aggregate when starved to form a migrating slug. We describe a previously unknown cell type in the social amoeba, which appears to provide detoxification and immune-like functions and which we term sentinel (S) cells. S cells were observed to engulf bacteria and sequester toxins while circulating within the slug, eventually being sloughed off. A Toll/interleukin-1 receptor (TIR) domain protein, TirA, was also required for some S cell functions and for vegetative amoebae to feed on live bacteria. This apparent innate immune function in social amoebae, and the use of TirA for bacterial feeding, suggest an ancient cellular foraging mechanism that may have been adapted to defense functions well before the diversification of the animals.