Fully endoscopic lumbar spinal surgery: Is it time to change?

Spinal surgery is becoming safer and progressively less invasive with advances in optical and instrument technology. The amount of decompression achievable with endoscopic approaches is now comparable to open approaches with the advantage of much less tissue trauma. This review aims at examining the status of endoscopic approaches in lumbar decompressive surgery.

Fully endoscopic cervical spine surgery: What does the future hold?

Fully endoscopic cervical spine surgery is an emerging novel approach to address cervical spinal pathology. Techniques, both anterior and posterior have been adapted to address various cervical pathologies. The primary goal of these procedures like other open techniques is to surgically decompress the canal centrally and/or along the foramen. The narrative review aims to provide the reader an overview of the rapidly advancing field of endoscopic cervical spinal surgery and evaluate whether these newer approaches could potentially reduce the cost and the risk associated with instrumented cervical fusion.

Carvacrol attenuates Campylobacter jejuni colonization factors and proteome critical for persistence in the chicken gut.

Campylobacter jejuni is a major foodborne pathogen that causes gastroenteritis in humans. Chickens act as the reservoir host for C. jejuni, wherein the pathogen asymptomatically colonizes the ceca leading to contamination of carcasses during slaughter. The major colonization factors in C. jejuni include motility, intestinal epithelial attachment, acid/bile tolerance, and quorum sensing. Reducing the expression of the aforementioned factors could potentially reduce C. jejuni colonization in chickens. This study investigated the efficacy of subinhibitory concentration (SIC; compound concentration not inhibiting bacterial growth) of carvacrol in reducing the expression of C. jejuni colonization factors in vitro. Moreover, the effect of carvacrol on the expression of C. jejuni proteome was investigated using liquid chromatography-tandem mass spectrometry. The motility assay was conducted at 42°C, and the motility zone was measured after 24 h of incubation. For the adhesion assay, monolayers of primary chicken enterocytes (∼105 cells/well) were inoculated with C. jejuni (6 log cfu/well) either in the presence or absence of carvacrol, and the adhered C. jejuni were enumerated after 90 min of incubation at 42°C. The effect of carvacrol on C. jejuni quorum sensing and susceptibility to acid/bile stress was investigated using a bioluminescence assay and an acid-bile survival assay, respectively. The SIC (0.002%) of carvacrol reduced the motility of C. jejuni strains S-8 and NCTC 81-176 by ∼50 and 35%, respectively (P < 0.05). Carvacrol inhibited C. jejuni S-8 and NCTC 81-176 adhesion to chicken enterocytes by ∼0.8 and 1.5 log cfu/mL, respectively (P < 0.05). Moreover, carvacrol reduced autoinducer-2 activity and increased the susceptibility of C. jejuni to acid and bile in both the strains (P < 0.05). Liquid chromatography-tandem mass spectrometry revealed that the SIC of carvacrol reduced the expression of selected C. jejuni colonization proteins critical for motility (methyl-accepting chemotaxis protein), adhesion (GroL), growth and metabolism (AspA, AcnB, Icd, Fba, Ppa, AnsA, Ldh, Eno, PurB-1), and anaerobic respiration (NapB, HydB, SdhA, NrfA) (P < 0.05). Results suggest the mechanisms by which carvacrol could reduce C. jejuni colonization in chickens.

Spinal cord injury-The role of surgical treatment for neurological improvement.

Acute spinal cord injury (ASCI) is common and no consensuses have been reached regarding timing of surgical decompression. This article highlights the main issues regarding surgical management of ASCI patients. The importance of timing of surgery along with physiological stability of the cord, and indications for surgery has been discussed to facilitate better understanding of the condition. The importance of the type of injury to the spinal column, besides the cord injury, is also discussed. A brief review of relevant literature has been done to try and answer the question whether early or late surgical treatment for ASCI is better than conservative management, reflecting the ethos of treatment for these problems in Robert Jones And Agnes Hunt Orthopaedic Hospital in Oswestry.

Review of MRSA screening and antibiotics prophylaxis in orthopaedic trauma patients; The risk of surgical site infection with inadequate antibiotic prophylaxis in patients colonized with MRSA.

AIMS: The primary aim of this study was to determine whether orthopaedic trauma patients receive appropriate antibiotic prophylaxis keeping in view the results of their MRSA screening. The secondary aim was to analyse the risk of developing MRSA surgical site infection with and without appropriate antibiotic prophylaxis in those colonized with MRSA. PATIENTS AND METHODS: We reviewed 400 consecutive orthopaedic trauma patient episodes. Preoperative MRSA screening results, operative procedures, prophylactic antibiotics and postoperative course were explored. In addition to these consecutive patients, the hospital MRSA database over the previous 5 years identified 27 MRSA colonized acute trauma patients requiring surgery. RESULTS: Of the 400 consecutive patient episodes, 395(98.7%) had MRSA screening performed on admission. However, in 236 (59.0%) cases, the results were not available before the surgery. Seven patient episodes (1.8%) had positive MRSA colonization. Analysis of 27 MRSA colonized patients revealed that 20(74%) patients did not have the screening results available before the surgery. Only 5(18.5%) received Teicoplanin and 22(81.4%) received cefuroxime for antibiotic prophylaxis before their surgery. Of those receiving cefuroxime, five (22.73%) patients developed postoperative MRSA surgical site infection (SSI) but none of those (0%) receiving Teicoplanin had MRSA SSI. The absolute risk reduction for SSI with Teicoplanin as antibiotic prophylaxis was 22.73% (CI=5.22%-40.24%) and NNT (Number Needed to Treat) was 5 (CI=2.5-19.2) CONCLUSION: Lack of available screening results before the surgery may lead to inadequate antibiotic prophylaxis increasing the risk of MRSA surgical site infection. Glycopeptide (e.g.Teicoplanin) prophylaxis should be considered when there is history of MRSA colonization or MRSA screening results are not available before the surgery.

ROCK2/rasHa co-operation induces malignant conversion via p53 loss, elevated NF-κB and tenascin C-associated rigidity, but p21 inhibits ROCK2/NF-κB-mediated progression.

To study ROCK2 activation in carcinogenesis, mice expressing 4-hydroxytamoxifen (4HT)-activated ROCK2 (K14.ROCKer) were crossed with mice expressing epidermal-activated rasHa (HK1.ras1205). At 8 weeks, 4HT-treated K14.ROCKer/HK1.ras1205 cohorts exhibited papillomas similar to HK1.ras1205 controls; however, K14.ROCKer/HK1.ras1205 histotypes comprised a mixed papilloma/well-differentiated squamous cell carcinoma (wdSCC), exhibiting p53 loss, increased proliferation and novel NF-κB expression. By 12 weeks, K14.ROCKer/HK1.ras1205 wdSCCs exhibited increased NF-κB and novel tenascin C, indicative of elevated rigidity; yet despite continued ROCK2 activities/p-Mypt1 inactivation, progression to SCC required loss of compensatory p21 expression. K14.ROCKer/HK1.ras1205 papillomatogenesis also required a wound promotion stimulus, confirmed by breeding K14.ROCKer into promotion-insensitive HK1.ras1276 mice, suggesting a permissive K14.ROCKer/HK1.ras1205 papilloma context (wound-promoted/NF-κB+/p53-/p21+) preceded K14.ROCKer-mediated (p-Mypt1/tenascin C/rigidity) malignant conversion. Malignancy depended on ROCKer/p-Mypt1 expression, as cessation of 4HT treatment induced disorganized tissue architecture and p21-associated differentiation in wdSCCs; yet tenascin C retention in connective tissue extracellular matrix suggests the rigidity laid down for conversion persists. Novel papilloma outgrowths appeared expressing intense, basal layer p21 that confined endogenous ROCK2/p-Mypt1/NF-κB to supra-basal layers, and was paralleled by restored basal layer p53. In later SCCs, 4HT cessation became irrelevant as endogenous ROCK2 expression increased, driving progression via p21 loss, elevated NF-κB expression and tenascin C-associated rigidity, with p-Mypt1 inactivation/actinomyosin-mediated contractility to facilitate invasion. However, p21-associated inhibition of early-stage malignant progression and the intense expression in papilloma outgrowths, identifies a novel, significant antagonism between p21 and rasHa/ROCK2/NF-κB signalling in skin carcinogenesis. Collectively, these data show that ROCK2 activation induces malignancy in rasHa-initiated/promoted papillomas in the context of p53 loss and novel NF-κB expression, whereas increased tissue rigidity and cell motility/contractility help mediate tumour progression.

Nutritional Supplement of Hatchery Eggshell Membrane Improves Poultry Performance and Provides Resistance against Endotoxin Stress.

Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM) as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS) challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase resistance of chickens to endotoxin.

Early failure with the Journey-Deuce bicompartmental knee arthroplasty.

The aim of this study was to evaluate the early functional outcome and survivorship of a bicompartmental knee arthroplasty implant (Journey-Deuce) in a cohort of patients with combined medial and patellofemoral degenerative osteoarthritis. Fifteen patients with a mean age of 57 years were followed up prospectively and evaluated with clinical examination, Oxford knee score and radiology imaging. Poor pain scores, concerns about the tibial fixation, early aseptic loosening of the tibial component and a revision rate of 60 % at a minimum follow-up of 54 months are reported. Implantation of this prosthesis was stopped at our institution well before the first revision due to an unfavourable early clinical response. This was further endorsed by an unacceptable revision rate. The outcome of the Journey-Deuce bicompartmental knee replacement was considerably worse than the published outcome of total knee replacement.

Protein profiles of hatchery egg shell membrane.

BACKGROUND: Eggshells which consist largely of calcareous outer shell and shell membranes, constitute a significant part of poultry hatchery waste. The shell membranes (ESM) not only contain proteins that originate from egg whites but also from the developing embryos and different contaminants of microbial and environmental origins. As feed supplements, during post hatch growth, the hatchery egg shell membranes (HESM) have shown potential for imparting resistance of chickens to endotoxin stress and exert positive health effects. Considering that these effects are mediated by the bioactive proteins and peptides present in the membrane, the objective of the study was to identify the protein profiles of hatchery eggshell membranes (HESM). METHODS: Hatchery egg shell membranes were extracted with acidified methanol and a guanidine hydrochloride buffer then subjected to reduction/alkylation, and trypsin digestion. The methanol extract was additionally analyzed by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). The tryptic digests were analyzed by liquid chromatography and tandem mass spectrometry (LC-MS-MS) to identify the proteins. RESULTS: Our results showed the presence of several proteins that are inherent and abundant in egg white such as, ovalbumin, ovotransferrin, ovocleidin-116, and lysozyme, and several proteins associated with cytoskeletal, cell signaling, antimicrobial, and catalytic functions involving carbohydrate, nucleic acid, and protein metabolisms. There were some blood derived proteins most likely originating from the embryos and several other proteins identified with different aerobic, anaerobic, gram positive, gram negative, soil, and marine bacterial species some commensals and others zoonotic. CONCLUSION: The variety of bioactive proteins, particularly the cell signaling and enzymatic proteins along with the diverse microbial proteins, make the HESM suitable for nutritional and biological application to improve post hatch immunity of poultry.

Poultry Femoral Head Separation and Necrosis: A Review.

Femoral head separation (FHS) is a degenerative skeletal problem in fast-growing poultry wherein the growth plate of the proximal femur separates from its articular cartilage. At its early phase, FHS may remain asymptomatic but lead to epiphyseal breakage, infection, and femoral head necrosis (FHN). Healthy femoral head is viewed as a positive trait for genetic selection. However, the etiology of FHS is poorly understood for use in noninvasive diagnosis and genetic selection. Focal cell death and atrophic changes are likely associated with separation of tissues and necrotic changes. Fibrotic thickening of the articular surface can also impair free movement of the proximal epiphysis in the acetabulum, leading to FHS, under strain. The major limitation to understanding the pathophysiology of FHN is the lack of suitable experimental models and biomarkers to diagnose the problem. In this review, we discuss the possible etiologic factors, anatomic features of the chicken femoral head, biomarkers, and molecular mechanisms relevant to FHN.

Ascorbic Acid Differentially Affects Stress Response and Resistance to Colibacillosis in Turkeys from Genetic Lines Differing in Growth Rate.

Genetic selection for fast growth can affect the ability of male turkeys to cope with stressors, resulting in decreased immunity to opportunistic bacterial infection. The purpose of the current study was to compare the effects of ascorbic acid (AA) on the stress response and resistance to Escherichia coli challenge of birds selected for increased 16-wk body weight (BW; F-line) with their random-bred parent line (RBC2). Male turkeys were raised in duplicate floor pens in a two line×two AA treatment×two stress challenge (SC) design. At 5 wk of age, AA (1200 ppm) was provided in drinking water for a 24-hr period, during which all birds were weighed. After AA treatment, the SC group was subjected to a transport stress protocol. Six hours after the start of transport, SC birds were also inoculated in the thoracic air sac with 1×10(4) colony-forming units of E. coli. The following morning four birds from each pen were bled, and all birds were weighed and necropsied 2 days later. BW and gain after SC were decreased in the F-line but not the RBC2 line, and there were no AA effects on BW. The weight of the bursa of Fabricius relative to BW was higher in the RBC2 line than in the F-line, was decreased by SC, and was not affected by AA. The heterophil∶lymphocyte ratio was higher in the SC F-line as compared to the SC RBC2 and was decreased by AA only in the SC F-line. Corticosterone (C) levels were increased by SC only in the F-line, and AA decreased C levels only in the RBC2 line. Airsacculitis scores were increased in the F-line SC birds. The challenge strain of E. coli was only detected in the air sac and liver of the AA-treated F-line SC birds and in the liver of the no-AA F-line birds. These results suggest that SC at 5 wk of age had a more deleterious effect on the fast-growing F-line than on its parent line and that AA may have increased susceptibility to colibacillosis in the SC F-line birds.

Effect of dexamethasone on bacteriostatic activity of turkey monocytes and implications for food safety.

Stress has been shown to affect the immune system of turkeys making them more susceptible to bacterial infections. Five-week-old male and female turkeys were treated with 3 intra-muscular injections of dexamethasone (Dex) at 0, 0.5 and 2.0mg/kg body weight. Twenty-four hours after the third injection birds were bled and white blood cell (WBC) differentials and bacteriostatic activity of monocytes were measured. Dex at both 0.5 and 2.0mg/kg decreased phagocytic activity in females only. Bacteriostatic activity was decreased at both concentrations of Dex at 8 and 16 h post-infection in both sexes and was lower in males as compared to females. Total WBC counts were increased in females at both concentrations of Dex whereas male total WBC counts were unaffected. Both males and females had an increase in the heterophil to lymphocyte ratio. Within the same study, replicate pens of turkeys were challenged with intra-air sac inoculation of 100 cfu of Escherichia coli. Isolation of E. coli was significantly increased by both Dex and E. coli challenge, but there were no differences between sexes. These results suggest that stress can compromise the bacteriostatic activity of turkey monocytes and increase bacterial colonization of blood and tissues, potentially affecting food safety.

Nutritional effects of egg shell membrane supplements on chicken performance and immunity.

Eggshell membranes (ESM) contain a variety of proteins and peptides which help in the development of embryo and provide protection to it. Many of the peptides and proteins associated with ESM have antimicrobial, immune-modulatory, and adjuvant properties. We hypothesized that the membrane byproducts from egg, provided as posthatch nutritional supplements to chickens, may improve their performance and immunity. To explore its effect, we fed 3 groups of broiler chicks with feed containing 0, 0.2, and 0.4% ESM from d 1 posthatch through 14 d and regular feed thereafter. The birds were individually weighed at the onset of the study and at weekly intervals until the termination at third wk when they were bled and euthanized. The relative weights of liver, spleen, bursa, and heart, hematology profiles, and clinical chemistry variables including serum IgM, IgG, and corticosterone concentrations were measured. The chickens in the ESM treated groups showed a statistically significant increase in BW with no impact on relative organ weights. Compared with controls, the WBC and lymphocyte percentage increased in chickens fed 0.4% ESM whereas the monocyte percentage decreased at both levels of ESM. Except for the serum protein which increased in ESM fed birds no other metabolic clinical chemistry variables showed any significant change. Both IgM and IgG(Y) levels were elevated and corticosterone levels reduced in chickens fed ESM supplemented diets. Our results suggest that ESM supplements during the early phases of growth may improve immunity and stress variables, and enhance their growth performance without any detrimental effect on other physiological parameters.

Efficacy of a novel prebiotic and a commercial probiotic in reducing mortality and production losses due to cold stress and Escherichia coli challenge of broiler chicks 1.

Prebiotics consisting of resistant starch may alter intestinal ecology, thus modulating inflammation and increasing intestinal health through increased cecal production of short-chain fatty acids (SCFA). Probiotics may directly alter the intestinal microbiome, resulting in the same effects. We hypothesize that adding prebiotics and probiotics to feed may protect the gut of young chicks under stress. Studies 1, 2, and 3 evaluated treatments in a cold stress (CS) and Escherichia coli (EC) oral challenge to 430 day-old broiler chicks for 3 wk. In study 1, prebiotics were administered as 15% of the diet during the first week only and consisted of the following: Hi-Maize resistant starch (HM), potato starch (PS), or raw potato (RP). In studies 2 and 3, the PS treatment was identical to study 1, and an additional probiotic treatment (PRO) was administered in feed and water. In study 1, PS protected BW during the first week and decreased the mortality of CS/EC-challenged birds during the first week and wk 3, while RP decreased the mortality of warm-brooded birds challenged with EC during the first week. In study 2, PS decreased and PRO increased the main effect mean (MEM) of the first week BW. PS and PRO numerically decreased the feed conversion ratio (FCR) by 23 and 29 points, respectively, in CS/EC-challenged birds with no effects on mortality. In study 3, PS decreased and PRO increased the first week and wk 3 MEM BW. PS numerically increased FCR by 16 points, while PRO decreased FCR by 2 points. Both PS and PRO tended to increase overall mortality, and PRO significantly increased mortality in the CS/EC challenge. These results suggest that the effects of PS may be too variable in this challenge model for further study; however, the PRO treatment improved production values and may have potential as an alternative to antibiotics during the first weeks after hatch.

Effect of butyrate on immune response of a chicken macrophage cell line.

Butyric acid is a major short chain fatty acid (SCFA), produced in the gastrointestinal tract by anaerobic bacterial fermentation, that has beneficial health effects in many species including poultry. To understand the immunomodulating effects of butyrate on avian macrophage, we treated a naturally transformed line of chicken macrophage cells named HTC with Na-butyrate in the absence or presence of Salmonella typhimurium lipopolysaccharide (LPS) or phorbol-12-myristate-13-acetate (PMA), a metabolic activator, evaluating its various functional parameters. The results demonstrate that, butyrate by itself had no significant effect on variables such as nitric oxide (NO) production and the expression of genes associated with various inflammatory cytokines but it inhibited NO production, and reduced the expression of cytokines such as IL-1ß, IL-6, IFN-γ, and IL-10 in LPS-stimulated cells. Butyrate decreased the expression of TGF-ß3 in the presence or absence of LPS, while it had no effect on IL-4, Tß4, and MMP2 gene expression. In addition, butyrate augmented PMA induced oxidative burst indicated by DCF-DA oxidation and restored LPS induced attenuation of tartrate resistant acid phosphatase (TRAP) activity. Although butyrate had no significant effect on phagocytosis or matrix metalloproteinase (MMP) activities of resting macrophages, it significantly suppressed the effects induced by their respective stimulants such as LPS induced phagocytosis and PMA induced MMP expression. These results suggest that butyrate has immunomodulatory property in the presence of agents that incite the cells thus, has potential to control inflammation and restore immune homeostasis.

Environmental augmentation with bacteriophage prevents colibacillosis in broiler chickens.

Bacteriophages are viruses that kill bacteria. They are plentiful in nature; are safe, having no known activity to human or animal cells; and are an attractive alternative to antibiotics. The objectives of this research were to establish an experimental model of colibacillosis induced by indirect exposure to Escherichia coli and to determine if bacteriophage could protect the birds from developing colibacillosis. In study 1 there were 6 treatments with 2 replicate pens of 25 birds. The treatments were control warm brooded; control cold stressed; litter inoculated with E. coli, warm brooded; litter inoculated with E. coli, cold stressed; seeder birds (5 per pen) challenged with E. coli, warm brooded; and seeder birds (5 per pen), cold stressed. The study concluded when the birds were 3 wk of age. Body weights at 1, 2, and 3 wk of age were significantly decreased (P ≤ 0.05) by cold stress, decreased at 1 and 2 wk of age by both the litter and seeder bird treatments compared with the control treatment and by the seeder bird treatment at 3 wk of age. Study 2 consisted of 8 treatments with 2 replicate pens of 20 birds per treatment. The treatments were control, warm brooded; control, cold stressed; litter inoculated with E. coli, cold stressed; and seeder birds (5/pen) challenged with E. coli, cold stressed with and without bacteriophage treatment. In the bacteriophage treatments the bacteriophages were sprayed on the litter. The study was concluded at 3 wk of age. Body weights at 1 wk of age were significantly (P ≤ 0.05) decreased from the control treatment by the seeder bird treatment and were significantly (P ≤ 0.05) higher in all the bacteriophage treatments compared with their matched untreated treatments, except in the control cold stressed treatment. Mortality was significantly (P ≤ 0.05) decreased by bacteriophage in the litter challenged treatment. These data suggest that augmentation of the environment with bacteriophage is a practical and efficacious way to prevent colibacillosis in broiler chickens.

Isolation and characterization of chicken bile matrix metalloproteinase.

Avian bile is rich in matrix metalloproteinases (MMP), the enzymes that cleave extracellular matrix proteins such as collagens and proteoglycans. Changes in bile MMP expression have been correlated with hepatic and gall bladder pathologies, but the significance of their expression in normal, healthy bile is not understood. We hypothesized that the MMP in bile may aid the digestion of native collagens that are resistant to conventional gastric proteases. Hence, the objective of this study was to characterize the bile MMP and check its regulation in association with dietary factors. We used substrate zymography, azocoll protease assay, and gelatin affinity chromatography to identify and purify the MMP from chicken bile. Using zymography and SDS PAGE, 5 bands at 70, 64, 58, 50, and 42 kDa were detected. The bands corresponding to 64, 50, and 42 kDa were identified as MMP2 using trypsin in-gel digestion and matrix-assisted laser desorption time-of-flight mass spectrometry and peptide mass fingerprinting. Chickens fed diets containing gelatin supplements showed higher levels of MMP expression in the bile by both azocoll assay and zymography. We conclude that the bile MMP may be associated with the digestion of collagens and other extracellular matrix proteins in avian diets.

Fast, multi-channel real-time processing of signals with microsecond latency using graphics processing units.

Fast, digital signal processing (DSP) has many applications. Typical hardware options for performing DSP are field-programmable gate arrays (FPGAs), application-specific integrated DSP chips, or general purpose personal computer systems. This paper presents a novel DSP platform that has been developed for feedback control on the HBT-EP tokamak device. The system runs all signal processing exclusively on a Graphics Processing Unit (GPU) to achieve real-time performance with latencies below 8 µs. Signals are transferred into and out of the GPU using PCI Express peer-to-peer direct-memory-access transfers without involvement of the central processing unit or host memory. Tests were performed on the feedback control system of the HBT-EP tokamak using forty 16-bit floating point inputs and outputs each and a sampling rate of up to 250 kHz. Signals were digitized by a D-TACQ ACQ196 module, processing done on an NVIDIA GTX 580 GPU programmed in CUDA, and analog output was generated by D-TACQ AO32CPCI modules.

In situ "artificial plasma" calibration of tokamak magnetic sensors.

A unique in situ calibration technique has been used to spatially calibrate and characterize the extensive new magnetic diagnostic set and close-fitting conducting wall of the High Beta Tokamak-Extended Pulse (HBT-EP) experiment. A new set of 216 Mirnov coils has recently been installed inside the vacuum chamber of the device for high-resolution measurements of magnetohydrodynamic phenomena including the effects of eddy currents in the nearby conducting wall. The spatial positions of these sensors are calibrated by energizing several large in situ calibration coils in turn, and using measurements of the magnetic fields produced by the various coils to solve for each sensor's position. Since the calibration coils are built near the nominal location of the plasma current centroid, the technique is referred to as an "artificial plasma" calibration. The fitting procedure for the sensor positions is described, and results of the spatial calibration are compared with those based on metrology. The time response of the sensors is compared with the evolution of the artificial plasma current to deduce the eddy current contribution to each signal. This is compared with simulations using the VALEN electromagnetic code, and the modeled copper thickness profiles of the HBT-EP conducting wall are adjusted to better match experimental measurements of the eddy current decay. Finally, the multiple coils of the artificial plasma system are also used to directly calibrate a non-uniformly wound Fourier Rogowski coil on HBT-EP.

Blebs produced by actin-myosin contraction during apoptosis release damage-associated molecular pattern proteins before secondary necrosis occurs.

Apoptosis is a fundamental homeostatic mechanism essential for the normal growth, development and maintenance of every tissue and organ. Dying cells have been defined as apoptotic by distinguishing features, including cell contraction, nuclear fragmentation, blebbing, apoptotic body formation and maintenance of intact cellular membranes to prevent massive protein release and consequent inflammation. We now show that during early apoptosis limited membrane permeabilization occurs in blebs and apoptotic bodies, which allows release of proteins that may affect the proximal microenvironment before the catastrophic loss of membrane integrity during secondary necrosis. Blebbing, apoptotic body formation and protein release during early apoptosis are dependent on ROCK and myosin ATPase activity to drive actomyosin contraction. We identified 231 proteins released from actomyosin contraction-dependent blebs and apoptotic bodies by adapted SILAC (stable isotope labeling with amino acids in cell culture) combined with mass spectrometry analysis. The most enriched proteins released were the nucleosomal histones, which have previously been identified as damage-associated molecular pattern proteins (DAMPs) that can initiate sterile inflammatory responses. These results indicate that limited membrane permeabilization occurs in blebs and apoptotic bodies before secondary necrosis, leading to acute and localized release of immunomodulatory proteins during the early phase of active apoptotic membrane blebbing. Therefore, the shift from apoptosis to secondary necrosis is more graded than a simple binary switch, with the membrane permeabilization of apoptotic bodies and consequent limited release of DAMPs contributing to the transition between these states.