HEP Group Research Articles

Mindfulness-Based Stress Reduction and Autonomic Modulation in Chronic Kidney Disease

Intro: Chronic kidney disease (CKD) is characterized by chronic overactivation of the sympathetic nervous system (SNS) that leads to increased blood pressure and cardiovascular disease risk. Mindfulness-Based Stress Reduction (MBSR) is an 8-week mindfulness meditation training program that has previously been shown to reduce blood pressure and improve autonomic balance in healthy and some clinical populations; however, MBSR has never previously been tested in CKD and the long-term effects of mindfulness mediation on SNS activity are unknown. Thus, we tested the hypothesis that 8 weeks of MBSR reduces SNS activity and reactivity in CKD patients. Method: In a randomized controlled trial, participants with CKD stages III-IV were randomized to 8 weeks of MBSR (n=17, 64±8 yr, 88% males) or a health education program (HEP, n=12, 63±10 yr, 83% males) that served as the active control intervention. MBSR was comprised of eight 2.5-hour group meetings and a retreat session focused on mindfulness training. HEP was administered in a similar structure and focused on health education. Direct intraneural measures of SNS activity directed to muscle (MSNA) was recorded via microneurography with concomitant beat-to-beat blood pressure and EKG monitoring at rest and during mental stress before and after the 8-week intervention. MSNA reactivity during mental arithmetic-induced stress (3 minutes) was compared between baseline and end of study (EOS) in the MBSR and HEP groups. Results. Baseline characteristics, kidney function, and comorbidities were comparable between groups. There was a significant Group (MBSR vs. HEP) by Time (baseline vs. EOS) interaction in the change in MSNA reactivity to mental stress. The MBSR group had a significant reduction in MSNA reactivity during the 1st, 2nd and 3rd minute of mental arithmetic at EOS (p<0.05 for all), while no change was observed within the HEP group. Resting MSNA and reactivity in blood pressure and heart rate during mental arithmetic remained unchanged in both groups from baseline to EOS. Conclusion. Patients with CKD had an amelioration of sympathetic reactivity during mental stress following 8-weeks of MBSR but not after HEP (control intervention). Our findings demonstrate that mindfulness training is feasible and may have potential beneficial effects on autonomic function in CKD. NIH R01HL135183; NIH R61AT10457; NIH KL2TR002381; VA Merit I01CX001065. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Identification of a gene expression signature associated with brain metastasis in colorectal cancer

PurposeBrain metastasis (BM) in colorectal cancer (CRC) is a rare event with poor prognosis. Apart from (K)RAS status and lung and bone metastasis no biomarkers exist to identify patients at risk. This study aimed to identify a gene expression signature associated with colorectal BM.MethodsThree patient groups were formed: 1. CRC with brain metastasis (BRA), 2. exclusive liver metastasis (HEP) and, 3. non-metastatic disease (M0). RNA was extracted from primary tumors and mRNA expression was measured using a NanoString Panel (770 genes). Expression was confirmed by qPCR in a validation cohort. Statistical analyses including multivariate logistic regression followed by receiver operating characteristic (ROC) analysis were performed.ResultsEMILIN3, MTA1, SV2B, TMPRSS6, ACVR1C, NFAT5 and SMC3 were differentially expressed in BRA and HEP/M0 groups. In the validation cohort, differential NFAT5, ACVR1C and SMC3 expressions were confirmed. BRA patients showed highest NFAT5 levels compared to HEP/M0 groups (global p = 0.02). High ACVR1C expression was observed more frequently in the BRA group (42.9%) than in HEP (0%) and M0 (7.1%) groups (global p = 0.01). High SMC3 expressions were only detectable in the BRA group (global p = 0.003). Only patients with BM showed a combined high expression of NFAT5, ACVR1C or SMC3 as well as of all three genes. ROC analysis revealed a good prediction of brain metastasis by the three genes (area under the curve (AUC) = 0.78).ConclusionsThe NFAT5, ACVR1C and SMC3 gene expression signature is associated with colorectal BM. Future studies should further investigate the importance of this biomarker signature.

Different feeding patterns affect meat quality of Tibetan pigs associated with intestinal microbiota alterations.

This study aimed to investigate the effects of different feeding patterns on meat quality, gut microbiota and its metabolites of Tibetan pigs. Tibetan pigs with similar body weight were fed the high energy diets (HEP, 20 pigs) and the regular diets (RFP, 20 pigs), and free-ranging Tibetan pigs (FRP, 20 pigs) were selected as the reference. After 6 weeks of experiment, meat quality indexes of semitendinosus muscle (SM) and cecal microbiota were measured. The results of meat quality demonstrated that the shear force of pig SM in FRP group was higher than that in HEP and RFP groups (p < 0.001); the pH-value of SM in HEP pigs was higher at 45 min (p < 0.05) and lower at 24 h (p < 0.01) after slaughter than that in FRP and RFP groups; the SM lightness (L* value) of FRP pigs increased compared with RFP and HEP groups (p < 0.001), while the SM redness (a* value) of FRP pigs was higher than that of RFP group (p < 0.05). The free fatty acid (FA) profile exhibited that the total FAs and unsaturated FAs of pig SM in HEP and RFP groups were higher than those in FRP group (p < 0.05); the RFP pigs had more reasonable FA composition with higher n-3 polyunsaturated FAs (PUFAs) and lower n-6/n-3 PUFA ratio than HEP pigs (p < 0.05). Based on that, we observed that Tibetan pigs fed high energy diets (HEP) had lower microbial α-diversity in cecum (p < 0.05), and distinct feeding patterns exhibited a different microbial cluster. Simultaneously, the short-chain FA levels in cecum of FRP and RFP pigs were higher compared with HEP pigs (p < 0.05). A total of 11 genera related to muscle lipid metabolism or meat quality, including Alistipes, Anaerovibrio, Acetitomaculun, etc., were identified under different feeding patterns (p < 0.05). Spearman correlation analysis demonstrated that alterations of free FAs in SM were affected by the genera Prevotellaceae_NK3B31_group, Prevotellaceae UCG-003 and Christensenellaceae_R-7_group (p < 0.05). Taken together, distinct feeding patterns affected meat quality of Tibetan pigs related to gut microbiota alterations.

Neural Signatures of Pain Modulation in Short-Term and Long-Term Mindfulness Training: A Randomized Active-Control Trial.

Mindfulness-based interventions are widely used to target pain, yet their neural mechanisms of action are insufficiently understood. The authors studied neural and subjective pain response in a randomized active-control trial of mindfulness-based stress reduction (MBSR) alongside long-term meditation practitioners. Healthy participants (N=115) underwent functional neuroimaging during a thermal acute pain task before and after random assignment to MBSR (N=28), an active control condition (health enhancement program [HEP]) (N=32), or a waiting list control condition (N=31). Long-term meditators (N=30) completed the same neuroimaging paradigm. Pain response was measured via self-reported intensity and unpleasantness, and neurally via two multivoxel machine-learning-derived signatures: the neurologic pain signature (NPS), emphasizing nociceptive pain processing, and the stimulus intensity independent pain signature-1 (SIIPS1), emphasizing stimulus-independent neuromodulatory processes. The MBSR group showed a significant decrease in NPS response relative to the HEP group (Cohen's d=-0.43) and from pre- to postintervention assessment (d=-0.47). The MBSR group showed small, marginal decreases in NPS relative to the waiting list group (d=-0.36), and in SIIPS1 relative to both groups (HEP group, d=-0.37; waiting list group, d=-0.37). In subjective unpleasantness, the MBSR and HEP groups also showed modest significant reductions compared with the waiting list group (d=-0.45 and d=-0.55). Long-term meditators reported significantly lower pain than nonmeditators but did not differ in neural response. Within the long-term meditator group, cumulative practice during intensive retreat was significantly associated with reduced SIIPS1 (r=-0.65), whereas daily practice was not. Mindfulness training showed associations with pain reduction that implicate differing neural pathways depending on extent and context of practice. Use of neural pain signatures in randomized trials offers promise for guiding the application of mindfulness interventions to pain treatment.

Long-term Effects of a Brief Mindfulness Intervention Versus a Health Enhancement Program for Treating Depression and Anxiety in Patients Undergoing Hemodialysis: A Randomized Controlled Trial

Background:Depression and anxiety affect approximately 50% of patients with kidney failure receiving hemodialysis and are associated with decreased quality of life and increased risk of hospitalization and mortality. A Brief Mindfulness Intervention (BMI) may be promising in treating depressive and anxiety symptoms in this population, but the long-term sustainability of the intervention’s effects is unknown.Objective:We previously conducted a randomized controlled trial (RCT; n = 55) comparing an 8-week BMI with an active control (Health Enhancement Program [HEP]) for patients receiving dialysis, with depression and/or anxiety. Here, we examine the 6-month follow-up data to determine the long-term sustainability of BMI versus HEP in reducing (1) depressive symptoms, (2) anxiety symptoms, and (3) the efficacy of BMI versus HEP in reducing the likelihood of hospitalization.Design:In this study, we analyzed 6-month follow-up data from an 8-week assessor-blinded parallel RCT, which evaluated the efficacy of a BMI against an active control, HEP, in patients receiving hemodialysis with symptoms of depression and/or anxiety.Setting:The study took place at hemodialysis centers in 4 tertiary-care hospitals in Montreal, Canada.Participants:Participants included adults aged ≥18 years who were receiving in-center hemodialysis 3 times per week and had symptoms of depression and/or anxiety as indicated by a score ≥6 on the Patient Health Questionnaire–9 (PHQ-9) and/or the General Anxiety Disorder–7 (GAD-7).Methods:Participants were randomized to the treatment arm (BMI) or the active control arm (HEP) and completed assessments at baseline, 8 weeks, and 6-month follow-up. Depression was assessed using the PHQ-9, and anxiety was assessed by the GAD-7. Hospitalization rates were assessed using medical chart information.Results:We observed significant decrease in depression scores over 6 months in both BMI and HEP groups, with no significant difference between groups. Anxiety scores significantly decreased over 6 months, but only in the BMI group. Brief Mindfulness Intervention and Health Enhancement Program were comparable in terms of hospitalization rates.Limitations:The limitations of our study include the modest sample size and lack of a third arm such as a waitlist control.Conclusions:Our results suggest that the beneficial effects of BMI and HEP for improving mood disorder symptoms in patients receiving dialysis persist at 6-month follow-up. Both interventions showed sustained effects for depressive symptoms, but BMI may be more useful in this population given its efficacy in reducing anxiety symptoms as well.Trial registration:Prior to recruitment, the trial had been registered (ClinicalTrials.gov Identifier: NCT03406845).

RECOGNIZING SYMPTOMS OF CRISIS BY ANALYZING FINANCIAL RESULTS – CASE OF ELEKTROPROMET d.d.

Up until 2010 Elektropromet d.d. was one of Croatian market leaders that supplied major Government companies, such as HEP Group (Croatian national energy company) or Croatian railways. Company had more than 600 employees in the 90’s, working in 4 divisions. Global and local market changes led to revenue decrease and questionable profitability. Company structure did not change in time, and market share was lost, bit by bit, although there was time for changes. The company did not have a quality control system and they did not recognize the symptoms of crisis. Those symptoms of crisis were visible from financial results in period from 2003 up to 2006 and the company prepared for restructuring in 2007 and 2008. Since the restructuring program was not applied, the company continued with the same business model. Results in that period, and the following years without making any changes led to decreasing business trend with negative result. The goal of this study is to analyze financial results from 2003 up to 2013 and confirm the hypothesis that crisis symptoms in Elektropromet d.d. were visible from financial result analysis. Financial results were measured and observed using different profitability formulas providing insight for crisis symptoms and providing answers on how to prevail business crisis. The results were analized using Year-over-year (YoY) result comparisons. Just by using YoY comparisons it was clear that the results are not profitable. Financial results from 2003 and 2004 provide an insight to early crisis. Profit was not generated from commercial business and regular sales, because expenses were higher than revenue and earnings. Calculating EBIT provides an answer on how profit was made and that early crisis symptoms were visible. After that it would be easy to define crisis stage and work on a model and define phases how to prevail the crisis. Management did not decide to recognize the crisis in that early period and the company continued with the same business model for next few years. The restructuring was prepared for 2007 and 2008, but it came too late and was canceled in early stage. Since Elektropromet is a stock corporation it was up to supervisory board to warn the shareholders that the board members are not leading the company in the right direction. The result and conclusion should assist corporations, medium size and small companies to recognize crisis symptoms and help them use financial management and controlling experience to avoid possible revenue and profitability loss.

The improvement in aerobic capacity, disease activity, and function in patients with rheumatoid arthritis following cardiac rehabilitation program: A single-center, controlled study.

This study aims to evaluate the effect of cardiac rehabilitation (CR) on disease activity, functional status, fitness, and modified cardiovascular disease (CVD) risk factors in patients with rheumatoid arthritis (RA) with a moderate disease activity. This single-center, controlled study included a total of 60 female RA patients (mean age 57.5 years; range, 50 to 64 years) with moderate disease activity according to the Disease Activity Score-28 (DAS28) between January 2014 and June 2015. The patients were divided into two equal groups as those receiving CR program (n=30) and those receiving home exercise program (HEP; n=30). The patients were evaluated at baseline, at Weeks 10 and 24 using exercise tolerance test (i.e., The Metabolic Equivalent of Task [MET] and maximal oxygen uptake [VO2 max]), Six-Minute Walk Test (6MWT), DAS28, Health Assessment Questionnaire (HAQ), modified Systematic Coronary Risk Evaluation (mSCORE), Short Form Health Survey-36 (SF-36), and Beck Depression Inventory (BDI). There was a significant improvement in the VO2 max (p<0.001), MET (p<0.001), DAS28 (p<0.001), HAQ (p<0.001), BDI (p=0.005), SF-36 physical function (p=0.039), pain (p<0.001) and vitality subscale scores (p=0.008), and 6MWT (p<0.001), after the initial and repetitive exercise programs in the CR group compared to the HEP group. At the end of Week 24, full compliance with HEP was higher in the patients with CR group, compared to the HEP group (p<0.001). There was no significant effect of supervised exercise program on the mSCORE, although systolic blood pressure (p=0.033) and resting heart rate (p=0.049) were significantly improved in the CR group versus HEP group. Based on our study results, supervised exercise program cannot reduce CVD risk as assessed by the mSCORE, although it improves physical fitness, disease activity, and functional outcomes which may reduce traditional CVD risk factors in patients with RA.

The effects of aerobic exercise on sleep quality measures and sleep-related biomarkers in individuals with Multiple Sclerosis: A pilot randomised controlled trial.

Sleep disturbances are highly prevalent in people with multiple sclerosis(MS), and are associated with pain, fatigue, depression, and reduced quality of life (QoL). Importantly, sleep has been considered a critical brain state for motor learning and memory consolidation. Therefore, interventions that target sleep managementin people with MS are needed. To explore the effects of a six weeks moderate-intensity aerobic exercise intervention on sleep characteristics and sleep-related biomarkers specifically serotonin, melatoninand cortisol in people with MS using a pilot randomized controlled trial. Participants were randomly allocated to either a moderate-intensity aerobic exercise program (MAE, n = 20) or a home exercise program (HEP, n = 20). Participants were assessed at baseline and follow-up. Subjective and objective measures were used to assess sleep quality. The Pittsburgh Sleep Quality Index (PSQI) and Insomnia Severity Index (ISI) were used to subjectively assess sleep. While Actigraphy was used to objectivelyassess sleep. Blood samples were collected for measurement of cortisol, melatonin and serotonin from MS participants in both groups at 8:00 am±1 hour. Seventeen participants in the MAE and 13 in the HEP group completed the study. Compared to the HEP group, people with MS who participated in a moderate-intensity aerobic exercise experienced significant improvements (P < 0.05) on the PSQI, ISI, and several objective sleep parameters measured using actigraphy. Only the serotonin levels increased significantly over the six-week period in the MAE group compared to the HEP group. The change score in serotonin (from baseline to follow up assessment) was significantly correlatedwith the change score in PSQI (r = -0.97, p < 001) and the change score in ISI (r = -0. 56, p = 0.015) only in the MAE group but not the HEP group. Exercise may be a non-pharmacological, inexpensive, safe method to improve sleep quality in people with MS. The improvement in the serotonin level due to aerobic exercisemight explain one of the physiologic mechanisms driving these improvements.

Diet selection and trade-offs between condensed tannins and nutrients in parasitized sheep

Foraging behavior by parasitized herbivores can be interpreted as a decision-making process where individuals are faced with trading-off the ingestion of nutrients with the ingestion of potentially medicinal -and toxic- plant secondary compounds. We determined how parasitized sheep prioritize selection of crude protein, energy and a medicinal plant secondary compound (quebracho tannins-QT). Foraging preferences were tested in 40 lambs before experiencing a parasitic infection (Phase 1), during an infection (Phase 2; 10,000 L3Haemonchus contortus per lamb) and after chemotherapy (Phase 3). Lambs were assigned to four groups (10 lambs/group) such that animals in Group 1 (Control) could choose between foods of high (HEP) or low (LEP) energy to protein ratios. The other groups received the same choice, but QT were added (4%) to HEP (Group 2), to LEP (Group 3) or to both foods (Group 4). All groups under a parasitic infection (Phase 2) increased their preference for HEP (from 0.44 to 0.66 ± 0.042; P < 0.05) and intake of digestible energy (from 0.106 to 0.126 ± 0.007 Mcal/kg BW; P < 0.05) relative to Phase 1, a pattern that remained during Phase 3. Only lambs receiving QT in HEP increased their intake of QT from Phase 1 to Phase 2 (P < 0.05). Fecal egg counts and blood parameters revealed a parasitic infection (P < 0.05) in Phase 2 that subsided in Phase 3, although no differences were detected among groups (P > 0.05). The importance of protein nutrition on parasitized animals has been highlighted before, but these results suggest that lambs prioritized the ingestion of energy-dense over protein-dense foods or medicinal condensed tannins when challenged by gastrointestinal parasitism. Consumption of medicinal tannins represented a side-effect of the preference manifested for energy-dense foods during testing.

Quasi-online accounting and monitoring system for distributed clouds

The HEP group at the University of Victoria operates a distributed cloud computing system for the ATLAS and Belle II experiments. The system uses private and commercial clouds in North America and Europe that run OpenStack, Open Nebula or commercial cloud software. It is critical that we record accounting information to give credit to cloud owners and to verify our use of commercial resources. We want to record the number of CPU-hours of the virtual machine. We continuously collect the CPU usage and an estimate of the HEPSpec06 units of the VM obtained during the boot of the VM and uploads it into an Elastic Search database. The information is processed and published as soon as it is available. The data is published in tables and plots in Kibana and as a cross check in ROOT. We have found the system to be useful beyond gathering accounting information and can be used for monitoring and diagnostic purposes. For example, we can use it to detect if the payload jobs are stuck in a waiting state for external information. We will report on the design and performance of the system, and show how it provides important accounting and monitoring information on a large distributed system.

Next Generation Monitoring: Tier 2 Experience

Monitoring IT infrastructure is essential for maximizing availability and minimizing disruption by detecting failures and developing issues. The HEP group at Liverpool have recently updated our monitoring infrastructure with the goal of increasing coverage, improving visualization capabilities, and streamlining configuration and maintenance. Here we present a summary of Liverpool’s experience, the monitoring infrastructure, and the tools used to build it. In brief, system checks are configured in Puppet using Hiera, and managed by Sensu, replacing Nagios. Centralised logging is managed with Elasticsearch, together with Logstash and Filebeat. Kibana provides an interface for interactive analysis, including visualization and dashboards. Metric collection is also configured in Puppet, managed by collectd and stored in Graphite, with Grafana providing a visualization and dashboard tool. The Uchiwa dashboard for Sensu provides a web interface for viewing infrastructure status. Alert capabilities are provided via external handlers. A custom alert handler is in development to provide an easily configurable, extensible and maintainable alert facility.

Comparison between Saline Solution Containing Heparin versus Saline Solution in the Lock of Totally Implantable Catheters

There are only 3 studies comparing the efficacy of 2 different types of lock used in totally implantable catheters regarding occlusion or reflux dysfunction. The present study contains the largest published casuistry (862 patients) and is the only one that analyzes 3 parameters: occlusion, reflux dysfunction, and flow dysfunction. This was a retrospective study of patients operated at a large oncology center and followed up in the outpatient clinic between 2007 and 2015. The patients were divided into 2 groups according to the type of lock: the Hep group (heparine), whose lock was composed of saline solution 0.9% with heparin (100 IU/mL) and the SS group (saline solution), whose lock was composed of saline solution 0.9%. The Hep group was composed of 270 patients (31%) and the SS group of 592 patients (69%). Regarding occlusion, there were 8 cases in the Hep group (2.96%) and 8 in the SS group (1.35%; P=0.11); in relation to reflux dysfunction, there were 8 cases in the Hep group (2.96%) and 8 in the SS group (1.35%; P=0.11); in relation to flow dysfunction, there was 1 case in the Hep group (0.37%) and 4 cases in the SS group (0.68%; P=1). There was no statistically significant difference between the groups regarding occlusion, reflux dysfunction, and flow dysfunction.

Analysis of tumor-infiltrating immune cells in gastrointestinal stromal tumors (GIST) after tyrosine kinase inhibitor therapy.

11042 Background: There is initial evidence that immune-infiltrates of tumor may correlate with the clinical course of patients with GIST beyond the response to tyrosine kinase inhibitors (TKI) according to the mutational status. We were interested to analyze the composition of tumor-infiltrating immune cells in GIST tumor tissues after different TKI regimens vs untreated controls. Methods: From 40 GIST patients who were treated with imatinib alone (neoadjuvant group) and other TKIs (M1 HEP group, progressing on more than ≥2 TKI inhibitors) prior to tumor removal, surgical specimens were available at ten 10 cases each,. They were compared with 20 untreated primary tumors graded for malignant behaviour (low vs high risk ) acc. to Miettinen&amp;Lasota 2006. 2-μm sections of formalin-fixed, paraffin-embedded tissue samples were used for IHC-staining anti-CD68 (Dako); anti-CD11c, anti-CD11b, anti-FOXP3 (all Abcam); anti-CD163, anti-CD4 , anti-CD8 (all Leica). Evaluation was performed with Dako REAL EnVision Detection System (K5007). The phenotypes of immune cells were compared in the defined GIST groups. In the neoadjuvant group we used recurrence-free survival (RFS) for correlation, and overall survival (OS) after TKI failure in the M1 HEP group. Results: Foxp3+ Tregs, CD163+ M2 macrophages and CD11b+ myeloid cells are significantly higher in TKI resistance or neoadjuvant (viable cells &gt; 30%) tissues compared with neoadjuvant (viable cells &lt; 10%) cases. The rate of CD11c+ M1 macrophages is higher in low-risk GISTs compared to high risk GISTs, but CD11b+ myeloid cells are exactly the opposite. Kaplan-Meier curves show high CD163+ M2 macrophages or CD11b+ myeloid cells to correlate significantly with worse RFS in after neoadjuvant IM (p 0.01). In the TKI resistant condition, patients with high CD11c+ M1 macrophages show better median OS (p = 0.07, ns). Conclusions: Our study reveals dynamic changes of tumor-infiltrating immune cell protagonists in GISTs after different TKI therapeutic regimen and after neoadjuvant treatment. We could not detect a different immune infiltrate in untreated primary tumors of different risk categories.

146 HEPARAN SULFATE IS INVOLVED IN NUCLEAR SPERM DECONDENSATION AFTER FERTILIZATION IN BOVINE

Reduced glutathione (GSH) is an endogenous disulfide bond reducer present in mammalian oocytes. It plays a critical role in sperm decondensation following fertilization, disrupting the protamine bonds that sustain the hypercondensed state of sperm DNA. However, disulfide bond reduction needs to be followed by protamine removal to achieve male pronuclear formation. In humans, heparan sulfate (HS) has been shown to exert this role (Romanato et al. 2008 Hum. Reprod. 23, 1145–1450). Although there are no reports in bovine, we recently demonstrated the presence of HS in cow oocytes by indirect immunofluorescence, using a specific anti-HS monoclonal antibody (Canel et al. 2015, Proc. SSR 48th Annual Meeting). Heparinases are known to cleave HS chains selectively, leading to its depolymerization. In the present work, we analysed the possible role of HS as protamine acceptor after fertilization in cattle. To this aim, we directly injected heparinase into the cytoplasm of IVF presumptive zygotes, and analysed its effect on pronuclei formation. Cumulus-oocyte complexes were collected from slaughtered cow ovaries and matured in vitro under standard conditions (Canel et al. 2012 Cell. Div. 7, 23–33). After 21 h, IVF was performed following Brackett and Oliphant’s protocol (1975 Biol. Reprod. 12, 260–274), using frozen–thawed semen from 1 or 2 bulls at a final concentration of 15 × 106 spermatozoa/mL (5 replicates). After 5 h of incubation, cumulus cells and sperm bound to zona pellucidae were removed from presumptive zygotes. Heparinase III solution (H8891, Sigma, St. Louis, MO, USA) was diluted in 50% (vol/vol) polyvinylpyrrolidone solution in PBS-(polyvinylpyrrolidone) at a final concentration of 50 U mL−1 and ~30 pL was mechanically injected into the cytoplasm of each IVF presumptive zygote (Hep group) using a 9-μm inner diameter injection pipette. A group of zygotes was injected with the same volume of 10% polyvinylpyrrolidone (sham), whereas others were not subjected to injection (control). All zygotes were cultured for 16 h from the beginning of IVF in SOF medium (Holm et al. 1999 Theriogenology 52, 693–700). For pronuclear formation assessment, presumptive zygotes were permeabilized with 0.2% Triton X-100 for 15 min at room temperature, and their DNA content was stained with 5 µg mL−1 propidium iodide and observed under an epifluorescence microscope. Zygotes showing 2 pronuclei (PN) were considered as synchronically fertilized, whereas those showing one PN and one condensed sperm head were considered as asynchronically fertilized. Data were analysed by Fisher’s exact test (P &lt; 0.05). The rate of IVF zygotes showing 2 PN was lower for the Hep group (60.3%, n = 131) than those from sham (94.1%, n = 119) and control groups (98%, n = 101), which did not differ between them (P &lt; 0.05). In conclusion, our results show for the first time that HS is involved in bull chromatin sperm decondensation and allow us to propose HS as a putative protamine acceptor during male pronucleus formation after IVF in cattle. Given the high frequency of sperm decondensation failure observed in bovine after intracytoplasmic sperm injection, this work provides new insights for the development of novel sperm/egg treatments that might improve intracytoplasmic sperm injection outcomes in cattle.

Molecular characterization, alternative splicing and expression analysis of ACSF2 and its correlation with egg-laying performance in geese.

ACSF2 (encoded by acyl-CoA synthetase family member 2) belongs to the acyl-CoA synthetase (ACS) family, activating fatty acids by forming a thioester bond with CoA. In our previous study, a SNP residing in the intron of ACSF2 was identified to be linked to goose egg-laying performance. But the structure of goose ACSF2 as well as its role in reproduction remains unknown. In this study, we cloned and characterized ACSF2 in Yangzhou geese. A total of four alternative splice variants, designated as ACSF2-1, ACSF2-2, ACSF2-3 and ACSF2-4 respectively, were identified in the ovary. The coding regions of the four variants are 1770, 1692, 1599 and 1917bp in length, respectively encoding 589, 563, 532 and 638 amino acids with conserved AMP-binding sites. All ACSF2 variants were widely expressed in 11 tested tissues in geese, except that the ACSF2-2 transcript was not detected in hypothalamus, pituitary gland and granulosa cells. Subcellular localization revealed that ACSF2 is a mitochondrial matrix protein. ACSF2 mRNA level was compared between high egg production (HEP; n=8) and low egg production (LEP; n=10) groups and showed a lower (P<0.05) mRNA level in the HEP group. Further experiments indicated that overexpressing ACSF2 resulted in a significant increase of caspase-3 mRNA levels and that ACSF2 knockdown triggered a decrease in the caspase-3 mRNA level in granulosa cells. Similarly, the lower caspase-3 mRNA levels were identified in ovaries of the HEP group with lower ACSF2 mRNA levels. The research showed that the ACSF2 mRNA levels had a positive correlation with caspase-3 mRNA levels invivo (R=0.86, P<0.01). Our results suggest that lower ACSF2 expression promotes the laying performance of goose possibly by inhibiting granulosa cell apoptosis and facilitating follicular development.

Abstract WP10: Peri-procedural Heparin Use in Stent-retriever Thrombectomy

Background and Objectives: The use of periprocedural heparin has been reported safe and potentially beneficial during thrombectomy with older generation devices. We aim to investigate the impact of heparin in the era of stent-retrievers. Methods: A TREVO2 trial post-hoc analysis was performed aiming to compare baseline characteristics and clinical outcomes between patients who received [HEP(+)] versus patients that did not receive periprocedural heparin [HEP(-)]. Results: Out of 173 patients treated in the trial, 58 received a mean 3999 units of periprocedural heparin (n=42 received only pre-procedural bolus for a mean of 2952 units). Baseline characteristics amongst HEP(+) and HEP(-) were similar, with the exception of lower NIHSS (17±5 vs 18±6; p=0.04), IV t-PA use (37 vs 64%; p&lt;0.01), and higher baseline ASPECTS (7.6±1.3 vs 7.1±1.7; p=0.02) in the HEP(+) patients. Occlusion site varied amongst HEP(+) and HEP(-) groups: vertebrobasilar 10 vs 5%; ICA 8 vs 21%; MCA-M1 70 vs 55%; MCA-M2 10 vs 17%; p=0.04, respectively. Time from symptom-onset to puncture was similar, while procedure duration was longer in HEP(+) patients (98±47 vs 82±42min; p&lt;0.01). TICI2b-3 reperfusion rate was comparable (65 vs 62%; p=0.74), as well as embolization to unaffected territory, and access-site complications. Hemorrhagic changes were similar (ECASS HI1/HI2 27 vs 24%; p=0.71 and PH1/PH2 22 vs 25%; p=0.85). Good outcome (mRS0-2) and mortality at 90-days were 40 vs 25% (p=0.07) and 29 vs 27% (p=0.85), respectively. Multivariate logistic regression for good outcome identified the following independent predictors: use of heparin bolus (OR:5.30; 95%CI 1.70-0.16.48; p&lt;0.001), intubation (OR:0.10; 95%CI 0.03-0.33; p&lt;0.001), TICI2b-3 reperfusion (OR:6.56; 95%CI 2.29-18.83; p&lt;0.001), diabetes mellitus (OR:0.11; 95%CI 0.03-0.39; p&lt;0.001), NIHSS (OR:0.84; 95%CI 0.75-0.93; p&lt;0.001), stent-retriever device (OR:3.54; 95%CI 1.28-9.03; p&lt;0.001), time from symptom-onset to puncture (OR:0.64; 95%CI 0.45-0.89; p&lt;0.001), and CHF (OR:0.23; 95%CI 0.06-0.83; p&lt;0.001). Conclusions: The use of periprocedural heparin in stent-retriever thrombectomy is safe and may have a positive impact on outcomes.

Non-operative management of femoroacetabular impingement: A prospective, randomized controlled clinical trial pilot study.

To determine the efficacy of physical therapy on pain and physical function in patients with femoroacetabular impingement. Randomized, participant- and assessor blinded controlled trial pilot study. This trial was registered at ClinicalTrials.gov (NCT01814124) and reported according to Consolidated Standards of Reporting Trials (CONSORT) requirements. Patients were randomly assigned to receive either a combination of manual therapy and supervised exercise (MTEX), plus advice and home exercise or advice and home exercise alone (Ad+HEP) over six weeks. Primary outcomes were average pain (Visual Analog Scale) and physical function (Hip Outcome Score) at week seven. Fifteen patients, mean age 33.7 (SD 9.5, 73% female) satisfied the eligibility criteria and completed week seven measurements. The between group differences for changes in pain or physical function were not significant. Both groups showed statistically significant improvements in pain: the MTEX group improved a mean of 17.6mm and the Ad+HEP group, 18.0mm. The results of this pilot study provide preliminary evidence that symptomatic femoroacetabular impingement may be amenable to conservative treatment strategies however further full-scale randomized controlled trials are required to demonstrate this. In this small pilot study, supervised manual therapy and exercise did not result in greater improvement in pain or function compared to advice and home exercise in patients with symptomatic femoroacetabular impingement.

Simultaneous colorectal and hepatic procedures for colorectal cancer result in increased morbidity but equivalent mortality compared with colorectal or hepatic procedures alone: outcomes from the National Surgical Quality Improvement Program

BackgroundSimultaneous colorectal and hepatic surgery for colorectal cancer (CRC) is increasing as surgery becomes safer and less invasive. There is controversy regarding the morbidity associated with simultaneous, compared with separate or staged, resections. MethodsData for 2005–2008 from the National Surgical Quality Improvement Program (NSQIP) were used to compare morbidity after 19 925 colorectal procedures for CRC (CR group), 2295 hepatic resections for metastatic CRC (HEP group), and 314 simultaneous colorectal and hepatic resections (SIM group). ResultsAn increasing number of simultaneous resections were performed per year. Fewer major colorectal and liver resections were performed in the SIM than in the CR and HEP groups. Patients in the SIM group had a longer operative time and postoperative length of stay compared with those in either the CR or HEP groups. Simultaneous procedures resulted in higher rates of postoperative morbidity and major morbidity than CR procedures, but not HEP procedures. This difference was driven by higher rates of wound and organ space infections, and a greater incidence of septic shock. Mortality rates did not differ among the groups. ConclusionsHospitals in the NSQIP are performing more simultaneous colonic and hepatic resections for CRC. These procedures are associated with increases in operative time, length of stay and rate of perioperative complications. Simultaneous procedures do not, however, increase perioperative mortality.

Association between Interferon-Inducible Protein 6 (IFI6) Polymorphisms and Hepatitis B Virus Clearance

CD8+ T cells are key factors mediating hepatitis B virus (HBV) clearance. However, these cells are killed through HBV-induced apoptosis during the antigen-presenting period in HBV-induced chronic liver disease (CLD) patients. Interferon-inducible protein 6 (IFI6) delays type I interferon-induced apoptosis in cells. We hypothesized that single nucleotide polymorphisms (SNPs) in the IFI6 could affect the chronicity of CLD. The present study included a discovery stage, in which 195 CLD patients, including chronic hepatitis B (HEP) and cirrhosis patients and 107 spontaneous recovery (SR) controls, were analyzed. The genotype distributions of rs2808426 (C > T) and rs10902662 (C > T) were significantly different between the SR and HEP groups (odds ratio [OR], 6.60; 95% confidence interval [CI], 1.64 to 26.52, p = 0.008 for both SNPs) and between the SR and CLD groups (OR, 4.38; 95% CI, 1.25 to 15.26; p = 0.021 and OR, 4.12; 95% CI, 1.18 to 14.44; p = 0.027, respectively). The distribution of diplotypes that contained these SNPs was significantly different between the SR and HEP groups (OR, 6.58; 95% CI, 1.63 to 25.59; p = 0.008 and OR, 0.15; 95% CI, 0.04 to 0.61; p = 0.008, respectively) and between the SR and CLD groups (OR, 4.38; 95% CI, 1.25 to 15.26; p = 0.021 and OR, 4.12; 95% CI, 1.18 to 14.44; p = 0.027, respectively). We were unable to replicate the association shown by secondary enrolled samples. A large-scale validation study should be performed to confirm the association between IFI6 and HBV clearance.

Influence of chitosan nanofiber scaffold on porcine endogenous retroviral expression and infectivity in pig hepatocytes

To investigate the influence of chitosan nanofiber scaffold on the production and infectivity of porcine endogenous retrovirus (PERV) expressed by porcine hepatocytes. Freshly isolated porcine hepatocytes were cultured with or without chitosan nanofiber scaffold (defined as Nano group and Hep group) for 7 d. The daily collection of culture medium was used to detect reverse transcriptase (RT) activity with RT activity assay kits and PERV RNA by reverse transcription-polymerase chain reaction (PCR) and real time PCR with the PERV specific primers. And Western blotting was performed with the lysates of daily retrieved cells to determine the PERV protein gag p30. Besides, the in-vitro infectivity of the supernatant was tested by incubating the human embryo kidney 293 (HEK293) cells. The similar changing trends between two groups were observed in real time PCR, RT activity assay and Western blotting. Two peaks of PERV expression at 10H and Day 2 were found and followed by a regular decline. No significant difference was found between two groups except the significantly high level of PERV RNA at Day 6 and PERV protein at Day 5 in Nano group than that in Hep group. And in the in-vitro infection experiment, no HEK293 cell was infected by the supernatant. Chitosan nanofiber scaffold might prolong the PERV secreting time in pig hepatocytes but would not obviously influence its productive amount and infectivity, so it could be applied in the bioartificial liver without the increased risk of the virus transmission.