Temporal Changes in Innate and Adaptive Immunity During Sepsis as Determined by ELISpot.

Background: The inability to evaluate host immunity in a rapid quantitative manner in patients with sepsis has severely hampered development of novel immune therapies. The ELISpot assay is a functional bioassay that measures the number of cytokine-secreting cells and the relative amount of cytokine produced at the single-cell level. A key advantage of ELISpot is its excellent dynamic range enabling a more precise quantifiable assessment of host immunity. Herein, we tested the hypothesis on whether the ELISpot assay can detect dynamic changes in both innate and adaptive immunity as they often occur during sepsis. We also tested whether ELISpot could detect the effect of immune drug therapies to modulate innate and adaptive immunity. Methods: Mice were made septic using sublethal cecal ligation and puncture (CLP). Blood and spleens were harvested serially and ex vivo IFN-γ and TNF-α production were compared by ELISpot and ELISA. The capability of ELISpot to detect changes in innate and adaptive immunity due to in vivo immune therapy with dexamethasone, IL-7, and arginine was also evaluated. Results: ELISpot confirmed a decreased innate and adaptive immunity responsiveness during sepsis progression. More importantly, ELISpot was also able to detect changes in adaptive and innate immunity in response to immune-modulatory reagents, for example dexamethasone, arginine, and IL-7 in a readily quantifiable manner, as predicted by the reagents known mechanisms of action. ELISpot and ELISA results tended to parallel one another although some differences were noted. Conclusion: ELISpot offers a unique capability to assess the functional status of both adaptive and innate immunity over time. The results presented herein demonstrate that ELISpot can also be used to detect and follow the in vivo effects of drugs to ameliorate sepsis-induced immune dysfunction. This capability would be a major advance in guiding new immune therapies in sepsis.

Binocular rivalry from luminance and contrast.

Binocular rivalry is the phenomenon that when two incompatible images are simultaneously presented, one to each eye, the two images compete with each other to be the dominant percept. Studying the underlying neural mechanisms of binocular rivalry is useful for understanding the mechanisms of interocular inhibition. Levelt's Propositions, a set of four propositions that were originally published over fifty years ago, are not only useful for characterizing the perceptual dynamics of binocular rivalry, but can also provide a metric for assessing the common or differential neural mechanisms of binocular rivalry when diverse stimulus types are used. In the present study, we conducted a series of psychophysics experiments, where we compared the rivalry dynamics of two quite different types of stimuli. Orthogonal gratings, a classic type of rivalry stimulus, were contrasted with luminance patches, a type of rivalry stimulus that is relatively less studied. Our results showed that, similar to the orthogonal gratings, the alternate percepts in luminance-only rivalry were described by the modified Levelt's Propositions, despite the clearly slower alternation rates for luminance patches. However, unlike the mixed percepts observed during transitions between oriented gratings, fusion percepts during luminance rivalry were common, could be lustrous, and obeyed the same Propositions, suggesting a regime of tri-stability. Overall, both types of rivalry are consistent with recent models that posit separate binocular and monocular channels embedded within neural circuits that also accomplish contrast normalization. Finally, luminance rivalry is discussed in the contexts of binocular summation and suppression, as well as Fechner's paradox.

Biomarker analyses in REGARD gastric/GEJ carcinoma patients treated with VEGFR2-targeted antibody ramucirumab.

BACKGROUND: Angiogenesis inhibition is an important strategy for cancer treatment. Ramucirumab, a human IgG1 monoclonal antibody that targets VEGF receptor 2 (VEGFR2), inhibits VEGF-A, -C, -D binding and endothelial cell proliferation. To attempt to identify prognostic and predictive biomarkers, retrospective analyses were used to assess tumour (HER2, VEGFR2) and serum (VEGF-C and -D, and soluble (s) VEGFR1 and 3) biomarkers in phase 3 REGARD patients with metastatic gastric/gastroesophageal junction carcinoma. METHODS: A total of 152 out of 355 (43%) patients randomised to ramucirumab or placebo had ⩾1 evaluable biomarker result using VEGFR2 immunohistochemistry or HER2, immunohistochemistry or FISH, of blinded baseline tumour tissue samples. Serum samples (32 patients, 9%) were assayed for VEGF-C and -D, and sVEGFR1 and 3. RESULTS: None of the biomarkers tested were associated with ramucirumab efficacy at a level of statistical significance. High VEGFR2 endothelial expression was associated with a non-significant prognostic trend toward shorter progression-free survival (high vs low HR=1.65, 95% CI=0.84,3.23). Treatment with ramucirumab was associated with a trend toward improved survival in both high (HR=0.69, 95% CI=0.38, 1.22) and low (HR=0.73, 95% CI=0.42, 1.26) VEGFR2 subgroups. The benefit associated with ramucirumab did not appear to differ by tumoural HER2 expression. CONCLUSIONS: REGARD exploratory analyses did not identify a strong potentially predictive biomarker of ramucirumab efficacy; however, statistical power was limited.

Effect of premedication regimen on infant pain and stress response to endotracheal intubation.

OBJECTIVE: (1) Evaluate the effect of different medications on pain and stress in neonates during nonemergent endotracheal intubation; (2) determine whether gestational age affects medication use; (3) determine whether better sedation results in a decrease in the number of attempts and/or total time for the procedure. STUDY DESIGN: Prospective observational study. Infant responses were measured using a clinical pain scale and blood glucose, a biochemical marker of acute stress. RESULT: A total of 166 infants were included, with adjusted gestational ages 24 to 44 weeks at the time of procedure. Premedication regimens included no medication ('none,' 27%), morphine (19%), morphine+midazolam (11%), fentanyl (14%), fentanyl+midazolam (19%) and midazolam alone (10%). Fentanyl+midazolam resulted in lower pain scores and less increase in blood glucose (both P<0.0001). No other regimen was different from 'none'. The most immature infants were less likely to receive premedication (P=0.023), although their pain scores and blood glucose responses were similar to more mature infants. None of the medication regimens reduced the total procedure time (P=0.55) or the number of attempts (P=0.145). CONCLUSION: Only fentanyl+midazolam significantly attenuated both the clinical pain score and the increase in blood glucose. Less mature infants had responses similar to those of more mature infants, but were less likely to receive premedication. None of the regimens decreased the time or number of attempts required for successful intubation.

Standards for vision science libraries: 2014 revision.

OBJECTIVE: This Association of Vision Science Librarians revision of the "Standards for Vision Science Libraries" aspires to provide benchmarks to address the needs for the services and resources of modern vision science libraries (academic, medical or hospital, pharmaceutical, and so on), which share a core mission, are varied by type, and are located throughout the world. METHODS: Through multiple meeting discussions, member surveys, and a collaborative revision process, the standards have been updated for the first time in over a decade. RESULTS: While the range of types of libraries supporting vision science services, education, and research is wide, all libraries, regardless of type, share core attributes, which the standards address. CONCLUSIONS: The current standards can and should be used to help develop new vision science libraries or to expand the growth of existing libraries, as well as to support vision science librarians in their work to better provide services and resources to their respective users.

Inhibition of the cannabinoid 2 receptor in CNS-injury induced immunodeficiency syndrome.

Central nervous system (CNS) injury is classified as an independent factor, increasing patients' susceptibility to infections. The concept of infection susceptibility and impaired immune function is referred to as CNS-injury induced immunodeficiency syndrome (CIDS). The endocannabinoid system, an important homeostatic system that can modulate immune function, contributes to the consequences of an acute CNS injury. The actions of the endocannabinoid system are mediated via cannabinoid receptors, cannabinoid 1 (CB1R) and cannabinoid 2 (CB2R), the latter of which are highly expressed by immune cells and upregulated as a result of infectious and non-infectious stressors. While the role of the CB2R in CNS immunity is primarily anti-inflammatory, focusing on the inhibition of the CB2R pathways may be of benefit for therapeutic targeting of the immunosuppression in CIDS. We hypothesize that inhibition of the CB2R will result in a decrease in the immunosuppression seen in CIDS, providing the patient protection against common infections such as pneumonia and urinary tract infections. However, due to the high variability of the patients' immune status during and after an acute CNS injury, identifying the most effective therapeutic window and CB2R antagonist dosage for effective immunostimulation is pivotal.

Sci-Fri AM: Imaging - 07: Symmetric geometric transfer matrix partial volume correction technique for emission tomography: Principle, validation, and robustness.

Partial volume correction (PVC) is often needed to correct for limited spatial resolution in quantitative Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) studies. In conventional region-based PVC methods, spill over between regions segmented from coregistered computed tomography (CT) or magnetic resonance (MR) images is accounted for by calculating regional spread functions (RSFs) in a geometric transfer matrix (GTM) framework. This paper describes a new analytically derived symmetric GTM (sGTM) method that considers spill over between RSFs rather than between regions. The sGTM is mathematically equivalent to Labbe's method, however it is region-based rather than voxel-based and it avoids handling large matrices. The sGTM method was validated using an MR-based 3D digital brain phantom and a physical phantom containing spheres 5 mm to 30 mm in diameter. The sGTM method was compared to the GTM method in terms of accuracy, precision, noise propagation, and robustness, i.e. effects of mis-registration or point spread function (PSF) estimation errors. The results showed that the sGTM method has accuracy similar to that of the GTM method, and within 5% of the true value. However, the sGTM method showed better precision and noise propagation than the GTM method, especially for spheres smaller than 13 mm. Moreover, the sGTM method was more robust than the GTM method when misregistration or errors in estimates of PSF occurred. In conclusion, the sGTM method was analytically derived and validated and shown to exhibit better noise characteristics and robustness compared to the GTM method.

Combined liver kidney transplantation: critical analysis of a single-center experience.

Combined liver kidney transplantation (LKT) can be successfully performed on patients with liver and renal failure; however, outcomes are inferior to liver transplantation alone (OLT). Our aim was to determine the indications for and outcome of LKT and whether patients with longer wait times required more frequent LKT versus OLT alone. We included 18/93 adults who underwent LKT from August 2007 to August 2010 for hepatitis C virus (HCV, n = 7), alcohol (n = 5), nonalcoholic steatohepatitis (n = 2), primary biliary sclerosis, polycystic kidney disease with liver involvement, hepatic adenomatosis, and ischemic hepatitis. Eleven were originally listed for LKT and 7 required listing for-kidney transplantation while awaiting OLT. Eight were on dialysis when first listed and 10 had a low glomerular filtration rate or known kidney disease. The mean calculated Model for End-Stage Liver Disease (MELD) score for LKT was 31.2 ± 3.54. Seven had hepatocellular carcinoma in explants. Two patients had acute cellular kidney rejection that responded to treatment. Recurrence of HCV was documented in 5 patients within 6 months of LKT; 2/5 received HCV therapy (interferon and ribavirin) without renal allograft rejection. One-year liver graft/patient survival was 94% after LKT. One patient died at 6 months post LKT due to severe HCV recurrence. Last mean serum creatinine level was 1.35 ± 0.28 mg/dL for LKT patients. LKT is a safe procedure with favorable outcomes even in patients with a high MELD score. Transplantation of patients with a high MELD score due to regional variations in organ allocation results in additional use of kidneys by OLT patients. Improved organ allocation algorithms in OLT would help to reduce combined transplants, sparing more kidneys.

Successful treatment of fibrosing cholestatic hepatitis after liver transplantation.

BACKGROUND: A minority of liver transplant (OLT) recipients with hepatitis C virus (HCV) develop fibrosing cholestatic hepatitis (FCH), a severe form of HCV recurrence associated with early graft failure and death. There are few reports of successful salvage strategies. In this retrospective study, we sought to determine the characteristics and outcomes for patients with FCH at our transplant center. METHODS: All cases of HCV-positive OLT recipients from July 2007 through July 2010 were reviewed. Patient demographics, donor characteristics, and the post-OLT clinical course were analyzed. Tacrolimus-based immunosuppression was used. FCH was treated by conversion to cyclosporine A (CsA) and aggressive treatment with pegylated interferon (IFN) alpha2A and ribavirin (RBV). Liver biopsies and HCV RNA were obtained frequently per protocol or for cause. RESULTS: The rate of FCH during the study period was 13.5% (5/37). Of the 5 patients with FCH (4 males, 4 Caucasian), mean age was 51 (± 4.8) years and the Model for End-Stage Liver Disease (MELD) score at listing was 26.6 (± 10). Three of the 5 received liver and kidney (L/K) transplants (60%); the rate of L/K transplant in non-FCH patients was 12.5%. HCV RNA levels ranged from 5 to 6.69 log IU/mL pre-OLT; none were on anti-HCV therapy at the time of OLT. Mean ischemic time was 385 (± 152) minutes; donor age was 34.4 (± 13.7) years. No CMV infections developed postoperatively. Time to histologic HCV recurrence was 2 (± 2.23) months (range, 1-6); FCH occurred at 2.2 (± 2.2) months. Patients were converted from tacrolimus to CsA and treated with IFN and RBV; 2 were changed to consensus IFN. HCV RNA increased post-OLT in all, but responded to therapy in 4 of 5. None of the L/K recipients experienced renal graft rejection during treatment. Four of 5 had clinical and histologic improvement; 1 progressed to cirrhosis with minimal inflammation. One-year patient survival after OLT in this group was 80%. Liver allograft rejection occurred in 60% at 4.7 (± 5.5) months and was treated by CsA and prednisone dosage adjustments. In this cohort of patients undergoing OLT for HCV, FCH occurred early after OLT but responded to aggressive management with conversion from tacrolimus to CsA and treatment with pegylated IFN or consensus IFN/RBV. There was a higher rate of combined L/K transplants in the FCH group compared with the non-FCH group. Liver allograft rejection occurred in 60% of cases, but responded to treatment in all; no renal graft rejection occurred in the 3 with L/K transplants while on IFN. One-year graft and patient survival was 80%. CONCLUSION: Better survival with FCH is possible with early initiation of IFN/RBV therapy with close monitoring of biopsies and viral load, and conversion from tacrolimus to CsA. Treatment can be performed even in L/K transplantation recipients, although it is associated with a higher incidence of treatable liver allograft rejection.

Interviewer effects in public health surveys.

Interviewer effects can have a substantial impact on survey data and may be particularly operant in public health surveys, where respondents are likely to be queried about racial attitudes, sensitive behaviors and other topics prone to socially desirable responding. This paper defines interviewer effects, argues for the importance of measuring and controlling for interviewer effects in health surveys, provides advice about how to interpret research on interviewer effects and summarizes research to date on race, ethnicity and gender effects. Interviewer effects appear to be most likely to occur when survey items query attitudes about sociodemographic characteristics or respondents' engagement in sensitive behaviors such as substance use. However, there is surprisingly little evidence to indicate whether sociodemographic interviewer-respondent matching improves survey response rates or data validity, and the use of a matched design introduces possible measurement bias across studies. Additional research is needed to elucidate many issues, including the influence of interviewers' sociodemographic characteristics on health-related topics, the role of within-group interviewer variability on survey data and the simultaneous impact of multiple interviewer characteristics. The findings of such research would provide much-needed guidance to public health professionals on whether or not to match interviewers and respondents on key sociodemographic characteristics.

Developing a methodology for three-dimensional correlation of PET-CT images and whole-mount histopathology in non-small-cell lung cancer.

BACKGROUND: Understanding the three-dimensional (3D) volumetric relationship between imaging and functional or histopathologic heterogeneity of tumours is a key concept in the development of image-guided radiotherapy. Our aim was to develop a methodologic framework to enable the reconstruction of resected lung specimens containing non-small-cell lung cancer (NSCLC), to register the result in 3D with diagnostic imaging, and to import the reconstruction into a radiation treatment planning system. METHODS AND RESULTS: We recruited 12 patients for an investigation of radiology-pathology correlation (RPC) in nsclc. Before resection, imaging by positron emission tomography (PET) or computed tomography (CT) was obtained. Resected specimens were formalin-fixed for 1-24 hours before sectioning at 3-mm to 10-mm intervals. To try to retain the original shape, we embedded the specimens in agar before sectioning. Consecutive sections were laid out for photography and manually adjusted to maintain shape. Following embedding, the tissue blocks underwent whole-mount sectioning (4-mum sections) and staining with hematoxylin and eosin. Large histopathology slides were used to whole-mount entire sections for digitization. The correct sequence was maintained to assist in subsequent reconstruction. Using Photoshop (Adobe Systems Incorporated, San Jose, CA, U.S.A.), contours were placed on the photographic images to represent the external borders of the section and the extent of macroscopic disease. Sections were stacked in sequence and manually oriented in Photoshop. The macroscopic tumour contours were then transferred to MATLAB (The Mathworks, Natick, MA, U.S.A.) and stacked, producing 3D surface renderings of the resected specimen and embedded gross tumour. To evaluate the microscopic extent of disease, customized "tile-based" and commercial confocal panoramic laser scanning (TISSUEscope: Biomedical Photometrics, Waterloo, ON) systems were used to generate digital images of whole-mount histopathology sections. Using the digital whole-mount images and imaging software, we contoured the gross and microscopic extent of disease. Two methods of registering pathology and imaging were used. First, selected pet and ct images were transferred into Photoshop, where they were contoured, stacked, and reconstructed. After importing the pathology and the imaging contours to MATLAB, the contours were reconstructed, manually rotated, and rigidly registered. In the second method, MATLAB tumour renderings were exported to a software platform for manual registration with the original pet and ct images in multiple planes. Data from this software platform were then exported to the Pinnacle radiation treatment planning system in DICOM (Digital Imaging and Communications in Medicine) format. CONCLUSIONS: There is no one definitive method for 3D volumetric RPC in nsclc. An innovative approach to the 3D reconstruction of resected nsclc specimens incorporates agar embedding of the specimen and whole-mount digital histopathology. The reconstructions can be rigidly and manually registered to imaging modalities such as ct and pet and exported to a radiation treatment planning system.

Incidence of hepatocellular carcinoma among individuals with hepatitis B virus infection identified using an automated data algorithm.

The purpose of this study was to develop an algorithm for identifying patients with chronic hepatitis B virus (HBV) using automated data sources from two US health systems and evaluate the algorithm's performance by quantifying the incidence of hepatocellular carcinoma (HCC) among chronic HBV patients. To allow comparisons with estimates from automated databases that may not contain all data elements used in this algorithm, we created three definitions of chronic HBV infection and used these definitions to create three overlapping cohorts. We compared the incidence of HCC in each cohort with the incidence of HCC in a matched general population comparison cohort with no evidence of HBV. Patients who met the most stringent criteria for chronic HBV infection (based on the standard definition of 6 months of infection using repeat laboratory tests and record review) were 146 times more likely to develop HCC than matched comparison patients (adjusted hazard ratio = 146.5, 95% CI: 74.0-289.8). Those not meeting the stringent criteria, but who met the criterion of at least one positive hepatitis B surface antigen test were 30 times more likely to develop HCC than comparison patients (adjusted hazard ratio = 29.8, 95% CI: 16.5-53.6). Finally, patients who met the criterion based on at least one HBV diagnosis were 38 times more likely to develop HCC than matched comparison patients (adjusted hazard ratio = 37.8, 95% CI: 25.9-55.1). The magnitude of the relative increase in HCC risk seen using different criteria used to define HBV infection indicate that these automated data algorithms can identify patients with chronic HBV infection.

White blood cell telomerase activity and incident respiratory illness among community-dwelling elderly vaccinated against seasonal influenza.

Immune cell telomerase activity may impact vaccine response in the elderly. Fifty persons aged 60-100 years were tested for post-influenza vaccination telomerase RNA expression (TERT) in peripheral blood mononuclear cells to assess for an association with influenza antibody levels and influenza-like illness or incident respiratory infection (IRI) in the year following vaccination. High rates of seroprotective influenza antibody (> or = 1:40 titers) were observed post-vaccination (86-92% to vaccine viral strains), with no association to TERT. No IRI occurred among persons in the top quartile of TERT expression, whereas the IRI rate was 33% in the lower three quartiles (Kaplan-Meier P=0.028). TERT expression was also IRI significantly higher in those who did not experience IRI than those who did in the follow-up period (0.845 vs. 0.301, P=0.024). These data suggest that telomerase expression may correlate with immune capacity for vaccine response in the elderly and could represent a target for recognizing risk for vaccine failure.

Differential DNA methylation patterns of small B-cell lymphoma subclasses with different clinical behavior.

Non-Hodgkin's lymphoma (NHL) is a group of malignancies of the immune system with variable clinical behaviors and diverse molecular features. Despite the progress made in classification of NHLs based on classical methods, molecular classifications are a work in progress. Toward this goal, we used an array-based technique called differential methylation hybridization (DMH) to study small B-cell lymphoma (SBCL) subtypes. A total of 43 genomic DMH experiments were performed. From these results, several statistical methods were used to generate a set of differentially methylated genes for further validation. Methylation of LHX2, POU3F3, HOXC10, NRP2, PRKCE, RAMP, MLLT2, NKX6.1, LRP1B and ARF4 was validated in cell lines and patient samples and demonstrated subtype-related preferential methylation patterns. For LHX2 and LRP1B, bisulfite sequencing, real-time reverse transcriptase-polymerase chain reaction and induction of gene expression following treatment with the demethylating agent, 5'-aza-2'-deoxycytidine, were confirmed. This new epigenetic information is helping to define molecular portraits of distinct subtypes of SBCL that are not recognized by current classification systems and provides valuable potential insights into the biology of these tumors.