Nucleosomes in serum as a marker for cell death

The concentration of nucleosomes is elevated in blood of patients with diseases which are associated with enhanced cell death. In order to detect these circulating nucleosomes, we used the Cell Death Detection-ELISA(Plus) (CDDE) from Roche Diagnostics (Mannheim, Germany) (details at http:\textbackslash{}\textbackslash{}biochem.roche.com). For its application in liquid materials we performed various modifications: we introduced a standard curve with nucleosome-rich material, which enabled direct quantification and improved comparability of the values within (CVinterassay:3.0-4.1%) and between several runs (CVinterassay:8.6-13.5%), and tested the analytical specificity of the ELISA. Because of the fast elimination of nucleosomes from circulation and their limited stability, we compared plasma and serum matrix and investigated in detail the pre-analytical handling of serum samples which can considerably influence the test results. Careless venipuncture producing hemolysis, delayed centrifugation and bacterial contamination of the blood samples led to false-positive results; delayed stabilization with EDTA and insufficient storage conditions resulted in false-negative values. At temperatures of -20 degreesC, serum samples which were treated with 10 mM EDTA were stable for at least 6 months. In order to avoid possible interfering factors, we recommend a schedule for the pre-analytical handling of the samples. As the first stage, the possible clinical application was investigated in the sera of 310 persons. Patients with solid tumors (n = 220; mean = 361 Arbitrary Units (AU)) had considerably higher values than healthy persons (n = 50; mean = 30 AU; P = 0.0001) and patients with inflammatory diseases (n = 40; mean = 296 AU; p = 0.096). Within the group of patients with tumors, those in advanced stages (UICC 4) showed significantly higher values than those in early stages (UICC 1-3) (P = 0.0004)

Hypnosis for acute procedural pain: a critical review

Clinical evidence for the effectiveness of hypnosis in the treatment of acute procedural pain was critically evaluated based on reports from randomized controlled clinical trials (RCTs). Results from the 29 RCTs meeting inclusion criteria suggest that hypnosis decreases pain compared to standard care and attention control groups and that it is at least as effective as comparable adjunct psychological or behavioral therapies. In addition, applying hypnosis in multiple sessions prior to the day of the procedure produced the highest percentage of significant results. Hypnosis was most effective in minor surgical procedures. However, interpretations are limited by considerable risk of bias. Further studies using minimally effective control conditions and systematic control of intervention dose and timing are required to strengthen conclusions

Dependence of TIMP-1 plasma levels on preanalytical specimen handling

Background: Tissue inhibitor of metalloproteinases-1 (TIMP-1) in blood might be a helpful biomarker in various diseases. However, various authors report that TIMP-1 is dependent on preanalytical procedures. Our study was performed to determine how storage conditions and time to centrifugation influence TIMP-1. Materials and Methods: Twenty-six blood specimens were collected from each of 20 volunteers. Two specimens from each person were centrifuged/measured within 1 h after venipuncture and frozen at -80 degrees C. They were thawed once or twice within 72 h. Eight specimens were stored at 20 degrees C in daylight, 8 at 20 degrees C covered and 8 at 4 C in daylight. Four of each of these 8 specimens were mixed once a day until centrifugation. A mixed and an unmixed specimen of each group was centrifuged/measured after 3, 6, 24 and 72 h. Results: TIMP-1 increased after freeze/ thaw (p < 0.001). Mixing blood specimens more than once caused increased TIMP-1 (p < 0.001). TIMP-1 increased within 3 h of storage (p < 0.001). The increase was lower in specimens covered and refrigerated (p < 0.001). Conclusion: TIMP-1 is unstable and has to be evaluated carefully. Blood should be centrifuged directly after venipuncture. For routine application, specimen handling must be standardized and carefully followed. Research should be done on specimens handled identically. Copyright (C) 2008 S. Karger AG, Basel

Stratification of THROMBOTIC RISK in patients that are undergoing to PICC implantation. The “CoRa-MaPiCC STUDY”

Background: The PICC is a central venous system inserted in the peripherally vein. The central peripheral catheter catheter system (PICC=peripherally inserted central catheters) is a widely used procedure in clinical practice by medical and nurse team. PICC has potential complications that are rare but not entirely irrelevant. It is used for continuous and discontinuous treatment and, after a correct insertion procedure, it can be used for a period of time between 1 week to 3 months. The last-generation dispositive has an high-biocompatibility due to the high quality of the materials (silicone or polyurethane), a total length of 40-60 cm and a diameter between 16 to 25 Gauge or 2 to 5 French. Aims: The aim of our study is to evaluate the trombotic risk related to the positioning of the device in relation to the individual risk factors. Therefore we have elaborated the CoRaMaPicc protocol that allows, based on the preliminary data evaluation, to reduce the trombotic risk in patients with high cardiovascular risk profile. Methods: The study has started in July 2016. We have enrolled 30 patients (18 males and 12 females with range age of 30-99 yrs and mean age of 82+13,5 yrs). Enrolled patients have Body mass Index or (BMI) with range from 16,3-26,5 Kg/mq and mean BMI value of 23,5+2,5 Kg/mq. Enrolled patients were subjected to laboratory assessment of cardiovascular risk factors like laboratory assessment of Methyl-Tetra-Hydro-Folate-Reductase or (MTHFR) and Homocysteine, Fibrinogen, D-Dimer, International Normalized Ratio (INR), Prothrombin Time (PT), activated Partial Thromboplastin Time (aPTT), Platelets (PLT), trombotic risk evaluation (CHA2DS2-VASc Score), and bledding risk evaluation (HAS-BLED Score). Before the insertion procedure of the PICC, single or duble lumen (Groshong or Power PICC type), the patient were subjected to a preliminary ultrasound and echo-doppler vein evaluation to underline the presence of atipical anatomical state of the patients veins. After the procedure the patients were subjected to second ultrasound and echo-doppler vein evaluation and then to a radiographic examination to verify the correct position of the PICC. All patients were enrolled for follow- Up to evaluate trombotic complications. Statistical analysis was performed using the Survival Analysis test to evaluate the patients during the follow-up period in relation to thrombotic events. The data obtained from our study have been described taking also into account the data present in the international literature. Setting: The study was performed using a specific setting in a room of the Internal Medicine Department of the Alfredo Fiorini Hospital (Terracina, Italy), guarantee an aseptic procedure , the right comfort for the patients and the appropriate privacy. Discussion: More data in the literature underline the presence of non-eliminable risk factors like anatomic state of the patients vein that can play a variable role of procedural risk factor and can predispose to procedural or post-procedural complication after the implantation procedure. In specific cases it is difficult to asses an echo-doppler examination of heart for meteorism or ascites and radiographic exams for the presence of pleural effusion. The CoRaMaPiCC protocol use different technique to explore the post-insertion period and the follow-up time at 12 week. In case of meteorism or ascites the ultrasound examination of the heart cameras was replaced by the radiographyc evaluation while in case of pleural effusion, the evaluation of the device position was deduced using the only ultrasound approach (when the vein system is explorable). Procedural algorithm utilized in our implant protocol allows to improve the diagnostic capabilities of the conditions that can predispose to thrombotic complications. During the study period (1 year), the PICCs were removed for death (7 patients; 4 patients for irreversible heart disease, 1 patient for cardiogenic shock 1 patient for septic shock and 1 patient for Creutzfeld-Jacob disease), for spontaneously remotion (1 patient for low compliance of the patient) and for end of therapy (22 patients). The presence of cardiovascular risk factors alone is not sufficient for determining statistically significant changes in the thrombotic risk. As reported historically by the description of the wirkow triad, the presence of three clinical signs, and in particular the presence of endothelial damage (functio lesa) is a sine qua non condition for the occurrence of thrombotic episodes. Conclusion: The CoRaMaPicc protocol allows, based on the evaluation of the data, to minimize endothelial damage and therefore to reduce the incidence of the wirchof's triade related with an high trombotic risk. The data were elaborated considering the different thrombotic risk linked to the presence of the homozygous MTHR mutation responsible for the increase of homocysteine values and therefore of the further thrombotic risk. Further studies are needed to expand the number of enrolled patients and increase the follow-up period. Our data give comfortable results but further evaluation are needed to have conclusive results

Development of the Innate Immune Response in NestlingTree Swallows (Tachycineta bicolor)

The innate immune system provides an immediate, short term, first line of defense from pathogens; its appearance early in development in vertebrates is evidence of its critical importance. Even so, few studies have investigated the development of the immune response as juveniles transition into adults. Ultimately, the ability to respond to pathogens confers fitness benefits in terms of health, survival, and reproductive success, and it follows that functions such as rapid growth cannot be fully met simultaneously since energy is a limiting resource. As a result, defense mechanisms are compromised at an early age due to energy allocation to rapid growth; therefore, immunity should increase as individuals mature. I studied the development of innate immunity in nestling Tree Swallows using microbicidal assays which were conducted in vitro to assess the ability of the immune system to kill E. coli via lysis. This research may provide insight into patterns of disease susceptibility, which in turn influence evolutionary fitness and population dynamics

Improving Surveillance of Hepatitis C Infections Among Patients Receiving Medication Treatment for Opioid Use Disorder

Background: Despite harm reduction efforts, Hepatitis C virus (HCV) infections among individuals with opioid use disorder tripled between 2010 and 2015 in the US. Shifting disease burden from chronic to acute HCV, could result in $12 billion cumulative Medicaid savings in two years. Patients receiving medication for OUD (mOUD) remain at risk for HCV, but only receive screening upon admission. This project aims to implement a HCV re-screening protocol in a mOUD program, determine protocol sustainability, and secondarily evaluate need for point of care testing. Methods: Patients with a negative screening result upon admission were prompted to opt in/out of re-screening. Participants were offered onsite venipuncture. Patients declining, were asked if they’d reconsider if blood sample were obtained via finger stick. Stakeholders provided feedback regarding sustainability via online surveys. Results: Of the 496 eligible patients, 429[PMV1] (86%) completed the survey with 124 (29%) interested in rescreening, and 52 (41%) completed a venipuncture. Of these, 9 (17%) tested positive for HCV antibody and 7 (14%) had detectable HCV RNA. Of patients declining re-screening, n=185[PMV2] (52%), endorsed amenability to finger stick. Stakeholders (n=24) agreed that rescreening was easy to implement and appropriate to integrate into practice. Conclusions: Provision of less invasive screening modalities may increase rescreening uptake. Introducing opportunities for HCV re-screening leads to earlier detection and subsequent treatment initiation, and increases identification rates overall. Integration of HCV screening services at mOUD treatment programs provides a sustainable approach to HCV surveillance, and has the potential reduce the burden of infectious disease

Pain assessment in children undergoing venipuncture: the Wong–Baker faces scale versus skin conductance fluctuations

The aim of this study was to evaluate the efficacy of the subjective Wong–Baker faces pain rating scale (WBFS) and of the objective skin conductance fluctuation (SCF) test in assessing pain in children undergoing venipuncture. One-hundred and fifty children (aged 5–16 years) entered the study. All underwent venipuncture at the antecubital fossa to collect blood specimens for routine testing in the same environmental conditions. After venipuncture, the children indicated their pain intensity using the WBFS, whereas the number of SCFs was recorded before, during and after venipuncture. So, pain level was measured in each child with WBFS and SCF. We found that the level of WBFS-assessed pain was lower in all children, particularly those above 8 years of age, than SCF-assessed pain (p < 0.0001). Moreover, the number of SCFs was significantly higher during venipuncture than before or after venipuncture (p < 0.0001). At multivariate regression analysis, age and previous experience of venipuncture influenced the WBFS (β = −1.81, p < 0.001, and β = −0.86, p < 0.001, respectively) but not SCFs. In conclusion, although both procedures can be useful for research and clinical practice, our findings show that WBFS was affected by age and previous venipuncture, whereas SCF produced uniform data. If verified in other studies, our results should be taken into account when using these tools to evaluate pain in children

Validation of the Medonic CA620/530 Vet 20-ml microcapillary sampler system for hematology testing of feline blood

The aim of the current study was to compare feline hematologic variables in blood collected in microcapillary tubes (20 ml) and conventional blood tubes with the Medonic CA620/530 Vet in-house hematologic analyzer. A comparison of results obtained in 60 cats presented at the clinics of the veterinary school showed that the correlations between the 2 methods were 0.97 for white blood cell, 0.95 for red blood cell, and 0.93 for platelet counts; 0.92 for hemoglobin concentration; and 0.99 for mean corpuscular volume. No clinically relevant differences between the 2 blood sampling techniques were observed for any variable, which suggests that both techniques are interchangeable in cats. Moreover, microcapillary tubes would allow easier repeated sampling in the same cat and would likely be useful in other small species

Testosterone Trajectories and Reference Ranges in a Large Longitudinal Sample of Male Adolescents

Pubertal dynamics plays an important role in physical and psychological development of children and adolescents. We aim to provide reference ranges of plasma testosterone in a large longitudinal sample. Furthermore, we describe a measure of testosterone trajectories during adolescence that can be used in future investigations of development.We carried out longitudinal measurements of plasma testosterone in 2,216 samples obtained from 513 males (9 to 17 years of age) from the Avon Longitudinal Study of Parents and Children. We used integration of a model fitted to each participant's testosterone trajectory to calculate a measure of average exposure to testosterone over adolescence. We pooled these data with corresponding values reported in the literature to provide a reference range of testosterone levels in males between the ages of 6 and 19 years.The average values of total testosterone in the ALSPAC sample range from 0.82 nmol/L (Standard Deviation [SD]: 0.09) at 9 years of age to 16.5 (SD: 2.65) nmol/L at 17 years of age; these values are congruent with other reports in the literature. The average exposure to testosterone is associated with different features of testosterone trajectories such as Peak Testosterone Change, Age at Peak Testosterone Change, and Testosterone at 17 years of age as well as the timing of the growth spurt during puberty.The average exposure to testosterone is a useful measure for future investigations using testosterone trajectories to examine pubertal dynamics