Delirium Screening and Pharmacotherapy in the ICU: The Patients Are Not the Only Ones Confused.
Background: Delirium is difficult to measure in the Intensive Care Unit (ICU). It is possible that by considering the rate of screening, incidence, and rate of treatment with antipsychotic medications (APMs) for suspected delirium, a clearer picture can emerge. Methods: A retrospective, observational study was conducted at two ICUs in Australia, between April and June of 2020. All adult ICU patients were screened; those who spoke English and did not have previous neurocognitive pathology or intracranial pathology were included in the analysis. Data were collected from the hospitals' electronic medical records. The primary outcome was incidence of delirium based on the use of the Confusion Assessment Method for ICU (CAM-ICU). Secondary outcomes included measures of screening for delirium, treatment of suspected delirium with APMs, and identifying clinical factors associated with both delirium and the use of APMs. Results: From 736 patients that were screened, 665 were included in the analysis. The incidence of delirium was 11.3% (75/665); on average, the Richmond Agitation and Sedation Scale (RASS) was performed every 2.9 h and CAM-ICU every 40 h. RASS was not performed in 8.4% (56/665) of patients and CAM-ICU was not performed in 40.6% (270/665) of patients. A total of 17% (113/665) of patients were prescribed an APM, with quetiapine being the most used. ICU length of stay (LOS), APACHE-III score, and the use of alpha-2 agonists were associated with the presence of delirium, while ICU LOS, the use of alpha-2 agonists, and the presence of delirium were associated with patients receiving APMs. Conclusions: The incidence of delirium was lower than previously reported, at 11.3%. The rate of screening for delirium was low, while the use of APMs for delirium was higher than the incidence of delirium. It is possible that the true incidence is higher than what was measured. Critical prospective assessment is required to optimize APM indications in the ICU.


[Test equipment for relative spectral response of CCD].
The auto-test equipment for relative spectral response of charge coupled devices (CCD) was designed and realized. The equipment covered the range of 400 to 950 nm. Firstly, testing theory of relative spectral response for detectors was introduced. Secondly, taking the high sensitive and broad spectral scientific spectrometer-QE65000 as reference, auto-test equipment for relative spectral response was built up by direct comparison method in the same radiation field. Uncertainty analysis showed that the maximum uncertainty of the equipment was less than 6.21%. This auto-test equipment can be used in the CCD assessment and its photoelectrical parameters testing.


Detection of collagen by multi-wavelength photoacoustic analysis as a biomarker for bone health assessment.
Collagen is an important biomarker of osteoporosis progression. Noninvasive, multispectral, photoacoustic (PA) techniques use pulsed laser light to induce PA signals to facilitate the visualization of chemical components that are strongly related to tissue health. In this study, the feasibility of multi-wavelength PA (MWPA) measurement of the collagen in bone, using the wavelength range of 1300-1800 nm, was investigated. First, the feasibility of this approach for detecting the collagen content of bone was demonstrated by means of numerical simulation. Then, ex vivo experiments were conducted on both animal and human bone specimens with different bone densities using the MWPA method. The relative collagen content was extracted and compared with the results of micro-computed tomography (micro-CT) and histology. The results showed that the "relative collagen content" parameter obtained using the MWPA approach correlated well with the bone volume ratio obtained from micro-CT images and histological analysis results. This study highlights the potential of the proposed PA technique for determining the collagen content of bones as a biomarker for bone health assessment.


Effect of an Early Resuscitation Protocol on In-hospital Mortality Among Adults With Sepsis and Hypotension: A Randomized Clinical Trial.
The effect of an early resuscitation protocol on sepsis outcomes in developing countries remains unknown. To determine whether an early resuscitation protocol with administration of intravenous fluids, vasopressors, and blood transfusion decreases mortality among Zambian adults with sepsis and hypotension compared with usual care. Randomized clinical trial of 212 adults with sepsis (suspected infection plus ≥2 systemic inflammatory response syndrome criteria) and hypotension (systolic blood pressure ≤90 mm Hg or mean arterial pressure ≤65 mm Hg) presenting to the emergency department at a 1500-bed referral hospital in Zambia between October 22, 2012, and November 11, 2013. Data collection concluded December 9, 2013. Patients were randomized 1:1 to either (1) an early resuscitation protocol for sepsis (n = 107) that included intravenous fluid bolus administration with monitoring of jugular venous pressure, respiratory rate, and arterial oxygen saturation and treatment with vasopressors targeting mean arterial pressure (≥65 mm Hg) and blood transfusion (for patients with a hemoglobin level <7 g/dL) or (2) usual care (n = 105) in which treating clinicians determined hemodynamic management. The primary outcome was in-hospital mortality and the secondary outcomes included the volume of intravenous fluid received and receipt of vasopressors. Among 212 patients randomized to receive either the sepsis protocol or usual care, 3 were ineligible and the remaining 209 completed the study and were included in the analysis (mean [SD] age, 36.7 [12.4] years; 117 men [56.0%]; 187 [89.5%] positive for the human immunodeficiency virus). The primary outcome of in-hospital mortality occurred in 51 of 106 patients (48.1%) in the sepsis protocol group compared with 34 of 103 patients (33.0%) in the usual care group (between-group difference, 15.1% [95% CI, 2.0%-28.3%]; relative risk, 1.46 [95% CI, 1.04-2.05]; P = .03). In the 6 hours after presentation to the emergency department, patients in the sepsis protocol group received a median of 3.5 L (interquartile range, 2.7-4.0 L) of intravenous fluid compared with 2.0 L (interquartile range, 1.0-2.5 L) in the usual care group (mean difference, 1.2 L [95% CI, 1.0-1.5 L]; P < .001). Fifteen patients (14.2%) in the sepsis protocol group and 2 patients (1.9%) in the usual care group received vasopressors (between-group difference, 12.3% [95% CI, 5.1%-19.4%]; P < .001). Among adults with sepsis and hypotension, most of whom were positive for HIV, in a resource-limited setting, a protocol for early resuscitation with administration of intravenous fluids and vasopressors increased in-hospital mortality compared with usual care. Further studies are needed to understand the effects of administration of intravenous fluid boluses and vasopressors in patients with sepsis across different low- and middle-income clinical settings and patient populations. clinicaltrials.gov Identifier: NCT01663701.


Predicting mechanism of biphasic growth factor action on tumor growth using a multi-species model with feedback control.
A large number of growth factors and drugs are known to act in a biphasic manner: at lower concentrations they cause increased division of target cells, whereas at higher concentrations the mitogenic effect is inhibited. Often, the molecular details of the mitogenic effect of the growth factor are known, whereas the inhibitory effect is not. Hepatoctyte Growth Factor, HGF, has recently been recognized as a strong mitogen that is present in the microenvironment of solid tumors. Recent evidence suggests that HGF acts in a biphasic manner on tumor growth. We build a multi-species model of HGF action on tumor cells using different hypotheses for high dose-HGF activation of a growth inhibitor and show that the shape of the dose-response curve is directly related to the mechanism of inhibitor activation. We thus hypothesize that the shape of a dose-response curve is informative of the molecular action of the growth factor on the growth inhibitor.


A clinical study of collagen meniscus implants to restore the injured meniscus.
The meniscus performs critical functions within the knee, and its loss frequently leads to osteoarthritis and irreversible joint damage. Because prosthetic replacement of the meniscus has proven ineffective, the authors used tissue engineering techniques to develop a resorbable collagen scaffold (collagen meniscus implant) that supports ingrowth of new tissue and eventual regeneration of the lost meniscus. Eight patients underwent arthroscopic placement the collagen meniscus implant to reconstruct and restore the irreparably damaged medial meniscus of one knee. Seven patients had one or more prior meniscectomies, and one patient had an acute meniscus injury. Patients were observed with frequent clinical, serologic, radiographic, and magnetic resonance imaging examinations for at least 24 months (range, 24-32 months). All patients underwent relook arthroscopy and biopsy of the implant regenerated tissue at either 6 or 12 months after implantation. All patients improved clinically from preoperatively to 1 and 2 years postoperatively based on pain, Lysholm scores, Tegner activity scale, and self assessment. Relook arthroscopy revealed tissue regeneration in all patients with apparent preservation of the joint surfaces based on visual observations. Histologic analysis confirmed new fibrocartilage matrix formation. Radiographs confirmed no progression of degenerative joint disease. The collagen meniscus implant is implantable, biocompatible, resorbable, and supports new tissue regeneration as it is resorbed. This tissue seems to function similar to meniscus tissue by protecting the chondral surfaces.


Renalase Protects against Renal Fibrosis by Inhibiting the Activation of the ERK Signaling Pathways.
Renal interstitial fibrosis is a common pathway for the progression of chronic kidney disease (CKD) to end-stage renal disease. Renalase, acting as a signaling molecule, has been reported to have cardiovascular and renal protective effects. However, its role in renal fibrosis remains unknown. In this study, we evaluated the therapeutic efficacy of renalase in rats with complete unilateral ureteral obstruction (UUO) and examined the inhibitory effects of renalase on transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) in human proximal renal tubular epithelial (HK-2) cells. We found that in the UUO model, the expression of renalase was markedly downregulated and adenoviral-mediated expression of renalase significantly attenuated renal interstitial fibrosis, as evidenced by the maintenance of E-cadherin expression and suppressed expression of α-smooth muscle actin (α-SMA), fibronectin and collagen-I. In vitro, renalase inhibited TGF-β1-mediated upregulation of α-SMA and downregulation of E-cadherin. Increased levels of Phospho-extracellular regulated protein kinases (p-ERK1/2) in TGF-β1-stimulated cells were reversed by renalase cotreatment. When ERK1 was overexpressed, the inhibition of TGF-β1-induced EMT and fibrosis mediated by renalase was attenuated. Our study provides the first evidence that renalase can ameliorate renal interstitial fibrosis by suppression of tubular EMT through inhibition of the ERK pathway. These results suggest that renalase has potential renoprotective effects in renal interstitial fibrosis and may be an effective agent for slowing CKD progression.


Role of nitric oxide in gut ischemia-reperfusion-induced hepatic microvascular dysfunction.
The overall objective of this study was to assess the contribution of an altered bioavailability of nitric oxide (NO) to the leukocyte adhesion and hypoxic stress elicited in the liver by gut ischemia-reperfusion (I/R). The accumulation of leukocytes, number of nonperfused sinusoids (NPS), and NADH autofluorescence were monitored (by intravital microscopy) in mouse liver after 15 min of superior mesenteric artery occlusion and 60 min of reperfusion. Leukostasis, NPS, and NADH autofluorescence (indicating hypoxia) were all increased in the liver at 60 min after gut I/R. The NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) exaggerated the liver leukostasis elicited by gut I/R, responses that were prevented by coadministration of L-arginine. The NO donor diethylenetriamine-NO (DETA-NO) and L-arginine were both effective in attenuating the gut I/R-induced leukostasis and increased NADH autofluorescence, whereas neither DETA nor D-arginine exerted a protective action. These findings indicate that NO is an important determinant of the liver leukostasis, impaired sinusoidal perfusion, and tissue hypoxia elicited by gut I/R.


Are spasticity, weakness, selectivity, and passive range of motion related to gait deviations in children with spastic cerebral palsy? A statistical parametric mapping study.
This study aimed to identify the relationships between clinical impairments and gait deviations in children with cerebral palsy (CP). A retrospective convenience sample of 367 children with CP was selected (3-18 years old) and divided in two groups based on clinical symptomatology [unilateral (uCP) / bilateral CP (bCP), (n = 167/200)]. All children underwent a three-dimensional gait analysis and a standardized clinical examination. Gait was inspected on a vector level (all sagittal motions combined), and an individual joint level (pelvis, hip, knee and ankle joint motions). Statistical non-parametric mapping was applied to identify specific parts of the gait cycle displaying relationships between the gait deviations of both groups and the impairment scores of spasticity, weakness, selectivity, and passive range of motion. Impairment scores were summarized in two ways: a) composite impairment scores (e.g. combined spasticity of all assessed muscles acting around the hip, knee and ankle joints) and b) joint specific impairment scores (e.g. spasticity of the muscles acting around the knee joint). Results showed that the vector and most of the individual motions were related to the composite scores. Direct and carry-over relationships were found between certain individual motions and joint impairment scores (around the same or neighboring joints, respectively). All correlations were more prominent for children with bCP compared to uCP, especially regarding the relationships of gait deviations with weakness and reduced selectivity. In conclusion, this study enabled the mapping of relationships between clinical impairments and gait deviations in children with CP, by identifying specific parts of the gait cycle that are related to each of these impairments. These results provide a comprehensive description of these relationships, while simultaneously highlighting the differences between the two CP groups. Integration of these findings could lead to a better understanding of the pathophysiology of gait deviations and, eventually, support individualized treatment planning.


Effect of androgen deprivation therapy on the contractile properties of type I and type II skeletal muscle fibres in men with non-metastatic prostate cancer.
The contractile properties of vastus lateralis muscle fibres were examined in prostate cancer (PrCa) patients undergoing androgen deprivation therapy (ADT) and in age- and activity-matched healthy male subjects (Control). Mechanically-skinned muscle fibres were exposed to a sequence of heavily Ca2+ -buffered solutions at progressively higher free [Ca2+ ] to determine their force-Ca2+ relationship. Ca2+ -sensitivity was decreased in both type I and type II muscle fibres of ADT subjects relative to Controls (by -0.05 and -0.04 pCa units, respectively, P < .02), and specific force was around 13% lower in type I fibres of ADT subjects than in Controls (P = .02), whereas there was no significant difference in type II fibres. Treatment with the reducing agent dithiothreitol slightly increased specific force in type I and type II fibres of ADT subjects (by ~2%-3%, P < .05) but not in Controls. Pure type IIx fibres were found frequently in muscle from ADT subjects but not in Controls, and the overall percentage of myosin heavy chain IIx in muscle samples was 2.5 times higher in ADT subjects (P < .01). The findings suggest that testosterone suppression can negatively impact the contractile properties by (i) reducing Ca2+ -sensitivity in both type I and type II fibres and (ii) reducing maximum specific force in type I fibres.


Comparative Evaluation of Shear Bond Strength of Brackets Bonded with self Etch Primer/Adhesive and Conventional Etch/Primer and Adhesive System.
The shear bond strength obtained with an acid primer might not be clinically reliable. The present study evaluated the use of a new self-etch primer as compared with the conventional bonding procedure. The findings indicated that the use of a self-etch primer to bond orthodontic brackets to the namel surface provided lower, but clinically accept-able, shear bond forces (mean, 7.1 ± 4.4 MPa). Hence, this study has been designed to evaluate the shear bond strength of orthodontic brackets bonded with a self-etching primer system, and compare it with the conventional acid etching priming system. This study was designed to compare the shear bond strength of orthodontic brackets bonded with conventional acid etching- priming and a self-etching primer. For this purpose, one hundred and twenty noncarious, not subjected to any pretreatment chemical agents (e.g. hydrogen peroxide), no cracks caused by the presence of the extraction forceps, no caries and unrestored human maxillary 1st and 2nd premolars, freshly extracted for orthodontic treatment were collected and stored in distilled water before bonding. Separate molds of cold-cure acrylic were prepared and the extracted premolar teeth were embedded in the cold cure acrylic up to the cement enamel junction. For the control group, the mean shear bond strength was 9.38 ± 6.02 MPa and for the experimental group, it is 6.91 ± 3.58 MPa. Group comparison using Chi-square test for the self-etching primer group adhesive remnant index (ARI) scores shows that more than half of the adhesive left over the tooth surface (score 3 and 4). For the conventional group, ARI scores show that there was no or slight amount of adhesive left over the tooth surface (score 5). The proportion of ARI score is significantly (P < 0.001) more in conventional (68.33%) as compared to self-etch primer (SEP) (28.33%). The proportion of ARI scores 4 and less is significantly more in SEP (71.67%) as compared to Conventional (6.67%) with P = 0.004 as per the Chi-square test. The bond strength of SEP is comparatively lesser than the conventional acid etching technique, but is definitely more or less to the required bond strength to resist masticatory forces. Majority of the studies conducted on self-etching primer are in vitro studies. Results of these cannot be extrapolated to the intraoral environment. Hence, further long-term clinical studies need to be carried out before actually recommending the use of self-etching primer in routine orthodontic bonding.


Krill Oil Treatment Increases Distinct PUFAs and Oxylipins in Adipose Tissue and Liver and Attenuates Obesity-Associated Inflammation via Direct and Indirect Mechanisms.
The development of obesity is characterized by the metabolic overload of tissues and subsequent organ inflammation. The health effects of krill oil (KrO) on obesity-associated inflammation remain largely elusive, because long-term treatments with KrO have not been performed to date. Therefore, we examined the putative health effects of 28 weeks of 3% (w/w) KrO supplementation to an obesogenic diet (HFD) with fat derived mostly from lard. The HFD with KrO was compared to an HFD control group to evaluate the effects on fatty acid composition and associated inflammation in epididymal white adipose tissue (eWAT) and the liver during obesity development. KrO treatment increased the concentrations of EPA and DHA and associated oxylipins, including 18-HEPE, RvE2 and 14-HDHA in eWAT and the liver. Simultaneously, KrO decreased arachidonic acid concentrations and arachidonic-acid-derived oxylipins (e.g., HETEs, PGD2, PGE2, PGF2α, TXB2). In eWAT, KrO activated regulators of adipogenesis (e.g., PPARγ, CEBPα, KLF15, STAT5A), induced a shift towards smaller adipocytes and increased the total adipocyte numbers indicative for hyperplasia. KrO reduced crown-like structures in eWAT, and suppressed HFD-stimulated inflammatory pathways including TNFα and CCL2/MCP-1 signaling. The observed eWAT changes were accompanied by reduced plasma leptin and increased plasma adiponectin levels over time, and improved insulin resistance (HOMA-IR). In the liver, KrO suppressed inflammatory signaling pathways, including those controlled by IL-1β and M-CSF, without affecting liver histology. Furthermore, KrO deactivated hepatic REL-A/p65-NF-κB signaling, consistent with increased PPARα protein expression and a trend towards an increase in IkBα. In conclusion, long-term KrO treatment increased several anti-inflammatory PUFAs and oxylipins in WAT and the liver. These changes were accompanied by beneficial effects on general metabolism and inflammatory tone at the tissue level. The stimulation of adipogenesis by KrO allows for safe fat storage and may, together with more direct PPAR-mediated anti-inflammatory mechanisms, attenuate inflammation.


Glucose intolerance (diabetes and IGT) in a selected South Indian population with special reference to family history, obesity and lifestyle factors--the Chennai Urban Population Study (CUPS 14).
The aim of the study was to assess the impact of family history of diabetes, obesity and lifestyle factors particularly physical activity on glucose intolerance in a selected south Indian population. The Chennai Urban Population Study (CUPS) is an epidemiological study involving two residential areas in Chennai in South India representing the middle and lower socio-economic group. Of the total of 1399 eligible subjects (age > or = 20 years), 1262 (90.2%) participated in the study. A detailed questionnaire was used to collect details on medical history, family history of diabetes, family income and physical activity. All the study subjects underwent a glucose tolerance test (GTT) and were categorized as having normal glucose tolerance (NGT), impaired glucose tolerance (IGT) or diabetes using WHO consulting group criteria. Obesity and abdominal obesity were defined using the new Asia Pacific guidelines. The overall prevalence of diabetes in the study population was 12.0%, (age-standardized -9.3%), which included 7.2% of known diabetic subjects and 4.8% undiagnosed diabetic subjects, while the prevalence of impaired glucose tolerance was 5.9% (age-standardized prevalence 5.0%). The prevalence of glucose intolerance (Diabetes + IGT) was significantly higher among subjects with both parents diabetic (55%) compared to those with one parent diabetic (22.1%, p = 0.005) and those with no family history (15.6%, p < 0.0001). Prevalence of glucose intolerance was significantly higher among subjects who had light grade physical activity (23.2%) compared to moderate (17.5%, p = 0.04) and heavy grade activity (8.1 % p < 0.00001). Subjects belonging to higher socio-economic status (SES) and who also had a positive family history of diabetes had five times greater prevalence of glucose intolerance compared to subjects from lower socioeconomic status and no family history (p < 0.0001). Regression analysis revealed age (p < 0.0001), waist circumference (p < 0.0001), body mass index (p < 0.0001), waist-hip ratio (p < 0.0001), systolic blood pressure (p < 0.0001), diastolic blood pressure (p < 0.0001), family history of diabetes (p < 0.0001), higher SES (p < 0.0001), moderate (p = 0.001) and light (p < 0.001) grade physical activity to be associated with glucose intolerance. Multiple logistic regression analysis showed that even after adjusting for variables like age and family history of diabetes, physical activity showed a significant association with glucose intolerance The prevalence of glucose intolerance is high in this selected urban south Indian population. Lifestyle factors and family history have a synergistic effect on increasing the risk for diabetes in this population.


Hysteroscopic removal of foreign bodies and its method of monitoring.
To evaluate transcervical removal of foreign bodies (TCRF) and to estimate the effectiveness of its monitoring methods. One hundred and thirteen women were identified as having residual intrauterine devices (IUD), residual pregnancy products, unabsorbed strings and broken hooks, which were not removed during routine curettage or IUD removal. All patients were monitored using B ultrasonography while TCRF was performed. Four cases were monitored by laparoscopy simultaneously. One case was monitored by laparoscopic ultrasonography. Foreign bodies of one hundred and nine patients were taken out by TCRF. Uterine bleeding, amenorrhoea, discharge, abdominal pain, micturition and hematuria disappeared postoperatively. Fetal bones embedded into intramural uterin in four cases were not removed completely. Of these four, one became pregnant 4 months later after TCRF and term delivered. One case encountered uterine perforation that was sutured by laparoscopy. TCRF is safe and efficient. Sufficient cervical canal distension, selection of equipment and methods to be used is important for successful TCRF. As a non-invasive and effective monitoring method, B ultrasonography is the first choice to monitor for TCRF. For patients with high risk factors for uterine perforation, laparoscopic monitoring should be done simultaneously. Laparoscopic ultrasonography monitoring has both the advantages of B ultrasonography and laparoscopy monitoring, but is invasive and expensive.


A reaction-coordinate perspective of magnetic relaxation.
Understanding and utilizing the dynamic quantum properties of metal ions is the frontier of many next generation technologies. One property in particular, magnetic relaxation, is a complicated physical phenomenon that is scarcely treated in undergraduate coursework. Consequently, principles of magnetic relaxation are nearly impenetrable to starting synthetic chemists, who ultimately design the molecules that fuel new discoveries. In this Tutorial Review, we describe a new paradigm for thinking of magnetic relaxation in metal complexes in terms of a simple reaction-coordinate diagram to facilitate access to the field. We cover the main mechanisms of both spin-lattice (T1) and spin-spin (T2) relaxation times within this conceptual framework and how molecular and environmental design affects these times. Ultimately, we show that many of the scientific methods used by inorganic chemists to study and manipulate reactivity are also useful for understanding and controlling magnetic relaxation. We also describe the cutting edge of magnetic relaxation within this paradigm.


Phenotype-based drug screening in primary ovarian carcinoma cultures identifies intracellular iron depletion as a promising strategy for cancer treatment.
Primary cultures of patient tumor cells (PCPTC) have been used for prediction of diagnosis-specific activity and individual patient response to anticancer drugs, but have not been utilized as a model for identification of novel drugs in high throughput screening. In the present study, ovarian carcinoma cells from three patients were tested in response to a library of 3000 chemically diverse compounds. Eight hits were retrieved after counter screening using normal epithelial cells, and one of the two structurally related hit compounds was selected for further preclinical evaluation. This compound, designated VLX 50, demonstrated a broad spectrum of activity when tested in a panel of PCPTCs representing different forms of leukemia and solid tumors and displayed a high tumor to normal cell activity. VLX 50 induced delayed cell death with some features of classical apoptosis. Significant in vivo activity was confirmed on primary cultures of human ovarian carcinoma cells in mice using the hollow fiber model. Mechanistic exploration was performed using gene expression analysis of drug exposed tumor cells to generate a drug-specific signature. This query signature was analyzed using the Gene Set Enrichment Analysis and the Connectivity Map database. Strong connections to hypoxia inducible factor 1 and iron chelators were retrieved. The mechanistic hypothesis of intracellular iron depletion leading to hypoxia signaling was confirmed by a series of experiments. The results indicate the feasibility of using PCPTC for cancer drug screening and that intracellular iron depletion could be a potentially important strategy for cancer therapy.


Cost-effectiveness in extracorporeal life support in critically ill adults in the Netherlands.
Extracorporeal life support (ECLS) is used to support the cardiorespiratory function in case of severe cardiac and/or respiratory failure in critically ill patients. According to the ELSO guidelines ECLS should be considered when estimated mortality risk approximates 80%. ECLS seems an efficient therapy in terms of survival benefit, but no undisputed evidence is delivered yet. The aim of the study is to assess the health-related quality of life after ECLS treatment and its cost effectiveness. We will perform a prospective observational cohort study. All adult patients who receive ECLS in the participating centers will be included. Exclusion criteria are patients in whom the ECLS is only used to bridge a procedure (like a high risk percutaneous coronary intervention or surgery) or the absence of informed consent. Data collection includes patient characteristics and data specific for ECLS treatment. Severity of illness and mortality risk is measured as precisely as possible using measurements for the appropriate age group and organ failure. For analyses on survival patients will act as their own control as we compare the actual survival with the estimated mortality on initiation of ECLS if conservative treatment would have been continued. Survivors are asked to complete validated questionnaires on health related quality of life (EQ5D-5 L) and on medical consumption and productivity losses (iMTA/iPCQ) at 6 and 12 months. Also the health related quality of life 1 month prior to ECLS initiation will be obtained by a questionnaire, if needed provided by relatives. With an estimated overall survival of 62% 210 patients need to be recruited to make a statement on cost effectiveness for all ECLS indications. If our hypothesis that ECLS treatment is cost-effective is confirmed by this prospective study this could lead to an even broader use of ECLS treatment. The trial is registered at ( NCT02837419 ) registration date July 19, 2016 and with the Dutch trial register, http://www.trialregister.nl/trialreg/admin/rctview.asp?TC=6599.


Effect on the duration of mechanical ventilation of identifying patients capable of breathing spontaneously.
Prompt recognition of the reversal of respiratory failure may permit earlier discontinuation of mechanical ventilation, without harm to the patient. We conducted a randomized, controlled trial in 300 adult patients receiving mechanical ventilation in medical and coronary intensive care units. In the intervention group, patients underwent daily screening of respiratory function by physicians, respiratory therapists, and nurses to identify those possibly capable of breathing spontaneously; successful tests were followed by two-hour trials of spontaneous breathing in those who met the criteria. Physicians were notified when their patients successfully completed the trials of spontaneous breathing. The control subjects had daily screening but no other interventions. In both groups, all clinical decisions, including the decision to discontinue mechanical ventilation, were made by the attending physicians. Although the 149 patients randomly assigned to the intervention group had more severe disease, they received mechanical ventilation for a median of 4.5 days, as compared with 6 days in the 151 patients in the control group (P=0.003). The median interval between the time a patient met the screening criteria and the discontinuation of mechanical ventilation was one day in the intervention group and three days in the control group (P<0.001). Complications -- removal of the breathing tube by the patient, reintubation, tracheostomy, and mechanical ventilation for more than 21 days -- occurred in 20 percent of the intervention group and 41 percent of the control group (P=0.001). The number of days of intensive care and hospital care was similar in the two groups. Total costs for the intensive care unit were lower in the intervention group (median, $15,740, vs. $20,890 in the controls, P=0.03); hospital costs were lower, though not significantly so (median, $26,229 and $29,048, respectively; P=0.3). Daily screening of the respiratory function of adults receiving mechanical ventilation, followed by trials of spontaneous breathing in appropriate patients and notification of their physicians when the trials were successful, can reduce the duration of mechanical ventilation and the cost of intensive care and is associated with fewer complications than usual care.


Personalized lifetime prediction of survival and treatment benefit in patients with heart failure with reduced ejection fraction: The LIFE-HF model.
Although trials have proven the group-level effectiveness of various therapies for heart failure with reduced ejection fraction (HFrEF), important differences in absolute effectiveness exist between individuals. We developed and validated the LIFEtime-perspective for Heart Failure (LIFE-HF) model for the prediction of individual (lifetime) risk and treatment benefit in patients with HFrEF. Cox proportional hazards functions with age as the time scale were developed in the PARADIGM-HF and ATMOSPHERE trials (n = 15 415). Outcomes were cardiovascular death, heart failure (HF) hospitalization or cardiovascular death, and non-cardiovascular mortality. Predictors were age, sex, New York Heart Association class, prior HF hospitalization, diabetes mellitus, extracardiac vascular disease, systolic blood pressure, left ventricular ejection fraction, N-terminal pro-B-type natriuretic peptide, and glomerular filtration rate. The functions were combined in life-tables to predict individual overall and HF hospitalization-free survival. External validation was performed in the SwedeHF registry, ASIAN-HF registry, and DAPA-HF trial (n = 51 286). Calibration of 2- to 10-year risk was adequate, and c-statistics were 0.65-0.74. An interactive tool was developed combining the model with hazard ratios from trials to allow estimation of an individual's (lifetime) risk and treatment benefit in clinical practice. Applying the tool to the development cohort, combined treatment with a mineralocorticoid receptor antagonist, sodium-glucose cotransporter 2 inhibitor, and angiotensin receptor-neprilysin inhibitor was estimated to afford a median of 2.5 (interquartile range [IQR] 1.7-3.7) and 3.7 (IQR 2.4-5.5) additional years of overall and HF hospitalization-free survival, respectively. The LIFE-HF model enables estimation of lifelong overall and HF hospitalization-free survival, and (lifetime) treatment benefit for individual patients with HFrEF. It could serve as a tool to improve the management of HFrEF by facilitating personalized medicine and shared decision-making.


[Determination of mineral elements in soybean from different producing areas by ICP-AES].
Soybeans from Jiangxi, Heilongjiang, Jilin and Anhui were digested by nitric acid. The contents of mineral elements K, P, Mg, Ca, Fe, Al, Zn, Mn and Na in four samples were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). The differences between samples were determined by Duncan's multiple-range test, and evaluations were based on a significance level of P < 0.05. The results show that soybeans are rich in mineral elements that human body needs. The contents of K, P, Mg, Ca and Fe are higher than the others, and the order from high to low does not change with the producing area. The order of Mn, Zn, Al, Na varied with the producing area. Producting area does not affect the contents of elements K, P and Mg significantly (P > 0.05). It can be concluded that the demands for major mineral elements K, P and Mg are stable in the growth of soybeans with the same variety. However, producing area has a great influence on the contents of elements Al, Ca, Fe, Na, Mn and Zn in soybean (P < 0.05). The content of Fe in Anhui soybean (70.93 mg x kg(-1)) and the content of Mn in Jiangxi soybean (32.69 mg x kg(-1)) are significantly higher than the other three samples. The results suggested that the contents of Al, Ca,Fe, Na, Mn and Zn were easily affected by growth conditions, such as water, soil and so on.


Immunohistochemical evaluation of molecular radiotherapy target expression in neuroblastoma tissue.
Neuroblastoma may be treated with molecular radiotherapy, 131I meta-Iodobenzylguanidine and 177Lu Lutetium DOTATATE, directed at distinct molecular targets: Noradrenaline Transporter Molecule (NAT) and Somatostatin Receptor (SSTR2), respectively. This study used immunohistochemistry to evaluate target expression in archival neuroblastoma tissue, to determine whether it might facilitate clinical use of molecular radiotherapy. Tissue bank samples of formalin fixed paraffin embedded neuroblastoma tissue from patients for whom clinical outcome data were available were sectioned and stained with haematoxylin and eosin, and monoclonal antibodies directed against NAT and SSTR2. Sections were examined blinded to clinical information and scored for the percentage and intensity of tumour cells stained. These data were analysed in conjunction with clinical data. Tissue from 75 patients was examined. Target expression scores varied widely between patients: NAT median 45%, inter-quartile range 25% - 65%; and SSTR2 median 55%, interquartile range 30% - 80%; and in some cases heterogeneity of expression between different parts of a tumour was observed. A weak positive correlation was observed between the expression scores of the different targets: correlation coefficient = 0.23, p = 0.05. MYCN amplified tumours had lower SSTR2 scores: mean difference 23% confidence interval 8% - 39%, p < 0.01. Survival did not differ by scores. As expression of both targets is variable and heterogeneous, imaging assessment of both may yield more clinical information than either alone. The clinical value of immunohistochemical assessment of target expression requires prospective evaluation. Variable target expression within a patient may contribute to treatment failure.


Polymorphic GGC repeats in the androgen receptor gene are associated with hereditary and sporadic prostate cancer risk.
Androgen receptor (AR) has long been hypothesized to play an important role in prostate cancer etiology. Two trinucleotide repeat polymorphisms (CAG and GGC repeats in exon 1 of the AR gene) have been investigated as risk factors for prostate cancer in several studies. However, the results are inconclusive, probably because of the variations of study designs, characteristics of study samples, and choices of analytical methods. In this study, we evaluated evidence for linkage and association between the two AR repeats and prostate cancer by using the following comprehensive approaches: (1) a combination of linkage and association studies, (2) a test for linkage by parametric analysis and the male-limited X-linked transmission/disequilibrium test (XLRC-TDT), (3) a test for association by using both population-based and family-based tests, and (4) a study of both hereditary and sporadic cases. A positive but weak linkage score (HLOD=0.49, P=0.12) was identified in the AR region by parametric analysis; however, stronger evidence for linkage in the region, especially at the GGC locus, was observed in the subset of families whose proband had < or = 16 GGC repeats (HLOD=0.70, P=0.07) or by using XLRC-TDT ( z'=2.65, P=0.008). Significantly increased frequencies of the < or = 16 GGC repeat alleles in 159 independent hereditary cases (71%) and 245 sporadic cases (68%) cases compared with 211 controls (59%) suggested that GGC repeats were associated with prostate cancer ( P=0.02). Evidence for the association between the < or = 16 GGC repeats and prostate cancer risk was stronger with XLRC-TDT ( z'=2.66, P=0.007). No evidence for association between the CAG repeats and prostate cancer risk was observed. The consistent results from both linkage and association studies strongly implicate the GGC repeats in the AR as a prostate cancer susceptibility gene. Further studies on this polymorphism in other independent data sets and functional analysis of the GGC repeat length on AR activity are warranted.


Chronic treatment with fluoxetine for more than 6 weeks decreases neurogenesis in the subventricular zone of adult mice.
Recent studies indicate that chronic treatment with serotonergic antidepressants upregulates adult neurogenesis of the dentate gyrus (DG). In contrast, some studies claimed that there was very little alteration of neurogenesis in the subventricular zone (SVZ) by the antidepressants. Since almost all of those studies treated animals with drugs for 2 to 4 weeks as chronic treatment models of antidepressants, it is possible that antidepressant treatments for longer periods would affect adult neurogenesis in the SVZ. In the present study, we examined the effects of long-term (up to 9 weeks) administration of fluoxetine (FLX), a selective serotonin reuptake inhibitor, on cell proliferation and survival in the DG and the SVZ of adult mice. As reported previously, in the DG of mice treated with FLX for 3, 6, or 9 weeks that were also injected with 5-bromodeoxyuridine (BrdU) in the last 3 days before perfusion, the numbers of Ki67- and BrdU-positive cells, which are cell proliferation markers, were significantly upregulated even at 3 weeks after the onset of the FLX treatments, and these increases were sustained in mice treated with FLX for 9 weeks. On the other hand, in the SVZ, we found a small, insignificant decrease in the numbers of Ki67- and BrdU-positive cells at 3 weeks, followed by highly significant decreases in the numbers of Ki67- and BrdU-positive cells at both 6 and 9 weeks. Furthermore, among olfactory newly generated cells that survived for 3 weeks after BrdU injection, the number of new cells was decreased at 9 weeks of FLX treatment. These results demonstrate that long-term (more than 6 weeks) treatment with FLX has the opposite effect on neurogenesis in the SVZ than it does in the DG. The results also suggest that the decrease in neurogenesis in the SVZ might be involved in some aspects of the drugs' therapeutic effects on depression. In addition, our findings raise the possibility that some of the side effects of antidepressants might be mediated by decreased adult neurogenesis in the SVZ.


Yolk carotenoids and stable isotopes reveal links among environment, foraging behavior and seabird breeding success.
Nutrients that are limited in availability, such as carotenoids, are potentially involved in trade-offs between homeostasis and reproduction. Despite their importance, factors that affect the capacity of female birds to meet their carotenoid requirements are poorly understood. We used delta(15)N stable isotope analysis to relate foraging behavior to yolk carotenoid deposition in two seabirds, Cassin's auklet (Ptychoramphus aleuticus) and rhinoceros auklet (Cerorhinca monocerata), during each of five years. As expected from their narrower trophic range, Cassin's auklets produced yolks with fewer carotenoid types than did rhinoceros auklets (one vs. three). Cassin's auklets also fed on a lower trophic level diet richer in carotenoids, yet had lower total yolk carotenoid levels, which suggests a role for species-specific adaptations for carotenoid uptake and utilization. Within both species, lower trophic-level feeding was linked to higher yolk carotenoid levels, but through different mechanisms. In Cassin's auklets, it was due to a population-wide response to environmental variation: in warm-water years, all females fed at a low trophic level and produced carotenoid-rich yolks. In rhinoceros auklets, it was due to individual differences similarly expressed in all years: females fed across a wide trophic range, and those that fed at a low trophic level produced carotenoid-rich yolks. Rhinoceros auklets bred more successfully in years when their yolks were rich in carotenoids, probably due to a correlated response to stronger marine primary production. Our results are novel because they link avian yolk carotenoid deposition to behavioral and environmental variations.


Severe scoliosis, torticollis and short stature in a woman with Wildervanck Syndrome (WS).
Wildervanck syndrome (WS) combines features of Klippel-Feil syndrome (KFS), sixth nerve palsy, and deafness. This is a case of a 23 year old woman, diagnosed with KFS (a triad of short neck, low posterior hairline and restricted neck movements) at the age of 20 days. The manifestations of the WS in this patient are severe: she has torticollis, and an extremely severe scoliosis. In addition, she is short (-3 SD; parental target height + 0.8SD) and has mixed sensorineural and conductive deafness. She also has ptosis, strabismus and a high myopia. Radiologically, there are multiple coalitions of cervical vertebrae. Intelligence is unaffected (IQ 95), but deafness, strabismus and high myopia forced her early out of school. Karyotype is 46, XX. In brief, this is a patient with severe WS and additional anomalies. Short and/or reduced parental target height is a part of WS.


Clubfoot correction using the Ilizarov technique.
Idiopathic clubfeet in children three years of age or older which have not been treated previously or which have recurred, can be treated successfully by using the Ilizarov technique. The authors report on four feet in three children which were treated by this method. Follow-up examinations at 11-24 months found all corrections successfully maintained.


Outcome of orthoses intervention in the rheumatoid foot.
This study was carried out to determine the effect of foot orthoses on pain, gait, and energy expenditure in patients with rheumatoid arthritis. Eighteen patients were evaluated for these parameters. Each patient was given a foot insert or shoe modification suitable for his or her foot deformity. Following 3 months of orthosis use, a significant difference was found in regards to pain (p < .05), step length and stride length (p < .05), and physiological cost index (p < .05). The results suggest that foot orthoses are an important feature in the rehabilitation of the rheumatoid foot.


Brain single photon emission computed tomographic evaluation of patients with childhood absence epilepsy.
This study was performed to determine the utility of 99mTc-hexamethylpropylenamine oxime (HMPAO) brain single photon emission computed tomography (SPECT) in evaluating patients with childhood absence epilepsy. Twenty-three patients (13 female, 10 male), aged 7 to 15 years (mean age 10.3 +/- 2.2), were studied. All patients underwent a detailed neurologic examination, interictal and ictal electroencephalography (EEG), computed tomography, and/or magnetic resonance imaging, and SPECT. The baseline study was performed during the interictal period and the activation study was performed on a separate day while the patients were having seizures provoked by hyperventilation. Their EEGs were monitored at the same time. Transaxial, sagittal, and coronal slices were obtained for both studies. The mean counts per pixel were calculated on 11 regions of interest on three representative transaxial slices. Count density was calculated for each region. Region-to-occipital cortex ratios were obtained. For each region, normalized ratios were used to obtain a side-to-side percent asymmetry index between baseline and activation studies. Visual interpretation of the baseline study showed that 10 of the 23 patients had a detectable abnormality in regional cerebral blood flow during the interictal period. These abnormalities consisted of relative hypoperfusion in the frontal lobes that could involve neighboring parietal and temporal regions. The activation study revealed that 13 of 23 patients had relative hyperperfusion in these brain regions that were relatively hypoperfused in the baseline study. These hyperperfused regions occupied larger areas than baseline hypoperfused regions. All patients had global increased perfusion in the ictal study. The side-to-side asymmetry indexes for these visually interpreted regional cerebral blood flow abnormalities ranged from 2 to 6%. The relatively consistent pattern of frontal regional cerebral blood flow alterations suggests that altered frontal lobe functions can be implicated in patients with childhood absence epilepsy.


A calcium-initiated signaling pathway propagated through calcineurin and cAMP response element-binding protein activates proenkephalin gene transcription after depolarization.
Essential components of a signal-transduction pathway regulating activity-dependent neuropeptide gene transcription have been identified. Proenkephalin (PEnk) gene activation after depolarization of chromaffin cells with 40 mM KCl was blocked by the voltage-sensitive calcium-channel blocker methoxyverapamil (D600) (30 microM) and by calcineurin inhibition with 100 nM cyclosporin A or ascomycin but not by inhibiting new protein synthesis with 0.5 microg/ml cycloheximide. KCl-induced elevation of PEnk mRNA was distinct from activation of the PEnk gene by either cAMP or protein kinase C. Twenty-five micromolar forskolin- and 100 nM phorbol 12-myristate 13-acetate-induced elevations of PEnk mRNA were cycloheximide-sensitive and were not blocked by cyclosporin A or ascomycin. KCl stimulated Ser-133 phosphorylation of cAMP response element-binding protein (CREB) in chromaffin cells, and CREB phosphorylation was blocked by both ascomycin and D600. A reporter gene containing 193 bases of the PEnk gene 5' flank driving luciferase gene expression (pENK12-Luc) transfected into chromaffin cells was transcriptionally activated by KCl depolarization. Activation was blocked by both ascomycin and D600 and required an intact CREB binding site (ENKCRE2). An oligonucleotide containing the PEnk cAMP response element-2 was gel-shifted by both unstimulated and potassium-stimulated chromaffin cell nuclear extracts into a prominent complex supershifted by CREB antibodies. Finally, stimulation of transcription of the pENK12-Luc reporter by KCl in chromaffin cells was blocked by coexpression of the CREB antagonist A-CREB but not by the AP-1 antagonist A-Fos. Stimulus-transcription coupling after depolarization in chromaffin cells occurs via calcineurin-dependent activation of CREB, a pathway distinct from cAMP- or protein kinase C-initiated signaling and independent of immediate early gene regulation.


Interview with Alison Goate.
Alison M Goate is the Samuel & Mae S Ludwig Professor of Genetics in Psychiatry, Professor of Genetics and Professor of Neurology at Washington University School of Medicine in St Louis (MO, USA). Dr Goate studied for her undergraduate degree in biochemistry at the University of Bristol (UK) and received her graduate training at Oxford University (UK). She performed postdoctoral studies with Professor Theodore Puck, Professor Louis Lim and Dr John Hardy before receiving a Royal Society University Research Fellowship to support her independent research program at St Mary's Hospital Medical School in London. In 1991, Dr Goate and colleagues reported the first mutation linked to an inherited form of Alzheimer's disease, in the amyloid precursor protein (APP) gene on chromosome 21. The mutation was found to be linked to inherited cases of early-onset Alzheimer's disease. In 1992, Dr Goate moved to Washington University as an Associate Professor in Genetics and Psychiatry. Dr Goate and colleagues have since identified mutations in four other genes, including two that cause Alzheimer's disease and two that cause the related dementia frontotemporal dementia. In addition to her work on dementia, Dr Goate's laboratory also studies the genetics of alcohol and nicotine dependence. Dr Goate has received numerous awards including the Potamkin Award from the American Academy of Neurology, the Zenith Award from the Alzheimer's Association, the Senior Investigator Award from the Metropolitan Life Foundation, the St Louis Academy of Science Innovation Award and the Carl and Gerty Cori Faculty Achievement Award at Washington University. Dr Goate has been a member of many scientific Review Boards and currently serves on the Editorial Boards of several journals.


Transition C2718T in the AR gene, resulting in generation of a termination codon and truncated form of the androgen receptor, causes complete androgen insensitivity syndrome.
The action of testosterone and 5 alpha-dihydrotestosterone are essential to the development of the male phenotype. Patients with karyotype 46,XY, resistant to these hormones, exhibit a wide spectrum of phenotypes: from phenotypic female, through a range of incomplete masculinization, to under-virilized, infertile man. These disturbances are caused by mutations in the androgen receptor gene (AR). We studied a 46,XY fenotypic female with typical symptoms of Complete Androgen Insensitivity Syndrome (CAIS). Multiple temperature single-stranded conformation polymorphism (MSSCP) and sequence analysis of exon 6 of the AR gene in a patient revealed a C2718T transition causing R786X mutation in the loop between helices VII and VIII of the LBD of the androgen receptor. The R786X mutation has been described in a patient with CAIS only once and no such mutations have been described in Eastern Europe.


Associations between patient factors and medication adherence: A Jordanian experience.
To explore the effect of patient characteristics and health beliefs on their medication adherence. Patients (n=167) with chronic conditions (mean age 58.9; SD=13.54, 53% males) were recruited from March 2009- to March 2010 using a cross sectional study design. Data collected included patients' demographics, medical conditions, medications therapeutic regimen, frequency of physician visits and health beliefs. Patient self-reported adherence to medications was assessed by the researcher using a validated and published scale. Treatment related problems (TRPs) were evaluated for each patient by competent clinical pharmacists. Associations between patient characteristics/health beliefs with adherence were explored. About half of the patients (46.1%) were non-adherent. A significant association was found between lower adherence and higher number of disease states (p<0.001), higher number of medications (p=0.001), and higher number of identified TRPs (p = 0.003). Patient adherence was positively affected by older age, higher educational level, and higher number of physician visits per month, while it was negatively affected by reporting difficulties with getting prescription refills on time. This study identified different factors that may negatively affect adherence, including higher number of medications and disease states, higher number of identified TRPs and inability to getting prescription refills on time. Hence, more care needs to be provided to patients with complex therapeutic regimens in order to enhance adherence.


Interactions of human myeloid cells with natural killer cell subsets in vitro and in vivo.
In both human and mouse it has been recently realized that natural killer (NK) cells do not emerge from the bone marrow with full functional competence but rather acquire functions in interaction with antigen-presenting cells (APCs), primarily dendritic cells (DCs). Here we review the mechanisms and the consequences of this NK-cell preactivation, as well as discuss new experimental models that now allow investigating these interactions for human NK cells and their response to human pathogens in vivo. These investigations will allow harnessing NK cells during vaccination for improved innate and adaptive immunity.


Propagation of dysbindin-1B aggregates: exosome-mediated transmission of neurotoxic deposits.
Given the detection of aggregated deposits in chronic mental diseases (CMD), the disturbance of proteostasis in those diseases is receiving increasing attention. The study of aggregated proteins can contribute to our understanding of the chronic and progressive condition of such diseases. Dysbindin, encoded by the schizophrenia susceptibility gene DTNBP1, has been reported to co-aggregate with DISC1. However, there has been no evidence to date on the aggregation tendency of dysbindin. Therefore, we investigated the isoform-specific aggregation of dysbindin. We found that dysbindin-1B aggregated into cell-invasive deposits in mice. Because of the efficient propagation of dysbindin-1B, we further studied the mechanism of propagation and identified it as exosome-mediated transmission of the aggregates. In addition, aggregates of dysbindin-1B were toxic. Through exosome-mediated propagation, the deposits of dysbindin-1B exerted toxic effects on recipient neurons a long distance away from the initial aggregation site in mice brain. The rapid long distance propagation of neurotoxic deposits of dysbindin-1B in affected neuronal circuitry indicates a possible mechanism for the progressive deterioration of neurons and cognitive function in CMD.


Immunoassay Interference on Thyroid Function Tests During Treatment with Nivolumab.
Background: Immune checkpoint inhibitors (ICIs) are associated with several endocrine side effects. In particular, the use of programmed cell death protein-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) inhibitors is related to a higher incidence of thyroid dysfunction. Patient Findings: An 85 years-old patient, diagnosed with a metastatic melanoma treated with nivolumab, presented to our hospital with severe ICI-related thyrotoxicosis. Diagnosis was complicated by a biochemical interference on thyroid hormones assay, probably induced by nivolumab. Summary: Baseline laboratory examination conducted before onset of anticancer therapy showed normal thyroid function test (TFTs). A few days after receiving the second nivolumab administration, the patient developed a severe thyrotoxicosis. According to destructive thyroiditis, in a short period thyrotropin (TSH) levels normalized and rapidly increased, but free thyroxine (fT4) levels were inappropriately elevated and did not decrease as expected. The sample was processed by using a Siemens Centaur® immunoassay. We reanalyzed the same sample at another laboratory and with a different immunoassay method (Roche Elecsys®). The results obtained from this assay confirmed severe hypothyroidism with appropriately low fT4 levels. We suspected a possible nivolumab-associated interference on the fT4 assay. Therefore, we subjected the same sample to a polyethylene glycol (PEG) 6000 precipitation, a simple method for the removal of macromolecules, before assaying for fT4 levels. Evaluation of the post-PEG-precipitation sample (Siemens Centaur immunoassay) revealed appropriately low fT4 levels. The patient was started on levothyroxine (LT4) therapy, with monthly TFT monitoring using the Roche immunoassay. Approximately 9 months after starting nivolumab therapy, the patient was advised treatment cessation. A month later, the TFTs were retested on a Siemens Centaur immunoassay, and appropriate fT4 levels were observed in accordance with normal TSH levels on adequate LT4 replacement therapy. Conclusions: We report a possible novel nivolumab-induced biochemical interference on assays of fT4 levels. The hypothesis of a biochemical drug-induced interference is further supported by the disappearance of the interference after the withdrawal of nivolumab. Further studies are needed to prove the biochemical mechanisms of this interference.


[Watermelon stomach: a cause of chronic blood loss in the elderly].
Watermelon stomach, or gastric vascular ectasia, is a relatively new, distinct clinical entity. This rare vascular abnormality of the stomach causes severe chronic blood loss and iron-deficiency anemia, particularly in the elderly. Endoscopically it is characterized by longitudinal antral folds which contain visible vessels radiating from the pylorus, and resembling the dark stripes on the surface of some watermelons. This endoscopic picture may easily be misinterpreted as antral hemorrhagic gastritis. The specific histologic features which establish the diagnosis on endoscopic biopsy are dilation and thrombosis of mucosal capillaries and fibromuscular hyperplasia of the lamina propria. Associated diseases, including achlorhydria, pernicious anemia and chronic liver disease have been reported. We recently encountered 2 cases of watermelon stomach associated with pernicious anemia and hypothyroidism. This entity should be considered in the differential diagnosis of iron-deficiency anemia.


Debate: Innovations in mindfulness for young people need to take more account of their unique needs if they are to live up to their potential.
While the current popularity of mindfulness suggests that it will foster various social-emotional, personal, and academic benefits, current claims for the benefits of mindfulness for children and young people is premature according to Butterfield, Roberts, Feltis, and Kocovski (2020) who argue that 'despite the media hype and the increase in peer-reviewed publications the evidence base for mindfulness programs for children is not robust' (p. 194). A recent report from a large scale mindfulness project conducted by a number of highly experienced mindfulness researchers and involving more than 28,000 young people outlines that: 'For the most part the young people did not engage with the mindfulness training' and 'our results suggested mindfulness training might work for some children and not for others, and under some conditions, but not others' (https://myriadproject.org/what-we-did/). The MYRIAD project carefully tested the effects of a brief mindfulness intervention for early teens and found it to have no impact on preventing mental health problems or promoting well-being. Researchers in this project found that since the young participants were generally not particularly engaged overall in mindfulness a more engaging accessible and effective format and teaching format and mode of delivery may have been needed. Myriad researchers included students' feedback comments: 'didn't find it useful'; found it 'boring'; had 'no choice'; 'couldn't do it; didn't do it'. More concerning is their finding of potentially harmful effects leading the researchers to state that: future research and innovations should carefully consider the unique needs and developmental stage of young people (Montero-Marin et al., 2022, p. 123). I argue in this short 'debate' piece that there is a pressing need for more nuanced, broad ranging and creative qualitative research that can help us understand young people's rich subjective experience of mindfulness, why they might not engage in mindfulness and how to better tailor it for their individual needs and contexts.


How does abstract and concrete garbage classification signage influence waste sorting behavior?
Despite the issuance of standardized garbage classification signage, the rate of garbage classification in China remains low. We conducted a pair of laboratory experiments to explore the cognitive processing differences between abstract (including recyclables, hazardous garbage, and food signs) and concrete (including paper, plastic, glass, metal, textiles, batteries, household chemicals, tubes, and food signs) classification signs. We tested a nudging strategy to enhance garbage classification behavior. In Experiment 1, we divided garbage classification signs into two conditions: an abstract condition (comprising abstract signs) and a concrete condition (comprising concrete signs). The Go/No Go task was used to simulate garbage classification behavior. Participants were instructed to press a key when the garbage stimulus matched the classification signs (Go condition) and to refrain from pressing the key when there was a mismatch (No Go condition). The results showed that responses under the concrete condition were expedited compared to those under the abstract condition. This suggests that concrete signage requires less cognitive exertion, thereby enhancing the efficiency of waste classification. In Experiment 2, we optimized the existing bin signage, which predominantly featured abstract signs (traditional condition), and transformed it into a bin signage that emphasized concrete classification signs. These concrete signs were strategically positioned on the upper part of the bins to draw attention (nudging condition). The results suggested that the nudging condition required fewer cognitive resources than the traditional condition, which in turn increased the efficiency of processing garbage classification. This study not only validates the effects of concreteness in garbage classification but also provides effective nudge strategies to complement existing garbage classification management policy tools in a realistic Chinese context.


Physical mapping and cloning of bacteriophage T4 anti-restriction endonuclease gene.
We have proposed that the ability of T4 to produce non-glucosylated progeny after a single cycle of growth on a galU rglA rglB+ mutant of Escherichia coli is due to the initiation of the rglB+ function by a phage-coded, anti-restriction endonuclease protein. Based on this hypothesis, we screened T4 deletion mutants for failure to give a burst in this host. The absence of an arn gene in phage mutants lacking the 55.5- to 58.4-kilobase region is verified by their inability to protect secondary infecting non-glucosylated phage from rglB-controlled cleavage. A functional arn gene was cloned on plasmid pBR325, and the 0.8-kilobase insert DNA was shown to be homologous to the DNA missing in the arn deletion phage.


Mechanisms underlying the antiarrhythmic and arrhythmogenic actions of quinidine in a Purkinje fiber-ischemic gap preparation of reflected reentry.
The effects of therapeutic levels of quinidine were studied in an ischemic gap preparation of reflected reentry. The preparation consisted of a Purkinje fiber mounted in a three-compartment chamber. A narrow central compartment was perfused with a solution prepared to mimic the extracellular milieu at a site of ischemia. Quinidine in concentrations that exert little effect on normal Purkinje tissue, 1 to 2 micrograms/ml, greatly impaired conduction and markedly prolonged refractoriness across the ischemic gap. The drug effected these changes by (1) extending the inexcitable zone within the depressed region, (2) decreasing the amplitude of the input signal entering this zone, and (3) decreasing the excitability of the tissue beyond the depressed zone (evaluated by current clamp techniques). These actions of the drug produced both antiarrhythmic and proarrhythmic effects. When the initial level of conduction impairment was high, quinidine totally suppressed reflected reentry at all frequencies by precipitating complete anterograde conduction block. At intermediate levels of block, the drug generally caused a prominent shift of the frequency dependence of reentrant activity to lower stimulation rates. Finally, when conduction was relatively less impaired, quinidine created the conditions for reflected reentry to occur. Our results suggest that the heart rate dependence of reentrant arrhythmias might be of prognostic value in the administration of antiarrhythmic drugs.


The effect of phytase enzyme and level on nutrient extraction by broilers.
Three experimental phytase enzyme preparations derived from the same Escherichia coli gene but produced in Saccharomyces cerevisiae (A), Pichia pastoris (B), and Pseudomonas fluorescens (C) were compared with a commercial enzyme preparation by addition to wheat-soybean meal diets fed to broiler chicks. A positive control diet contained sufficient available phosphorus for normal broiler growth and a negative control diet was phosphorus deficient. The 4 enzymes were added to the negative control diet at 3 levels each (150, 450, and 1,250 U/kg), and all diets were pelleted above 80 degrees C. Broiler chicks were fed experimental diets from 4 to 21 d. Chick performance and nutrient digestibility showed that the pel leting process inactivated enzymes A and C and the commercial enzyme. When added to the negative control diet, enzyme B had positive effects on broiler performance and calcium and phosphorus digestibility, and increasing levels of enzyme had greater positive effects. Enzyme B also increased the AME and protein digestibility over those of either control diet. These results suggest that enzyme B was not inactivated by pelleting above 80 degrees C, whereas the other enzymes were.


[Death and dying in the nurse formation process].
Death is one of the great mysteries of life. We live in a death denying-society. This aim of the present qualitative study was to see if undergraduate Nursing students are prepared to face death and dying during training. Seven students enrolled in the discipline "Supervised Training II", the last discipline of the course were interviewed using a semi-structured script, and the collected data were submitted to content analysis. The following categories were discussed: facing death in daily life, reaction to death, and dying in training. It was observed that the process of death and dying is not approached in a manner that supplies the students' needs in routine situations that require them to provide dying patients humane conditions to die.


Further studies on the cardiomegaly induced by beta-adrenoceptor agonists.
Rats received continuous infusions of beta-adrenoceptor agonists by means of Alzet osmotic minipumps implanted subcutaneously. After 7 days of isoprenaline infusion (2 mg/kg per day) the heart/body weight ratio increased about 40 per cent compared with placebo treatment. The difference persisted after freeze-drying indicating a true hypertrophy and not merely an oedema. When terbutaline (20 mg/kg per day) was substituted for isoprenaline, the increase in wet weight ratio reached only about 5 per cent. Procaterol (2 mg/kg per day) and pirbuterol (20 mg/kg per day) had no effect on the heart weight. It is concluded that in doses expected to cause comparable stimulation of beta 2-adrenoceptors the unselective agonist isoprenaline is able, more than the beta 2-selective agonists terbutaline, procaterol and pirbuterol to cause cardiac hypertrophy thus indicating the involvement of beta 1-adrenoceptors.


DICER regulates the expression of major satellite repeat transcripts and meiotic chromosome segregation during spermatogenesis.
Constitutive heterochromatin at the pericentric regions of chromosomes undergoes dynamic changes in its epigenetic and spatial organization during spermatogenesis. Accurate control of pericentric heterochromatin is required for meiotic cell divisions and production of fertile and epigenetically intact spermatozoa. In this study, we demonstrate that pericentric heterochromatin is expressed during mouse spermatogenesis to produce major satellite repeat (MSR) transcripts. We show that the endonuclease DICER localizes to the pericentric heterochromatin in the testis. Furthermore, DICER forms complexes with MSR transcripts, and their processing into small RNAs is compromised in Dicer1 knockout mice leading to an elevated level of MSR transcripts in meiotic cells. We also show that defective MSR forward transcript processing in Dicer1 cKO germ cells is accompanied with reduced recruitment of SUV39H2 and H3K9me3 to the pericentric heterochromatin and meiotic chromosome missegregation. Altogether, our results indicate that the physiological role of DICER in maintenance of male fertility extends to the regulation of pericentric heterochromatin through direct targeting of MSR transcripts.


Soluble immunoglobulin A in breast milk is inversely associated with atopic dermatitis at early age: the PASTURE cohort study.
The role of breastfeeding for the development of atopic diseases in childhood is contradictory. This might be due to differences in the composition of breast milk and levels of antimicrobial and anti-inflammatory components. The objective of this study was to examine whether levels of total immunoglobulin A (IgA) or transforming growth factor-β1 (TGF-β1) in breast milk were associated with the risk of developing atopic dermatitis (AD), atopic sensitization or asthma at early age taking breastfeeding duration into account. The birth cohort study PASTURE conducted in Finland, France, Germany and Switzerland provided 610 breast milk samples collected 2 months after delivery in which soluble IgA (sIgA) and TGF-β1 levels were measured by ELISA. Duration of breastfeeding was assessed using weekly food frequency diaries from month 3 to month 12. Data on environmental factors, AD and asthma were collected by questionnaires from pregnancy up to age 6. Atopic status was defined by specific IgE levels in blood collected at the ages of 4 and 6 years. Multivariate logistic regression models were used for statistical analysis. Soluble IgA and TGF-β1 levels in breast milk differed between countries, and sIgA levels were associated with environmental factors related to microbial load, for example, contact to farm animals or cats during pregnancy, but not with raw milk consumption. sIgA levels were inversely associated with AD up to the of age 2 years (P-value for adjusted linear trend: 0.005), independent of breastfeeding duration. The dose of sIgA ingested in the first year of life was associated with reduced risk of AD up to the age of 2 (aOR, 95% CI: 0.74; 0.55-0.99) and 4 years (0.73; 0.55-0.96). No clear associations between sIgA and atopy or asthma up to age 6 were observed. TGF-β1 showed no consistent association with any investigated health outcome. IgA in breast milk might protect against the development of AD.


Multi-institutional, prospective, randomized, double-blind, placebo-controlled phase IIb trial of the tumor lysate, particle-loaded, dendritic cell (TLPLDC) vaccine to prevent recurrence in high-risk melanoma patients: A subgroup analysis.
Checkpoint inhibitors (CPI) in combination with cell-based vaccines may produce synergistic antitumor immunity. The primary analysis of the randomized and blinded phase IIb trial in resected stage III/IV melanoma demonstrated TLPLDC is safe and improved 24-month disease-free survival (DFS) in the per treatment (PT) analysis. Here, we examine efficacy within pre-specified and exploratory subgroups. Stage III/IV patients rendered disease-free by surgery were randomized 2:1 to TLPLDC vaccine versus placebo. The pre-specified PT analysis included only patients completing the primary vaccine/placebo series at 6 months. Kaplan-Meier analysis was used to compare 24-month DFS among subgroups. There were no clinicopathologic differences between subgroups except stage IV patients were more likely to receive CPI. In stage IV patients, 24-month DFS was 43% for vaccine versus 0% for placebo (p = 0.098) in the ITT analysis and 73% versus 0% (p = 0.002) in the PT analysis. There was no significant difference in 24-month DFS when stratified by use of immunotherapy or CPI. For patients with resected recurrent disease, 24-month DFS was 88.9% versus 33.3% (p = 0.013) in the PT analysis. All benefit from vaccination was in the PT analysis; no benefit was found in patients receiving up to three doses. The TLPLDC vaccine improved DFS in patients completing the primary vaccine series, particularly in the resected stage IV patients. The efficacy of the TLPLDC vaccine will be confirmed in a phase III study evaluating adjuvant TLPLDC + CPI versus Placebo + CPI in resected stage IV melanoma patients.


A water-soluble fragment of the thyroid-stimulating hormone receptor which binds both thyroid-stimulating hormone and thyroid-stimulating hormone receptor antibodies.
Previous studies have shown that freezing and thawing of human thyroid homogenates releases a water-soluble substance which reversibly binds to TSH-receptor antibodies. This substance has been designated long-acting thyroid stimulator absorbing activity (LAA). We now describe a new method for measuring LAA based on the TSH-receptor assay and application of the technique to the study of LAA. Our results indicate that LAA is a heat-labile glycoprotein which co-elutes with haemoglobin on gel filtration. Furthermore, LAA is retarded by columns of Sepharose-TSH but not by Sepharose coupled to human chorionic gonadotrophin, normal immunoglobulin G or bovine serum albumin, suggesting that LAA contains a binding site for TSH as well as for TSH-receptor antibodies. It would seem therefore that LAA is a water-soluble fragment of the TSH receptor possibly resulting from proteolytic cleavage of the receptor at a site close to the cell surface.


The southpaw advantage? Lateral preference in mixed martial arts.
Performers with a left-orientation have a greater likelihood of obtaining elite levels of performance in many interactive sports. This study examined whether combat stance orientation was related to skill and success in Mixed Martial Arts fighters. Data were extracted for 1468 mixed martial artists from a reliable and valid online data source. Measures included fighting stance, win percentage and an ordinal measure of skill based on number of fights. The overall analysis revealed that the fraction of fighters using a southpaw stance was greater than the fraction of left-handers in the general population, but the relationship between stance and hand-preference is not well-understood. Furthermore, t-tests found no statistically significant relationship between laterality and winning percentage, although there was a significant difference between stances for number of fights. Southpaw fighters had a greater number of fights than those using an orthodox stance. These results contribute to an expanding database on the influence of laterality on sport performance and a relatively limited database on variables associated with success in mixed martial arts.


Reducing barriers associated with delivering health care services to migratory agricultural workers.
Between one and two million migratory agricultural workers (MAWs), primarily from Mexico and Central America, leave their homes each year to plant, cultivate, harvest and pack fruits, vegetables, and nuts in the USA. While in the USA, most lack health insurance, a permanent residence, and a regular healthcare provider. Publications over the past two decades in the USA have reported that a majority of MAWs encounter barriers to receiving medical services. Migratory agricultural workers experience high rates of occupational illness and injury. Poor access to medical care continues to exacerbate health problems among members of this population related to their working environments. In most studies concerning healthcare access issues for this population, researchers collected their information from healthcare service providers; rarely have they included input from migratory agricultural workers. This study was different in that opinions about healthcare access issues were collected directly from MAWs. The primary purpose of this study was to describe issues related to barriers associated with the delivery of healthcare services to migratory agricultural workers. A secondary purpose was to suggest strategies for reducing these barriers. In this study, data from focus group sessions were used to develop a survey questionnaire. Four certified bilingual interpreters were trained to administer the questionnaire. A total of 157 usable questionnaires were returned from MAWs living in employer-provided camps in Northwest Ohio. The statistical analyses were primarily descriptive. The most significant barriers hampering access to medical services among the 157 respondents were cost (n=113; 72.0%), crop demands (n=102; 65.0%), the lack of an interpreter (n=98; 62.4%), travel distance (n=88; 56.1%) and transportation (n=82; 52.2%). Approximately half (n=82; 52.2%) said that they had access to transportation for traveling to a medical clinic. As a group, respondents were willing to travel an average of 29.1 km (18.1 miles) (range 0-129 km [0-80 miles]) to obtain medical services. Female heads of households had significantly less access to transportation compared with male heads of households (t=2.35; df=74; p<0.05). Three general categories of barriers to health care for MAWs surfaced in this study: (1) work environment; (2) migratory agricultural worker resources; and (3) healthcare clinic practices. Work environment issues relate mostly to the employers. Resources are barriers for MAWs because they are poor and have limited funds for the cost of transportation to clinics and the fees associated with accessing health care. Most of the barriers identified related to healthcare clinic practices. Some strategies to address healthcare clinic practice barriers were developed by the group conducting the study. By listening to what MAWs described as barriers to health care, providers can help improve access which can reduce the use of high cost hospital emergency room care.


Contribution of asparagine 346 residue to the carbapenemase activity of CMY-2 β-lactamase.
Only a few plasmid-borne AmpC (pAmpC) β-lactamases, such as CMY-2, can account for carbapenem resistance in Enterobacteriaceae in combination with outer membrane impermeability. The aim of this study was to elucidate the contribution of Asn-346, which is well conserved among carbapenem-hydrolyzing pAmpCs, to the hydrolysis spectrum of CMY-2. Site-directed mutagenesis experiments were carried out to replace Asn-346 with glycine, alanine, valine, glutamate, aspartate, serine, threonine, glutamine, tyrosine, isoleucine, lysine, and histidine. The recombinant plasmids were transferred into wild-type and porin-deficient Escherichia coli strains. Asn-346 replacement reduced significantly the MICs of all β-lactams, except the Asn-346-Ile substitution that increased the MICs of cephalosporins, whereas it decreased those of carbapenems. The biochemical characterization, along with a molecular modeling study, showed that the size and the polarity of the side chain at position 346 assisted substrate binding and turnover. This study shows for the first time that the amino acid at position 346 contributes to the β-lactamase activity of cephalosporinases. Asparagine and isoleucine residues, which are well conserved at position 346 among AmpC-type enzymes, modulate their hydrolysis spectrum in an opposing sense. Ile-346 confers higher level of cephalosporins resistance, whereas Asn-346 confers carbapenem resistance in combination with outer membrane impermeability.


Objective antihistamine side effects are mitigated by evening dosing of hydroxyzine.
First-generation antihistamines have potency, pharmacokinetic, and cost advantages compared with nonsedating second-generation antihistamines. Bedtime dosing of hydroxyzine was investigated as a dosing strategy to minimize reaction time degradation and adverse subjective symptoms previously documented for hydroxyzine in divided doses. Hydroxyzine, 50 mg qhs, was compared with terfenadine, 60 mg bid, in this double-blind, placebo-controlled crossover study of 15 healthy, asymptomatic adults. Computer-based eye-hand reaction time tests of simple reaction time (SRT) and choice reaction time (CRT) were not statistically different among the three drugs. Drowsiness, dry mouth, and irritability were significant for hydroxyzine (P = .0001, .001 and .02, respectively) compared with terfenadine or placebo, but less than seen in a previous study of hydroxyzine, 25 mg bid. Symptom scores with terfenadine were comparable to placebo. Histamine skin test wheal and flare were both significantly and comparably suppressed by hydroxyzine and terfenadine (P = .0001). While wheal suppression by hydroxyzine was universal, four of the 15 subjects showed little or no suppression with terfenadine (P = .03). Although bedtime dosing of hydroxyzine did not eliminate subjective symptoms, it maintained skin H1-receptor antagonism the following morning and alleviated the prolongation of reaction times previously reported with hydroxyzine in divided doses. The significant adverse subjective symptoms and psychomotor performance degradations caused by first-generation antihistamines can be mitigated by creative dosing schedules.


InfoEvolve: moving from data to knowledge using information theory and genetic algorithms.
InfoEvolve is a unified suite of data mining and empirical modeling tools capable of discovering low-bias and low-variance solutions to complex processes. The method is based on a common set of principles involving information theory and genetic algorithms. InfoEvolve can also discover multiple strategies embedded in complex data sets for achieving a desired target or goal. This latter aspect may prove to be very useful in drug design. The paper analyzes the following: InfoEvolve from a theoretical standpoint; a conceptual overview of InfoEvolve with a short description of the modeling method; the method using the example of homogeneous identification of DNA from an analysis of its melting curve behavior; and key learnings and additional applications of the technology for both drug design and genome analysis.


Selection of variables using 'independence Bayes' in computer-aided diagnosis of upper gastrointestinal bleeding.
In this paper two problems of computer-aided diagnosis with 'independence Bayes' were investigated: selection of variables and monotonicity in performance as the number of measurements is increased. Using prospective data from patients with upper gastrointestinal bleeding, the stepwise forward selection approach maximizing the apparent diagnostic accuracy was analysed with respect to different kinds of bias in estimation of the true diagnostic accuracy and to the stability of the number and type of variables selected. The results of this study suggest first that the selection of variables should be evaluated against the estimated true diagnostic accuracy obtained using all variables, and secondly that the results of a single selected sequence may be severely biased.


Phage display against corneal epithelial cells produced bioactive peptides that inhibit Aspergillus adhesion to the corneas.
Dissection of host-pathogen interactions is important for both understanding the pathogenesis of infectious diseases and developing therapeutics for the infectious diseases like various infectious keratitis. To enhance the knowledge about pathogenesis infectious keratitis, a random 12-mer peptide phage display library was screened against cultured human corneal epithelial cells (HCEC). Fourteen sequences were obtained and BLASTp analysis showed that most of their homologue counterparts in GenBank were for defined or putative proteins in various pathogens. Based on known or predicted functions of the homologue proteins, ten synthetic peptides (Pc-A to Pc-J) were measured for their affinity to bind cells and their potential efficacy to interfere with pathogen adhesion to the cells. Besides binding to HCEC, most of them also bound to human corneal stromal cells and umbilical endothelial cells to different extents. When added to HCEC culture, the peptides induced expression of MyD88 and IL-17 in HCEC, and the stimulated cell culture medium showed fungicidal potency to various extents. While peptides Pc-C and Pc-E inhibited Aspergillus fumigatus (A.f) adhesion to HCEC in a dose-dependent manner, the similar inhibition ability of peptides Pc-A and Pc-B required presence of their homologue ligand Alb1p on A.f. When utilized in an eyeball organ culture model and an in vivo A.f keratitis model established in mouse, Pc-C and Pc-E inhibited fungal adhesion to corneas, hence decreased corneal disruption caused by inflammatory infiltration. Affinity pull-down of HCEC membrane proteins with peptide Pc-C revealed several molecules as potential receptors for this peptide. In conclusion, besides proving that phage display-selected peptides could be utilized to interfere with adhesion of pathogens to host cells, hence could be exploited for managing infectious diseases including infectious keratitis, we also proposed that the phage display technique and the resultant peptides could be used to explore host-pathogen interactions at molecular levels.


Decoding the regulation of mast cell exocytosis by networks of Rab GTPases.
Exocytosis is a key event in mast cell functions. By this process, mast cells release inflammatory mediators, contained in secretory granules (SGs), which play important roles in immunity and wound healing but also provoke allergic and inflammatory responses. The mechanisms underlying mast cell exocytosis remained poorly understood. An essential step toward deciphering the mechanisms behind exocytosis is the identification of the cellular components that regulate this process. Because Rab GTPases regulate specific trafficking pathways, we screened 44 Rabs for their functional impacts on exocytosis triggered by the FcεRI or combination of Ca ²⁺ ionophore and phorbol ester. Because exocytosis involves the continuous reorganization of the actin cytoskeleton, we also repeated our screen in the presence of cytochalasin D that inhibits actin polymerization. In this paper, we report on the identification of 30 Rabs as regulators of mast cell exocytosis, the involvement of 26 of which has heretofore not been recognized. Unexpectedly, these Rabs regulated exocytosis in a stimulus-dependent fashion, unless the actin skeleton was disrupted. Functional clustering of the identified Rabs suggested their classification as Rabs involved in SGs biogenesis or Rabs that control late steps of exocytosis. The latter could be further divided into Rabs that localize to the SGs and Rabs that regulate transport from the endocytic recycling compartment. Taken together, these findings unveil the Rab networks that control mast cell exocytosis and provide novel insights into their mechanisms of action.


Single High-Sensitivity Cardiac Troponin I to Rule Out Acute Myocardial Infarction.
This study examined the performance of single high-sensitivity cardiac troponin I (hs-cTnI) measurement strategies to rule out acute myocardial infarction. This was a prospective, observational study of consecutive patients presenting to the emergency department (n = 1631) in whom cTnI measurements were obtained using an investigational hs-cTnI assay. The goals of the study were to determine 1) negative predictive value (NPV) and sensitivity for the diagnosis of acute myocardial infarction, type 1 myocardial infarction, and type 2 myocardial infarction; and 2) safety outcome of acute myocardial infarction or cardiac death at 30 days using hs-cTnI less than the limit of detection (LoD) (<1.9 ng/L) or the High-STEACS threshold (<5 ng/L) alone and in combination with normal electrocardiogram (ECG). Acute myocardial infarction occurred in 170 patients (10.4%), including 68 (4.2%) type 1 myocardial infarction and 102 (6.3%) type 2 myocardial infarction. For hs-cTnI<LoD (27%), the NPV and sensitivity for acute myocardial infarction were 99.6% (95% confidence interval 98.9%-100%) and 98.8 (97.2%-100%). For hs-cTnI<5 ng/L (50%), the NPV and sensitivity for acute myocardial infarction were 98.9% (98.2%-99.6%) and 94.7% (91.3%-98.1%). In combination with a normal ECG, 1) hs-cTnI<LoD had an NPV of 99.6% (98.9%-100%) and sensitivity of 99.4% (98.3%-100%); and 2) hs-cTnI<5 ng/L had an NPV of 99.5% (98.8%-100%) and sensitivity of 98.8% (97.2%-100%). The NPV and sensitivity for the safety outcome were excellent for hs-cTnI<LoD alone or in combination with a normal ECG, and for hs-cTnI<5 ng/L in combination with a normal ECG. Strategies using a single hs-cTnI alone or in combination with a normal ECG allow the immediate identification of patients unlikely to have acute myocardial infarction and who are at very low risk for adverse events at 30 days.


Melatonin administration to wild-type mice and nontreated NLRP3 mutant mice share similar inhibition of the inflammatory response during sepsis.
The NLRP3 inflammasome is involved in the innate immune response during inflammation. Moreover, melatonin blunts the NF-κB/NLRP3 connection during sepsis. Thus, we compared the roles of the NLRP3 inflammasome and/or melatonin treatment in the septic response of wild-type and NLRP3-/- mice. Mouse myocardial tissue was used for this purpose. The nuclear turnover of NF-κB was enhanced during sepsis, with an increase in TNFα, iNOS, and pro-IL-1β. The lack of inflammasome in NLRP3-/- mice significantly reduced that response and blunted IL-1β maturation due to the lack of caspase-1. Clock and Bmal1 did not change in both mouse strains, enhancing Chrono expression in mutants. RORα, which positively regulates Bmal1, was enhanced at a similar extend in both mouse strains, whereas the expression of the Bmal1 repressor, Rev-Erbα, increased in WT but was depressed in NLRP3-/- mice. Nampt, transcriptionally controlled by Bmal1, increased in WT mice together with Sirt1, whereas they remained unchanged in NLRP3-/- mice. Melatonin treatment reduced the septic response in a comparable manner as did the lack of NLRP3, but unlike the latter, it normalized the clock genes turnover through the induction of RORα and repression of Rev-Erbα and Per2, leading to enhanced Nampt and Sirt1. The lack of NLRP3 inflammasome converts sepsis to a moderate inflammatory disease and identifies NLRP3 as a main target for the treatment of sepsis. The efficacy of melatonin in counteracting the NLRP3 inflammasome activation further confirms the indoleamine as a useful therapeutic drug against this serious condition.


Nur77 influences immunometabolism to regulate the release of proinflammatory cytokines and the formation of lipid bodies during Mycobacterium tuberculosis infection of macrophages.
Infection of macrophages with Mycobacterium tuberculosis induces innate immune responses designed to clear the invading bacterium. However, bacteria often survive within the intracellular environment by exploiting these responses triggered by macrophages. Here, the role of the orphan nuclear receptor Nur77 (Nr4a1) in regulating the response of macrophages infected with M. tuberculosis (Mtb) has been delineated. Nur77 is induced early during infection, regulates metabolism by binding directly at the promoter of the TCA cycle enzyme, isocitrate dehydrogenase 2 (IDH2), to act as its repressor, and shifts the balance from a proinflammatory to an anti-inflammatory phenotype. Depletion of Nur77 increased transcription of IDH2 and, consequently, the levels of intracellular succinate, leading to enhanced levels of the proinflammatory cytokine IL-1β. Further, Nur77 inhibited the production of antibacterial nitric oxide and IL-1β in a succinate dehydrogenase (SDH)-dependent manner, suggesting that its induction favors bacterial survival by suppressing bactericidal responses. Indeed, depletion of Nur77 inhibited the intracellular survival of Mtb. On the other hand, depletion of Nur77 enhanced lipid body formation, suggesting that the fall in Nur77 levels as infection progresses likely favors foamy macrophage formation and long-term survival of Mtb in the host milieu.


Sequence analysis of the haemagglutinin (HA) of influenza A (H1N1) viruses present in clinical material and comparison with the HA of laboratory-derived virus.
We used the polymerase chain reaction to amplify the HA1 coding region of influenza A (H1N1) viruses present in clinical material from recent cases of influenza in the U.K. Previously, we have demonstrated that isolation of human influenza viruses in embryonated hens' eggs selects variants which have amino acid substitutions in their haemagglutinin (HA) clustering around the receptor-binding site. Such egg-selected variants are often antigenically distinct from each other and from corresponding viruses isolated on mammalian cells. Since in general the virus used for vaccine production is an egg-adapted virus, it is important to determine the extent to which these variants are present in the natural virus which causes disease in man. To achieve this, amplified products from clinical material were cloned and many individual clones sequenced. Our results indicate that the HA of the naturally occurring virus is relatively homogeneous and represented by virus isolated in the laboratory on MDCK cells, whereas the variants isolated in eggs are present only at low levels in clinical material.


Moderate drought causes dramatic floral transcriptomic reprogramming to ensure successful reproductive development in Arabidopsis.
Drought is a major constraint that leads to extensive losses to agricultural yield worldwide. The potential yield is largely determined during inflorescence development. However, to date, most investigations on plant response to drought have focused on vegetative development. This study describes the morphological changes of reproductive development and the comparison of transcriptomes under various drought conditions. The plants grown were studied under two drought conditions: minimum for successful reproduction (45-50% soil water content, moderate drought, MD) and for survival (30-35%, severe drought, SD). MD plants can produce similar number of siliques on the main stem and similar number of seeds per silique comparing with well-water plants. The situation of SD plants was much worse than MD plants. The transcriptomes of inflorescences were further investigated at molecular level using microarrays. Our results showed more than four thousands genes with differential expression under severe drought and less than two thousand changed under moderate drought condition (with 2-fold change and q-value < 0.01). We found a group of genes with increased expression as the drought became more severe, suggesting putative adaptation to the dehydration. Interestingly, we also identified genes with alteration only under the moderate but not the severe drought condition, indicating the existence of distinct sets of genes responsive to different levels of water availability. Further cis-element analyses of the putative regulatory sequences provided more information about the underlying mechanisms for reproductive responses to drought, suggesting possible novel candidate genes that protect those developing flowers under drought stress. Different pathways may be activated in response to moderate and severe drought in reproductive tissues, potentially helping plant to maximize its yield and balance the resource consumption between vegetative and reproductive development under dehydration stresses.