Nutmeg intoxication in Texas, 1998-2004.
Nutmeg is a spice that contains volatile oils comprised of alkyl benzene derivatives (myristicin, elemicin, safrole, etc.), terpenes and myristic acid. Nutmeg has a long history of abuse. This study describes the nutmeg ingestion calls received by Texas poison centers from 1998 to 2004. There were 17 calls involving nutmeg ingestion, of which 64.7% involved intentional abuse. When abuse and non-abuse ingestions were compared, abuse ingestions were more likely to involve males (100 versus 66.7%) and adolescents (55.6 versus 16.7%). The majority of both abuse and non-abuse calls were managed outside of health care facilities (54.5 and 66.7%, respectively). None of the ingestions resulted in more than moderate clinical effects or death.


Catalysis of lysine 48-specific ubiquitin chain assembly by residues in E2 and ubiquitin.
Protein ubiquitination is catalyzed by ubiquitin-conjugating enzymes (E2s) in collaboration with ubiquitin-protein ligases (E3s). This process depends on nucleophilic attack by a substrate lysine on a thioester bond linking the C terminus of ubiquitin to a cysteine in the E2 active site. Different E2 family members display specificity for lysines in distinct contexts. We addressed the mechanistic basis for this lysine selectivity in Ubc1, an E2 that catalyzes the ubiquitination of lysine 48 (K48) in ubiquitin, leading to the formation of K48-linked polyubiquitin chains. We identified a cluster of polar residues near the Ubc1 active site, as well as a residue in ubiquitin itself, that are required for catalysis of K48-specific ubiquitin ligation, but not for general activity toward other lysines. Our results suggest that the active site of Ubc1, as well as the surface of ubiquitin, contains specificity determinants that channel specific lysines to the central residues involved directly in catalysis.


A thioacetamide-induced hepatic encephalopathy model in C57BL/6 mice: a behavioral and neurochemical study.
Hepatic encephalopathy (HE) is a neuropsychiatric syndrome resulting from liver failure. In the present study, we aimed to standardize an animal model of HE induced by thioacetamide (TAA) in C57BL/6 mice evaluating behavioral symptoms in association with liver damage and alterations in neurotransmitter release. HE was induced by an intraperitoneal single dose of TAA (200 mg/kg, 600 mg/kg or 1,200 mg/kg). Behavioral symptoms were evaluated using the SHIRPA battery. Liver damage was confirmed by histopathological analysis. The glutamate release was measured using fluorimetric assay. The neuropsychiatric state, motor behavior and reflex and sensory functions were significantly altered in the group receiving 600 mg/kg of TAA. Biochemical analysis revealed an increase in the glutamate release in the cerebral cortex of HE mice. HE induced by 600 mg/kg TAA injection in C57BL/6 mice seems to be a suitable model to investigate the pathogenesis and clinical disorders of HE.


DART-MS for rapid, preliminary screening of urine for DMAA.
Dimethylamylamine (DMAA) is a sympathomimetic amine found in weight-loss/workout supplements or used as an appetite suppressant. DMAA is a stimulant that is banned by the World Anti-Doping Agency (WADA). Adverse health effects as well as fatalities have been implicated with its use. Direct analysis in real time mass spectrometry (DART-MS) is an ambient ionization method that was employed to rapidly identify the presence of DMAA in various samples without any extraction or preparations whatsoever. DMAA was first identified in supplements, sampled directly in their solid forms. Furthermore, DMAA was detected directly in urine over 48 h as a means of indicating recent abuse of the substance. DART-MS analysis is instantaneous, and coupled with the high mass accuracy associated with the time-of-flight mass analyzer, results in unequivocal identification of the presence of DMAA. These features demonstrate DART-MS as an attractive potential alternative screening method for the presence of drugs and medications or for toxicological investigations.


Metallopharmaceuticals based on silver(I) and silver(II) polydiguanide complexes: activity against burn wound pathogens.
The in vitro pharmacodynamics of silver(I) and silver(II) complexes of a polydiguanide ligand, chlorhexidine, were assayed to examine the value of the bactericidal endpoint as an alternative means of evaluating their antibacterial activities against burn wound pathogens. Synthesis of silver(I) chlorhexidine [Ag(I)CHX] was accomplished by in situ precipitation of the complex from a feebly acidic or neutral aqueous solution of AgNO(3) and chlorhexidine, whereas silver(II) chlorhexidine [Ag(II)CHX] was synthesized by oxidation of Ag(I), followed by complexation of the oxidized metal with chlorhexidine. Their antibacterial potencies were assessed in vitro by determining the MICs and MBCs for four Gram-positive and four Gram-negative bacteria. Time-kill assays using three different concentrations of these agents were also performed. The MICs of Ag(I)CHX and Ag(II)CHX were much lower than those of chlorhexidine, AgNO(3) and silver sulfadiazine. The time-kill study provided quantitative information on actual times required to reach the bactericidal endpoint using a particular concentration of the active agent. The lethality rates of Ag(I)CHX and Ag(II)CHX against the tested bacteria were 2× to 8× faster than those of chlorhexidine or AgNO(3) at a concentration equal to or 4× MIC. Ag(I)CHX and Ag(II)CHX showed superior antibacterial activity and faster killing kinetics compared with chlorhexidine and AgNO(3). These complexes may serve as new-generation antibacterial agents in wound care.


Governing the Lebanese health system: strengthening the national response to the burden of Syrian refugees.
Lebanon is providing sanctuary to an estimated 1.5 million Syrian refugees, with potential consequences for its health system. Here, we analyse how it has responded to this challenge, identify sensitive areas where a strong national governance system is needed and explore how it might be implemented. An effective response to the Syrian refugee crisis requires concerted international action. Nonetheless, geography dictates that the Lebanese health system must play a central role. We identify some areas where a strengthened stewardship role of the Ministry of Public Health is urgently required. We argue that the Ministry is well placed to take a lead, with its detailed knowledge of the Lebanese health system and its legitimacy to formulate a national health response. Finally, we suggest that this crisis could be a catalyst for the strengthening of the Lebanese health system, based on evidence-informed policies that would benefit refugees and the Lebanese population alike.


GM-CSF and the impaired pulmonary innate immune response following hyperoxic stress.
We have previously demonstrated that mice exposed to sublethal hyperoxia (an atmosphere of >95% oxygen for 4 days, followed by return to room air) have significantly impaired pulmonary innate immune response. Alveolar macrophages (AM) from hyperoxia-exposed mice exhibit significantly diminished antimicrobial activity and markedly reduced production of inflammatory cytokines in response to stimulation with LPS compared with AM from control mice in normoxia. As a consequence of these defects, mice exposed to sublethal hyperoxia are more susceptible to lethal pneumonia with Klebsiella pneumoniae than control mice. Granulocyte/macrophage colony-stimulating factor (GM-CSF) is a growth factor produced by normal pulmonary alveolar epithelial cells that is critically involved in maintenance of normal AM function. We now report that sublethal hyperoxia in vivo leads to greatly reduced alveolar epithelial cell GM-CSF expression. Systemic treatment of mice with recombinant murine GM-CSF during hyperoxia exposure preserved AM function, as indicated by cell surface Toll-like receptor 4 expression and by inflammatory cytokine secretion following stimulation with LPS ex vivo. Treatment of hyperoxic mice with GM-CSF significantly reduced lung bacterial burden following intratracheal inoculation with K. pneumoniae, returning lung bacterial colony-forming units to the level of normoxic controls. These data point to a critical role for continuous GM-CSF activity in the lung in maintenance of normal AM function and demonstrate that lung injury due to hyperoxic stress results in significant impairment in pulmonary innate immunity through suppression of alveolar epithelial cell GM-CSF expression.


Independent control of presynaptic inhibition by reticulospinal and sensory inputs at rest and during rhythmic activities in the cat.
To be functionally relevant during movement, the transmission from primary afferents must be efficiently controlled by presynaptic inhibition. Sensory feedback, central pattern generators, and supraspinal structures can all evoke presynaptic inhibition, but we do not understand how these inputs interact during movement. Here, we investigated the convergence of inputs from the reticular formation and sensory afferents on presynaptic inhibitory pathways and their modulation at rest and during two fictive motor tasks (locomotion and scratch) in decerebrate cats. The amplitude of primary afferent depolarization (PAD), an estimate of presynaptic inhibition, was recorded in individual afferents with intra-axonal recordings and in a mix of afferents in lumbar dorsal rootlets (dorsal root potential [DRP]) with bipolar electrodes. There was no spatial facilitation between inputs from reticulospinal and sensory afferents with DRPs or PADs, indicating an absence of convergence. However, spatial facilitation could be observed by combining two sensory inputs, indicating that convergence was possible. Task-dependent changes in the amplitude of responses were similar for reticulospinal and sensory inputs, increasing during fictive locomotion and decreasing during fictive scratch. During fictive locomotion, DRP and PAD amplitudes evoked by reticulospinal inputs were increased during the flexion phase, whereas sensory-evoked DRPs and PADs showed maximal amplitude in either flexion or extension phases. During fictive scratch, the amplitudes of DRPs and PADs evoked by both sources were maximal in flexion. The absence of spatial facilitation and different phase-dependent modulation patterns during fictive locomotion are consistent with independent presynaptic inhibitory pathways for reticulospinal and sensory inputs.


A 2D mechanistic model of breast ductal carcinoma in situ (DCIS) morphology and progression.
Ductal carcinoma in situ (DCIS) of the breast is a non-invasive tumor in which cells proliferate abnormally, but remain confined within a duct. Although four distinguishable DCIS morphologies are recognized, the mechanisms that generate these different morphological classes remain unclear, and consequently the prognostic strength of DCIS classification is not strong. To improve the understanding of the relation between morphology and time course, we have developed a 2D in silico particle model of the growth of DCIS within a single breast duct. This model considers mechanical effects such as cellular adhesion and intra-ductal pressure, and biological features including proliferation, apoptosis, necrosis, and cell polarity. Using this model, we find that different regions of parameter space generate distinct morphological subtypes of DCIS, so elucidating the relation between morphology and time course. Furthermore, we find that tumors with similar architectures may in fact be produced through different mechanisms, and we propose future work to further disentangle the mechanisms involved in DCIS progression.


Israeli dietitians' professional stigma attached to obese patients.
The aim of this study was to explore and define the different dimensions of professional stigma attached to obese patients by dietitians. Four focus groups were conducted with 23 Israeli dietitians. Findings showed that while treating obese patients, dietitians underwent a stigmatization process involving cognitive, emotional, and behavioral phases. Obese patients with an internal locus of control, who took responsibility for their failure to diet, triggered positive feelings (e.g., pity and empathy), whereas obese patients with an external locus of control, who blamed others for their failure, triggered negative feelings (e.g., anger and frustration). Participants' emotional rejection of obese patients was manifested in three behavioral dimensions: instrumental avoidance (e.g., shorter sessions); professional avoidance (e.g., less energy and effort); and interpersonal avoidance (negative tone and evasive verbal and body language). Continuing education for dietitians is recommended to assist them in dealing with their negative feelings and behaviors toward resistant obese patients.


Probing Immune-Mediated Clearance of Acute Middle Ear Infection in Mice.
Acute otitis media (AOM) is commonly caused by bacterial pathobionts of the nasopharynx that ascend the Eustachian tube to cause disease in the middle ears. To model and study the various complexities of AOM, common human otopathogens are injected directly into the middle ear bullae of rodents or are delivered with viral co-infections which contribute to the access to the middle ears in complex and partially understood ways. Here, we present the novel observation that Bordetella bronchiseptica, a well-characterized respiratory commensal/pathogen of mice, also efficiently ascends their Eustachian tubes to colonize their middle ears, providing a flexible mouse model to study naturally occurring AOM. Mice lacking T and/or B cells failed to resolve infections, highlighting the cooperative role of both in clearing middle ear infection. Adoptively transferred antibodies provided complete protection to the lungs but only partially protected the middle ears, highlighting the differences between respiratory and otoimmunology. We present this as a novel experimental system that can capitalize on the strengths of the mouse model to dissect the molecular mechanisms involved in the generation and function of immunity within the middle ear.


Growth and biologic properties of karyotypically defined subcategories of adult acute lymphocytic leukemia in mice with severe combined immunodeficiency.
Modest progress has been achieved over the past two decades in the treatment of adult acute lymphocytic leukemia (ALL). With modern therapy, response rates are 70% to 80%, but cure rates only average 25% to 30%. Improved in vivo models are needed to investigate the biology of adult ALL and to test new treatment concepts. Fresh leukemia samples from children with ALL have been successfully transplanted into mice with severe combined immunodeficiency (SCID), but no experience exists for adult ALL. We treated SCID mice with 2 mg cyclophosphamide 24 hours before intravenously injecting 20 x 10(6) viable leukemia cells obtained from 13 patients with newly diagnosed adult ALL within five defined phenotype/karyotype subcategories. Ten (76%) of 13 injected leukemia specimens representing all five categories engrafted. The median survival duration of mice was 20 weeks from the time of leukemia cell injection. The rate of engraftment by ALL subset was as follows: two of two T-cell, two of three t(11q23), two of two hyperdiploid, two of three t(9;22), and two of three diploid ALL. The pattern of organ involvement by leukemia in the mice was similar to that of the human disease. Immunohistochemistry and flow cytometry documented the stability of each leukemic phenotype after passage through SCID mice. Cells transplanted from the spleen and bone marrow of mice engrafted with ALL into recipient mice resulted in consistent engraftment. The survival duration in passage groups was similar to that in groups injected with primary cells. The high frequency of engraftment, availability of frozen original specimens, and successful passages in SCID mice provide an in vivo model of adult ALL suitable for further studies of the disease biology and for design of drug studies for the different subtypes of previously untreated adult ALL.


Intraluminal infusion of Penta-Galloyl Glucose reduces abdominal aortic aneurysm development in the elastase rat model.
An abdominal aortic aneurysm (AAA) is a progressive chronic dilatation of the abdominal aorta with terminally rupture when the aortic wall is so weakened that aortic wall stress exceeds wall strength. No effective medical treatment exists so far. We aimed to test whether intraluminal admission of Penta-Galloyl Glucose (PGG) treatment in a rodent AAA model could hold the potential to inhibit aneurysmal progression. Male Sprague Dawley rats had either intraluminal elastase infused for AAA induction or saline to serve as controls. In two independent experimental series, elastase was used to induce AAA followed by an intraluminal PGG (directly or by a drug eluting balloon) treatment. All rats were followed for 28 days and euthanized. In both series, maximal infrarenal aortic diameter was measured at baseline and at termination as a measure of AAA size. In series 2, maximal internally AAA diameter was followed by ultrasound weekly. AAA tissues were analyzed for elastin integrity by millers stain, collagen deposition by masson trichrome staining. In other AAA tissue samples the mRNA level of CD45, lysyloxidase (LOX), lysyloxidase like protein 1 (LOXL1) were determined by qPCR. Direct administration of PGG significantly reduced AAA expansion when compared to controls. PGG treatment resulted in a higher number and more preserved elastic fibers in the aneurysmal wall, while no significant difference was seen in the levels of CD45 and LOX mRNA levels. The drug eluting balloon (DEB) experiment showed no significant difference in AAA size observed neither macroscopically nor ultrasonically. Also the aneurysmal mRNA levels of CD45, LOX and LOXL1 were unchanged between groups. A significant reduced expansion of AAAs was observed in the PGG group, suggesting PGG as a drug to inhibit aneurysmal progression, while administration through a DEB did not show a promising new way of administration.


Antioxidant activity of prune (Prunus domestica L.) constituents and a new synergist.
Ethanol extract of prune was separated into hexane-soluble and H(2)O-soluble fractions, and the H(2)O-soluble fraction was further separated into a methanol (MeOH) eluate and an H(2)O eluate by Diaion HP-20 column chromatography. The MeOH eluate exhibited the strongest antioxidant activity among the separated fractions evaluated by oxygen radical absorbance capacity (ORAC). Further purification of the MeOH eluate led to isolation of a novel compound, 4-amino-4-carboxychroman-2-one, together with four known compounds (p-coumaric acid, vanillic acid beta-glucoside, protocatechuic acid, and caffeic acid), structures of which were identified by NMR and MS analyses. The ORAC values of these isolated compounds showed 0.15-1.43 units (micromol of Trolox equiv)/micromol, and the new compound showed a remarkable synergistic effect on caffeoylquinic acid isomers. The antioxidant activity of the MeOH eluate was highly dependent on the major prune components, caffeoylquinic acid isomers, with a contribution from the new synergist.


Conditions determining the local stability of populations.
The conditions that determine the local stability classification of an equilibrium population configuration are analyzed. The population investigated is age-structured and density-dependent, where density is determined by an age-weighted population size. 2 demographic parameters are introduced: the marginal birth rate and the marginal death rate, which describe the marginal density-dependence of the birth and death rates of the equilibrium population. Certain necessary and/or sufficient conditions determining stability are developed, most of them involving the net reproduction rate of the population, and examples illustrating these conditions are presented.


Host-Pathogen Interactions: VII. Plant Pathogens Secrete Proteins which Inhibit Enzymes of the Host Capable of Attacking the Pathogen.
The results presented demonstrate that microbial pathogens of plants have the ability to secrete proteins which effectively inhibit an enzyme synthesized by the host; an enzyme whose substrate is a constituent of the cell wall of the pathogen. The system in which this was discovered is the anthracnose-causing fungal pathogen (Colletotrichum lindemuthianum) and its host, the French bean (Phaseolus vulgaris). An endo-beta-1, 3-glucanase present in the bean leaves is specifically inhibited by a protein secreted by C. lindemuthianum. The cell walls of C. lindemuthianum are shown to be composed largely of a 1, 3-glucan.


Grant writing: Moving from generating ideas to applying to grants that matter.
There is an extraordinary burden placed upon the healthcare system and people as a result of health disparities that exist within the United States. If there is going to be a concerted effort to develop innovative strategies to reduce health disparities, input from the community and behavioral scientists can and should be included in this approach and narrative. Grant writing provides one vehicle to express the narrative and to provide a means to fund research and programs within clinic-based and community settings. This paper describes a four-step inquiry process to guide healthcare professionals with varying degrees of clinical and scholarship interests through the grant writing process. They include: (1) Why write grants (motivations), (2) what is the area of focus? (Interests), (3) whom should be on the project? (partnerships), and (4) what needs to happen next to move the idea forward? (actions) The complexity of psychosocial issues means that behavioral science is well suited to develop both hypotheses-driven and phenomenological research to understand bio-psycho-social health issues. Grant writing does not need to be mysterious or daunting. It can provide a means to an end, not only to fund research but also as a means to an end of health disparities.


Zika virus NS3 protease induces bone morphogenetic protein-dependent brain calcification in human fetuses.
The most frequent fetal birth defect associated with prenatal Zika virus (ZIKV) infection is brain calcification, which in turn may potentially affect neurological development in infants. Understanding the mechanism could inform the development of potential therapies against prenatal ZIKV brain calcification. In perivascular cells, bone morphogenetic protein (BMP) is an osteogenic factor that undergoes maturation to activate osteogenesis and calcification. Here, we show that ZIKV infection of cultivated primary human brain pericytes triggers BMP2 maturation, leading to osteogenic gene expression and calcification. We observed extensive calcification near ZIKV+ pericytes of fetal human brain specimens and in vertically transmitted ZIKV+ human signal transducer and activator of transcription 2-knockin mouse pup brains. ZIKV infection of primary pericytes stimulated BMP2 maturation, inducing osteogenic gene expression and calcification that were completely blocked by anti-BMP2/4 neutralizing antibody. Not only did ZIKV NS3 expression alone induce BMP2 maturation, osteogenic gene expression and calcification, but purified NS3 protease also effectively cleaved pro-BMP2 in vitro to generate biologically active mature BMP2. These findings highlight ZIKV-induced calcification where the NS3 protease subverts the BMP2-mediated osteogenic signalling pathway to trigger brain calcification.


Assessment of bone marrow changes in postmenopausal women with varying bone densities: magnetic resonance spectroscopy and diffusion magnetic resonance imaging.
Recent studies suggest that bone marrow adipose tissue might play a role in the pathogenesis of osteoporosis. There are inconsistent findings on the relationship among marrow fat content, bone mineral density and apparent diffusion coefficient (ADC). This study aimed to prospectively explore the efficacy of MR spectroscopy (MRS) and diffusion-weighted MR imaging (DWI) in detecting vertebral marrow changes in postmenopausal women with varying bone densities. Both MRS and DWI of the lumber spine were performed in 102 postmenopausal women (mean age, (67.3 +/- 6.5) years; range, 55 - 83 years), who underwent dual X-ray absorptiometry. Marrow fat content and ADC were compared and correlated among three groups: 24 with normal bone density, 31 with osteopenia and 47 with osteoporosis. Vertebral marrow fat content was significantly increased in the osteoporotic group ((65.60 +/- 7.68)%, P < 0.001) and the osteopenic group ((57.68 +/- 6.45)%, P < 0.001), when compared with the normal bone density group ((51.67 +/- 3.27)%). ADC values were significantly decreased in the osteoporotic group ((0.39 +/- 0.03) x 10(-3)mm(2)/s, P < 0.001) and in the osteopenic group ((0.42 +/- 0.02) x 10(-3)mm(2)/s, P < 0.001), when compared with the normal bone density group ((0.47 +/- 0.03) x 10(-3)mm(2)/s). The marrow fat content negatively correlated with both bone density (r = -0.731, P < 0.001) and marrow ADC (r = -0.572, P < 0.001). The bone density positively correlated with the ADC values (r = 0.802, P < 0.001). Postmenopausal women experience a corresponding increase in vertebral marrow fat content as the bone density decreases. Marrow fat content and ADC correlate to the bone density. MRS and DWI may indirectly assess the early bone marrow changes in postmenopausal women.


The role of local radiotherapy following rituximab-containing chemotherapy in patients with transformed indolent B-cell lymphoma.
This study aimed to evaluate the outcomes of local radiotherapy (LRT) in patients with histologic transformation (HT) following rituximab-containing chemotherapy. We retrospectively analysed 92 patients with biopsy-confirmed HT undergoing rituximab-containing chemotherapy at our institution between 2003 and 2015. Of the 36 patients with limited-stage disease at diagnosis of HT, 29 (78%) received LRT. The estimated 5-year progression-free survival (PFS) rate was significantly better in patients who underwent LRT than in those who did not (93% and 42%, respectively; P < 0.05). Multivariate analyses employing age, sex, performance status, LRT and treatment response demonstrated that LRT was an independent prognostic factor for PFS (hazard ratio [HR]: 11.8; 95% confidence interval [CI]: 1.28-108.1; P < 0.05). Of the 32 patients who underwent LRT for HT lesion treatment, 31 (97%) did not show disease progression within radiation fields; among them, 27 patients (84%) survived without disease progression during the follow-up period. One patient developed hypothyroidism due to LRT; the others had no acute or late-onset complications of LRT. Our data support the recommendation of LRT for HT lesion treatment following rituximab-containing chemotherapy in select patients with localised HT, as a rational treatment approach with potentially limited toxicity.


Effect of breathing pattern on esophageal pressure gradients in humans.
This study examines the hypothesis that respiratory muscle action alters regional pleural pressures. Seven normal volunteers were studied in the seated posture with three esophageal balloons positioned in the upper, middle, and lower esophagus. The intraesophageal pressure swings were recorded during natural breathing, enhanced rib cage, and enhanced diaphragmatic breathing. The pressures were also recorded at three frequencies ranging from 12 to 60 breaths/min. Data were processed with an ensemble average technique that largely removes cardiogenic artifact. During quiet natural breathing, the pressure swings in the lower esophagus were, on the average, 30% larger than the pressure swings in the upper esophagus but became nearly equal as the frequency increased to 60 breaths/min. Rib cage breathing nearly abolished the intraesophageal dynamic pressure differences, whereas abdominal breathing preserved the observed pressure difference even at 60 breaths/min. We concluded that muscle action can affect intrathoracic pressures in a regional way depending on the groups of muscles that are active. These results are discussed in terms of the topographical distribution of ventilation.


Distinct phenotypic and functional characteristics of human natural killer cells obtained by rapid interleukin 2-induced adherence to plastic.
Human natural killer (NK) cells can be functionally subdivided into adherent (A) and non-adherent (NA) subpopulations. In the presence of 22 nM of interleukin 2(IL2), a substantial proportion of resting (R)-NK cells developed adherence to plastic as early as after 5 min of IL2 incubation, and by 1-5 hr of IL2 induction, 16% (range, 4-30%) of NK cells were adherent. Optimal concentration of IL2 for adherence of NK cells was 2-22 nM. This adherence was blocked completely by antibody to IL2 receptor (IL2R)-beta and, partially, by antibodies to beta 1 or beta 2 integrins, ICAM-1, CD2 or LFA3, but not by antibodies to the IL2R-alpha or CD56 antigen. A-NK cells separated from NA-NK cells after 5 hr of incubation in the presence of IL2 were significantly (P < 0.05) enriched in CD56dimCD16dim or -IL2Rp55+ and IL2Rp75+ cells, but were depleted of CD56bright CD16- cells. While surface density of CD56 and CD16 antigens was lower, that of beta 2 integrins (CD18, CD11a, CD11b) was higher on A-NK than on NA-NK cells. In a single-cell cytotoxicity assay, 61% of A-NK vs 37% of NA-NK cells bound, and 24% of A-NK vs 11% of NA-NK cells killed, K562 targets. In 4-day cultures with 0.02 or 2.2 nM of IL2, A-NK cells developed lymphokine-activated killer (LAK) activity later than NA-NK cells. By autoradiography, three to eight times more A-NK than NA-NK cells incorporated [3H]TdR into cell nuclei between 48 and 96 hr of IL2 incubation. In 14-day cultures in the presence of 22 nM of IL2, A-NK cells, which were initially adherent but later grew as single-cell suspensions, proliferated better (30-fold; P < 0.03) and expressed lower membrane density of CD56 than NA-NK cells. In culture, A-NK cells had consistently higher cytotoxicity against K562 targets than NA-NK cells, but cytotoxicity against Daudi was similar for both subsets. The data indicate that short incubation (1-5 hr) of human NK cells in the presence of 22 nM of IL2 allows for selection of a subpopulation which differs from the rest of NK cells not only by properties of rapid adherence to plastic, but also by a characteristic phenotype (CD3-CD56dim or -CD16dim or -beta 2integrinsbrightIL2Rp75+), rapid expression of IL2R-alpha, higher NK activity, delayed development of LAK activity, and ability to respond optimally in the presence of 22 nM of IL2.(ABSTRACT TRUNCATED AT 400 WORDS)


Clinical success of compomer and amalgam restorations in primary molars. Follow up in 36 months.
The purpose of this study was to evaluate the three-year clinical performance of a compomer and an amalgam in primary molars and to determine fluoride content in the enamel of these teeth after exfoliation. Twenty patients aged six to eight years randomly received one compomer and one amalgam restoration. Restorations were evaluated at baseline, 12, 24 and 36 months according to modified USPHS criteria. In six pairs of exfoliated teeth fluoride content at 1 and 5 mm distance from the restorations was determined. Replicas were made to determine marginal adaptation (MA) by scanning electron microscopy (SEM). At occlusal margins compomer restorations had better MA than amalgam restorations (p < 0.01). Compomer and amalgam restorations did not lead to different fluoride uptake in adjacent enamel and had an overall clinically satisfactory performance. The study confirmed that the assessed compomer is a suitable restorative material for class I and class II restorations in primary molar teeth.


Mediating Roles of Intimate Partner Violence, Stress, and Social Support on Depressive Symptoms Among Thai Women.
Prior research has shown that intimate partner violence (IPV), stress, and social support are associated with depressive symptoms. However, a possible mediating role of these variables linking depressive symptoms has not been fully investigated in Thailand. This study was conducted to assess the mediating roles of IPV, stress, and social support in the relationships between childhood abuse (CA), number of sexual partners (NSP), and depressive symptoms among 400 Thai women attending gynecology clinics. Results indicated that IPV was significantly positively correlated with stress and depressive symptoms but negatively correlated with social support. CA and NSP were significantly positively correlated with IPV, stress, and depressive symptoms, but negatively correlated with social support. Structural equation models (SEMs) showed that not only did IPV severity exhibit significantly indirect effect on depressive symptoms (β = .178; p < .05), but it also had a significant, positive total effect on depressive symptoms (β = .252; p < .05). In particular, IPV severity had the fully mediating effect on depressive symptoms through social support (β = -.204; p < .05) and stress (β = .158; p < .05). CA severity and NSP exhibited the significant indirect effect on depressive symptoms through IPV severity, stress, and social support. All the models analyzed showed that stress had an important mediator role (β = .583; p < .05) on depressive symptoms. The model fitted very well to the empirical data and explained 53% of variance. Findings affirmed the important role of these mediators as well as the need to design interventions for reducing stress or increasing support for women experiencing IPV. Utilizing an empowerment approach among female outpatients to decrease stress and depressive symptoms is recommended.


[Neoplasms and angiogenesis].
Angiogenesis is a process of new blood vessels formation in many physiological and pathological conditions such as wound healing, cancer, ischaemic diseases (of heart or limb) and chronic inflammations. Neoplastic angiogenesis plays an important role in progression of the disease and development of metastases. The angiogenic process may be initiated by hypoxia in cancerous cells, mutations in suppressor genes or oncogenes. Cancer cells activate specific angiogenic factors mainly vasoendothelial growth factor (VEGF) that stimulate migration and division of vascular endothelial cells. The studies of several pro- and antiangiogenic molecules and factors, and mechanism of vessel growth, have revealed that therapeutic interferences with these processes offers a tool for clinical applications in various pathologies. Inhibition of angiogenesis can prevent diseases with excessive vessel growth such as, particularly, cancers. Stimulation of angiogenesis can be beneficial in the treatment of diseases such as coronary disease and limb ischaemia. In this article some aspects are presented of current knowledge of angiogenesis and its regulation.


Superior mesenteric artery syndrome in a patient with Charcot Marie Tooth disease.
The extrinsic compression of the third part of the duodenum as it passes through the aorto-mesenteric angle is known as the superior mesenteric artery syndrome (SMAS). This syndrome is a rare mechanical cause of upper intestinal obstruction, with a reported incidence of between 0.2% and 0.78%. Clinical manifestations of SMAS may be chronic or acute; chronic symptoms include intermittent gastric pain, fullness and occasional episodes of postprandial vomiting, while acute symptoms include incoercible vomiting, oral intolerance, mainly epigastric abdominal distension and abdominal pain. Surgery is recommended only when initial conservative treatment fails. Here, we report what appears to be the third published case of SMAS associated with hereditary motor and sensory neuropathy or Charcot Marie Tooth disease.


Entropy Theory and Glass Transition: A Test by Monte Carlo Simulation.
This article reviews the results of a test of the Gibbs-DiMarzio theory by Monte Carlo Simulation. The simulation employed the bond-fluctuation model on a simple cubic lattice. This model incorporates two kinds of interactions: the excluded volume interaction among all monomers of the melt and an internal energy of the chains, which favors large bonds and makes the chains stiffen with decreasing temperature. The stiffening of the chains leads to an increase of their volume requirements, which competes with the packing constraints at low temperatures. This competition strongly slows down the structural relaxation of the melt and induces the glassy behavior. The model therefore takes into account the main opposing forces which the Gibbs-DiMarzio theory makes responsible for the glass transition. For this model the entropy was calculated from the internal and the free energy (derived from the chemical potential and the single chain partition function) and compared with various theoretical predictions: the Gibbs-DiMarzio theory, a theory by Flory for semiflexible polymers and an extended theory by Wittmann considering Milchev's criticism on Flory's calculation. The latter extended theory provides the best description of the simulation data.


Could early tweet counts predict later citation counts? A gender study in Life Sciences and Biomedicine (2014-2016).
In this study, it was investigated whether early tweets counts could differentially benefit female and male (first, last) authors in terms of the later citation counts received. The data for this study comprised 47,961 articles in the research area of Life Sciences & Biomedicine from 2014-2016, retrieved from Web of Science's Medline. For each article, the number of received citations per year was downloaded from WOS, while the number of received tweets per year was obtained from PlumX. Using the hurdle regression model, I compared the number of received citations by female and male (first, last) authored papers and then I investigated whether early tweet counts could predict the later citation counts received by female and male (first, last) authored papers. In the regression models, I controlled for several important factors that were investigated in previous research in relation to citation counts, gender or Altmetrics. These included journal impact (SNIP), number of authors, open access, research funding, topic of an article, international collaboration, lay summary, F1000 Score and mega journal. The findings showed that the percentage of papers with male authors in first or last authorship positions was higher than that for female authors. However, female first and last-authored papers had a small but significant citation advantage of 4.7% and 5.5% compared to male-authored papers. The findings also showed that irrespective of whether the factors were included in regression models or not, early tweet counts had a weak positive and significant association with the later citations counts (3.3%) and the probability of a paper being cited (21.1%). Regarding gender, the findings showed that when all variables were controlled, female (first, last) authored papers had a small citation advantage of 3.7% and 4.2% in comparison to the male authored papers for the same number of tweets.


A School-Based Program for Overweight and Obese Adolescents: A Randomized Controlled Trial.
Given the dramatic increase in adolescent overweight and obesity, models are needed for implementing weight management treatment through readily accessible venues. We evaluated the acceptability and efficacy of a school-based intervention consisting of school nurse-delivered counseling and an afterschool exercise program in improving diet, activity, and body mass index (BMI) among overweight and obese adolescents. A pair-matched cluster-randomized controlled school-based trial was conducted in which 8 public high schools were randomized to either a 12-session school nurse-delivered cognitive-behavioral counseling intervention plus school-based after school exercise program, or 12-session nurse contact with weight management information (control). Overweight or obese adolescents (N = 126) completed anthropometric and behavioral assessments at baseline and 8-month follow-up. Main outcome measures included diet, activity, and BMI. Mixed effects regression models were conducted to examine differences at follow-up. At follow-up, students in intervention compared with control schools were not different in BMI, percent body fat, and waist circumference. Students reported eating breakfast (adjusted mean difference 0.81 days; 95% confidence interval [CI] 0.11-1.52) on more days/week; there were no differences in other behaviors targeted by the intervention. While a school-based intervention including counseling and access to an after-school exercise program is theoretically promising with public health potential, it was not effective in reducing BMI or key obesogenic behaviors. Our findings are important in highlighting that interventions targeted at the individual level are not likely to be sufficient in addressing the adolescent obesity epidemic without changes in social norms and the environment.


Nonlinear ultrasonic phased array imaging.
This Letter reports a technique for the imaging of acoustic nonlinearity. By contrasting the energy of the diffuse field produced through the focusing of an ultrasonic array by delayed parallel element transmission with that produced by postprocessing of sequential transmission data, acoustic nonlinearity local to the focal point is measured. Spatially isolated wave distortion is inferred without requiring interrogation of the wave at the inspection point, thereby allowing nonlinear imaging through depth.


Carrageenan-based hydrogels for the controlled delivery of PDGF-BB in bone tissue engineering applications.
One of the major drawbacks found in most bone tissue engineering approaches developed so far consists in the lack of strategies to promote vascularisation. Some studies have addressed different issues that may enhance vascularisation in tissue engineered constructs, most of them involving the use of growth factors (GFs) that are involved in the restitution of the vascularity in a damaged zone. The use of sustained delivery systems might also play an important role in the re-establishment of angiogenesis. In this study, kappa-carrageenan, a naturally occurring polymer, was used to develop hydrogel beads with the ability to incorporate GFs with the purpose of establishing an effective angiogenesis mechanism. Some processing parameters were studied and their influence on the final bead properties was evaluated. Platelet derived growth factor (PDGF-BB) was selected as the angiogenic factor to incorporate in the developed beads, and the results demonstrate the achievement of an efficient encapsulation and controlled release profile matching those usually required for the development of a fully functional vascular network. In general, the obtained results demonstrate the potential of these systems for bone tissue engineering applications.


Functional characterization of LePGT1, a membrane-bound prenyltransferase involved in the geranylation of p-hydroxybenzoic acid.
The AS-PT (aromatic substrate prenyltransferase) family plays a critical role in the biosynthesis of important quinone compounds such as ubiquinone and plastoquinone, although biochemical characterizations of AS-PTs have rarely been carried out because most members are membrane-bound enzymes with multiple transmembrane alpha-helices. PPTs [PHB (p-hydroxybenzoic acid) prenyltransferases] are a large subfamily of AS-PTs involved in ubiquinone and naphthoquinone biosynthesis. LePGT1 [Lithospermum erythrorhizon PHB geranyltransferase] is the regulatory enzyme for the biosynthesis of shikonin, a naphthoquinone pigment, and was utilized in the present study as a representative of membrane-type AS-PTs to clarify the function of this enzyme family at the molecular level. Site-directed mutagenesis of LePGT1 with a yeast expression system indicated three out of six conserved aspartate residues to be critical to the enzymatic activity. A detailed kinetic analysis of mutant enzymes revealed the amino acid residues responsible for substrate binding were also identified. Contrary to ubiquinone biosynthetic PPTs, such as UBIA in Escherichia coli which accepts many prenyl substrates of different chain lengths, LePGT1 can utilize only geranyl diphosphate as its prenyl substrate. Thus the substrate specificity was analysed using chimeric enzymes derived from LePGT1 and UBIA. In vitro and in vivo analyses of the chimeras suggested that the determinant region for this specificity was within 130 amino acids of the N-terminal. A 3D (three-dimensional) molecular model of the substrate-binding site consistent with these biochemical findings was generated.


Teenage prostitution--child abuse: a survey of the Ilorin situation.
In the light of civilization that has come to erode some of the sexual constraints in the traditional African society's opennes today toward sexuality and sexual expression, adolescents commence sexual activity at an increasing earlier age and the average marriage come much later. It is therefore not surprising that an increasing proportion of adolescents in Nigeria are engaging in teenage prostitution. This study is an attempt to conduct a general survey into the nature of teenage prostitution in Nigeria and to provide information on the prevalence, factors, trends and characteristics of teenage prostitution. A semi-structured questionnaire was administered to 150 known teenage prostitutes. Among other things the finding suggest that childhood sexual abuse perhaps leads to prostitution. Half of the sample used in the study had their first sexual experience early in life, and about the same number lived with their parents when they began prostituting. Implications of the findings were discussed and suggestions made for further research.


Reduced level of the spindle checkpoint protein BUB1B is associated with aneuploidy in colorectal cancers.
The majority of solid human malignancies demonstrate DNA aneuploidy as a consequence of chromosomal instability. We wanted to investigate whether Aurora A, Aurora B, BUB1B and Mad2 were associated with the development of aneuploidy in colorectal adenocarcinomas as suggested by several in vitro studies, and if their protein levels were related to alterations at the corresponding chromosomal loci. Expression levels of these spindle proteins were investigated by immunohistochemistry using tissue micro-arrays in a series of DNA aneuploid and diploid colorectal adenocarcinomas previously examined for genomic aberrations by comparative genomic hybridization. All proteins were overexpressed in malignant tissues compared to controls (P < 0.001 for all). BUB1B level was significantly reduced in aneuploid compared to diploid cancers (P = 0.001), whereas expression of the other proteins was not associated with DNA ploidy status. High levels of Aurora A (P = 0.049) and low levels of Aurora B (P = 0.031) were associated with poor prognosis, but no associations were revealed between protein expression and genomic aberration. A significant reduction of BUB1B level was detected in aneuploid compared to diploid colorectal cancers, consistent with earlier studies showing that loss of spindle checkpoint function may be involved in development of DNA aneuploidy. Our data also show that spindle proteins are overexpressed in colorectal cancers, and that expression of the Aurora kinases is associated with prognosis in colorectal cancer.


Sequencing of BRCA1/2-alterations using NGS-based technology: annotation as a challenge.
In this study, the molecular profile of different BRCA-associated tumor types was assessed with regard to the classification and annotation of detected BRCA1/2 variants. The aim was to establish guidelines in order to facilitate the interpretation of BRCA1/2 alterations in routine diagnostics. Annotation of detected variants was evaluated compared to background mutations found in normal tissue samples and manually reviewed according to distinct online databases. This retrospective study included 48 samples (45 tumors, three non-tumors), which were sequenced with the GeneReader (QIAGEN). Thereof ten samples were additionally analyzed with the Ion S5™ (Thermo Fisher) and 20 samples with the MiSeq™ (Illumina®) to compare the different NGS devices, as well as the sequencing results and their quality. The analysis showed that the individual NGS platforms detected different numbers of BRCA1/2 alterations in the respective tumor sample. In addition, the GeneReader revealed variability in the detection and classification of pathogenic alterations within the platform itself as well as in comparison with the other platforms or online databases. The study concluded that the Ion S5™ in combination with the Oncomine™ Comprehensive Assay v3 is most recommendable for current and prospective requirements of molecular analysis in routine diagnostics. In addition to the two BRCA1/2 genes, a broad number of other genes (BRCAness genes and genes involved in the repair pathway) is covered by the panel, which may open up new treatment options for patients depending on the respective eligibility criteria.


Evaluation of Health Surveillance in the Zona da Mata Mineira: from standards to practice.
Health Surveillance is of paramount importance when confronted with the current Brazilian epidemiological setting. This study aims to evaluate the level of Health Surveillance in the Zona da Mata Mineira region. This is an evaluative study, and data were collected in the first half of 2016. The research universe consisted of seven municipalities in the Zona da Mata Mineira complex. Municipalities were selected with reference to available operational epidemiological, environmental and sanitary surveillance. The subjects under analysis were the Surveillance coordinators (n = 21), Health Surveillance coordinators (n = 7) and Health secretaries (n = 7). We conducted interviews using a semi-structured and elaborated questionnaire based on the Donabedian triad: Structure, Process and Outcome. A score system was established in order to obtain the classification of performance, realm and sub-realms, in which a score was assigned for each criterion. We used incipient, intermediate or advanced cutoff points to classify the level of Health Surveillance of scores were below 5.99 points, between 6.0 and 7.99 points and between 8 and 10 points, respectively. The level of regional Health Surveillance performance level was classified as an intermediate.


Azathioprine or methotrexate in the treatment of patients with steroid-dependent or steroid-resistant ulcerative colitis: results of an open-label study on efficacy and tolerability in inducing and maintaining remission.
The role of azathioprine and methotrexate in inducing and maintaining remission in patients with ulcerative colitis is still controversial. To evaluate the efficacy and tolerability of these two drugs in a series of patients with steroid-dependent or steroid-resistant active ulcerative colitis. Forty-two patients were treated with a daily dose of azathioprine (2 mg/kg) and, if intolerant or not responding, with methotrexate (12.5 mg/week intramuscularly), and their efficacy was established by clinical, endoscopic and histological examinations at 6 months. Patients achieving clinical remission continued with treatment and were followed up. Of the 42 patients on azathioprine, 10 experienced early side-effects requiring withdrawal from treatment, 22 (69%) achieved complete remission, six (19%) achieved improvement and four (12%) obtained no substantial benefit. Methotrexate, administered to eight patients intolerant to and two patients resistant to azathioprine, induced complete remission in six patients (60%) and improvement in four (40%). During follow-up, a larger number of patients on azathioprine relapsed in comparison with patients on methotrexate [16/28 (57%) vs. 2/10 (20%), respectively; P < 0.05]. Only minor side-effects were observed on both treatments. Azathioprine is effective in patients with steroid-dependent or steroid-resistant ulcerative colitis. Methotrexate seems to be a good alternative in patients intolerant to or not responding to azathioprine.


Machine Learning Methods for Automated Quantification of Ventricular Dimensions.
Medaka (Oryzias latipes) and zebrafish (Danio rerio) contribute substantially to our understanding of the genetic and molecular etiology of human cardiovascular diseases. In this context, the quantification of important cardiac functional parameters is fundamental. We have developed a framework that segments the ventricle of a medaka hatchling from image sequences and subsequently quantifies ventricular dimensions.


The use of trigonometry in bloodstain analysis.
Bloodstain pattern analysis (BPA) is a valid forensic method which belongs to the category of biological methods using trigonomic models. Despite its development through the years, the method has been re-formulated a standard one and globally used, recognized in standard sheets. This method permits exact analysis of the dynamic and characteristic properties of bloodstains after impact on surfaces such as floors, walls, and ceilings, various exterior and interior items, and clothes. It is also possible to determine the characteristics of blood from the outer part of the body. According to the presence of blood and its quantity, it is also possible to use this method for verification of reconstruction of criminal acts, while being tested for its validity with primary conditions of preserved and readable traces of blood. Even though this method is not considered as the major one or the only one information obtained in this way can be used for judicial. In our research, we tested the validity of this method in an experimental model using firearms. We compared measurements of the lengths of trajectory of impact and the height of the blood sprayed upwards from a distance of 1, 3, 5 and 10 meters. The experiment was based on two main presumptions. The first was the knowledge of the value of the distance and the angle of impact of the bloodstain, the second, the ability of the blood to reach a certain height and the angle of its impact. In accordance with trigonometric formulas, both the impact of the selected distance of drops of blood, and the height of the selected bloodstain could be determined without any verification of the flight trajectory and the distance of bloodstains. The results indicate that the method for these requirements differs from the real values, while increasing the measurements with the indicated spot of the shot. Aside from the unique values which were calculated, other results of the impact of the distance of drops of bloodstain were considered of lower value, and the values concerning the height of the bloods stains after the shot higher than real values. In spite of the lack of total accuracy, we recommend using this method widely and more often for investigation and verification of individual acts in criminal and forensic practice.


Kindlin-2 regulates hemostasis by controlling endothelial cell-surface expression of ADP/AMP catabolic enzymes via a clathrin-dependent mechanism.
Kindlin-2, a widely distributed cytoskeletal protein, has been implicated in integrin activation, and its absence is embryonically lethal in mice. In the present study, we tested whether hemostasis might be perturbed in kindlin-2(+/-) mice. Bleeding time and carotid artery occlusion time were significantly prolonged in kindlin-2(+/-) mice. Whereas plasma concentrations/activities of key coagulation/fibrinolytic proteins and platelet counts and aggregation were similar in wild-type and kindlin-2(+/-) mice, kindlin-2(+/-) endothelial cells (ECs) showed enhanced inhibition of platelet aggregation induced by adenosine 5'-diphosphate (ADP) or low concentrations of other agonists. Cell-surface expression of 2 enzymes involved in ADP/adenosine 5'-monophosphate (AMP) degradation, adenosine triphosphate (ATP) diphosphohydrolase (CD39) and ecto-5'-nucleotidase (CD73) were increased twofold to threefold on kindlin-2(+/-) ECs, leading to enhanced ATP/ADP catabolism and production of adenosine, an inhibitor of platelet aggregation. Trafficking of CD39 and CD73 at the EC surface was altered in kindlin-2(+/-) mice. Mechanistically, this was attributed to direct interaction of kindlin-2 with clathrin heavy chain, thereby controlling endocytosis and recycling of CD39 and CD73. The interaction of kindlin-2 with clathrin was independent of its integrin binding site but still dependent on a site within its F3 subdomain. Thus, kindlin-2 regulates trafficking of EC surface enzymes that control platelet responses and hemostasis.


Impact of Preoperative Anemia on Outcomes After Posterior Spinal Fusion for Adolescent Idiopathic Scoliosis.
The aim of this study was to investigate the relationship of preoperative anemia and outcomes after posterior spinal fusion (PSF) for adolescent idiopathic scoliosis (AIS). A retrospective cohort study was performed using the American College of Surgeons National Surgical Quality Improvement Program-Pediatric database from 2016 to 2018. All pediatric patients (age 10-18 years) with AIS undergoing PSF were identified. Two cohorts were categorized into anemic and nonanemic cohorts based on age-based and sex-based criteria for anemia. Thirty-day outcomes and readmission rates were evaluated. A total of 4929 patients were identified, of whom 592 (12.0%) were found to have preoperative anemia. The anemic cohort had a greater prevalence of comorbidities and longer operative times. Compared with the nonanemic cohort, the anemic cohort experienced significantly higher rates of perioperative bleed/transfusion (nonanemic, 67.4% vs. anemic, 73.5%; P = 0.004) and required a greater total amount of blood transfused (nonanemic, 283.2 ± 265.5 mL vs. anemic, 386.7 ± 342.6 mL; P < 0.001). The anemic cohort experienced significantly longer hospital stays (nonanemic, 3.8 ± 2.2 days vs. anemic, 4.2 ± 3.9 days; P = 0.001), yet discharge disposition (P = 0.58), 30-day complication rates (P = 0.79) and unplanned reoperation rates (P = 0.90) were similar between cohorts. On multivariate analysis, anemia was found to be an independent predictor of perioperative bleed/transfusion (odds ratio, 1.36; 95% confidence interval, 1.12-1.66; P = 0.002) as well as a longer length of hospital stay (relative risk, 0.46; 95% confidence interval, 0.25-0.67; P < 0.001) but was not an independent predictor for postoperative complications (P = 0.85). Our study suggests that preoperative anemia may be a risk factor for a greater perioperative bleed/transfusion event and slightly longer length of stay; however, it was not associated with greater 30-day complication and readmission rates in patients with AIS undergoing PSF.


Introduction of a proline residue into position 31 of the loop of the dimeric 4-alpha-helical protein ROP causes a drastic destabilization.
The exchange of an alanine with a proline residue in position 31 of the loop region of the dimeric 4-alpha-helical-bundle protein ROP causes a reduction in the alpha-helix content of 7% and a reduction in stability of about 40% compared to the wild type parameters. The Gibbs energy of unfolding by denaturants extrapolated linearly to zero denaturant concentration, delta G0D (buffer, 25 degrees C), has been determined to be 43 kJ (mol dimer)-1. The corresponding ROPwt value is 72 kJ (mol dimer)-1 (Steif et al., 1993). The extrapolated delta G0D values obtained from urea and GdmHCI un- and refolding studies are identical within error limits. Deconvolution of the stability values into enthalpy and entropy terms resulted in the following parameters. At T1/2 = 43 degrees C (Cprotein = 0.05 mg.ml-1) the ROP A31P mutant is characterized by delta Hv.H.0 = 272 kJ (mol dimer)-1, delta Cp = 7.2 kJ (mol dimer)-1 K-1, delta S0 = 762 J (mol dimer)-1 K-1. These parameters are only approximately 50% as large as the corresponding values of ROPwt. We assume that the significant reduction in stability reflects the absence of at least one hydrogen bond as well as deformation of the protein structure. This interpretation is supported by the reduction in the change in heat capacity observed for the A31P mutant relative to ROPwt, by the increased aggregation tendency of the mutant and by the reduced specific CD absorption at 222 nm. All results support the view that in the case of ROP protein the loop region plays a significant role in the maintenance of native structure and conformational stability.


Granulosa cell tumor of ovary: A clinicopathological study of four cases with brief review of literature.
Adult granulosa cell tumor (GCT) is a rare ovarian malignancy having good prognosis in comparison with other epithelial tumors. The study aims to collect data of all granulosa cell tumors diagnosed in ESIC Medical College & PGIMSR, Rajajinagar, Bangalore over the last 3 years and to describe the patient profile, ultrasonographic and various histopathological features. A total of 4 granulosa cell tumors were diagnosed in ESIC Medical College & PGIMSR, Rajajinagar, Bangalore during the period from June 2010 to June 2013. The patient's age, clinical manifestations, radiological and histopathological findings were evaluated. All 4 patients were diagnosed as adult granulosa cell tumor, three of four cases were in premenopausal age group and one case was in perimenopausal age. The clinical manifestations were menorrhagia and abdominal pain. Ultrasonographically, 2 cases of granulosa cell tumors were both solid and cystic and one case each was either solid or cystic. Histologically, variety of patterns like diffuse, trabecular, cords, spindle and clear cells were noted. Both Call-Exner bodies and nuclear grooves were observed in all cases. All four cases showed simple hyperplasia without atypia endometrial findings. Follow up on all patients revealed no evidence of recurrence. Granulosa cell tumor of the ovary is a rare ovarian entity. The important prognostic factor is staging of the tumor. Staging and histopathology helps in prediction of survival. Also diligent endometrial pathology has to be sorted to rule out endometrial carcinoma.


Does Early Surgical Decompression in Cauda Equina Syndrome Improve Bladder Outcome?: Comments on a Recent Study by Srikandarajah et al.
This study by Srikandarajah et al is a retrospective cohort study of 200 CES patients. Although parts of the study appear to be well done we identified serious problems that impacted their conclusions. There is strong consensus that patients with incomplete cauda equina syndrome (CESI) should be operated upon expeditiously to prevent progression to CES with urinary retention (CESR). There is controversy concerning optimal timing of surgery in patients who have entered CESR. Some studies conclude urgent surgery for CESR does nothing to improve chance of recovery; others conclude surgery should be done within a 48 or 24-hours window after onset of bladder paralysis. Srikandarajah et al concluded that in CESI patients, decompressive surgery within 24 hours of onset of autonomic symptoms reduces bladder dysfunction at follow-up. In CESR patients, "no statistically significant difference in outcome was observed" regarding timing of surgery. We analyzed the methods used by Srikandarajah et al to collect and analyze their data. The primary problem with their study is that it does not consider deterioration to CESR that occurs in CESI patients over time. We also found serious problems with the methods, implementation (including numeric errors), and interpretation of the statistical analysis. The authors' conclusion that in CESR patients, "no statistically significant difference in outcome was observed" regarding timing of surgery is not justified as a representation of their findings, because absence of statistical significance does not mean acceptance of the null hypothesis of "no effect". Their numeric results do show elevated risk of delay. We do not want to detract from the basic importance of this study, because it emphasizes the importance of operating expeditiously on patients with CESI. However, the authors' conclusions are compromised by the methodological problems. N/A.


A novel, resorbable suture anchor: pullout strength from the human cadaver greater tuberosity.
The pullout strength of a collagen bone anchor that creates interference fixation as the result of radial swelling on hydration was compared with a Mitek rotator cuff anchor after insertion into the greater tuberosity of human cadaver humeri. Bones were fully hydrated at 37 degrees C. Stiffness, peak load, and the mode of failure were recorded. Real and apparent bone densities were measured. Peak load for the collagen anchor at 15 minutes (121.0N +/- 81.3N) was greater than at 2 minutes (60.5N +/- 38.5N) after insertion (P <.05). At between 5 and 60 minutes after insertion, peak loads for the Mitek and the collagen anchors did not differ. After 30 minutes from insertion, the mode of failure of the collagen anchor changed from pullout with minor body damage to pullout with major body damage. Peak load at pullout correlated with bone density for the Mitek (P <.05, r = 0.516) but for the collagen bone anchor appeared unaffected by bone density.


Striving to be heard and recognized: nurse solutions for improvement in the outpatient hemodialysis work environment.
One group of nurses not frequently studied but who have felt the burden of shortages of registered nurses (RNs) is that of nephrology nurses. Nurses' valuable insights must be solicited for the accurate assessment of current work environment issues and for their specific solutions. The purposes of this study were to explore nurse satisfaction and dissatisfaction with the work environment in outpatient hemodialysis facilities, to solicit nurses' insights on potential solutions, and to develop national strategies for improvement. Focus group qualitative methodology was utilized with focus groups and individual interviews, including a total of 101 RN participants from 45 outpatient hemodialysis facilities. The overall category of this study was Striving to be Heard and Recognized with the following four supportive categories: Maintaining Competency, Resolving Staffing and Resources, Strengthening Organizational Leadership, and Rebuilding Relationships. Nephrology nurse participants have identified strategies within each category for the improvement of their work environment. The voices of nephrology nurses must be heard and acted upon in a timely manner to improve workplace environment.


Comparison of clinical outcomes of different erythropoietin usage strategies.
There is no comprehensive study that compares the different usage strategies of recombinant human erythropoietin (rHuEPO) in platinum-induced anemia. In order to clarify this issue, we conducted a prospective clinical study. Seventy-seven patients were studied in three main groups. Group 1 (n = 17) consisted of cancer patients without anemia. These patients received rHuEPO starting from the first chemotherapy cycle. Group 2 (n = 26) consisted of patients whose hemoglobin (Hb) values decreased by at least 1 g/dL after the first cycle of chemotherapy. Group 3 (n = 34) consisted of patients whose Hb values dropped below 10.5 g/dL after the second chemotherapy cycle. Groups 2 and 3 were each divided into two subgroups. In groups 1, 2A and 3A rHuEPO (5000 U/day subcutaneously three times a week) treatment was continued until three weeks after the completion of chemotherapy. In groups 2B and 3B, rHuEPO was given for 12 weeks only. There were no prominent differences between the Hb values of these groups throughout the chemotherapy cycles. Transfusion rates and the number of patients who became anemic were also not different between groups. No rHuEPO usage strategies are superior to others in terms of Hb levels and transfusion requirements. The decision as to when rHuEPO is to be added to platinum-containing therapy should be tailored to the health conditions of individual patients.


The predictivity of serum biochemical markers in acute biliary pancreatitis.
Background and Aim. There are no accurate methods of differentiating acute biliary pancreatitis. Obstructions of biliary ducts, idiopathic pancreatitis may be related with biliary origin which needs identification for acute treatment. We searched for the predictivity of biochemical markers in early acute biliary pancreatitis. Patients and Methods. Serum levels of AST (Aspartate Transaminase),ALT (Alanine Transaminase), ALP (Alkaline Phosphatase), GGT (Gamma Glutamyl Transferase), total bilirubin, direct bilirubin, LDH (Lactate Dehydrogenase), amylase, lipase, CRP (C-Reactive Protein) and WBC (White Blood Cell) were measured in 157 patients with acute pancreatitis. Biliary and nonbiliary pancreatitis were differentiated by Magnetic Resonance Cholangiopancreatography (MRCP), Endoscopic Retrograde Cholangiopancreatography (ERCP), Intraoperative Cholangiopancreatography (IOC). Cut-off points of admission biochemical markers with sensitivity, specifity, positive predictive value and negative predictive value were determined after identification of significant variables. Receiver Operator Curves were plotted for each biochemical marker. Results. Serum Alkaline Phosphatase, total bilirubin, direct bilirubin, amylase and lipase levels were significantly higher in biliary pancreatitis with a positive predictive value of 80.8%, 83.9%, 81.6%, 78.8%, 79.7%. Conclusion. Increased Alkaline Phosphatase,total bilirubin, direct bilirubin, amylase and lipase levels may be used in prediction of biliary pancreatitis.


Left atrial appendage closure: A therapy uniquely suited for specific populations of patients with atrial fibrillation.
Atrial fibrillation (AF) is the most common clinically relevant arrhythmia and confers a fivefold increased risk for stroke. Cardioembolic stroke secondary to AF is a devastating event, but is largely preventable with appropriate oral anticoagulation (OAC). The PROTECT and PREVAIL trials demonstrated that the WATCHMAN left atrial appendage closure (LAAC) device in combination with short-term warfarin therapy is noninferior to long-term warfarin with respect to a composite endpoint of stroke, cardiovascular death, and systemic embolism. Importantly, the WATCHMAN confers a significant reduction in life-threatening bleeding compared to OAC. Although direct-acting oral anticoagulant (DOAC) are superior to warfarin in eligible patients, several important AF populations exist in whom left atrial appendage (LAA) closure may be preferable to DOAC. Populations warranting strong consideration of LAAC include patients with contraindications to DOAC, end-stage renal disease, prior intracranial hemorrhage, recurrent gastrointestinal bleeding, and patients undergoing transcatheter aortic valve replacement or left atrial electrical isolation. Device-related thrombosis is an important complication of LAAC, and DOAC may be preferential to warfarin for prevention and treatment of this complication remains unexplored. Prospective clinical trials comparing DOAC to LAAC in these unique populations are either ongoing or needed.


Presynaptic release probability and readily releasable pool size are regulated by two independent mechanisms during posttetanic potentiation at the calyx of Held synapse.
At the immature calyx of Held, the fast decay phase of a Ca(2+) transient induced by tetanic stimulation (TS) was followed by a period of elevated [Ca(2+)](i) for tens of seconds, referred to as posttetanic residual calcium (Ca(res)). We investigated the source of Ca(res) and its contribution to posttetanic potentiation (PTP). After TS (100 Hz for 4 s), posttetanic Ca(res) at the calyx of Held was largely abolished by tetraphenylphosphonium (TPP(+)) or Ru360, which inhibit mitochondrial Na(+)-dependent Ca(2+) efflux and Ca(2+) uniporter, respectively. Whereas the control PTP lasted longer than Ca(res), inhibition of Ca(res) by TPP(+) resulted in preferential suppression of the early phase of PTP, the decay time course of which well matched with that of Ca(res). TS induced significant increases in release probability (P(r)) and the size of the readily releasable pool (RRP), which were estimated from plots of cumulative EPSC amplitudes. TPP(+) or Ru360 suppressed the posttetanic increase in P(r), whereas it had little effect on the increase in RRP size. Moreover, the posttetanic increase in P(r), but not in RRP size, showed a linear correlation with the amount of Ca(res). In contrast, myosin light chain kinase (MLCK) inhibitors and blebbistatin reduced the posttetanic increase in RRP size with no effect on the increase in P(r). Application of TPP(+) in the presence of MLCK inhibitor peptide caused further suppression of PTP. These findings suggest that Ca(res) released from mitochondria and activation of MLCK are primarily responsible for the increase in P(r) and that in the RRP size, respectively.


Rolling circle amplification-driven encoding of different fluorescent molecules for simultaneous detection of multiple DNA repair enzymes at the single-molecule level.
DNA repair enzymes (e.g., DNA glycosylases) play a critical role in the repair of DNA lesions, and their aberrant levels are associated with various diseases. Herein, we develop a sensitive method for simultaneous detection of multiple DNA repair enzymes based on the integration of single-molecule detection with rolling circle amplification (RCA)-driven encoding of different fluorescent molecules. We use human alkyladenine DNA glycosylase (hAAG) and uracil DNA glycosylase (UDG) as the target analytes. We design a bifunctional double-stranded DNA (dsDNA) substrate with a hypoxanthine base (I) in one strand for hAAG recognition and an uracil (U) base in the other strand for UDG recognition, whose cleavage by APE1 generates two corresponding primers. The resultant two primers can hybridize with their respective circular templates to initiate RCA, resulting in the incorporation of multiple Cy3-dCTP and Cy5-dGTP nucleotides into the amplified products. After magnetic separation and exonuclease cleavage, the Cy3 and Cy5 fluorescent molecules in the amplified products are released into the solution and subsequently quantified by total internal reflection fluorescence (TIRF)-based single-molecule detection, with Cy3 indicating the presence of hAAG and Cy5 indicating the presence of UDG. This strategy greatly increases the number of fluorescent molecules per concatemer through the introduction of RCA-driven encoding of different fluorescent molecules, without the requirement of any specially labeled detection probes for simultaneous detection. Due to the high amplification efficiency of RCA and the high signal-to-ratio of single-molecule detection, this method can achieve a detection limit of 6.10 × 10-9 U mL-1 for hAAG and 1.54 × 10-9 U mL-1 for UDG. It can be further applied for simultaneous detection of multiple DNA glycosylases in cancer cells at the single-cell level and the screening of DNA glycosylase inhibitors, holding great potential in early clinical diagnosis and drug discovery.


The application of subliminal priming in lie detection: scenario for identification of members of a terrorist ring.
We studied a lie detection protocol immune to countermeasures. The 4 stimulus conditions were (1 and 2) supraliminal acquaintance name primed by subliminal acquaintance name (A-A) versus subliminal nonacquaintance name (N-A) and (3 and 4) supraliminal nonacquaintance name primed by subliminal acquaintance name (A-N) versus subliminal nonacquaintance name (N-N). In Experiment 1 and replication, principal components analysis-derived event-related potential components revealed significant differences between dishonestly answered supraliminal acquaintance conditions with differing primes (A-A vs. N-A). In Experiment 2 subjects were required to lie in A-N and N-N conditions, in contrast to Experiment 1, in which subjects lied in A-A and N-A conditions. No significant effects were found. In Experiment 3, the lying task was removed and no significant differences were found. We conclude that subliminal primes modulate ERPs in conditions with supraliminal acquaintance name when the task involves lying.


Transferrin localizes in Bunina bodies in amyotrophic lateral sclerosis.
Transferrin, an iron-binding protein, plays an important role in the transport and delivery of circulating ferric iron to the tissues. Amyotrophic lateral sclerosis (ALS) is characterized by the presence of Bunina bodies, skein-like inclusions, Lewy body-like inclusions/round inclusions, and basophilic inclusions in the remaining anterior horn cells in the spinal cord. We examined transverse paraffin sections of lumbar spinal cords from 12 ALS cases including two ALS with dementia and two ALS with basophilic inclusions, using antibodies to human transferrin. The results demonstrated that transferrin localized in Bunina bodies and some of the basophilic inclusions. In contrast, skein-like inclusions and Lewy body-like inclusions or round inclusions did not show obviously detectable transferrin immunoreactivities. Our findings suggest that although the mechanisms underlying transferrin accumulation in Bunina bodies and basophilic inclusions are unknown, transferrin could be involved in forming these inclusions. Furthermore, following cystatin C, transferrin is the second protein that localizes in the Bunina bodies.


Cutting edge: retrobulbar inflammation, adipogenesis, and acute orbital congestion in a preclinical female mouse model of Graves' orbitopathy induced by thyrotropin receptor plasmid-in vivo electroporation.
Graves' orbitopathy (GO) is a complication in Graves' disease (GD) but mechanistic insights into pathogenesis remain unresolved, hampered by lack of animal model. The TSH receptor (TSHR) and perhaps IGF-1 receptor (IGF-1R) are considered relevant antigens. We show that genetic immunization of human TSHR (hTSHR) A-subunit plasmid leads to extensive remodeling of orbital tissue, recapitulating GO. Female BALB/c mice immunized with hTSHR A-subunit or control plasmids by in vivo muscle electroporation were evaluated for orbital remodeling by histopathology and magnetic resonance imaging (MRI). Antibodies to TSHR and IGF-1R were present in animals challenged with hTSHR A-subunit plasmid, with predominantly TSH blocking antibodies and were profoundly hypothyroid. Orbital pathology was characterized by interstitial inflammation of extraocular muscles with CD3+ T cells, F4/80+ macrophages, and mast cells, accompanied by glycosaminoglycan deposition with resultant separation of individual muscle fibers. Some animals showed heterogeneity in orbital pathology with 1) large infiltrate surrounding the optic nerve or 2) extensive adipogenesis with expansion of retrobulbar adipose tissue. A striking finding that underpins the new model were the in vivo MRI scans of mouse orbital region that provided clear and quantifiable evidence of orbital muscle hypertrophy with protrusion (proptosis) of the eye. Additionally, eyelid manifestations of chemosis, including dilated and congested orbital blood vessels, were visually apparent. Immunization with control plasmids failed to show any orbital pathology. Overall, these findings support TSHR as the pathogenic antigen in GO. Development of a new preclinical model will facilitate molecular investigations on GO and evaluation of new therapeutic interventions.


Malatya and Other Criteria for Liver Transplantation in Hepatocellular Carcinoma.
Liver transplantation is a curative treatment option for hepatocellular carcinoma. In this review, we aimed to review liver transplantation criteria for hepatocellular carcinoma and patient survivals. We reviewed literature in terms of liver transplant criteria for hepatocellular carcinoma. Patient eligibility criteria, post-transplant survivals, tumor recurrence and expansion of Milan criteria rates were analyzed. The Milan criteria, after being published in 1996, have become for deceased donor liver transplantation in hepatocellular carcinoma worldwide. Later, many transplant centers published their own liver transplant criteria. Most of the criteria consisted of morphological tumor characteristics based on tumor size and number. The newest published one is Malatya criteria. The 5-year overall survival according the all of the criteria is greater than 50%. There were just one paper which compare criteria according to survival and Malatya criteria were the best amongst extended criteria with 5-year OS 79.7% in that study. It is clear that morphological criteria consisting only of tumor size and number are insufficient in patient selection for liver transplantation and should thus be combined with biological, inflammatory, radiological, pathological and genetic markers that predict the biological behavior of the tumor. Efforts to find the best criteria are still ongoing and 5-year overall survival should be greater than 60%.


Effects of protein concentration and source on nutrient digestibility by mature steers limit-fed high-concentrate diets.
Five ruminally fistulated 3-yr-old mature Holstein steers (average BW 691+/-23 kg) were used in a 5 x 5 Latin square experiment with a 2 x 2 + 1 fact orial arrangement of treatments. Effects of protein concentration and protein source on nutrient digestibility, excretion of DM and fecal N, ruminal fluid volume and dilution rate, ruminal characteristics, and in situ DM disappearance of whole shelled corn, ground corn, and orchardgrass hay were measured in steers limit-fed high-concentrate diets at 1.5% of BW. A negative control basal diet (NC; 9% CP) was supplemented to achieve either 11 or 14% CP; supplemental CP was either from soybean meal (11 and 14% SBM) or a 50:50 ratio of CP from urea and soybean meal (11 and 14% U). Dry matter and OM digestibilities were 5% greater (P < .07) for steers fed the SBM diets than for those fed the U diets. Starch digestibility did not differ (P > .10) among steers fed any of the diets. Nitrogen source did not affect (P > .10) apparent N digestibility or fecal N excretion; however, steers fed the NC diet had the lowest (P < .10) apparent N digestibility compared with those fed all other diets. Ruminal fluid volume was lower (P < .06) when steers were fed the NC diet compared with all other diets; there were no differences (P > .74) among diets for ruminal fluid dilution rate. In general, ruminal ammonia N and VFA molar proportions were not affected by protein source or concentration. Although CP concentration affected (P < .06) in situ DM disappearance of ground corn, CP concentration did not (P > .48) affect total tract digestion of DM or OM. This indicates that CP concentration may have affected site of digestion, but not extent of digestion. When mature ruminants were limit-fed a corn-based diet to meet primarily a maintenance function, protein source and concentration had little effect on measures of nutrient digestion.


[Value of cardiac pacing in hypertrophic obstructive cardiomyopathy refractory to medical treatment].
Recent studies have shown that dual-chamber cardiac pacing could improve patients with hypertrophic obstructive cardiomyopathy resistant to medical treatment. After a brief historical review, the authors report the present status of this original therapeutic option. The results of different studies are in agreement and show that the functional improvement is associated with a large reduction of the intraventricular pressure gradient with permanent DDD pacing. Although the exact mechanism of the improvement remains unknown, all groups underline the importance of complete and permanent ventricular capture. This requires precise regulation of the atrioventricular delay which in most cases must be programmed at a value inferior to 90 ms, which compromises ventricular filling which is already abnormal under basal conditions. The inefficacy of pacing in some cases is explained by the necessity of selecting a very short atrioventricular delay which alters diastolic function and finally masks the benefits of ventricular capture. In this particular situation, the authors propose improving atrioventricular synchrony by pharmacological prolongation of the PR interval, or, when unsuccessful, by ablation of the atrioventricular junction. This attitude was adopted in 5 cases when pacing with an AVD programmed between 20 and 70 ms, provided only a partial haemodynamic result. Improved atrioventricular synchrony allowed selection of an AVD of 100 to 150 ms, so reducing the gradient by more than 70%, compared with 30% under the initial programming parameters. The authors conclude that a positive and sustained benefit of cardiac pacing can only be obtained by an individualised regulation of pacing parameters to optimize atrioventricular synchrony which seems to be the key element of success.


Incorporation of β-actin loading control into zymography.
Gelatin zymography and immunoblot are widely used gel electrophoresis techniques to study matrix metalloproteinases-2 and -9. Each method has its advantages and disadvantages. Zymography is exquisitely sensitive but offers no loading control to ensure equal sample loading. Immunoblot is a 100-1000-fold less sensitive, but allows for the probing of a sample loading control such as β-actin to ensure accurate protein loading. In this report, we describe two simple protocols that combine gelatin zymography to study MMP-2 and -9 levels with an in-gel β-actin immunoblot loading control, thus combining sensitivity and accuracy in a single assay. The protocols incorporate the loading of molecular weight markers to demarcate MMP-2/-9 from the β-actin. The first protocol utilizes the overlay of a 10% zymography gel over a 5% Tris-Glycine separating gel from which the β-actin is transferred. The second protocol involves the direct transfer of the β-actin from a single 10% zymography gel.


A retrospective analysis of incidence and management of palatal fistula.
Cleft palate repair may be compromised by a number of complications, most commonly the development of a fistula. Fistulas may cause hypernasal speech, articulation problems and food or liquid regurgitation from the nose. The study determines the incidence and management of cleft palatal fistulas in a series of primary cleft palate repair surgeries. It is a retrospective analysis of total 185 palatal fistula cases operated at our hospital from the year 2004 to 2016. Of 185 palatal fistulas, 132 cases had been operated at our institute for primary palatoplasty, and the rest 53 were the outside-operated cases. The patients with bilateral as well as unilateral cleft lip and palate were included. Isolated cleft palate patients were also included in the study. Palatal fistulas were subdivided into three types depending on their size. Anterior palatal fistulas were mostly treated by using tongue flap (65.57%), followed by local flaps (34.43%). Middle and posterior palatal fistulas were mostly treated by von Langenbeck Palatoplasty. One patient (>5 mm fistula) was treated using free radial forearm flap. Anterior palatal fistulas (65.57%) were most commonly reported, followed by middle (24.86%) and posterior (9.18%). Most commonly, the size of the fistulas ranged from 2 mm to 5 mm. The complication rate was reported to be 3.75% in case of tongue flap and 11.9% complications were reported in case of local flaps. Tongue flap remains the flap of choice for managing very difficult and challenging anterior palatal fistulas compared to local flaps.


Hemoglobin trigger and approach to red blood cell transfusions during veno-venous extracorporeal membrane oxygenation: the international TRAIN-ECMO survey.
Optimal red blood cell transfusion practice during veno-venous extracorporeal membrane oxygenation (VV ECMO) is still under debate. This survey aimed to assess the Hb trigger (also comparing with other critically ill patients) and major physiologic determinants considered for transfusions during veno-venous extracorporeal membrane oxygenation. Voluntary Web-based survey, endorsed by the European Society of Intensive Care Medicine, conducted among VV ECMO pratictioners worldwide. A total of 447 respondents worldwide answered the questionnaire: 277 (61.9%) from Europe, 99 (22.1%) from North America, 36 (8.2%) from Asia and Oceania, and 35 (7.8%) from Central and South America. Among the respondents, 59.2% managed less than 12 venous extracorporeal membrane oxygenation runs/year, 19.4% between 12 and 24 runs/year, and 21.4% more than 24 runs/year. Of the respondents, 54.4% do not use a predefined Hb trigger in veno-venous extracorporeal membrane oxygenation, and, while the rate of adoption of a defined trigger varied worldwide, the effective value of Hb did not differ significantly among macro-regions. In patients on veno-venous extracorporeal membrane oxygenation, the Hb trigger to initiate red blood cell transfusion, was higher than in other critically ill patients: 9.1 ± 1.8 g/dL versus 8.3 ± 1.7 g/dL, p < 0.01. The Hb trigger was lower in centers with more than 24 venous extracorporeal membrane oxygenation runs/year (8.4 mg/dL (95% CI: 7.7-8.9)); (8.9 mg/dL (95% CI: 8.2-9.7)) in centers with between 12 and 24 venous extracorporeal membrane oxygenation runs/year; and (9.6 mg/dL (95% CI: 9.1-10.0)) in centers with fewer than 12 venous extracorporeal membrane oxygenation runs/year (p < 0.01). Several and variable adjunctive parameters are considered in cases of uncertainty for transfusion: the principal are hemodynamic status, SvO2, lactates, and fluid balance. Although the use of a predefined Hb trigger is still under-adopted among centers with low or median extracorporeal membrane oxygenation case volume, the majority of respondents use a higher Hb trigger for veno-venous extracorporeal membrane oxygenation patients compared with other critically ill patients. Higher volume centers tolerate lower Hb levels.


Complex regional pain syndrome type 1. Some treatments assessed versus placebo, limited efficacy.
(1) Complex regional pain syndrome type 1 generally occurs after trauma and usually affects a limb; (2) How is complex regional pain syndrome type 1 diagnosed? What is its natural course? How safe and effective are available treatments? To answer these questions, we reviewed the literature using the standard Prescrire methodology; (3) Diagnosis is mainly based on clinical features, including pain disproportionate to the initial trauma, associated with cutaneous vasomotor, trophic and sweating disorders; (4) Some clinical signs call for additional examinations to help rule out another vascular, neurological, infectious or rheumatic disorder. Radiological evidence of bone demineralisation supports the diagnosis, but radiography, magnetic resonance imaging (MRI) and scintigraphy generally contribute little to the diagnosis of complex regional pain syndrome; (5) Some patients recover spontaneously after a few weeks, while others develop chronic pain or even severe disability after a period of years; (6) The results of small placebo-controlled trials suggest that corticosteroids are effective during the initial phase of this syndrome; (7) A very high oral dose of alendronic acid provided sustained pain relief in a randomised trial. Other studies suggest that bisphosphonates have some impact. The adverse effects of alendronic acid given at such high doses are poorly known; (8) Calcitonin, antiepileptics, antidepressants and opiates have no proven efficacy; (9) Transcutaneous neurostimulation is rapidly effective and safe, but its efficacy also diminishes rapidly. Therefore, the sessions have to take place at increasingly shorter intervals. (10) Spinal neurostimulation with implanted electrodes has been assessed in a comparative trial in 54 patients. Some efficacy was observed, but one-third of patients had complications requiring further surgery; (11) Various substances have been given intravenously with the goal of achieving regional anaesthesia, but none was found to have any tangible efficacy; (12) Sympathectomy has a negative risk-benefit balance; (13) Non-drug approaches such as physiotherapy, relaxation and biofeedback seem to help some patients, with little risk of adverse effects. These methods have not been comparatively evaluated; (14) In practice, there is no truly effective treatment for complex regional pain syndrome. The few beneficial treatments have not been directly compared with one another. The advantages and disadvantages of the various treatment options must be discussed with each patient.


Impact of arch reobstruction and early hypertension on late hypertension after coarctation repair.
Late hypertension after coarctation repair is associated with high mortality, but its risk factors remain unclear. This study aims to determine early and late postoperative risk factors for late hypertension after coarctation repair. A cross-sectional study including transthoracic echocardiogram and 24-h blood pressure (BP) monitoring was performed in 144 patients aged ≥10 years with previous coarctation repair. Median age at repair was 39 days (interquartile range 0-3 years). Early postoperative hypertension was evaluated by calculating the mean of BP measurements taken on the same day before hospital discharge or the need for antihypertensives prior to discharge. Multivariable analyses were performed to adjust for gender, surgical age, and follow-up age. After a mean follow-up period of 22 ± 7 years, 59% (84/142) of patients were hypertensive: 58% (82/142) on 24-h BP monitoring and 1% (2/142) on antihypertensives. Early postoperative hypertension was present in 58% (73/126): 39% (49/126) on BP measurements and 19% (24/126) on antihypertensives. Early and late arch reobstruction (transthoracic echocardiogram peak gradient ≥25 mmHg) was present in 37% (23/62) and 23% (33/144), respectively. On multivariable logistic analysis, early postoperative hypertension and maximum descending arch velocity on echocardiography were associated with late hypertension on 24-h BP monitoring (odds ratio 2.21, 95% confidence interval 1.05-4.66, P = 0.04; and odds ratio 2.28, 95% confidence interval 1.08-4.81, P = 0.03; respectively). There is a high prevalence of late hypertension after coarctation repair. Arch reobstruction may be a major determinant of late hypertension. Early postoperative hypertension may identify very early in life those at risk of developing late hypertension.


Cardiac safety of afatinib: a review of data from clinical trials.
Afatinib is an oral irreversible ErbB family blocker that targets epidermal growth factor receptor (EGFR/ErbB1), human epidermal growth factor receptor 2 (HER2/ErbB2), and HER4 (ErbB4) and is approved for the first-line treatment of advanced non-small cell lung cancer (NSCLC) with certain sensitizing EGFR mutations. As anti-HER2 therapies have been associated with cardiac dysfunction, we report cardiac safety data for afatinib. Cardiac data were analyzed from phase III trials of afatinib 40 mg in treatment-naive patients with EGFR mutation-positive NSCLC (LUX-Lung 3 [LL3]; n = 229 afatinib, n = 111 chemotherapy) and afatinib 50 mg in EGFR tyrosine kinase inhibitor-pretreated NSCLC patients (LUX-Lung 1 [LL1]; n = 390 afatinib, n = 195 placebo). Additional pooled data from 49 trials (n = 3865 afatinib-treated patients) is reported. Cardiac failure adverse events (CF-AEs), including symptomatic cardiac failure and depressed left ventricular ejection fraction (LVEF), were analyzed. Time at risk-adjusted CF-AE rates (events/100 patient-years) were similar for afatinib versus placebo in LL1 (2.40 vs 2.23) and versus chemotherapy in LL3 (2.28 vs 2.92); the pooled afatinib CF-AE rate (2.88) was consistent with that for both trials. The frequency of clinically significant LVEF reductions was higher for chemotherapy in LL3 (2/15 [13.3 %], afatinib 13/208 [6.3 %]; p = 0.267) and similar to placebo in LL1 (5/122 [4.1 %], afatinib 14/304 [4.6 %]; p = 1.000). Afatinib was not associated with cardiac failure or LVEF reductions in the afatinib clinical trial program.


The binding of cupric ions to bovine pancreatic ribonuclease studies with diligand metal-ion buffers.
A procedure has been developed for the use of metal-ion buffers that depends on the formation of 2:1 complexes between suitable chelators and metal ions. beta-Alanine has been used as the chelator for Cu(2+) ions in a study of Cu(2+) binding by bovine pancreatic ribonuclease by the equilibrium-dialysis technique at pH7.0, 6.1 and 5.2. The results indicated the presence of two avid binding sites, the more avid group being implicated in the inhibition of enzyme activity by Cu(2+) ions. The binding constants of the more avid site were 2.97x10(7), 7.97x10(5) and 1.25x10(4) at pH7.0, 6.1 and 5.2 respectively, and the binding constants of the less avid site were 5.27x10(6) and 1.71x10(5) at pH7.0 and 6.1 respectively. The data show that the Cu(2+) is chelated to the protein through at least two ligand groups on the ribonuclease molecule.


24 h Accelerometry: impact of sleep-screening methods on estimates of sedentary behaviour and physical activity while awake.
Although accelerometers can assess sleep and activity over 24 h, sleep data must be removed before physical activity and sedentary time can be examined appropriately. We compared the effect of 6 different sleep-scoring rules on physical activity and sedentary time. Activity and sleep were obtained by accelerometry (ActiGraph GT3X) over 7 days in 291 children (51.3% overweight or obese) aged 4-8.9 years. Three methods removed sleep using individualised time filters and two methods applied standard time filters to remove sleep each day (9 pm-6 am, 12 am-6 am). The final method did not remove sleep but simply defined non-wear as at least 60 min of consecutive zeros over the 24-h period. Different methods of removing sleep from 24-h data markedly affect estimates of sedentary time, yielding values ranging from 556 to 1145 min/day. Estimates of non-wear time (33-193 min), wear time (736-1337 min) and counts per minute (384-658) also showed considerable variation. By contrast, estimates of moderate-to-vigorous activity (MVPA) were similar, varying by less than 1 min/day. Different scoring methods to remove sleep from 24-h accelerometry data do not affect measures of MVPA, whereas estimates of counts per minute and sedentary time depend considerably on which technique is used.