Cardiac glycosides induce cell death in human cells by inhibiting general protein synthesis.
Cardiac glycosides are Na(+)/K(+)-pump inhibitors widely used to treat heart failure. They are also highly cytotoxic, and studies have suggested specific anti-tumor activity leading to current clinical trials in cancer patients. However, a definitive demonstration of this putative anti-cancer activity and the underlying molecular mechanism has remained elusive. Using an unbiased transcriptomics approach, we found that cardiac glycosides inhibit general protein synthesis. Protein synthesis inhibition and cytotoxicity were not specific for cancer cells as they were observed in both primary and cancer cell lines. These effects were dependent on the Na(+)/K(+)-pump as they were rescued by expression of a cardiac glycoside-resistant Na(+)/K(+)-pump. Unlike human cells, rodent cells are largely resistant to cardiac glycosides in vitro and mice were found to tolerate extremely high levels. The physiological difference between human and mouse explains the previously observed sensitivity of human cancer cells in mouse xenograft experiments. Thus, published mouse xenograft models used to support anti-tumor activity for these drugs require reevaluation. Our finding that cardiac glycosides inhibit protein synthesis provides a mechanism for the cytotoxicity of CGs and raises concerns about ongoing clinical trials to test CGs as anti-cancer agents in humans.


Guarding quality in challenging times. Interview by Emily Friedman.
As Dennis S. O'Leary, MD, FACPE, reaches the 18-month mark in his tenure as president of the Joint Commission on Accreditation of Hospitals, Chicago, the organization's future is a foremost concern. In this concluding article of a two-part series, he discusses his vision of where the Commission is going, and how and why. Among key issues that will affect the Joint Commission's future, he says, are changing determinants of quality, evaluation of nonhospital organizations, the interplay of the increasingly diverse individuals and groups that participate in Joint Commission activities, and Joint Commission relations with key health professions and other interest groups, including consumers. The first article, which focused on the Joint Commission's "clinical indicators" project, was published in the July-August issue of Physician Executive.


Mean Platelet Volume and Platelet Distribution Width Indicate that Platelets Remain Small for Most of Their Lifespans in Patients with Essential Thrombocythemia.
Platelets are large when young and immature and shrink as they age. The mean platelet volume (MPV) and platelet distribution width (PDW) reflect the volume and distribution of platelets, respectively. We compared the MPVs and PDWs of patients with immune thrombocytopenic purpura (ITP) or essential thrombocythemia (ET) to those of healthy individuals to test whether these values can reflect the pathomechanisms of these diseases. Platelet counts, MPVs, and PDWs were measured in 153 healthy individuals and in 20 and 34 patients with ITP and ET, respectively, using an XN-3000 instrument. The MPVs and PDWs were significantly higher in ITP patients than in healthy individuals or ET patients. The MPVs were significantly lower in ET patients than in both ITP patients and healthy individuals; however, the PDWs in ET patients were similar to those in healthy individuals. The MPVs and PDWs did not correlate with platelet count in ITP or ET patients. Our data suggested an increased production of young, large platelets and nondiscriminatory destruction of circulating platelets irrespective of their ages in ITP. Decreased MPVs and unelevated PDWs in ET suggest that clonal process-produced platelets remain small throughout their lifespans.


Mechanism of suppression of lymphocyte proliferation by Concanavalin A-activated mononuclear cells.
In this study, the mechanism by which Concanavalin A (Con A) pre-treated lymphocytes suppress mitogen-induced proliferation of responder cells was investigated. Responder-cell proliferation could only be suppressed when these cells were co-cultured with the Con A pre-treated cells but not with their supernatants, nor in chambers where the suppressor cells were separated from the responder cells by a millipore filter. Suppression could not be mediated by heat-killed Con A-activated cells or lysates from these cells. Trypsinization of the Con A-induced suppressor cells resulted in loss of suppressor activity which could be restored if the cells were allowed to recover overnight. Trypsinization of the responder cells, however, before their co-culture with the Con A pre-treated cells did not affect suppression. Addition of cytochalasin B to the co-culture resulted in reduced suppression and cycloheximide treatment of the suppressor cells abolished their activity. The results indicate that for optimal suppression to occur, cell-to-cell contact is required and viable, intact Con A-inducible suppressor cells, actively synthesizing protein are essential. Furthermore, suppression may be mediated via a membrane receptor on the suppressor cell as manipulations of normal membrane function may abolish suppression.


Stimulation of luteinizing hormone beta gene promoter activity by the orphan nuclear receptor, steroidogenic factor-1.
The orphan nuclear receptor, steroidogenic factor-1 (SF-1), is expressed in the pituitary and in the gonadotrope precursor cell line, alphaT3-1, where it is believed to enhance expression of the common gonadotropin alpha-subunit gene through transactivation of the gonadotrope-specific element (GSE). Sequence analysis of the rat luteinizing hormone beta-subunit (LH beta) gene promoter revealed the presence of a consensus GSE at -127 to -119 (TGACCTTGT). We have demonstrated the ability of SF-1 to bind specifically to this putative GSE sequence by electrophoretic mobility shift assay, utilizing both alphaT3-1 nuclear extracts and in vitro translated SF-1. In addition, mutation of the putative LHbeta-GSE (TGAAATTGT) eliminated specific DNA binding. To examine the ability of SF-1 to enhance LHbeta promoter activity, CV-1 cells, which lack endogenous SF-1, were cotransfected with an SF-1-containing expression vector and an LHbeta-luciferase reporter construct. When cotransfected with -209/+5 of the LHbeta promoter, SF-1 increased luciferase activity by 56-fold. SF-1 responsiveness was markedly diminished with loss of the putative GSE region in deletion constructs and in the presence of a two base pair mutation, analogous to the mutation which eliminated DNA binding. Finally, the LHbeta-GSE was able to confer SF-1 responsiveness on a heterologous minimal growth hormone promoter, GH50 (57-fold). We conclude that SF-1 both binds to and transactivates the rat LHbeta promoter. These data suggest that SF-1 may participate in the expression of the LHbeta gene by the gonadotrope.


Catalytic cycloisomerization of enynes by using a nickel-zinc-acid system.
Catalytic cycloisomerization of enynes has been accomplished in the presence of an Ni0-PPh3-Zn-carboxylic acid or -ZnCl2 system. A nickel(I)-hydride complex, thought to be generated by reduction of the protonated nickel(II) complex with Zn, is proposed as the catalytic species. This cycloisomerization shows reactivity behavior that is different from that of a conventional metal-catalyzed reaction. In particular, in the reaction with (E)-enynes, the catalytic system has a selectivity that favors the formation of the 1,3-diene over the 1,4-diene. In addition, this catalytic system has been applied to the domino cyclization of dienynes for the construction of tricyclic compounds.


Caveolin-1 ablation reduces the adverse cardiovascular effects of N-omega-nitro-L-arginine methyl ester and angiotensin II.
Caveolae are the major cellular membrane structure through which extracellular mediators transmit information to intracellular signaling pathways. In vascular tissue (but not ventricular myocardium), caveolin-1 (cav-1) is the main component of caveolae; cav-1 modulates enzymes and receptors, such as the endothelial nitric oxide synthase and the angiotensin II (AngII) type 1 receptor. Evidence suggests that AngII and aldosterone (ALDO) are important mediators of ventricular injury. We have described a model of biventricular damage in rodents that relies on treatment with N-omega-nitro-l-arginine methyl ester (L-NAME (nitric oxide synthase inhibitor)) and AngII. This damage initiated at the vascular level and was observed only in the presence of ALDO and an activated mineralocorticoid receptor (MR). We hypothesize that cav-1 modulates the adverse cardiac effects mediated by ALDO in this animal model. To test this hypothesis, we assessed the ventricular damage and measures of inflammation, in wild-type (WT) and cav-1 knockout (KO) mice randomized to either placebo or L-NAME/AngII treatment. Despite displaying cardiac hypertrophy at baseline and higher blood pressure responses to L-NAME/AngII, cav-1 KO mice displayed, as compared with WT, decreased treatment-induced biventricular damage as well as decreased transcript levels of the proinflammatory marker plasminogen activator inhibitor-1. Additionally, L-NAME/AngII induced an increase in cardiac MR levels in WT but not cav-1-ablated mice. Moreover and despite similar circulating ALDO levels in both genotypes, the myocardial damage (as determined histologically and by plasminogen activator inhibitor-1 mRNA levels) was less sensitive to ALDO levels in cav-1 KO vs. WT mice, consistent with decreased MR signaling in the cav-1 KO. Thus, we conclude that the L-NAME/AngII-induced biventricular damage is mediated by a mechanism partially dependent on cav-1 and signaling via MR/ALDO.


A discrepancy between clinical course and magnetic resonance imaging in a case of non-herpetic acute limbic encephalitis.
We report the case of a 64-year old man who presented memory disturbance, low-grade fever, weight loss, and bilateral hand tremors for three months. He was diagnosed with non-herpetic acute limbic encephalitis (NHALE). Follow-up magnetic resonance imaging (MRI) revealed new lesions after symptomatic improvement following steroid pulse therapy. This may indicate that there is a time lag between the disturbance or recovery of neurons and astrocytes. Thus, other lesions might occasionally appear during convalescence in patients with NHALE, even if only minimal lesions were found on the initial MRI.


[Progress of clinical diagnosis and treatment in fungal keratitis].
Fungal keratitis is an important cause of corneal blindness in China. Delayed diagnosis and treatment contribute to its poor prognosis. In recent years, with the advancement of laboratory test techniques, imaging diagnostic techniques and treatment methods, the diagnosis and treatment of fungal keratitis has constantly improved. This article reviews the recent progress of the laboratory and imaging diagnosis, medicine and surgical treatment in fungal keratitis. It may be helpful to promote the application of the new technologies in China and to improve the prognosis of fungal keratitis.(Chin J Ophthalmol, 2020, 56: 631-636).


Does a school snack make a difference? An evaluation of the World Food Programme emergency school feeding programme in Lebanon among Lebanese and Syrian refugee children.
To investigate the effects of providing a daily healthy school snack on children's nutritional, social and educational outcomes and explore stakeholders' perceptions of an emergency school feeding programme (SFP). Convergence triangulation mixed-methods study design. Associations were examined between receiving the school feeding intervention and children's outcomes using multivariable regression models. Quantitative data were complemented with interviews and focus group discussions with parents and staff. In vulnerable communities in Lebanon, the World Food Programme has implemented an emergency SFP targeting Lebanese (attending morning sessions) and Syrian refugee children (attending afternoon sessions) in public schools. Children from ten intervention schools (morning n 403; afternoon n 379) and ten matched control schools (morning n 399; afternoon n 401), as well as twenty-nine parents and twenty-two school staff members. Diet diversity was higher in intervention schools as compared with control with a significantly higher consumption of dairy products, nuts and fruit in both sessions. Child-reported food insecurity experience was lower in children attending the afternoon session of intervention v. control schools. The SFP intervention was associated with higher school engagement and sense of school community in the morning session only. While the SFP was significantly associated with higher attendance for children in afternoon sessions only, it was significantly associated with school retention of children in both sessions. A daily healthy snack potentially acts as an incentive to improve children's nutritional outcomes, school engagement, sense of belonging, equality between students and improvement in children's attendance and retention in public schools.


Validating cancer drug targets through chemical genetics.
Targeted therapies for cancer promise to revolutionize treatment by specifically inactivating pathways needed for the growth of tumor cells. The most prominent example of such therapy is imatinib (Gleevec), which targets the BCR-ABL kinase and provides an effective low-toxicity treatment for chronic myelogenous leukemia. This success has spawned myriad efforts to develop similarly targeted drugs for other cancers. Unfortunately, the high expectations of these efforts have not yet been realized, likely due to the genetic diversity among and within tumors, as well as the complex and largely unpredictable interactions of drug-like compounds with innumerable targets that affect cellular and organismal metabolism. While improvements in sequencing technologies are beginning to address the first problem, solving the second problem requires methods for linking specific features of the cancer genome to their optimally targeted therapies. One approach, referred to as chemical genetics, accomplishes this by genetic control of chemical susceptibility. Chemical genetics is a crucial tool for the rational development of cancer drugs.


GENITAL TRACT SCREENING FINDS WIDESPREAD INFECTION WITH MUSTELID GAMMAHERPESVIRUS 1 IN THE EUROPEAN BADGER ( MELES MELES).
: Sexually transmitted diseases (STDs) can be important drivers of population dynamics because of their negative effects on reproduction. However, screening for STDs, especially in wildlife populations, is widely neglected. Using the promiscuous, polygynandrous European badger ( Meles meles) as a model, we investigated the presence and prevalence of herpesviruses (HVs) in a wild, high-density population and assessed potential differences in somatic fitness and female reproductive condition between infected and uninfected individuals. We collected n=98 genital swabs from 71 females (51 adults and 20 cubs) and 27 males (26 adults and 1 cub) during spring and summer 2015. Using a PCR specific for a mustelid α-HV, all genital-swab samples tested negative. In a panherpes PCR, a γ-HV was found in 55% (54/98; 39 adults and 15 cubs), identified as mustelid gammaherpesvirus 1 (MusGHV-1) using DNA sequencing. This contrasts with the results of a previous study, which reported MusGHV-1 in 98% (354/361) of blood samples taken from 218 badgers in the same population using PCR. The detection of MusHV-1 in the female reproductive tract strongly indicates the potential for a horizontal and, likely also a vertical, route of transmission. Our results suggest a potential linkage of genital HVs and impaired future reproductive success in females, but because reproductive failure can have many reasons in badgers, the causative link of this negative relationship remains to be investigated.


Pulmonary Calcifications: Is it an Entity in the New World Dialysis Patient?
Metastatic pulmonary calcification (MPC) has been described in the literature to affect up to 60% of dialysis patients. Several case series of MPC were described in 1960s and 1970s. Patients are generally asymptomatic or may present with acute respiratory distress. This entity is associated with up to 60% mortality. We hereby report a case of chronic kidney disease on maintenance hemodialysis who presented with unexplained recurrent dyspnea despite adequate hemodialysis. She was evaluated and found to have a rare presentation of calciphylaxis.


Theca-lutein ovarian cysts associated with homozygous alpha-thalassemia.
We present a case of theca-lutein cysts associated with homozygous alpha-thalassemia. Theca-lutein cysts have been associated with various types of nonimmune hydrops fetalis but have not been reported previously in association with homozygous alpha-thalassemia.


Conduction pattern of excitation in the amphibian atrium assessed by multiple-site optical recording of action potentials.
Spontaneous action potentials were monitored from multiple sites in the bullfrog atrium using a voltage-sensitive merocyanine-rhodanine dye together with a 100-element photodiode matrix array, and we have assessed the spread of the excitation from the pacemaker. Isochrone curves of conduction were obtained by timing the initiation of the action potential-related optical signals: we constructed maps of the spread. Excitatory waves appeared to conduct radially from the pacemaking area over the atrium, and the conduction velocity in the left atrium exceeded that in the right atrium.


[Foreign DNA in developing embryos of the loach Misgurnus Fossilis L].
The fate of pAT153 DNA microinjected into the embryos of loach was examined. At the earliest stages of development (2 h) the high molecular weight transgenome consisting of pAT153 sequences is formed. The transgenome replicates intensively during the course of early development (until the blastula--early gastrula stages), but later the replication slows down which results in the elimination of the transgenome from the majority of embryos at more advanced stages. The analyses of the transgenome structure with the help of restriction endonucleases have shown that, after microinjection of linear DNA, the high molecular weight transgenome is formed by stochastic ligation of the sticky ends of injected molecules. Our data suggest that the transgenome formation from circular or supercoil DNA occurs after its linearization by host endonuclease.


Changes in body composition in response to challenges during aging in rats.
Body composition changes over the lifespan of Brown Norway rats, in patterns similar to those of humans. Young adults are lean, with little fat, much of which is intra-abdominal. As they age, rats exhibit linear growth, and both lean and fat mass increase until late middle to early old age. Fat mass continues to accumulate throughout the lifespan, both viscerally and subcutaneously; aging animals carry a higher proportion of their fat mass peripherally. After middle age, skeletal muscle mass begins to decline, and sarcopenia develops when animals reach senescence. Finally, in late old age, or senescence, body weights begin to decline, and both fat and lean mass are lost. Healthy aged rats generally respond to negative energy balance challenges less robustly than younger adult animals, although they do appropriately regulate adipose tissue stores and preserve lean mass. The response to a positive energy balance challenge (high fat feeding) is less well regulated in aging animals, and dietary-induced obesity develops rapidly in aged animals. Here we present a summary of several studies of body composition in response to challenges of energy balance in aging male Brown Norway rats, with special emphasis on adipose tissue partitioning.


[A case of retrosplenial amnesia].
A case of retrosplenial amnesia was reported. The patient was an 81-year-old right-handed male. He developed amnesic syndrome following cerebral infarction situated left retrosplenial region. His immediate memory was preserved. Recent memory for both verbal and nonverbal modalities was disturbed. He also showed retrograde amnesia for 2 years. The is the first report of retrosplenial amnesia in Japan. We should take into account of the retrosplenial region as a causative site of amnesic syndrome.


Essential amino acids regulate fatty acid synthase expression through an uncharged transfer RNA-dependent mechanism.
To better understand the regulation of gene expression by amino acids, we studied the effects of these macronutrients on fatty acid synthase (FAS), an enzyme crucial for energy storage. When HepG2 cells were fed serum-free media selectively deficient in each amino acid, the omission of any single classic essential amino acid as well as Arg or His (essential in some rapidly growing cells) resulted in FAS mRNA levels that were about half of those in complete medium. Control message levels were unaffected and omission of nonessential amino acids did not alter FAS expression. FAS mRNA levels peaked 12-16 h after feeding complete and Ser (nonessential)-deficient media but did not increase in cells fed Lys (essential)-deficient medium. With Lys, FAS mRNA increased over the physiologic concentration range of 15-150 microM, and low concentrations of lysine decreased FAS but not apoB protein mass. Transcription inhibitors mimicked treatment with Lys-deficient media, and nuclear run-off assays showed that Lys-deficient media abolished FAS but not apoB transcription. After treatment with Lys-deficient media, the intracellular Lys pool was rapidly depleted in association with an increase of uncharged (deacylated) tRNA Lys from < 1 to 64% of available tRNA Lys. Even in the presence of the essential amino acid His, increasing the level of uncharged tRNA His with histidinol, a competitive inhibitor of the histidinyl-tRNA synthetase, blocked FAS expression. Tyrosinol treatment did not alter FAS mRNA levels. These results suggest that essential amino acids regulate FAS expression by altering uncharged tRNA levels, a novel mechanism for nutrient control of gene expression in mammalian cells.


Cognitive and Brain Activity Changes After Mnemonic Strategy Training in Amnestic Mild Cognitive Impairment: Evidence From a Randomized Controlled Trial.
Background: Mnemonic strategy training (MST) has been shown to improve cognitive performance in amnestic mild cognitive impairment (a-MCI), however, several questions remain unresolved. The goal of the present study was to replicate earlier pilot study findings using a randomized controlled design and to evaluate transfer effects and changes in brain activation. Methods: Thirty patients with a-MCI were randomized into MST or education program. At baseline, participants completed clinical and neuropsychological assessments as well as structural and functional magnetic resonance imaging (fMRI). Interventions were administered individually and comprised four sessions, over 2 weeks. MST taught patients to use a three-step process to learn and recall face-name associations. Post-treatment assessment included fMRI, a separate face-name association task, neuropsychological tests, and measures of metamemory. Behavioral (i.e., non-fMRI) measures were repeated after one and 3-months. Results: Participants in the MST condition showed greater improvement on measures of face-name memory, and increased associative strategy use; effects that were accompanied by increased fMRI activation in the left anterior temporal lobe. While all participants reported greater contentment with their everyday memory following intervention, only the MST group reported significant improvements in their memory abilities. There was no clear indication of far-transfer effects to other neuropsychological tests. Conclusion: Results demonstrate that patients with a-MCI not only show stimulus specific benefits of MST, but that they appear capable of transferring training to at least some other cognitive tasks. MST also facilitated the use of brain regions that are involved in face processing, episodic and semantic memory, and social cognition, which are consonant with the cognitive processes engaged by training.


Short telomere length, myocardial infarction, ischemic heart disease, and early death.
We tested the hypothesis that short telomere length is associated with increased risk of myocardial infarction, ischemic heart disease, and early death. We measured leukocyte telomere length in 2 prospective studies of 19 838 Danish general population participants from the Copenhagen City Heart Study and the Copenhagen General Population Study. Participants were followed for up to 19 years for incident myocardial infarction (n=929), ischemic heart disease (n=2038), and death (n=4342). Follow-up was 100% complete. Telomere length decreased linearly with increasing age in women and men in both studies (P=7×10(-74) to P=3×10(-125)). Multifactorially adjusted hazard ratios were 1.10 (95% CI 1.01-1.19) for myocardial infarction, 1.06 (1.00-1.11) for ischemic heart disease, and 1.09 (1.05-1.13) for early death per 1000-base pair decrease in telomere length. The multifactorially adjusted hazard ratios for the shortest versus the longest decile of telomere length were 1.49 (1.07-2.07) for myocardial infarction, 1.24 (1.01-1.53) for ischemic heart disease, and 1.25 (1.07-1.46) for early death. Short telomere length is associated with only modestly increased risk of myocardial infarction, ischemic heart disease, and early death.


Caspase cleavage of Ets-1 p51 generates fragments with transcriptional dominant-negative function.
Ets-1 is a transcription factor that plays an important role in various physiological and pathological processes, such as development, angiogenesis, apoptosis and tumour invasion. In the present study, we have demonstrated that Ets-1 p51, but not the spliced variant Ets-1 p42, is processed in a caspase-dependent manner in Jurkat T-leukaemia cells undergoing apoptosis, resulting in three C-terminal fragments Cp20, Cp17 and Cp14 and a N-terminal fragment, Np36. In vitro cleavage of Ets-1 p51 by caspase 3 produces fragments consistent with those observed in cells undergoing apoptosis. These fragments are generated by cleavage at three sites located in the exon VII-encoded region of Ets-1 p51. This region is absent from the Ets-1 p42 isoform, which therefore cannot be cleaved by caspases. In Ets-1 p51, cleavage generates C-terminal fragments containing the DNA-binding domain, but lacking the transactivation domain. The Cp17 fragment, the major cleavage product generated during apoptosis, is devoid of transcriptional activity and inhibits Ets-1 p51-mediated transactivation of target genes by competing with Ets-1 p51 for binding to Ets-binding sites present in the target promoters. In the present study, we have demonstrated that caspase cleavage of Ets-1 within the exon VII-encoded region leads to specific down-regulation of the Ets-1 p51 isoform during apoptosis. Furthermore, our results establish that caspase cleavage generates a stable C-terminal fragment that acts as a natural dominant-negative form of the full-length Ets-1 p51 protein.


CB(1) and CB(2) receptor-mediated signalling: a focus on endocannabinoids.
The discovery that the major psychoactive component of marijuana activated two G-protein coupled receptors prompted the search for the endogenous cannabinoid ligands now termed endocannabinoids. To date three putative ligands have been isolated, all consisting of arachidonic acid linked to a polar head group. Both synthetic and endogenous cannabinoids have been the focus of extensive study over the past few years. The signalling events produced by endocannabinoids as compared with Delta(9) -THC and synthetic cannabinoids contain many similarities. However, as research focuses more on endogenous ligands the divergence between these classes of compounds grows. This review focuses upon the developments in endocannabinoid signal transduction from receptor-mediated activation of common G-protein linked effector pathways through downstream regulation of gene transcription.


Survival Predictors of Heart Rate Variability After Myocardial Infarction With and Without Low Left Ventricular Ejection Fraction.
Heart rate variability (HRV) and heart rate (HR) dynamics are used to predict the survival probability of patients after acute myocardial infarction (AMI), but the association has been established in patients with mixed levels of left ventricular ejection fraction (LVEF). We investigated whether the survival predictors of HRV and HR dynamics depend on LVEF after AMI. We studied 687 post-AMI patients including 147 with LVEF ≤35% and 540 with LVEF >35%, of which 23 (16%) and 22 (4%) died during the 25 month follow-up period, respectively. None had an implanted cardioverter-defibrillator. From baseline 24 h ECG, the standard deviation (SDNN), root mean square of successive difference (rMSSD), percentage of successive difference >50 ms (pNN50) of normal-to-normal R-R interval, ultra-low (ULF), very-low (VLF), low (LF), and high (HF) frequency power, deceleration capacity (DC), short-term scaling exponent (α1), non-Gaussianity index (λ25 s), and the amplitude of cyclic variation of HR (Acv) were calculated. The predictors were categorized into three clusters; DC, SDNN, α1, ULF, VLF, LF, and Acv as Cluster 1, λ25 s independently as Cluster 2, and rMSSD, pNN50, and HF as Cluster 3. In univariate analyses, mortality was best predicted by indices belonging to Cluster 1 regardless of LVEF. In multivariate analyses, however, mortality in patients with low LVEF was best predicted by the combinations of Cluster 1 predictors or Cluster 1 and 3 predictors, whereas in patients without low LVEF, it was best predicted by the combinations of Cluster 1 and 2 predictors. The mortality risk in post-AMI patients with low LVEF is predicted by indices reflecting decreased HRV or HR responsiveness and cardiac parasympathetic dysfunction, whereas in patients without low LVEF, the risk is predicted by a combination of indices that reflect decreased HRV or HR responsiveness and indicator that reflects abrupt large HR changes suggesting sympathetic involvement.


Codman Award paper--the experience of intensive care unit nurses providing care to the brain dead patient.
The brain dead patient is a by-product of advances in biomedical technology and the findings of this study confirm the view in the literature that caring for the brain dead patient is an emotionally laden process. This personal distress is directly related to two sources of inconsistencies. First, the nurse may possess coexisting beliefs, perceptions, values, opinions, knowledge and actions which are discrepant. Second, the nurse's personal values, knowledge, and behaviours may be in direct opposition to those of her nursing and medical colleagues and those of the family. The presence of the subjective tension results in the use of distancing tactics and/or the designation of a third person as the target of nursing care in an effort to reduce the personal and interpersonal dissonance.


Greenotyper: Image-Based Plant Phenotyping Using Distributed Computing and Deep Learning.
Image-based phenotype data with high temporal resolution offers advantages over end-point measurements in plant quantitative genetics experiments, because growth dynamics can be assessed and analysed for genotype-phenotype association. Recently, network-based camera systems have been deployed as customizable, low-cost phenotyping solutions. Here, we implemented a large, automated image-capture system based on distributed computing using 180 networked Raspberry Pi units that could simultaneously monitor 1,800 white clover (Trifolium repens) plants. The camera system proved stable with an average uptime of 96% across all 180 cameras. For analysis of the captured images, we developed the Greenotyper image analysis pipeline. It detected the location of the plants with a bounding box accuracy of 97.98%, and the U-net-based plant segmentation had an intersection over union accuracy of 0.84 and a pixel accuracy of 0.95. We used Greenotyper to analyze a total of 355,027 images, which required 24-36 h. Automated phenotyping using a large number of static cameras and plants thus proved a cost-effective alternative to systems relying on conveyor belts or mobile cameras.


Vitamin D Supplementation to Prevent COVID-19 Infections and Deaths-Accumulating Evidence from Epidemiological and Intervention Studies Calls for Immediate Action.
The COVID-19 pandemic poses an unprecedented threat to human health, health care systems, public life, and economy around the globe. The repertoire of effective therapies for severe courses of the disease has remained limited. A large proportion of the world population suffers from vitamin D insufficiency or deficiency, with prevalence being particularly high among the COVID-19 high-risk populations. Vitamin D supplementation has been suggested as a potential option to prevent COVID-19 infections, severe courses, and deaths from the disease, but is not widely practiced. This article provides an up-to-date summary of recent epidemiological and intervention studies on a possible role of vitamin D supplementation for preventing severe COVID-19 cases and deaths. Despite limitations and remaining uncertainties, accumulating evidence strongly supports widespread vitamin D supplementation, in particular of high-risk populations, as well as high-dose supplementation of those infected. Given the dynamics of the COVID-19 pandemic, the benefit-risk ratio of such supplementation calls for immediate action even before results of ongoing large-scale randomized trials become available.


PixR, a Novel Activator of Conjugative Transfer of IncX4 Resistance Plasmids, Mitigates the Fitness Cost of mcr-1 Carriage in Escherichia coli.
The emergence of the plasmid-borne colistin resistance gene mcr-1 threatens public health. IncX4-type plasmids are one of the most epidemiologically successful vehicles for spreading mcr-1 worldwide. Since MCR-1 is known for imposing a fitness cost to its host bacterium, the successful spread of mcr-1-bearing plasmids might be linked to high conjugation frequency, which would enhance the maintenance of the plasmid in the host without antibiotic selection. However, the mechanism of IncX4 plasmid conjugation remains unclear. In this study, we used high-density transposon mutagenesis to identify factors required for IncX4 plasmid transfer. Eighteen essential transfer genes were identified, including five with annotations unrelated to conjugation. Cappable-seq, transcriptome sequencing (RNA-seq), electrophoretic mobility shift assay, and β-galactosidase assay confirmed that a novel transcriptional regulator gene, pixR, directly regulates the transfer of IncX4 plasmids by binding the promoter of 13 essential transfer genes to increase their transcription. PixR is not active under nonmating conditions, while the expression of the pixR, pilX3-4, and pilX11 genes increased 3- to 6-fold upon contact with recipient Escherichia coli C600. Plasmid invasion and coculture competition assays revealed the essentiality of pixR for spreading and persistence of mcr-1-bearing IncX4 plasmids in bacterial populations. Effective conjugation is crucial for alleviating the fitness cost exerted by mcr-1 carriage. The existence of the IncX4-specific pixR gene increases plasmid transmissibility while promoting the invasion and persistence of mcr-1-bearing plasmids in bacterial populations, which helps explain their global prevalence. IMPORTANCE The spread of clinically relevant antibiotic resistance genes is often linked to the dissemination of epidemic plasmids. However, the underlying molecular mechanisms contributing to the successful spread of epidemic plasmids remain unclear. In this report, we shine a light on the transfer activation of IncX4 plasmids. We show how conjugation promotes the invasion and persistence of IncX4 plasmids within a bacterial population. The dissection of the regulatory network of conjugation helps explain the rapid spread of epidemic plasmids in nature. It also reveals potential targets for the development of conjugation inhibitors.


[Chemostimulation and catecholamine content of the rat adrenal medulla].
Unanaesthetized rats whose arterial chemoreceptors were stimulated by an one hour acute exposition to hypoxic gaseous mixtures with various carbon dioxide concentrations, presented depletion of the catecholamines content of their adrenal glands only when hypocapnia or increased pH was present (non compensated hypoxia). Moreover, exposition to simultaneous hypoxia and hypercapnia increased the epinephrine stock of the adrenal glands. No changes were found in the myocardium amine content in the same conditions. When anaesthetized rats were treated by iv injection of almitrine bismesylate, a peripheral chemoreceptors stimulating drug, adrenal catecholamines content was insignificantly reduced. In the myocardium, the amines remained at control levels. The most powerful factor related to catecholamines depletion in the adrenals seems to be the hypocapnia or the alkalosis induced by the hyperventilation provoked by glomic stimulation. No indication has been found in favor of an effective adrenergic stimulation caused directly by chemoreceptors stimulation.


The ACTH response to dexamethasone in Persian Gulf War veterans.
The basis of postdeployment health symptoms in Gulf War veterans remains poorly understood. Alterations in the feedback regulation of the hypothalamic-pituitary-adrenal (HPA) axis have been demonstrated in posttraumatic stress disorder (PTSD) and other bodily disorders related to stress. The objective of this article was to examine whether similar HPA axis alterations are related to Gulf War deployment, postdeployment health symptoms, or PTSD. Plasma adrenocorticotropic hormone (ACTH) was measured on consecutive mornings at 08:00 h before and after a low dose of oral dexamethasone (DEX) at 23:00 h in Gulf War veterans with PTSD (n = 14), Gulf War veterans without PTSD (n = 11), and healthy veterans never deployed to a war zone (n = 12). Both Gulf War veterans with PTSD and Gulf War veterans without PTSD had significantly lower post-DEX ACTH levels than the nonexposed veterans, in the absence of group differences in basal ACTH or DEX levels. Among Gulf War veterans, post-DEX ACTH levels were significantly associated with musculoskeletal symptoms. Gulf War deployment and postdeployment health symptoms appear to be associated with alterations in feedback regulation of the pituitary gland that suggests a possible common link between postdeployment health symptoms and other chronic stress-related conditions.


Evaluation of a new lithium colorimetric assay performed on the dade behring dimension X-pand system.
Lithium is the standard first line of treatment for bipolar disorder. Because of its potential toxicity, levels of the drug have to be constantly monitored. To this end, an accurate and rapid assay is thus required. Moreover, lithium measurement allows differential diagnosis in patients with hallucinations, dementia and amnesia. The lithium concentration in blood is usually evaluated by flame photometry, atomic absorption spectroscopy or ion selective electrode. The aim of this study was to evaluate the analytical performance of a new colorimetric assay for lithium. Within-day precision of a pool with a target value of 1.25 mmol/l was 2.18% (CV) and day-to-day precision was assessed using serum aliquots containing two concentrations of lithium, and CVs were 5.47 and 1.6% at 0.6 and 1.15 mmol/l, respectively. The lower detection limit (LLD) was <0.08 mmol/l. The assay seemed to be linear up to 5 mmol/l. Results of a comparison method using Deming regression were: y=1.161x-0.0075 (r=0.9879) for regression with the Flame Photometer and y=0.9729 x+0.0133 (r=0.9787) for regression with atomic absorption spectroscopy. Moreover, in this study we did not find any interference with drugs or other analytes tested. In conclusion, this assay may be a method of choice for measuring lithium blood concentrations, especially in emergency situations.


Protective effect of theaflavins on homocysteine-induced injury in HUVEC cells in vitro.
A model of homocysteine-induced injury in vascular endothelial cells was established to evaluate the protective role of theaflavins on homocysteine-injured human vascular endothelial cells (HUVECs). The cells were co-incubated with 3 concentrations of theaflavins (5, 10, or 20 mg/L) and 0.5 mM homocysteine for 24 hours. The morphology and viability of the cells were determined, and the DNA damage was detected by a comet assay. Superoxide dismutase, malondialdehyde, glutathione peroxidase, nitric oxide, nitric oxide synthase, and endothelin-1 were measured. The results showed that theaflavins can reduce the changes in and damage of homocysteine-injured HUVECs, increase the viability of homocysteine-injured HUVECs, and alleviate DNA damage induced by homocysteine. These results indicate that theaflavins can inhibit homocysteine-induced injury of HUVECs. Further studies showed that theaflavins may reduce the production of homocysteine-induced reactive oxygen species and partly modulate the secretory dysfunction of vascular endothelial cells caused by homocysteine. This finding indicates that the mechanism by which theaflavins inhibit homocysteine-induced injury may relate to their antioxidant activity and the regulation of the secretion of endothelium-derived factors. These findings suggest that theaflavins may be beneficial in the prevention of atherosclerosis and cardiovascular disease.


Development and internal validation of a Wasp Sting Severity Score to assess severity and indicate blood purification in persons with Asian wasp stings.
In recent years, the incidence of wasp sting has increased annually in China. Organ damage and high mortality due to mass wasp envenomation remain major challenges. Timely and appropriate medical intervention can improve survival. However, there are currently no normalized tools for early assessment of severity. The clinical data of wasp sting patients hospitalized from 2011 to 2019 were used as a training set. Logistic regression was used to explore major risk factors for the development of a severe case of wasp sting (SC). The Wasp Sting Severity Score (WSS) was determined considering these risk factors to identify SCs and was tested in a validation dataset that was prospectively collected in 2020. The data of 1131 wasp sting patients from 2011 to 2019 were included in the training set. Logistic regression analysis showed that tea-colored urine, number of stings, and lactate dehydrogenase and total bilirubin levels were risk factors for developing an SC. The WSS was developed considering these four risk factors, and the total possible WSS was 20 points. The WSS was tested using the validation dataset, comprising the data of 153 patients, in 2020, and we found that a WSS ≥3 points was an important indication for blood purification, with a sensitivity of 71.9%, specificity of 92.6% and an area under the curve of 0.918 (95% confidence interval 0.873-0.962). Among patients with more than 30 stings, mortality in those who underwent plasma exchange (PE) within 24 h after admission was significantly lower than that in those who did not receive PE treatment (14.3% versus 46.9%, P = 0.003). However, continuous venovenous hemofiltration (CVVH) (P = 0.317) and hemoperfusion (HP) (P = 0.869) did not significantly reduce mortality. Patients with WSS scores ≥3 should be considered for blood purification as early as possible in addition to routine treatment. In addition, PE is better than CVVH and HP at reducing mortality in patients suffering from severe wasp stings.


Ambipolar Phosphine Derivatives to Attain True Blue OLEDs with 6.5% EQE.
A family of new branched phosphine derivatives {Ph2N-(C6H4)n-}3P → E (E = O 1-3, n = 1-3; E = S 4-6, n = 1-3; E = Se 7-9, n = 1-3; E = AuC6F5 4-6, n = 1-3), which are the donor-acceptor type molecules, exhibit efficient deep blue room temperature fluorescence (λem = 403-483 nm in CH2Cl2 solution, λem = 400-469 nm in the solid state). Fine tuning the emission characteristics can be achieved varying the length of aromatic oligophenylene bridge -(C6H4)n-. The pyramidal geometry of central R3P → E fragment on the one hand disrupts π-conjugation between the branches to preserve blue luminescence and high triplet energy, while on the other hand provides amorphous materials to prevent excimer formation and fluorescence self-quenching. Hence, compounds 2, 3, 5, and 12 were used as emitters to fabricate nondoped and doped electroluminescent devices. The luminophore 2 (E = O, n = 2) demonstrates excellently balanced bipolar charge transport and good nondoped device performance with a maximum external quantum efficiency (EQEmax) of 3.3% at 250 cd/m(2) and Commission International de L'Eclairage (CIE) coordinates of (0.15, 0.08). The doped device of 3 (E = O, n = 3) shows higher efficiency (EQEmax of 6.5, 6.0 at 100 cd/m(2)) and high color purity with CIE (0.15, 0.06) that matches the HDTV standard blue. The time-resolved electroluminescence measurement indicates that high efficiency of the device can be attributed to the triplet-triplet annihilation to enhance generation of singlet excitons.


Mitochondrial Ca2+ dynamics in cells and suspensions.
Mitochondria play a significant role in shaping cytosolic Ca2+ signals. Thus, transfer of Ca2+ across the mitochondrial membrane is much studied, not only in intact cells but also in artificial systems such as mitochondrial suspensions or permeabilised cells. Observed rates of Ca2+ changes vary by at least one order of magnitude. In this work, we investigate the relationship between the Ca2+ dynamics observed in various experimental conditions using a computational model calibrated on experimental data. Results confirm that mitochondrial Ca2+ exchange fluxes through the mitochondrial Ca2+ uniporter (MCU) and the Na+ /Ca2+ exchanger obey the same basic kinetics in cells and in suspensions, and emphasise the important role played by the high Ca2+ levels reached in mitochondria-associated endoplasmic reticulum membranes in intact cells. Tissue specificity can be ascribed to the different modes of regulation of the MCU by Ca2+ , probably related to the specific levels of expression of the Ca2+ sensing regulator subunit of this channel. The model emphasises the importance of mitochondrial density and buffering in controlling the rate of Ca2+ exchanges with mitochondria, as verified experimentally. Finally, we show that heterogeneity between individual mitochondria can explain the large range of amplitudes and rates of rise in mitochondrial Ca2+ concentration that have been observed experimentally.


Depression, Hopelessness, and Complicated Grief in Survivors of Suicide.
Suicide often has a severe impact on the surviving family and friends. There is a need to better understand the psychological and psychopathological consequences of losing a significant other by suicide. The aim of the present study was to assess hopelessness, depression, suicide risk, complicated grief, intrusive memories, and avoidance in a sample of suicide survivors. In this observational study, 35 bereaved individuals were recruited at the Suicide Prevention Centre of Sant'Andrea Hospital in Rome. Individuals were administered a series of validated instruments: the Beck Depression Inventory II (BDI), the Beck Hopelessness Scale (BHS), the Inventory of Complicated Grief (ICG), the Impact of Event Scale (IES), the Subjective Happiness Scale (SHS), and the Satisfaction With Life Scale (SWLS). Most survivors (62.8%) obtained high scores on measures of complicated grief. Scores on the measure of complicated grief were associated with intrusiveness of thoughts and memories, attempts to prevent the thoughts and emotions related to the event, depressive symptoms and hopelessness, and lower scores for feelings of happiness and satisfaction with life. A subgroup of suicide survivors may be at risk of severe psychological distress and suicidal behavior. Identification of these survivors is a necessary step for providing appropriate counseling and psychotherapy.


Primary eye care in sub-Saharan African: do we have the evidence needed to scale up training and service delivery?
The models for addressing the delivery of an eye-care service in sub-Saharan Africa have seen considerable revision in the last 30 years, and the on-going challenges, as well as the future needs, will probably require many more changes and new systems. There is a need to assess the different models that are currently employed, in order to ensure that all potential contributions to the elimination of avoidable blindness are used; the evolving concept of primary eye care (PEC) requires such assessment. For the current review, the published literature on eye care provided by general front-line healthworkers was screened for articles that provided evidence of the impact of such PEC on the general delivery of eye care in sub-Saharan Africa. Of the 103 relevant articles detected, only three provided evidence of the effectiveness of PEC and the authors of all three of these articles suggested that such eye care was not meeting the needs or expectations of the target populations, the trainers, or programmes of eye care. Among the main problems identified were a lack of a clear definition of the scope of practice for PEC, the need for clarifying the specific skills that a front-line healthworker could perform correctly, and the changing needs and expectations for the delivery of an eye-care service in Africa. If PEC is to become adequately grounded in Africa, the generation of further evidence of the effectiveness and limitations of such care would be a prudent move.


Probing the chemistries of the substrate and flavin ring system of p-hydroxybenzoate hydroxylase by raman difference spectroscopy.
Details of the substrate, p-hydroxybenzoate, and substrate analog, p-aminobenzoate, binding to p-hydroxybenzoate hydroxylase have been elicited by Raman difference spectroscopy. Deep red (752 nm) excitation was used to avoid interference from a fluorescence background. The Raman data provide information on changes in the ligand upon binding as well as changes in the flavin ring system of the enzyme in the enzyme-substrate complex. For p-aminobenzoate, its three most intense Raman features, due to a phenyl mode (1607 cm-1) and carboxylate stretching (1383 cm-1) and scissoring (863 cm-1) motions, are little perturbed upon binding and show no changes in the pH range 6.5-8.5. However, changes in a number of spectral features associated with isoalloxazine modes in this pH range are evidence for a protonation/deprotonation event occurring in or near the active site. A feature in the difference spectrum of the complex at 1700 cm-1 is assigned to the stretch of the 4C&dbd;O group of the isoalloxazine; the relatively narrow profile of this feature is due to the ring being held in a rigid network of hydrogen bonds as demonstrated by the X-ray-derived structure [Schreuder, H. A., Prick, P. A. J., Wierenga, R. K., Vriend, G., Wilson, K. S., Hol, W. G. J., & Drenth, J. (1989) J. Mol. Biol. 208, 679-696]. The absence of a corresponding negative band in the spectrum near 1725 cm-1 shows that in the enzyme, in the absence of ligand, the 4C&dbd;O peak is "washed out" by a fluctuating series of hydrogen bonds to water molecules which penetrate to the flavin ring, resulting in a broad C&dbd;O stretching feature which escapes detection in the difference spectrum. For p-hydroxybenzoate, upon complexation, the -COO- symmetric stretch shifts 10 cm-1, which is ascribed to the formation of the salt bridge to the guanidinium of Arg 214, seen in the X-ray structure. This is in contrast with the results for the complex involving the p-amino analog where no shift in the carboxylate mode is detected and demonstrates an advantage of using vibrational spectroscopy as a fine probe of active site interactions, since the X-ray structures for the p-amino and p-hydroxy analog complexes indicate that the structures in the -COO- group guanidinium regions are the same. The Raman difference data for the substrate complex in the 1700 cm-1 region closely resemble those for the p-amino analog, indicating that in both cases the 4C&dbd;O group is participating in a rigid hydrogen bonding network in the complexes with ligand but is in a more dynamic hydrogen bonding environment involving water molecules in the unliganded enzyme. In order to measure the pKa of the -OH group in bound p-hydroxybenzoate, the substrate was labeled with 18O in both -COO- oxygen atoms. By subtracting the Raman spectrum of the complex with labeled substrate from that with unlabeled substrate, a simple difference spectrum was obtained with features involving the -COO- group alone. These features were used to measure the pKa of the ring hydroxyl group which was found to be 8.3. The value determined from absorption spectroscopy is 7.4, and possible reasons for the discrepancy are discussed. Both methods are in accord, however, in that they show that the pKa of the bound substrate is substantially below that for the free, a device which assists in the hydroxylation at the 3-position.


Ecological and socioeconomic correlates of fruit, juice, and vegetable consumption among African-American boys.
Investigators have reported that the availability of foods in local grocery stores correlated with consumption when using large geopolitical units of analysis, e.g., zip codes. Associations across smaller geopolitical units, e.g., census tracts, have not been tested, nor has this work focused on restaurant availability, child consumption, or specific ethnic groups. This study examined whether median family income and fruit, juice, and vegetable (FJV) availability in grocery stores, restaurants, and homes in 11 census tracts correlated with FJV consumption among 11- to 14-year-old African-American Boy Scouts. FJV consumption was measured in 90 scouts using two 24-h food recalls. Instruments were developed to measure the availability of FJV at area grocery stores, restaurants, and homes where troop members resided. Median household income (from 1990 census) was significantly correlated with restaurant fruit availability. Significant correlations were found between restaurant juice and vegetable availability and Boy Scout reported consumption of juice and vegetables. Census tract may be a useful unit when studying restaurant, but not grocery store, FJV availability. Within a census tract, restaurant FJV availability may be a significant target for community intervention and process evaluation.


Direct Exchange of PerfluoroCarbon Liquid for Silicone Oil: A Surgical Technique to Control Pressure and Avoid Retinal Slippage.
Perfluorocarbon heavy liquid (PFCL) is used in vitreoretinal surgery to flatten the unsupported detached retina before insertion of silicone oil in cases of giant retinal tear or relaxing retinectomy. Direct exchange of PFCL for silicone oil is recommended to reduce retinal slippage when compared with fluid-air exchange, but it is commonly regarded as a difficult procedure. We describe our technique for direct PFCL-silicone oil exchange using a 20-gauge drainage cannula, reliably avoiding the complications of retinal slippage and high intraoperative intraocular pressure. We present a consecutive case series of patients undergoing PFCL-oil exchange and explain, using Poiseuille's equation for laminar fluid flow through a cannula, the rationale for using a 20-gauge drainage cannula rather than smaller gauges to avoid high intraocular pressure. Twenty-six patients underwent PFCL-oil exchange from February 1, 2019, to September 30, 2019. There was no intraoperative retinal slippage or pressure-related complications. Postoperatively 20 patients underwent oil removal. Six suffered retinal redetachment, and 14 remained attached. The vision postoil removal ranged from 6/6 to hand movements. We are confident that the PFCL-oil exchange technique described here is reliable and safe. The use of a 20-gauge drainage cannula is recommended regardless of vitrectomy gauge.


Oncogramme, a new individualized tumor response testing method: application to colon cancer.
Colon cancer is the second leading cause of cancer-related death in industrialized countries. Many anti-cancer researches are consequently performed and individualized tumor response testing (ITRT) methods are now used to individualize patient chemotherapeutic administrations. Then, a new ITRT method, Oncogramme, was developed for colon cancer. Colon tumor fragments from different patients were dissociated and seeded in a defined culture medium. Cell preparation process as well as culture medium allowed high cell viability and a good primary culture success rate. After treatment of isolated tumoral cells by chemotherapeutics alone or in combination, cytotoxicity was determined by cell death assay allowing the Oncogramme establishment, which was validated by statistical analysis. Indeed, significant results were obtained such as different profile for each patient's cells with various drugs, and variability between patient's cells in the response to each drug. Procedure described here to obtain the Oncogramme is a new, fast and technically reliable ITRT method applied to colon cancer. For an individualized cancer treatment use, this test should be further validated by a phase I clinical trial.


[The diagnosis and treatment of primary thyroid lymphoma].
Primary thyroid lymphoma (PTL) refers to the malignant tumor of primary lymphoid tissue in the thyroid, accounting for approximately 5% of all thyroid malignancies and less than 3% of all extranodal lymphomas. PTL usually develops in females more than 50 years old. Most of PTL cases are non-Hodgkin's lymphomas. The major pathological subtypes are mucosa-associated lymphoid tissue lymphoma and diffuse large B-cell lymphoma. Treatment options for individual pathological subtypes are also different, so pathological diagnosis is very important. The main treatment options include surgical treatment, radiotherapy, chemotherapy, targeted therapy and combined therapy. The main aims of surgical treatment are to obtain a determined diagnosis and to relieve airway pressure, . The prognosis of PTL is generally excellent.


Uptake mechanism of oppositely charged fluorescent nanoparticles in HeLa cells.
The endocytotic mechanisms involved in the uptake of charged polystyrene nanoparticles into HeLa cells were investigated. Uptake experiments were done in the presence or absence of drugs known to inhibit various factors in endocytosis. Independent of the particle charge, endocytosis is highly dependent on dynamin, F-actin, and tyrosine-specific protein kinases, which suggests a dynamin-dependent and lipid raft-dependent mechanism. However, cholesterol depletion did not hinder particle uptake. Regarding positively charged particles, macropinocytosis, the microtubule network, and cyclooxygenases are also involved. The clathrin-dependent pathway plays a minor role.


SV40 DNA replication inhibition by the monofunctional DNA alkylator Et743.
Ecteinascidin 743 (Et743) is a highly cytotoxic anticancer agent isolated from the squirt Ecteinascidia turbinate, which alkylates DNA in the minor groove at GC-rich sequences resulting in an unusual bending toward the major groove. The ability of Et743 to block DNA replication was studied using the well-established simian virus (SV40) model for mammalian DNA replication in cells and cell-free extracts. Intracellular SV40 DNA isolated from Et743-treated BSC-1 cells was analyzed by native, two-dimensional agarose gel electrophoresis. A low frequency of Et743 adducts detected at 30-100 nM drug concentrations inhibited SV40 origin activity and induced formation of unusual DNA replication intermediates. Under cell-free conditions, only a high Et743 adduct frequency reduced SV40 DNA synthesis. Comparative studies involving related DNA alkylators, tomamycin and saframycin A, revealed inhibition of SV40 DNA replication in cells at concentrations approximately 10 times higher than Et743. Under cell-free conditions tomamycin- or saframycin-A-adducted DNA templates inhibited DNA synthesis similarly to Et743. Et743 appears to be unusual among other alkylators, because its adducts strongly inhibit intracellular SV40 DNA replication but are relatively weak as cis inhibitors as measured under cell-free conditions.


Effect of cationic liposomes on intracellular trafficking and efficacy of antisense oligonucleotides in mouse peritoneal macrophages.
We have investigated the intracellular fate and antisense effect of oligonucleotide/cationic liposome complexes using phosphorothioate oligonucleotides (S-Oligo) targeted to inducible nitric oxide synthase in mouse peritoneal macrophages. Confocal laser microscopic analysis revealed that, after application of fluorescein isothiocyanate (FITC)-labeled S-Oligo alone, the intracellular localization of fluorescence exhibited a punctate pattern in the cytoplasm, suggesting that the oligonucleotides were mainly confined to the endosomal and/or lysosomal compartments. In the case of complexation with Lipofectin and DMRIE-C liposomes, cellular uptake of FITC-S-Oligo was not greatly enhanced and the fluorescence localization in the cells was similar to that of FITC-S-Oligo alone. LipofectAMINE slightly enhanced cellular uptake of FITC-S-Oligo; however, the intracellular localization profile of FITC-S-Oligo remained largely unchanged. The antisense effect was slightly enhanced by LipofectAMINE under only very limited experimental conditions. It was concluded that cationic liposomes are not a potential carrier for S-Oligo in peritoneal macrophages because of their inability to promote the release of S-Oligo from the endosomal compartments to the cytosol over a non-toxic concentration range.


Violence prevention program targeting Connecticut adolescents: description and preliminary results.
Recent statistics show that adolescents are increasingly becoming both the perpetrators and the victims of violent crime in our nation. At our institution, we have developed a violence prevention program specifically targeting adolescents. This three-hour workshop utilizes a multimedia style of presenting information that we believe is well suited to young audiences. To date, over 1,000 adolescents from all over the state have participated in our program. In addition to describing the program, we also summarize the results of program evaluations and demographic profiles completed by audience members. Most participants report that the program was successful in educating them about the causes and consequences of violent injury and offered them alternatives to violence in resolving difficult situations. It is our hope that the education we provide will translate into fewer violent injuries within the state of Connecticut.


Picometer detection by adaptive holographic interferometry in a liquid-crystal light valve.
The large dispersive properties and the narrow frequency bandwidth of the two-wave mixing in a liquid-crystal light valve is used to realize an adaptive holographic interferometer in the Raman-Nath regime. We report experimental observation of picometer periodic displacements and estimate the theoretical signal-to-noise ratio and the minimum quantum-noise-limited detectable displacement.


[Hyperlipidemic pancreatitis during pregnancy--a case report].
We present a case of acute pancreatitis during pregnancy, associated with hyperlipidemia. The patient, 23 years old in 36 g. w., was hospitalized at the Clinic of Obstetrics and Gynecology with nausea, multiple vomitting, persistent abdominal pain and febrility. Because of the clinical apperance of acute pancreatitis Ceaserean Section was performed with subsequent revision of the abdominal cavity, necrectomy of the pancreas, laparostoma, lavage, drainage, nutritional yienostoma. Intubation with artificial pulmonary ventilation and high volume continuous veno-venous haemofiltration (HV CVVH) was performed because of the development of polyorgan insufficiency and ARDS. After the procedure the patient condition became stable. After four months of hospitalisation, she was discharged from hospital with stable vital signs.


Regulatory mechanisms controlling gene expression mediated by the antioxidant response element.
The expression of genes encoding antioxidative and Phase II detoxification enzymes is induced in cells exposed to electrophilic compounds and phenolic antioxidants. Induction of these enzymes is regulated at the transcriptional level and is mediated by a specific enhancer, the antioxidant response element or ARE, found in the promoter of the enzyme's gene. The transcription factor Nrf2 has been implicated as the central protein that interacts with the ARE to activate gene transcription constitutively or in response to an oxidative stress signal. This review focuses on the molecular mechanisms whereby the transcriptional activation mediated by the interaction between the ARE and NF-E2-related factor 2 (Nrf2) is regulated. Recent studies suggest that the sequence context of the ARE, the nature of the chemical inducers, and the cell type are important for determining the activity of the enhancer in a particular gene.


Attitudes Toward Submental Fat Among Adults in the United States.
Excess submental fat (SMF) can cause submental fullness resulting in negative perceptions of individuals. However, the impact of SMF on perceptions of social traits has not been well studied. To characterize the impact of SMF on external value judgments in adults in the United States. Respondents completed an online survey in which they reacted to statements about individuals with varying grades of SMF. Attributes were rated on a scale from 0 to 100 with higher scores for more positive attributes. Similar proportions of respondents (N = 1996) indicated that women and men with double chins were less attractive than those without (91% and 90%, respectively). A double chin was more likely to be noticed on a woman than on a man (78% of respondents). With increasing SMF, individuals were perceived as significantly less likeable, intelligent, happy, active, and easygoing. Those with greater amounts of SMF were rated as significantly less attractive than those with less SMF. For all attributes, male respondents rated all individuals lower than female respondents did. Results from this study provide further evidence of negative perceptions of individuals with SMF. Aesthetics of the submental area, especially SMF, likely impact the overall assessment of attractiveness and social attributes.


Donor substrate regulation of transketolase.
The influence of substrates on the interaction of apotransketolase with thiamin diphosphate was investigated in the presence of magnesium ions. It was shown that the donor substrates, but not the acceptor substrates, enhance the affinity of the coenzyme either to only one active center of transketolase or to both active centers, but to different degrees in each, resulting in a negative cooperativity for coenzyme binding. In the absence of donor substrate, negative cooperativity is not observed. The donor substrate did not affect the interaction of the apoenzyme with the inactive coenzyme analogue, N3'-pyridyl-thiamin diphosphate. The influence of the donor substrate on the coenzyme-apotransketolase interaction was predicted as a result of formation of the transketolase reaction intermediate 2-(alpha,beta-dihydroxyethyl)-thiamin diphosphate, which exhibited a higher affinity to the enzyme than thiamin diphosphate. The enhancement of thiamin diphosphate's affinity to apotransketolase in the presence of donor substrate is probably one of the mechanisms underlying the substrate-affected transketolase regulation at low coenzyme concentrations.


A tagged visual analog scale is a reliable method to assess keel bone deviations in laying hens from radiographs.
Laying hens often suffer from keel bone damage (KBD) that includes pathologies with different etiologies, like diverse forms of fractures and deviations. Since KBD is a problem in all countries and housing systems, methods for the assessment of deviations are urgently needed. Comparisons between genetic lines and between studies are important to detect underlying mechanisms. Field researchers often use palpation as a low-cost and feasible technique for the assessment of KBD. In contrast to palpation, radiography is effective and highly precise at least in detecting keel bone fractures. The aim of this study was to: i) develop a scoring system to assess keel bone deviations from radiographs, ii) to assess inter- and intra-observer reliability of this scoring system, and iii) to investigate whether fractures and deviations of the keel are correlated. In total, 192 hens were used for the investigation. Digital radiographs were taken and evaluated for all hens after slaughter. We developed a tagged visual analog scale with two extreme images as anchors and four intermediate tags, resulting in six images representing the range from "no deviation" to "highly deviated" on a 10 cm line. Eleven participants scored 50 radiographs of keels with varying degree of severity, whereas five images were scored twice to assess intra-observer reliability. Intraclass correlation coefficient for inter-observer reliability was 0.979 with a confidence interval of 0.968 < ICC < 0.987 (F49,268 = 54.2, p < 0.0001). Intraclass correlation coefficient for intra-observer reliability was 0.831 with a confidence interval of 0.727 < ICC < 0.898 (F54,55 = 10.8, p < 0.0001). Individual intra-observer reliability ranged from 0.6 to 0.949. The Spearman correlation showed a strong positive correlation of fractures and deviations (s roh= 0.803, p < 0.001). The tagged visual analog scale could be a reliable instrument for the scoring of keel bone deviations. Our results support the assumption that the majority of highly deviated keels suffer from fractures as well. Further research is needed to investigate the correlation of palpation scores with the evaluation on radiographs.


Postnatal changes in tryptophan hydroxylase and serotonin transporter immunoreactivity in multiple brainstem nuclei of the rat: implications for a sensitive period.
Previously, we found that the brainstem neuronal network in normal rats undergoes abrupt neurochemical, metabolic, and physiological changes around postnatal days (P) 12-13, a critical period when the animal's response to hypoxia is also the weakest. This has special implications for sudden infant death syndrome (SIDS), insofar as seemingly normal infants succumb to SIDS when exposed to respiratory stressors (e.g., hypoxia) during a narrow postnatal window. Because an abnormal serotonergic system has recently been implicated in SIDS, we conducted a large-scale investigation of the 5-HT-synthesizing enzyme tryptophan hydroxylase (TPH) and serotonin transporter (SERT) with semiquantitative immunohistochemistry in multiple brainstem nuclei of normal rats aged P2-21. We found that 1) TPH and SERT immunoreactivity in neurons of raphé magnus, obscurus, and pallidus and SERT in the neuropil of the pre-Bötzinger complex, nucleus ambiguus, and retrotrapezoid nucleus were high at P2-11 but decreased markedly at P12 and plateaued thereafter until P21; 2) SERT labeling in neurons of the lateral paragigantocellular nucleus (LPGi) and parapyramidal region (pPy) was high at P2-9 but fell significantly at P10, followed by a gradual decline until P21; 3) TPH labeling in neurons of the ventrolateral medullary surface was stable except for a significant fall at P12; and 4) TPH and SERT immunoreactivity in a number of other nuclei was relatively stable from P2 to P21. Thus, multiple brainstem nuclei exhibited a significant decline in TPH and SERT immunoreactivity during the critical period, suggesting that such normal development can contribute to a narrow window of vulnerability in postnatal animals.


Patient knowledge of medicines dispensed from Ghanaian community pharmacies.
One vital requirement for patient adherence to medicines is good patient knowledge of the medicines dispensed and this will invariably be linked to good labelling and counselling. The aim of this study was to evaluate the quality of labelling of medicines and determine patient knowledge of the administration of medicines dispensed from a community pharmacy in Ghana. From 6th to 29th January, 2010, dispensed prescriptions of 280 clients were purposely sampled to evaluate the quality of labelling. These clients were also interviewed about their knowledge of the last medicine received immediately after dispensing. A scoring system was employed by awarding a point for each attribute written on the package and each attribute stated by the patient. The dispensing attributes noted were name, dosage, frequency, duration, quantity and route of administration. Of the 280 patients interviewed, 157 (56%) were males. Thirty one (11%) had no education and 99(35%) were secondary school graduates. Antimalarials comprised 17.9% and analgesics, 15.4% of medicines dispensed. The name, quantity, dosage, frequency, duration of therapy and route of administration were written on the label in 98%, 99%, 55%, 54%, 6% and 2% respectively of the dispensed medicines. The mean labelling score was 3.096 (SD=1.05) out of 6. The corresponding patient knowledge values were 63%, 80%, 80%, 75%, 57% and 86%. The mean knowledge score was 4.375 (SD; 1.38) out of 6. The chi square test p-value for the effect of demographic characteristics (sex, educational background, location) on patient knowledge of medicines dispensed were p=0.454; p=0.000, and p=0.138 respectively. Patient knowledge of the administration of dispensed medicines was rated good; and this largely corresponded with the quality of labelling, except that the duration of therapy and route of administration was not frequently written and so labelling was rated just above average.


Relevance of pharmacokinetics and bioavailability of intranasal corticosteroids in allergic rhinitis.
Intranasal corticosteroids offer effective symptomatic treatment of allergic rhinitis in children and adults. When used in recommended doses and administration regimens side effect profiles are acceptable and the risk of serious systemic adverse reactions is small. In children, assessments with the sensitive measure of knemometry has found reassuring results. Rigorous comparisons of specific intranasal corticosteroids in clinical use are not available and we have no knowledge as to whether variations in pharmacokinetic properties or bioavailability may reflect significant clinical differences. When the evidence on pharmacokinetics, bioavailability, systemic activity, side effects and efficacy are taken together, however, the most recently launched compounds fluticasone propionate, mometasone furoate, fluticasone furoate and ciclesonide seem preferable to older products. The documented once-daily administration, the quick onset of action and the concurrent effect on eye symptoms of the new drugs support the suggestion. Evidence for intranasal corticosteroids in pre-school children has not been provided. In this paper, a brief review of recent patents for the treatment of allergic rhinitis has also been presented.


The medicare home health wage index. Is it time for change?
Of all of the elements and factors that make up the calculation of the payment rate under the Medicare home health benefit, the wage index may be the most important, yet overlooked, component.


[Value of 18F-FETNIM PET-CT for detection of tumor hypoxia in non-small-cell lung cancer].
To assess the feasibility of [(18)F]fluoroerythronitroimidazole ((18)F-FETNIM) with integrated positron emission tomography and computed tomography (PET-CT) imaging in detection of hypoxia in non-small-cell lung cancer (NSCLC) patients. Forty-two patients with newly diagnosed NSCLC underwent (18)F-FETNIM PET-CT before treatment. Nineteen patients rested for approximately 120 minutes before undergoing PET-CT, 23 patients underwent 2 sequential PET-CT scans at 60 minutes and 120 minutes after intravenous injection (18)F-FETNIM. (18)F-FETNIM uptake was quantified by calculating the maximum standardized uptake value in the tumor (SUVmax-T) and contralateral normal lung tissue (SUVmax-N). Regions of interest (ROIs) were drawn in the tumor and contralateral position and the radioactivity ratio of tumor to normal (T/N) was calculated. SUVmax-T (2.43 +/- 1.34) was significantly higher than SUVmax-N (0.87 +/- 0.46, P < 0.001) at 120 min. SUVmax-T (2.80 +/- 1.09) and SUVmax-N (1.16 +/- 0.56) at 60 min were significantly higher than SUVmax-T (2.61 +/- 1.10) and SUVmax-N (P < 0.01) at 120 min. T/N (2.56 +/- 0.71) at 60 min was higher than that at 120 min (2.48 +/- 0.60), but the difference between them was not significant (P = 0.324). Our results indicate that (18)F-FETNIM PET-CT may be a useful tool for evaluating hypoxia and may be a means to target specifically tumor cells resistant to conventional treatment before and during ongoing therapy in NSCLC.


Hepatitis C core and nonstructural 3 proteins trigger toll-like receptor 2-mediated pathways and inflammatory activation.
Recent evidence suggests that toll-like receptors (TLRs) recognize certain viruses. We reported that hepatitis C virus (HCV) core and nonstructural 3 (NS3) proteins activate inflammatory pathways in monocytes. The aim of this study was to investigate the role of TLRs in innate immune cell activation by core and NS3 proteins. Human monocytes, human embryonic kidney cells transfected with TLR2, and peritoneal macrophages from TLR2, MyD88 knockout, and wild-type mice were studied to determine intracellular signaling and proinflammatory cytokine induction by HCV proteins. HCV core and NS3 proteins triggered inflammatory cell activation via the pattern recognition receptor TLR2 and failed to activate macrophages from TLR2 or MyD88-deficient mice. HCV core and NS3 induced interleukin (IL)-1 receptor-associated kinase (IRAK) activity, phosphorylation of p38, extracellular regulated (ERK), and c-jun N-terminal (JNK) kinases and induced AP-1 activation. Activation of nuclear factor-kappaB by core and NS3 was associated with increased IkappaBalpha phosphorylation. TLR2-mediated cell activation was dependent on the conformation of core and NS3 proteins and required sequences in the regions of aa 2-122 in core and aa 1450-1643 in NS3. Although cellular uptake of core and NS3 proteins was independent of TLR2 expression, cell activation required TLR2. HCV core protein and TLR2 showed intracellular colocalization. The hyper-elevated TNF-alpha induction by TLR2 ligands in monocytes of HCV-infected patients was not due to increased TLR2 expression. HCV core and NS3 proteins trigger inflammatory pathways via TLR2 that may affect viral recognition and contribute to activation of the innate immune system.


Identification of Phase Separating Proteins With Distributed Reduced Alphabet Representations of Sequences.
Phase separation of proteins play key roles in cellular physiology including bacterial division, tumorigenesis etc. Consequently, understanding the molecular forces that drive phase separation has gained considerable attention and several factors including hydrophobicity, protein dynamics, etc., have been implicated in phase separation. Data-driven identification of new phase separating proteins can enable in-depth understanding of cellular physiology and may pave way towards developing novel methods of tackling disease progression. In this work, we exploit the existing wealth of data on phase separating proteins to develop sequence-based machine learning method for prediction of phase separating proteins. We use reduced alphabet schemes based on hydrophobicity and conformational similarity along with distributed representation of protein sequences and biochemical properties as input features to Support Vector Machine (SVM) and Random Forest (RF) machine learning algorithms. We used both curated and balanced dataset for building the models. RF trained on balanced dataset with hydropathy, conformational similarity embeddings and biochemical properties achieved accuracy of 97%. Our work highlights the use of conformational similarity, a feature that reflects amino acid flexibility, and hydrophobicity for predicting phase separating proteins. Use of such "interpretable" features obtained from the ever-growing knowledgebase of phase separation is likely to improve prediction performances further.


A Comparison of the Risk for Chronic Fascioliasis between Children 3 to 5 Years and Children 6 to 12 Years of Age in the Cusco Region of Peru.
School-age children bear the highest burden of fascioliasis in endemic countries. Few studies have addressed Fasciola in preschool children. We performed a secondary data analysis using two Fasciola databases from Cusco, Peru, comparing preschoolers with elementary school children. We included 2,630 children, 50% were female, the median age was 8.4 years (interquartile range [IQR] 6.1-10.5), and 15% (396/2,630) were < 5 years of age. Children < 5 years were less likely to be infected with Fasciola hepatica (P = 0.008) and Hymenolepis nana (P < 0.001) and more likely to have anemia (P < 0.001) and a lower median height for age Z (HAZ) score (P = 0.002). Fascioliasis was less common in younger children, but this group may be at higher risk for chronic complications caused by fascioliasis.


Interaction of Mycobacterium tuberculosis with human respiratory epithelial cells (HEp-2).
Mycobacterium tuberculosis (MTB) is known to adhere to, invade, multiply inside, and modulate respiratory epithelial cell functions. However, the mode and the molecules involved in the interaction of MTB with respiratory cells are not known. To study post-adherence morphological changes in human respiratory epithelial cells and identify MTB components involved in the binding to these cells. The mechanism of interaction of MTB human respiratory epithelial cells (HEp-2) was investigated by incubating MTB with HEp-2 cells, whereupon the morphological changes were examined under scanning electron microscopy. MTB components involved in the interaction with HEp-2 cells were identified by probing Western blots of mycobacterial sonicates with biotinylated HEp-2 extracts. MTB readily bound to the surface of HEp-2 cells. Adherence of tubercle bacilli on the surface caused membrane perturbation resulting in increased surface membrane projections, particularly towards the margins of the cells. Long thin epithelial membrane projections formed loops around the bacilli holding them on the surface. Membrane projections fused to form a web in which the bacilli were entrapped. Concomitant membrane cavitations seen beneath the site of attachment of the bacilli conceivably caused internalization of the organisms. At least five MTB proteins ( approximately 112, 35, 28, 21 and 15 kDa) specific for HEp-2 cells were identified. Of these, the 28 kDa protein was the predominant HEp-2-binding protein and was found to be heparin-binding hemagglutinin (HBHA). While M. bovis BCG strain showed the same five HEp-2-binding proteins, M. smegmatis, displayed only one (31 kDa) HEp-2-specific protein. MTB binds to HEp-2 cells through multiple proteins, and the binding causes membrane perturbations that result in internalization of the organisms.


Maternal perspectives: The needs of Saudi families of children with Down's syndrome.
Down syndrome (DS) is a chromosomal condition that is affecting an increasing number of infants born in Saudi Arabia. This study investigates the needs of families with children with DS in Saudi Arabia from mothers' perspectives and in a cultural and religious context. An exploratory qualitative research design was used, and semi-structured interviews were conducted with 14 mothers of children with DS. Interview data were thematically analyzed. The most important need was found to be financial or tangible support. An increase in benefits or monthly financial allowances and social security insurance would address these concerns. Societal support is also needed; mothers want DS to be perceived differently and require more support, including better rehabilitation and education services. Informational support is needed to improve the conditions of children with DS. Recommendations for improving the quality of life of children with DS are discussed.


A longitudinal study of hormonal reactions accompanying life events in recently resettled refugees.
Refugees constitute a growing section of the general population in many countries. It is therefore important to study which factors in everyday life are important to recently resettled refugees after they have been granted residence. Life events from a checklist, as well as categories derived from written responses to open-ended questions were analysed (repeated-measures ANOVA). Changes over time in mean serum/plasma concentrations of cortisol, thyroxine, prolactin, and dehydroepiandrosterone sulphate (DHEA-S) were compared in subjects with and without the reported events. Distress in significant others (close friends or first-degree relatives) and a perception of excessive demands in everyday life were associated with increases in serum cortisol. Events associated with decreased levels of prolactin were typically situations of strain in relation to authorities or individuals to which the subjects were in a position of dependency. DHEA-S changed in opposite directions in post-traumatic stress disorder (PTSD) and non-PTSD subjects. DHEA-S also changed with positive events. Distress in significant others and a too demanding everyday life lead to significant changes in the stress-responsive hormones cortisol, prolactin and DHEA-S. Since DHEA-S behaves differently in PTSD, this condition is a potential confounder in studies of DHEA-S with an unknown proportion of PTSD among participants.


Frailty Among Non-Elderly Patients Undergoing Cardiac Surgery.
Usually viewed as a characteristic of old age, frailty may also occur in non-elderly people, primarily in those suffering from chronic disease. Frailty may increase operative risk. To determine the prevalence of frailty patients undergoing coronary artery bypass (CABG) and/or heart valve replacement or reconstruction and/or heart valve surgery, as well as the influence of frailty on postoperative outcomes. Our study comprised 100 adults who underwent consecutive elective cardiac operations. Frailty was assessed using the Fried scale. Patients also performed a 6-minute walk test, and we measured maximal inspiratory and expiratory pressures. A p value <0.05 was considered significant. Of a cohort of 100 patients, based on the Fried frailty criteria, 17 patients (17%) were considered frail, 70 (70%) pre-frail and only 13 (13%) were non-frail. Among patients with valvular heart disease, 11 (18.6%) were considered frail and 43 (73%) pre-frail. Fifty three percent of the patients considered frail were less than 60 years old (median=48 years old). The differences in frailty phenotype between patients with valvular heart disease and coronary artery disease were not statistically significant (p=0.305). A comparison between non-frail, pre-frail, and frail patients showed no significant difference in the distribution of comorbidities and cardiac functional status, regardless of their cardiac disease. However, hospital mortality was significantly higher in frail patients (29.4%, p=0.026) than in pre-frail patients (8.6%) and non-frail patients (0%). Frailty is prevalent even among non-elderly patients undergoing CABG or valvular heart surgery and is associated with higher postoperative hospital mortality.


Influence of antibiotics and case exposure on hospital-acquired Clostridium difficile infection independent of illness severity.
Previous studies of the association between antibiotic exposure and risk of hospital-acquired Clostridium difficile-associated infection (CDI) have not fully accounted for patient severity of illness, and competing risks. To determine the potential effects of interventions on hospital-acquired CDI risk. All adults admitted to a teaching hospital between 2004 and 2014 for more than two days were included. Exposures to all antibiotics and cases of CDI were determined. Patients were followed until discharge from hospital, death, or acquisition of hospital-acquired CDI (defined as positive toxin assay in unformed stool >2 days following admission). Multivariable proportional hazards competing-risks modelling with time-dependent covariates was used, accounting for patient severity of illness using the Escobar model. In all, 208,104 patients were studied. Hospital-acquired CDI risk was 0.46 events per 1000 patient-days, decreasing significantly during the study period. Compared to the 5th percentile hospital death risk (0.02%), patients with a 50% risk of death in hospital had an adjusted hazard ratio (aHR) of hospital-acquired CDI of 5.5. Exposure to some antibiotics significantly increased hospital-acquired CDI risk, being highest for carbapenems (aHR: 1.47 after one week of continuous exposure) and intravenous vancomycin (aHR: 1.53). On the ward, sharing a room with other patients newly diagnosed with CDI significantly increased the risk of subsequent disease (aHR: 1.16 on CDI diagnosis day). The primary determinant of hospital-acquired CDI was patient severity of illness. Exposure to both antibiotics and other patients with CDI significantly increased the subsequent risk of hospital-acquired CDI but this risk was small relative to patient severity of illness.


[Neuromuscular block monitoring for optimisation of conditions for endotracheal intubation].
The choice of an appropriate moment for endotracheal intubation is essential to avoid serious motor and cardiovascular reactions during laryngoscopy and tube insertion.The purpose of the study was to compare the effects of intubation on laryngoscopy conditions and cardiovascular response, when choice of the moment for intubation was directed by either clinical or train-of-four assessment. Adult ASA I patients, scheduled for lumbar disc hernia surgery, who received 0.15 mg kg(-1) of cis-atracurium for muscle relaxation, were divided in two groups. Patients in group I were intubated when the attending anaesthesiologist assessed muscle relaxation to be adequate. Patients in group II were intubated when there was no visual response to train-of-four stimulation of the ulnar nerve. Forty-five patients were enrolled in the study. The mean time for intubation was 162.3+/-35 sec in group I and 339.3+/-73.7 sec in group II. Adequate and excellent conditions for intubation were achieved in all patients of group II, compared to only 53% of patients in group I. Heart rate and arterial blood pressure immediately after intubation were significantly lower (p<0.001) in group II. The objective assessment of neuromuscular relaxation priorto endotracheal intubation provides better conditions and minimization of cardiovascular reaction.


Micro-RNA Implications in Type-1 Diabetes Mellitus: A Review of Literature.
Type-1 diabetes mellitus (T1DM) is one of the most well-defined and complex metabolic disorders, characterized by hyperglycemia, with a constantly increasing incidence in children and adolescents. While current knowledge regarding the molecules related to the pathogenesis and diagnosis of T1DM is vast, the discovery of new molecules, such as micro ribonucleic acids (micro-RNAs, miRNAs), as well as their interactions with T1DM, has spurred novel prospects in the diagnosis of the disease. This review aims at summarizing current knowledge regarding miRNAs' biosynthesis and action pathways and their role as gene expression regulators in T1DM. MiRNAs follow a complex biosynthesis pathway, including cleaving and transport from nucleus to cytoplasm. After assembly of their final form, they inhibit translation or cause messenger RNA (mRNA) degradation, resulting in the obstruction of protein synthesis. Many studies have reported miRNA involvement in T1DM pathogenesis, mainly through interference with pancreatic b-cell function, insulin production and secretion. They are also found to contribute to β-cell destruction, as they aid in the production of autoreactive agents. Due to their elevated accumulation in various biological specimens, as well as their involvement in T1DM pathogenesis, their role as biomarkers in early preclinical T1DM diagnosis is widely hypothesized, with future studies concerning their diagnostic value deemed a necessity.


Predictors of Neonatal Tetanus Mortality in Katsina State, Northwestern Nigeria.
The mortality rate of neonatal tetanus (NNT) remains high in Nigeria. The study was guided by Mosley and Chen's model for the elements of child survival in developing countries. The goal of the study was to assess the associations between selected NNT risk factors, number of maternal tetanus toxoid injections, frequency of antenatal visits, place of delivery, and cord care with neonatal mortality as the outcome variable. The study is a retrospective record review using data from 332 NNT records and analyzed using a logistic regression model. Neonates whose mothers had 1 dose of tetanus toxoid vaccine were found to be 4% less prone to NNT mortality compared to neonates whose mothers did not have any dose of tetanus toxoid vaccine during pregnancy (P < .05, odds ratio = 4.12, 95% confidence interval = 1.04-16.29. Frequency of antenatal visits, place of delivery, and cord care were all not significant predictors of NNT mortality. The study shows that there is association between NNT risk factors and neonatal mortality, hence the need to further strengthen the NNT surveillance system for early detection of potential risk factors. This would help develop specific public health interventions aimed at improving the outcome of NNT. The identification and analysis of NNT mortality risk factors and promoting tetanus toxoid vaccination among pregnant women are effective strategies toward attaining NNT elimination goals in Nigeria.


The Cold War and Modern Memory: Veterans Reflect on Military Service.
This paper uses data from focused interviews to look at how veterans who served primarily during the peacetime Cold War portrayed the effects of military service. Most veterans described being a soldier, sailor, or airman as a neutral, transitional role. Veterans also described their service as having features that are consistent with views of such service as both a positive turning point and a negative disruption. However, only one veteran described military service as operating as a positive turning point in his own life, and just two described it has having been a disruption in their lives. In addition, veterans who served as officers described learning leadership and confidence in the armed forces, which may help explain an observed quantitative officer premium. This latter finding is consistent with a view of the armed forces as facilitating the accumulation of advantage.


Radiation therapy induces the DNA damage response in peripheral blood.
Stereotactic body radiation therapy (SBRT) is a radiotherapy modality that delivers highly conformal, ablative doses to a well-defined target. Here, using a semiquantitative multiplexed assay to analyze ATM and H2AX phosphorylation, we show that ATM kinase activity in peripheral blood mononuclear cells is induced following SBRT. This observation of a systemic ATM kinase-dependent DNA damage response in the peripheral blood is unprecedented and promotes the use of ATM serine-1981 phosphorylation as a predictive biomarker for DNA damaging modalities and ATM inhibitors.


Three-Dimensional Adipocyte Culture as a Model to Study Cachexia-Induced White Adipose Tissue Remodeling.
Cancer cachexia (CC) presents itself as a syndrome with multiple manifestations, causing a marked multi-organ metabolic imbalance. Recently, cachectic wasting has been proposed to be stimulated by several inflammatory mediators, which may disrupt the integrative physiology of adipose tissues and other tissues such as the brain and muscle. In this scenario, the tumor can survive at the host's expense. In recent clinical research, the intensity of depletion of the different fat deposits has been negatively correlated with the patient's survival outcome. Studies have also shown that various metabolic disorders can alter white adipose tissue (WAT) remodeling, especially in the early stages of cachexia development. WAT dysfunction resulting from tissue remodeling is a contributor to overall cachexia, with the main modifications in WAT consisting of morpho-functional changes, increased adipocyte lipolysis, accumulation of immune cells, reduction of adipogenesis, changes in progenitor cell population, and the increase of "niches" containing beige/brite cells. To study the various facets of cachexia-induced WAT remodeling, particularly the changes progenitor cells and beige remodeling, two-dimensional (2D) culture has been the first option for in vitro studies. However, this approach does not adequately summarize WAT complexity. Improved assays for the reconstruction of functional AT ex vivo help the comprehension of physiological interactions between the distinct cell populations. This protocol describes an efficient three-dimensional (3D) printing tissue culture system based on magnetic nanoparticles. The protocol is optimized for investigating WAT remodeling induced by cachexia induced factors (CIFs). The results show that a 3D culture is an appropriate tool for studying WAT modeling ex vivo and may be useful for functional screens to identify bioactive molecules for individual adipose cell populations applications and aid the discovery of WAT-based cell anticachectic therapy.


Odontogenic tumors in Port Harcourt: South-South geopolitical zone of Nigeria.
A retrospective study of odontogenic tumors (OTs) in Port Harcourt was undertaken to establish its prevalence and compare with known data in the literature from Nigeria and elsewhere. All pathologically diagnosed OTs between 2008 and 2013 at the archives of the Department of Oral Pathology and Oral Biology of the University of Port Harcourt/University of Port Harcourt Teaching Hospital were retrospectively studied and classified according to the 2005 WHO classification of OTs and allied diseases. These were recorded into a computer and analyzed using Statistical Package for Social Sciences (SPSS 21.0, Inc., Chicago, IL, USA). A total of sixty-three cases of OTs were recorded for the period under review. Fifty-two of these were cases of ameloblastoma (82.54%). This was followed by adenomatoid odontogenic tumour (AOT) 4 (6.35%) and odontogenic myxoma 3 (4.76%). Most lesions were seen within the second to fourth decades of life and mandible was most frequently affected. It is concluded that the pattern of occurrence of OTs in Port Harcourt followed a general pattern in Nigeria and other African countries but slightly differs from findings from other parts of the world.