Pharmacology of BMY 20064, a potent Ca2+ entry blocker and selective alpha 1-adrenoceptor antagonist.
BMY 20064 is a dihydropyridine Ca2+ entry blocker with potent and selective alpha 1-adrenoceptor antagonist properties. The drug was equal in potency to nifedipine as a Ca2+ entry blocker in depolarized smooth muscle preparations. It was less active than nifedipine in antagonizing Ca2+-induced contractions of isolated guinea pig papillary muscles paced at 0.2, 1.0, or 2 Hz. BMY 20064 was a potent (0.1-0.2 X prazosin) and selective alpha 1-adrenoceptor antagonist in radioligand binding assays and in ganglion-blocked anesthetized rats challenged with phenylephrine. BMY 20064 blocked both the K+ and alpha 1-adrenergic agonist-induced increases in 45Ca uptake into rabbit aortic rings. The drug was more effective than nifedipine, prazosin, or combinations of the drugs in preventing ATP depletion of the rat heart during global ischemia. BMY 20064 was a potent antihypertensive agent in normotensive rats and in SHR. BMY 20064 administered intraarterially (i.a.) dilated both femoral and coronary arterial beds of the dog. Hemodynamic changes elicited by BMY 20064 in anesthetized dogs were similar to those induced by nifedipine. BMY 20064 appears to be a more effective myocardial antiischemic agent than nifedipine, prazosin, or combinations of nifedipine and prazosin. A drug of this type may be more efficacious than dihydropyridines in the management of ischemic episodes.


Advances in immunotherapy for pediatric acute myeloid leukemia.
Achieving better disease control in patients diagnosed with acute myeloid leukemia (AML) has proven challenging. Overall survival has been impacted by addressing treatment related mortality with focused supportive care measures. Despite this improvement, it remains difficult to induce durable leukemia remissions despite aggressive chemotherapeutic regimens. The addition of hematopoietic stem cell transplants (HSCT) has allowed further treatment intensification and provided the benefit of graft-versus-leukemia (GVL) effect. However, HSCT carries the risk of transplant related morbidities, particularly GVHD, and anti-tumor responsiveness is still suboptimal. Thus, there is a need for alternate therapies. Immunotherapy has the potential to address this need. Areas covered: Expert opinion: The elusiveness of an ideal surface antigen target together with an immunosuppressive leukemic microenvironment add to the already difficult challenge in developing AML-targeted immunotherapies. Though many hurdles remain, recent translational discovery and progressive clinical advances anticipate exciting future developments. This review highlights promises and challenges to immune-based therapies for AML. It aims to summarize immunotherapeutic strategies trialed in AML patients to date, inclusive of: antibodies, vaccines, and cellular therapy. It emphasizes those being used in the pediatric population, but also includes adult clinical trials and translational science that may ultimately extend to pediatric patients.


Gene Expression Networks in the Murine Pulmonary Myocardium Provide Insight into the Pathobiology of Atrial Fibrillation.
The pulmonary myocardium is a muscular coat surrounding the pulmonary and caval veins. Although its definitive physiological function is unknown, it may have a pathological role as the source of ectopic beats initiating atrial fibrillation. How the pulmonary myocardium gains pacemaker function is not clearly defined, although recent evidence indicates that changed transcriptional gene expression networks are at fault. The gene expression profile of this distinct cell type in situ was examined to investigate underlying molecular events that might contribute to atrial fibrillation. Via systems genetics, a whole-lung transcriptome data set from the BXD recombinant inbred mouse resource was analyzed, uncovering a pulmonary cardiomyocyte gene network of 24 transcripts, coordinately regulated by chromosome 1 and 2 loci. Promoter enrichment analysis and interrogation of publicly available ChIP-seq data suggested that transcription of this gene network may be regulated by the concerted activity of NKX2-5, serum response factor, myocyte enhancer factor 2, and also, at a post-transcriptional level, by RNA binding protein motif 20. Gene ontology terms indicate that this gene network overlaps with molecular markers of the stressed heart. Therefore, we propose that perturbed regulation of this gene network might lead to altered calcium handling, myocyte growth, and contractile force contributing to the aberrant electrophysiological properties observed in atrial fibrillation. We reveal novel molecular interactions and pathways representing possible therapeutic targets for atrial fibrillation. In addition, we highlight the utility of recombinant inbred mouse resources in detecting and characterizing gene expression networks of relatively small populations of cells that have a pathological significance.


AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle.
5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR) has previously been reported to be taken up into cells and phosphorylated to form ZMP, an analog of 5'-AMP. This study was designed to determine whether AICAR can activate AMP-activated protein kinase (AMPK) in skeletal muscle with consequent phosphorylation of acetyl-CoA carboxylase (ACC), decrease in malonyl-CoA, and increase in fatty acid oxidation. Rat hindlimbs were perfused with Krebs-Henseleit bicarbonate containing 4% bovine serum albumin, washed bovine red blood cells, 200 microU/ml insulin, and 10 mM glucose with or without AICAR (0.5-2.0 mM). Perfusion with medium containing AICAR was found to activate AMPK in skeletal muscle, inactivate ACC, and decrease malonyl-CoA. Hindlimbs perfused with 2 mM AICAR for 45 min exhibited a 2.8-fold increase in fatty acid oxidation and a significant increase in glucose uptake. No difference was observed in oxygen uptake in AICAR vs. control hindlimb. These results provide evidence that decreases in muscle content of malonyl-CoA can increase the rate of fatty acid oxidation.


An examination of occupational medicine practices.
The traditional occupational health on-site utilization of physicians as part of an overall occupational health program has been in decline in recent times. The on-site practitioner is increasingly being replaced by physicians who practice at an off-site location. The effects of this change on the health care delivered to the work force are largely unknown. By utilizing data from the 4490 facilities surveyed during the National Occupational Exposure Survey, it was possible to perform analyses of the association between the on- and off-site practice of medicine and several industrial facility characteristics and services usually associated with occupational medicine. Examination of the data indicated that, in comparison with on-site activity, the off-site practice of occupational medicine appears to result in diminished provision of the screening tests and medical examinations for which data were available.


Building home care into managed long-term care: the VNS CHOICE model.
New approaches to home and community-centered long term care offer strategic opportunities for home health providers to thrive by providing expanded service options for the growing population of chronically ill, frail elders. The traditional expertise of home health providers makes a powerful combination with the flexibility of capitation financing to successfully meet both the needs of dually eligible older adults and government mandates to reduce overall expenditures. VNS CHOICE, a home-centered, managed long-term care program, is built on the principles and delivery system of home health care. By understanding the VNS CHOICE service delivery model and lessons learned from its first two years of operating experience, home care providers across the country may identify innovations to incorporate into their agency product lines.


Tibial Plateau Fractures: Addressing Commonly Encountered Challenges in Reduction and Fixation.
Tibial plateau fractures encompass a broad array of injuries. The mechanism of injury ranges from low to high energy. Devastating complications and associated injuries can occur in both low-energy and high-energy mechanisms; therefore, a careful and detailed physical examination and review of imaging must be performed. Furthermore, a mechanistic approach to fracture classification and morphology enhances the surgeon's understanding of the injury and the ability to predict associated injuries and develop effective strategies for management.


FiRe: Evaluation of a fidelity measure to promote implementation of evidence-based rehabilitation.
This article describes a measure developed to assess fidelity of working with the Boston University approach to Psychiatric Rehabilitation (BPR) in Dutch mental health care. The instrument is intended to measure and improve BPR adherence and clinician competence on an individual level and within individual rehabilitation processes. Rotating pairs of 4 BPR specialists used Fidelity of Rehabilitation (FiRe) to evaluate the fidelity of BPR as applied in 114 rehabilitation processes. The content of these processes was documented by the 27 rehabilitation practitioners who delivered the approach. Interrater reliability (.66) was good as were correlations between the first and second assessments (.74), also increasing over time. FiRe was able to distinguish those delivering the approach at an expert level from nonexperts, expressing concurrent validity. FiRe was also related to practitioner attitude toward rehabilitation, a possibly similar construct. In this study, the average level of BPR adherence according to our fidelity model was low. During the 10.5-month review period, overall FiRe scores improved slightly, but this effect was not significant. However, practitioners reported that feedback based on FiRe helped them to enhance their skills. This first version of FiRe is a valid and reliable instrument with which to gain insight into the quality and fidelity of individual practitioners' rehabilitation practices. The instrument used in this study will be further developed in line with study results. More importantly, FiRe raises questions about the definition of good BPR, a topic that should be explored more extensively in future research. (PsycINFO Database Record


A Marine Bacterium with Animal-Pathogen-Like Type III Secretion Elicits the Nonhost Hypersensitive Response in a Land Plant.
Active plant immune response involving programmed cell death called the hypersensitive response (HR) is elicited by microbial effectors delivered through the type III secretion system (T3SS). The marine bacterium Hahella chejuensis contains two T3SSs that are similar to those of animal pathogens, but it was able to elicit HR-like cell death in the land plant Nicotiana benthamiana. The cell death was comparable with the transcriptional patterns of H. chejuensis T3SS-1 genes, was mediated by SGT1, a general regulator of plant resistance, and was suppressed by AvrPto1, a type III-secreted effector of a plant pathogen that inhibits HR. Thus, type III-secreted effectors of a marine bacterium are capable of inducing the nonhost HR in a land plant it has never encountered before. This suggests that plants may have evolved to cope with a potential threat posed by alien pathogen effectors. Our work documents an exceptional case of nonhost HR and provides an expanded perspective for studying plant nonhost resistance.


Determinants of the bronchodilation response to salbutamol on histamine-induced bronchoconstriction.
Assessment of the bronchodilation response to short-acting beta2-adrenoreceptor agonists on pharmacologically induced bronchoconstriction has often been used to investigate airway smooth muscle beta2-adrenoreceptor function. However, little is known about factors affecting this response. In the present study, the bronchodilation response to 0.2 mg of salbutamol on histamine-induced bronchoconstriction was assessed in 101 steroid-naïve asthmatic subjects. The associations of the response with a wide range of challenge procedure-related variables, clinical asthma severity indicators, and blood markers of airway inflammation were investigated. The response was re-assessed after 6 and 12 weeks' therapy with inhaled budesonide. Baseline FEV1, final histamine concentration, and the maximal fall in FEV1 explained 35-59% of the total variation in the response to salbutamol, depending on the index chosen to express the response. Serum concentration of myeloperoxidase, an index of neutrophilic inflammation, was associated with a poor response. The preceding week daily PEF variation, rescue bronchodilator use, severity of asthmatic symptoms, blood eosinophil count, and serum eosinophilic cationic protein and eosinophilic protein X concentrations were not associated with the response. The salbutamol response seemed to diminish during budesonide treatment but when adjusted by the challenge procedure-related variables the treatment effect vanished. In conclusion, the bronchodilation response to salbutamol on histamine-induced bronchoconstriction is largely determined by challenge procedure-related variables. It seems to be unrelated to the clinical severity of asthma and is not affected by treatment with inhaled corticosteroids. Neutrophilic airway inflammation may be associated with a poor response.


Destabilization of bacteriophage T4 mRNAs by a mutation of gene 61.5.
We identified a novel gene of bacteriophage T4, gene 61.5, which appears to be involved in protein synthesis late in infection. Northern blot analysis revealed that a mutant of 61.5 accumulated truncated transcripts of representative late genes. Using a double mutant of genes 61.5 and 55, which prevents transcription of late genes, we demonstrate that even transcripts of middle genes, while full-length when initially expressed, are similarly truncated at later stages of infection. These results indicate that the abnormality in transcript length occurs late in infection, regardless of whether the transcript derives from a middle or a late gene. Primer-extension analysis revealed that the 5' ends of the late gene 23 transcripts that accumulated in gene 61.5 mutant-infected cells were located at internal discrete sites as well as at the expected transcription start site. Moreover, the decay rates of full-length transcripts from genes uvsY or 45 were more than twofold faster in the absence of a functional gene 61.5. These results suggest that mutation of gene 61.5 activates endonucleolytic cleavage of middle and late transcripts, probably by RNase M.


Immigration and suicidality in the young.
Little research has focused on the relation of immigration and suicidal behaviour in youth. Nevertheless, the impact of migration on the mental health of youth is an issue of increasing societal importance. This review aimed to present studies on the prevalence of suicidal behaviour in immigrant youth in various countries and to provide possible explanations for suicidal behaviour in immigrant youth, especially regarding acculturation. The review included a literature search to locate articles on the subject of suicidal behaviour in immigrant youth in the context of acculturation. Studies on suicidal behaviour in culturally diverse youth are few and most of the existing research does not differentiate ethnic minorities from immigrants. Studies on epidemiology and on specific risk factors were found regarding various immigrant youth including Hispanics in the United States, Asians in North America and Europe, as well as comparative studies between different immigrant groups in specific countries. The relation between immigration status and suicidal behaviours in youth appears to vary by ethnicity and country of settlement. Time spent in the new country as well as intergenerational communication and conflicts with parents have, in many of the studies, been related to suicidality in immigrant youth. Summing up, there is a clear and urgent need to further pursue the work in this field, to develop targeted public health interventions as well as psychosocial treatment for preventing suicide in these youth.


[Induction of direct mutations of intracellular phage cd exposed to O-methylhydroxylamine].
Intracellular development of DNA-containing cd phage in the presence of O-methylhydroxylamine (in vivo mutagenesis) results in 50-fold increase of mutants in the phage progeny. The main effect is due to the mutagen presence during replication of phage DNA (within 10-20 min after the infection). The presence of the mutagen both before and after DNA replication does not produce any considerable mutagenic effect. Comparison of the data obtained with kinetic reaction of O-methylhydroxylamine with nucleic acid components is due to enzymatic formation of modified precursors, N4-metoxycytidine and/or N6-metoxyadenosine derivatives, which have dual functional specificity, and to their incorporation into genome under DNA replication. The presence of O-methylhydroxylamine increases not only the number of mixed clones with a high content of mutants, but also the number of pure mutant clones. Recombinogenic activity of O-methylhydroxylamine is considered to be a possible cause of this effect.


The regulation of lipogenesis in vivo in the lactating mammary gland of the rat during the starved-refed transition. Studies wtih acarbose, a glucosidase inhibitor.
Depression of carbohydrate digestion by oral administration of acarbose, a glucosidase inhibitor, led to a 75% inhibition of the re-activation of lipogenesis in vivo in the mammary gland of 18 h-starved lactating rats refed with 5 g of chow diet. Rates of [1-14C]glucose incorporation in vitro into lipid and CO2 in mammary-gland acini isolated from refed animals were elevated compared with acini from starved rats, but acarbose treatment completely prevented this stimulation. Gastric intubation of glucose led to a large stimulation of lipogenesis in the mammary gland of starved lactating rats, similar to that induced by refeeding with chow diet; this was dependent on the amount of glucose given and the time elapsed between glucose administration and injection of 3H2O for the measurement of lipogenesis. The switch-on of lipogenesis in the mammary gland of starved lactating rats, by refeeding or by intubation of glucose, was associated with a decrease in the ratio of [glucose 6-phosphate]/[fructose 1,6-bisphosphate] in the gland, indicative of an increase in phosphofructokinase activity. A time-course study revealed that the ratio decreased rapidly over the first 30 min of chow refeeding, after which a large surge in lipogenesis was seen. Acarbose, given 25 min after the onset of refeeding, led to a stepwise increase in the ratio, in parallel with the observed decrease in lipogenic activity. It is concluded that the control of lipogenesis in the mammary gland is closely linked to the availability of dietary carbohydrate. An important site of regulation of lipogenesis in the gland appears to be at the level of phosphofructokinase. A possible role of insulin in the regulation of phosphofructokinase activity, and the acute modulation of insulin-sensitivity in the gland during the starved-refed transition, are discussed.


Ghrelin receptor antagonism of hyperlocomotion in cocaine-sensitized mice requires βarrestin-2.
The "brain-gut" peptide ghrelin, which mediates food-seeking behaviors, is recognized as a very strong endogenous modulator of dopamine (DA) signaling. Ghrelin binds the G protein-coupled receptor GHSR1a, and administration of ghrelin increases the rewarding properties of psychostimulants while ghrelin receptor antagonists decrease them. In addition, the GHSR1a signals through βarrestin-2 to regulate actin/stress fiber rearrangement, suggesting βarrestin-2 participation in the regulation of actin-mediated synaptic plasticity for addictive substances like cocaine. The effects of ghrelin receptor ligands on reward strongly suggest that modulation of ghrelin signaling could provide an effective strategy to ameliorate undesirable behaviors arising from addiction. To investigate this possibility, we tested the effects of ghrelin receptor antagonism in a cocaine behavioral sensitization paradigm using DA neuron-specific βarrestin-2 KO mice. Our results show that these mice sensitize to cocaine as well as wild-type littermates. The βarrestin-2 KO mice, however, no longer respond to the locomotor attenuating effects of the GHSR1a antagonist YIL781. The data presented here suggest that the separate stages of addictive behavior differ in their requirements for βarrestin-2 and show that pharmacological inhibition of βarrestin-2 function through GHSR1a antagonism is not equivalent to the loss of βarrestin-2 function achieved by genetic ablation. These data support targeting GHSR1a signaling in addiction therapy but indicate that using signaling biased compounds that modulate βarrestin-2 activity differentially from G protein activity may be required.


Rotaxanation as a sequestering template to preclude incidental metal insertion in complex oligochromophores.
The high yielding CuI-mediated click reaction is an effective procedure for the preparation of oligoporphyrinoid conjugates. However, the CuI catalyst leads to the adventitious and usually undesirable insertion of Cu ions into any non-metalated porphyrinoid centers during reaction. Here we report a "sacrificial rotaxane" strategy for the multifunctionalization of porphyrins with free base corroles without incidental copper insertion. This strategy can be considered a general method for implementing CuI-mediated click reactions with metal cation sequestration to avoid detrimental effects caused by the presence of copper cations.


Effects of the live attenuated measles-mumps-rubella booster vaccination on disease activity in patients with juvenile idiopathic arthritis: a randomized trial.
The immunogenicity and the effects of live attenuated measles-mumps-rubella (MMR) vaccination on disease activity in patients with juvenile idiopathic arthritis (JIA) are matters of concern, especially in patients treated with immunocompromising therapies. To assess whether MMR booster vaccination affects disease activity and to describe MMR booster immunogenicity in patients with JIA. Randomized, multicenter, open-label clinical equivalence trial including 137 patients with JIA aged 4 to 9 years who were recruited from 5 academic hospitals in The Netherlands between May 2008 and July 2011. Patients were randomly assigned to receive MMR booster vaccination (n=68) or no vaccination (control group; n=69). Among patients taking biologics, these treatments were discontinued at 5 times their half-lives prior to vaccination. Disease activity as measured by the Juvenile Arthritis Disease Activity Score (JADAS-27), ranging from 0 (no activity) to 57 (high activity). Disease activity in the year following randomization was compared between revaccinated patients and controls using a linear mixed model. A difference in JADAS-27 of 2.0 was the equivalence margin. Primary immunogenicity outcomes were seroprotection rates and MMR-specific antibody concentrations at 3 and 12 months. Of 137 randomized patients, 131 were analyzed in the modified intention-to-treat analysis, including 60 using methotrexate and 15 using biologics. Disease activity during complete follow-up did not differ between 63 revaccinated patients (JADAS-27, 2.8; 95% CI, 2.1-3.5) and 68 controls (JADAS-27, 2.4; 95% CI, 1.7-3.1), with a difference of 0.4 (95% CI, -0.5 to 1.2), within the equivalence margin of 2.0. At 12 months, seroprotection rates were higher in revaccinated patients vs controls (measles, 100% vs 92% [95% CI, 84%-99%]; mumps, 97% [95% CI, 95%-100%] vs 81% [95% CI, 72%-93%]; and rubella, 100% vs 94% [95% CI, 86%-100%], respectively), as were antibody concentrations against measles (1.63 vs 0.78 IU/mL; P = .03), mumps (168 vs 104 RU/mL; P = .03), and rubella (69 vs 45 IU/mL; P = .01). Methotrexate and biologics did not affect humoral responses, but low patient numbers precluded definite conclusions. Among children with JIA who had undergone primary immunization, MMR booster vaccination compared with no booster did not result in worse JIA disease activity and was immunogenic. Larger studies are needed to assess MMR effects in patients using biologic agents. clinicaltrials.gov Identifier: NCT00731965.


Pediatric extracorporeal shockwave lithotripsy: its efficiency at various locations in the upper tract.
Several reports demonstrated the effectiveness of extracorporeal shockwave lithotripsy (SWL) in the management of pediatric upper tract urolithiasis. The majority of these studies used the stone-free rate as the main indicator for treatment success. Efficacy Quotient (EQ) is an important outcome measure because it not only takes into consideration the stone-free rate, but also includes other undesirable outcomes that increase patient morbidity and utilizes hospital resources. Nevertheless, very few studies reported on the EQ of pediatric SWL at various locations in the upper tract. We retrospectively reviewed the records of 84 children (103 urinary calculi) who were treated with SWL in our institution between 1995 and 2005. There were 84 renal (pelvis, n = 27; upper calyx, n = 13; middle calyx, n = 8; lower calyx, n = 36) and 19 ureteric (upper third, n = 10; middle third, n = 5; lower third n = 4) stones. EQ for renal pelvic stones was 67% compared to 48% for caliceal stones (upper calyx: 52%, middle calyx: 46% and lower calyx: 47%). Overall, the EQ for renal stone was 54% and it was 59% for ureteric stones. This efficiency in the ureter was achieved at the expense of a higher number of shock waves delivered (3606 vs. 2592, P < 0.01). Overall, the stone-free rate was 82% (renal, 81%; ureteric calculi 84%). The only factor that affected the stone-free rate was the size of the stone (33% in stones >2 cm and 87% in stones <2 cm (P < 0.01). The overall retreatment rate was 41% (renal, 40% and ureteric 42%). However, the number of total treatments ranged from 1 to 5 sessions in renal stones and 1 to 2 sessions in ureteric stones. Auxiliary procedures were performed in 9 patients (stenting, n = 3; ureteroscopy, n = 4 and percutaneous nephrolithotomy, n = 2). Pediatric SWL appears to be more efficient in the renal pelvic stones compared to caliceal stones. The efficiency of SWL in the ureter is achieved with a higher number of shock waves delivered. A discussion on the possible future modifications of the definition of EQ, especially in children, is also provided.


Meeting calcium recommendations during middle childhood reflects mother-daughter beverage choices and predicts bone mineral status.
Longitudinal data regarding the influence of beverage intakes on calcium adequacy are lacking. This study evaluated calcium intake from ages 5 to 9 y as a function of mother-daughter beverage choices and as a predictor of bone mineral status. Intakes of energy, calcium, milk, sweetened beverages, fruit juices, and non-energy-containing beverages were measured with the use of three 24-h dietary recalls in 192 non-Hispanic white girls aged 5, 7, and 9 y and their mothers. Calcium intakes from ages 5 to 9 y were categorized as either meeting or falling below recommended adequate intakes (AIs). The girls' bone mineral status was assessed with dual-energy X-ray absorptiometry at age 9 y. The mean 5-y calcium intake was related to bone mineral density at age 9 y (beta = 0.27, P < 0.001). The girls who met the AI for calcium were not heavier (P = 0.83) but had higher energy intakes (P < 0.0001) than did the girls who consumed less than the AI. Compared with the girls who consumed less than the AI, the girls who met the AI consumed, on average, almost twice as much milk (P < 0.0001), had smaller decreases in milk intake (P < 0.01), and consumed 18% less sweetened beverages (P < 0.01) from ages 5 to 9 y; the 2 groups did not differ significantly in juice and non-energy-containing beverage intakes. The girls who met the AI were also served milk more frequently than were the girls who consumed less than the AI (P < 0.0001) and had mothers who drank milk more frequently (P < 0.01) than did the mothers of the girls who consumed less than the AI. These findings provide new longitudinal evidence that calcium intake predicts bone mineral status during middle childhood and reflects mother-daughter beverage choice patterns that are established well before the rapid growth and bone mineralization observed in adolescence.


An investigation into the attitudes and practices of intensive care nurses towards verbal communication with unconscious patients.
This study attempted to investigate the attitudes and practices of intensive care nurses towards verbal communication with unconscious patients. A sample of five staff nurses working in an intensive care unit in Northern Ireland formed the basis for the study. The research design was non-experimental and descriptive-exploratory in nature, incorporating 4-hourly observational periods and structured interviews. Qualitative and quantitative analysis indicated that intensive care nurses spend on average 5% of their time verbally communicating with unconscious patients. Most of this communication involves informing the patient of immediate procedural matters or providing reassuring statements. Most intensive care nurses claim that verbal communication with unconscious patients is very important, and some ambiguity is apparent as to the unconscious patient's level of awareness. Major factors influencing communication are the patient's level of consciousness, the amount of physical care being given and the presence of relatives.


A pragmatic cluster randomized clinical trial of diabetes prevention strategies for women with gestational diabetes: design and rationale of the Gestational Diabetes' Effects on Moms (GEM) study.
Women with gestational diabetes (GDM) are at high risk of developing diabetes later in life. After a GDM diagnosis, women receive prenatal care to control their blood glucose levels via diet, physical activity and medications. Continuing such lifestyle skills into early motherhood may reduce the risk of diabetes in this high risk population. In the Gestational Diabetes' Effects on Moms (GEM) study, we are evaluating the comparative effectiveness of diabetes prevention strategies for weight management designed for pregnant/postpartum women with GDM and delivered at the health system level. The GEM study is a pragmatic cluster randomized clinical trial of 44 medical facilities at Kaiser Permanente Northern California randomly assigned to either the intervention or usual care conditions, that includes 2,320 women with a GDM diagnosis between March 27, 2011 and March 30, 2012. A Diabetes Prevention Program-derived print/telephone lifestyle intervention of 13 telephonic sessions tailored to pregnant/postpartum women was developed. The effectiveness of this intervention added to usual care is to be compared to usual care practices alone, which includes two pages of printed lifestyle recommendations sent to postpartum women via mail. Primary outcomes include the proportion of women who reach a postpartum weight goal and total weight change. Secondary outcomes include postpartum glycemia, blood pressure, depression, percent of calories from fat, total caloric intake and physical activity levels. Data were collected through electronic medical records and surveys at baseline (soon after GDM diagnosis), 6 weeks (range 2 to 11 weeks), 6 months (range 12 to 34 weeks) and 12 months postpartum (range 35 to 64 weeks). There is a need for evidence regarding the effectiveness of lifestyle modification for the prevention of diabetes in women with GDM, as well as confirmation that a diabetes prevention program delivered at the health system level is able to successfully reach this population. Given the use of a telephonic case management model, our Diabetes Prevention Program-derived print/telephone intervention has the potential to be adopted in other settings and to inform policies to promote the prevention of diabetes among women with GDM.


Upregulation of IL-21 receptor on B cells and IL-21 secretion distinguishes novel 2009 H1N1 vaccine responders from nonresponders among HIV-infected persons on combination antiretroviral therapy.
Mechanisms underlying failure of novel 2009 H1N1 influenza vaccine-induced Ab responses in HIV-infected persons are poorly understood. This study prospectively evaluated 16 HIV-infected patients on combination antiretroviral therapy and eight healthy controls (HC) who received a single 15 μg dose of nonadjuvanted novel 2009 H1N1 influenza vaccine during the 2009 H1N1 epidemic. Peripheral blood was collected at baseline (T0) and at 7 d (T1) and 28 d (T2) postvaccination for evaluation of immune responses. Prevaccination hemagglutination inhibition Ab titer was <1:20 in all except one study participant. At T2, all HC and 8 out of 16 patients (50%) developed a vaccine-induced Ab titer of ≥ 1:40. Vaccine responder (R) and vaccine nonresponder patients were comparable at T0 in age, CD4 counts, virus load, and B cell immunophenotypic characteristics. At T2, HC and R patients developed an expansion of phenotypic and functional memory B cells and ex vivo H1N1-stimulated IgG Ab-secreting cells in an ELISPOT assay. The memory B cell response was preceded by a significant expansion of plasmablasts and spontaneous H1N1-specific Ab-secreting cells at T1. At T2, HC and R patients also exhibited significant increases in serum IL-21 levels and in the frequency and mean fluorescence intensity of IL-21R-expressing B cells, which correlated with serum H1N1 Ab titers. Vaccine nonresponder patients failed to develop the above-described vaccine-induced immunologic responses. The novel association of novel 2009 H1N1 vaccine-induced Ab responses with IL-21/IL-21R upregulation and with development of memory B cells and plasmablasts has implications for future research in vaccine design.


Knowledge and Attitude of Health Professionals toward Telemedicine in Resource-Limited Settings: A Cross-Sectional Study in North West Ethiopia.
In resource-limited environments, such as those categorized as underdeveloped countries, telemedicine becomes viewed as effective channel for utilizing the scarce medical resources and infrastructures. The aim of this study was to assess knowledge and attitude toward telemedicine among cross section of health professionals' working in three hospitals in North West Ethiopia. An institution-based cross-sectional study was conducted among 312 health professionals working in three different hospitals of North Gondar Administrative Zone during November 13 to December 10, 2017. Data were collected using structured self-administered questionnaires. Data entry and analysis were done using SPSS version 20. The mean, percentage, and standard deviation were calculated to describe the characteristics of respondents. The chi-square test was used as appropriate, to evaluate the statistical significance of the differences between the responses of the participants. A P value of < 0.05 was considered significant. A total of 312 study subjects were approached and included in the study from November 13 to December 10, and the response rate was 95.5%. The majority of respondents (195 (65.4%)) were male, and the majority of the respondents (66.1%) were in the age group of 21-29 years. A large number of respondents (224 (75%)) were bachelor's degree holders. Only 37.6% of the respondents had demonstrated good knowledge of telemedicine, of which 74.1% were male, 65.2% of them were in the age group of 20-29 years, and 63.4% of them had >5 years of work experience. 191 (64.0%) respondents had good attitude toward telemedicine. The findings of the study suggest that although the respondents' knowledge of telemedicine is limited, most of them have good attitude toward telemedicine. This study underlined the need of giving training on telemedicine in order to fill the knowledge gap.


Fact retrieval and memory consolidation for a movement sequence: bidirectional effects of 'unrelated' cognitive tasks on procedural memory.
The generation of long-term memory for motor skills can be modulated by subsequent motor experiences that interfere with the consolidation process. Recent studies suggest that even a non-motor task may adversely affect some aspects of motor sequence memory. Here we show that motor sequence memory can be either enhanced or reduced, by different cognitive tasks. Participants were trained in performing finger movement sequences. Fully explicit instructions about the target sequence were given before practice. The buildup of procedural knowledge was tested at three time-points: immediately before training (T1), after practice (T2), and 24 hours later (T3). Each participant performed the task on two separate occasions; training on a different movement sequence on each occasion. In one condition, interference, participants performed a non-motor task immediately after T2. Half the participants solved simple math problems and half performed a simple semantic judgment task. In the baseline condition there was no additional task. All participants improved significantly between T1 and T2 (within-session gains). In addition, in the baseline condition, performance significantly improved between T2 and T3 (delayed 'off-line' gains). Solving math problems significantly enhanced these delayed gains in motor performance, whereas performing semantic decisions significantly reduced delayed gains compared to baseline. Thus, procedural motor memory consolidation can be either enhanced or inhibited by subsequent cognitive experiences. These effects do not require explicit or implicit new learning. The retrieval of unrelated, non-motor, well established knowledge can modulate procedural memory.


Disparities in self-reported geriatric depressive symptoms due to sociodemographic differences: an extension of the bi-factor item response theory model for use in differential item functioning.
This study evaluates the measurement noninvariance, or differential item functioning (DIF), in the Center for Epidemiological Studies-Depression (CES-D) items attributable to age, sex, and race/ethnicity among community-dwelling older adults. Participants (N=2773) were from the New Haven site of the Established Populations for Epidemiologic Studies in the Elderly. Statistical analyses included exploratory factor analysis, bi-factor confirmatory factor analysis, and a bi-factor multiple indicator and multiple causes (MIMIC) model to address measurement noninvariance. Blacks, compared with whites, were more likely to endorse items loading on the interpersonal factor, which include "people dislike me" and "people are unfriendly." Women were less likely to endorse the interpersonal items and the "I felt like a failure" item (odds ratio [OR]=0.63, 95% confidence interval [CI]: 0.42, 0.94) than men. But women had a higher proportional odds than men for endorsing the "crying" item (OR=1.86, 95% CI: 1.17, 2.96). Those 75 years and older (relative to those aged 65-74) were less likely to endorse the "I felt like a failure" item (OR=0.65, 95% CI: 0.43, 0.97). However, measurement noninvariance found in both the "crying" and "failure" items were attributable to women and to those aged 75 and older were trivial after controlling for the underlying level of depressive symptomatology. Therefore, the interpersonal items showed measurement noninvariance attributable to sex and race. The bi-factor MIMIC model is useful for examining measurement noninvariance due to sociodemographics in a multidimensional depression instrument.


Review Article: Synthesis, properties, and applications of fluorescent diamond particles.
Diamond particles containing color centers-fluorescent crystallographic defects embedded within the diamond lattice-outperform other classes of fluorophores by providing a combination of unmatched photostability, intriguing coupled magneto-optical properties, intrinsic biocompatibility, and outstanding mechanical and chemical robustness. This exceptional combination of properties positions fluorescent diamond particles as unique fluorophores with emerging applications in a variety of fields, including bioimaging, ultrasensitive metrology at the nanoscale, fluorescent tags in industrial applications, and even potentially as magnetic resonance imaging contrast agents. However, production of fluorescent nanodiamond (FND) is nontrivial, since it requires irradiation with high-energy particles to displace carbon atoms and create vacancies-a primary constituent in the majority color centers. In this review, centrally focused on material developments, major steps of FND production are discussed with emphasis on current challenges in the field and possible solutions. The authors demonstrate how the combination of fluorescent spectroscopy and electron paramagnetic resonance provides valuable insight into the types of radiation-induced defects formed and their evolution upon thermal annealing, thereby guiding FND performance optimization. A recent breakthrough process allowing for production of fluorescent diamond particles with vibrant blue, green, and red fluorescence is also discussed. Finally, the authors conclude with demonstrations of a few FND applications in the life science arena and in industry.


Backaction-driven transport of Bloch oscillating atoms in ring cavities.
We predict that an atomic Bose-Einstein condensate strongly coupled to an intracavity optical lattice can undergo resonant tunneling and directed transport when a constant and uniform bias force is applied. The bias force induces Bloch oscillations, causing amplitude and phase modulation of the lattice which resonantly modifies the site-to-site tunneling. For the right choice of parameters a net atomic current is generated. The transport velocity can be oriented oppositely to the bias force, with its amplitude and direction controlled by the detuning between the pump laser and the cavity. The transport can also be enhanced through imbalanced pumping of the two counterpropagating running wave cavity modes. Our results add to the cold atoms quantum simulation toolbox, with implications for quantum sensing and metrology.


Successful Rituximab Monotherapy in Advanced-stage, Epstein-Barr Virus-positive Diffuse Large B-Cell Lymphoma in an Adolescent With Systemic Juvenile Idiopathic Arthritis.
Epstein-Barr virus-positive diffuse large B-cell lymphoma (EBV+ DLBCL) in pediatrics most commonly occurs as an iatrogenic immunodeficiency-associated lymphoproliferative disease. We report an 18-year-old female individual with refractory systemic juvenile idiopathic arthritis, treated with multiple immunosuppressive agents, who was diagnosed with stage III, EBV+ DLBCL. The patient achieved sustained complete remission after 4 weekly doses of rituximab monotherapy and reduction of immunosuppression. This case suggests that a post-transplant lymphoproliferative disease-like treatment approach can be a safe and effective therapy in a nontransplant, yet severely immunosuppressed, patient with EBV+ DLBCL.


The managed-care perspective for improving quality.
The role of managed-care organizations (MCOs) is to determine the existence of a benefit and how it is to be covered. Clinical guidelines and evidence-based medicine are critical parts of those decisions, and help drive much of the medical policies of MCOs. The goal is to identify and support medically appropriate and cost-effective interventions, and not cover treatments that do not have those characteristics. This will become increasingly important as more payers move to defined contribution plans, which place added responsibility for care decisions on the medical care consumer. Improving quality, creating consistency, and establishing the credibility of MCOs and their guidelines are imperative for integrating evidence-based medicine into coverage decisions. The ultimate outcome sought by MCOs and payers is improvement of the health of the populations they serve.


The Impact of the COVID-19 Pandemic on Immunization Campaigns and Programs: A Systematic Review.
The COVID-19 pandemic has had an impact on health service delivery, including immunization programs, and this review assesses the impact on vaccine coverage across the globe and identifies the potential underlying factors. A systematic search strategy was employed on PubMed, Embase, MedRxiv, BioRxiv, and WHO COVID-19 databases from December 2019 till 15 September 2020. Two review authors independently assessed studies for inclusion, assessed quality, and extracted the data (PROSPERO registration #CRD42020182363). A total of 17 observational studies were included. The findings suggest that there was a reduction in the vaccination coverage and decline in total number of vaccines administered, which led to children missing out on their vaccine doses. An approximately fourfold increase was also observed in polio cases in polio endemic countries. Factors contributing to low vaccine coverage included fear of being exposed to the virus at health care facilities, restriction on city-wide movements, shortage of workers, and diversion of resources from child health to address the pandemic. As the world re-strategizes for the post-2020 era, we should not let a crisis go to waste as they provide an opportunity to establish guidelines and allocate resources for future instances. High-quality supplementary immunization activities and catch-up programs need to be established to address gaps during the pandemic era.


[Induction of intracellular endonuclease synthesis in Serratia marcescens by agents suppressing DNA replication].
Endonuclease synthesis in Serratia marcescens was studied in the presence of agents selectively suppressing DNA biosynthesis: nalidixic acid, mitomycin, hydroxyurea and thymine limitation. All the agents suppressing DNA replication induced exocellular endonuclease biosynthesis irrespective of their action mechanism. The greatest inducing effect was exerted when the agents were added to cells in the late exponential phase. Endonuclease biosynthesis was induced 1-2 hours after adding the agent and was inhibited with chloramphenicol. The induction of exocellular endonuclease synthesis in Serratia marcescens by the classical inducing agents of a SOS response seems to be indicative of a Lex A regulated process.


Fibroblast growth factor 4 directs gap junction expression in the mesenchyme of the vertebrate limb Bud.
Pattern in the developing limb depends on signaling by polarizing region mesenchyme cells, which are located at the posterior margin of the bud tip. Here we address the underlying cellular mechanisms. We show in the intact bud that connexin 43 (Cx43) and Cx32 gap junctions are at higher density between distal posterior mesenchyme cells at the tip of the bud than between either distal anterior or proximal mesenchyme cells. These gradients disappear when the apical ectodermal ridge (AER) is removed. Fibroblast growth factor 4 (FGF4) produced by posterior AER cells controls signaling by polarizing cells. We find that FGF4 doubles gap junction density and substantially improves functional coupling between cultured posterior mesenchyme cells. FGF4 has no effect on cultured anterior mesenchyme, suggesting that any effects of FGF4 on responding anterior mesenchyme cells are not mediated by a change in gap junction density or functional communication through gap junctions. In condensing mesenchyme cells, connexin expression is not affected by FGF4. We show that posterior mesenchyme cells maintained in FGF4 under conditions that increase functional coupling maintain polarizing activity at in vivo levels. Without FGF4, polarizing activity is reduced and the signaling mechanism changes. We conclude that FGF4 regulation of cell-cell communication and polarizing signaling are intimately connected.


Biogenesis of an antitumor antibiotic protein, neocarzinostatin.
A study of the biogenesis of the antitumor protein antibiotic neocarzinostatin (NCS) was undertaken. The production of NCS, as well as the growth of Streptomyces carzinostaticus in a production medium, was sensitive to puromycin, chloramphenicol, and actinomycin D. However, when a 12-hr culture in production medium was transferred to a nongrowth medium consisting of a phosphate buffer with Mg(2+) and Ca(2+), rapid NCS synthesis and liberation occurred. NCS production in this medium was no longer sensitive to actinomycin D, but was sensitive to puromycin and chloramphenicol. The conversion of a precursor NCS to an active form was shown to occur in this medium. Subcellular analysis suggested that NCS synthesis occurred by a mechanism similar to that of protein synthesis by membrane polysomes.


Designing nanoparticle translocation through cell membranes by varying amphiphilic polymer coatings.
Nanoparticle (NP)-assisted drug delivery has been emerging as an active research area. Understanding and controlling the interaction of the coated NPs with cell membranes is key to the development of the efficient drug delivery technologies and to the management of nanoparticle-related health and safety issues. Cellular uptake of nanoparticles coated with mixed hydrophilic/hydrophobic polymer ligands is known to be strongly influenced by the polymer pattern on the NP surface and remains open for further exploration. To unravel the physical mechanism behind this intriguing phenomenon, here we perform dissipative particle dynamics simulations to analyze the forces and efficacy time as the copolymer-coated NPs pass through the lipid bilayer so as to provide better design of coated NPs for future drug delivery applications. Four characteristic copolymer ligands are constructed to perform the simulations: hydrophilic-hydrophobic (AB), hydrophobic-hydrophilic (BA), hydrophobic-hydrophilic-hydrophobic-hydrophilic (BABA), and a random pattern with hydrophilic and hydrophobic beads. We mainly study the critical force and potential of mean force required for entering inside of the lipid bilayer and penetration force to pass all the way through the cell membrane as well as the translocation time for these patterned NPs across the bilayer. Through copolymer ligand pattern designing, we find a suitable nanoparticle candidate with a specific polymer coating pattern for drug delivery. These findings provide useful guidelines for the molecular design of patterned NPs for controllable cell penetrability and help establish qualitative rules for the organization and optimization of copolymer ligands for desired drug delivery.


Using embryology screencasts: a useful addition to the student learning experience?
Although podcasting has been a well used resource format in the last few years as a way of improving the student learning experience, the inclusion of enhanced audiovisual formats such as screencasts has been less used, despite the advantage that they work well for both visual and auditory learners. This study examines the use of and student reaction to a set of screencasts introduced to accompany embryology lectures within a second year module at Brighton and Sussex Medical School. Five mini-lecture screencasts and one review quiz screencast were produced as digital recordings of computer screen output with audio narration and released to students via the managed learning environment (MLE). Analysis of server log information from the MLE showed that the screencasts were accessed by many of the students in the cohort, although the exact numbers were variable depending on the screencast. Students accessed screencasts at different times of the day and over the whole of the access period, although maximum downloads were predictably recorded leading up to the written examination. Quantitative and qualitative feedback demonstrated that most students viewed the screencasts favorably in terms of usefulness to their learning, and end-of-module written examination scores suggest that the screencasts may have had a positive effect on student outcome when compared with previous student attainment. Overall, the development of a series of embryology screencasts to accompany embryology lecture sessions appears to be a useful addition to learning for most students and not simply an innovation that checks the box of "technology engagement."


The PREMISE database of 20 Macaca fascicularis PET/MRI brain images available for research.
Non-human primate studies are unique in translational research, especially in neurosciences where neuroimaging approaches are the preferred methods used for cross-species comparative neurosciences. In this regard, neuroimaging database development and sharing are encouraged to increase the number of subjects available to the community, while limiting the number of animals used in research. Here we present a simultaneous positron emission tomography (PET)/magnetic resonance (MR) dataset of 20 Macaca fascicularis images structured according to the Brain Imaging Data Structure standards. This database contains multiple MR imaging sequences (anatomical, diffusion and perfusion imaging notably), as well as PET perfusion and inflammation imaging using respectively [15O]H2O and [11C]PK11195 radiotracers. We describe the pipeline method to assemble baseline data from various cohorts and qualitatively assess all the data using signal-to-noise and contrast-to-noise ratios as well as the median of intensity and the pseudo-noise-equivalent-count rate (dynamic and at maximum) for PET data. Our study provides a detailed example for quality control integration in preclinical and translational PET/MR studies with the aim of increasing reproducibility. The PREMISE database is stored and available through the PRIME-DE consortium repository.


Polyaxial locking plate fixation in distal femur fractures: a biomechanical comparison.
Uniaxial, first-generation locking plates have become increasingly popular for fixation of supracondylar femur fractures. Polyaxial plates are currently available, which allow for variable-angle screw insertion; however, the biomechanical integrity of these new locking constructs is yet unproven. This study compares the mechanical stability of a conventional locking plate with that of a new polyaxial design. A comminuted supracondylar femur fracture (AO/OTA33-A3) gap model was created in fourth-generation synthetic composite bones. Fixation was obtained with 2 different plate constructs: (1) a conventional locking plate (uniaxial screw heads threading directly into plate) and (2) a polyaxial locking plate (screw heads are captured and "locked" into a fixed angle using locking caps). Eight specimens of each type were then tested in axial, torsional, and cyclic axial modes on a material testing machine. The mean axial stiffness for the polyaxial locking plate was 24.4% greater than the conventional locking plate (168.2 vs 127.1 N/mm; P < 0.0001). The mean torsional stiffness was also greater for the polyaxial plate (2.78 vs 2.57 Nm/degree; P = 0.0226). Cyclic axial loading caused significantly less (P = 0.0034) mean irreversible deformation in the polyaxial plate (5.6 mm) than in the conventional plate (8.8 mm). The mean ultimate load to failure was significantly higher (P = 0.0005) for the polyaxial plate (1560 N) than for the conventional plate (1337 N). The tested plate construct with its polyaxial locking screw mechanism provides a biomechanically sound fixation option for supracondylar femur fractures. The frictional locking mechanism allows maintenance of angular stability while affording the option of variable screw placement.


Polarization control of optical transmission of a periodic array of elliptical nanoholes in a metal film.
The spectral dependences of polarized optical transmission of a metal film with a periodic array of elliptical nanoholes have been studied. Such nanostructured metal films exhibit enhanced broadband optical transmission that one can control by selecting the polarization of incident and (or) transmitted light.


Association between serum level of ubiquinol and NT-proBNP, a marker for chronic heart failure, in healthy elderly subjects.
Ubiquinone and ubiquinol represent the oxidized and reduced forms of Coenzyme Q10 (CoQ10). CoQ10 is present in membranes of almost all human tissues and organs, with highest concentration in the heart. In patients with heart failure, serum levels of the N-terminal pro-brain natriuretic peptide (NT-proBNP) are an indicator of disease severity. Here, we investigated the relationship between serum levels of CoQ10 and NT-proBNP in healthy volunteers of an elderly study population (mean age 52 years, n = 871). We found a negative association between serum levels of ubiquinol and NT-proBNP (P < 0.001). Accordingly, the CoQ10 redox state (% oxidized form of CoQ10) is positively associated with serum NT-proBNP level (P < 0.001). Compared to patients who survived a myocardial infarction (n = 21), healthy subjects have lower NT-proBNP level (500.39 ± 631.28 pg/ml vs. 76.90 ± 120.27 pg/ml, P < 0.001), higher ubiquinol serum level (0.43 ± 0.19 µmol/L vs. 0.71 ± 0.32 µmol/L; P < 0.001), and a lower CoQ10 redox state (27.6 ± 13.8% vs. 17.6 ± 10.1%; P < 0.001). Interestingly, ubiquinol supplementation (150 mg/day; 14 day; n = 53) slightly reduces the expression of CLCN6, a gene related to NT-proBNP level. In summary, higher serum levels of ubiquinol are associated with lower serum NT-proBNP levels in healthy elderly subjects. However, to what extent a high serum level of ubiquinol is a protective factor for heart failure remains to be elucidated in prospective studies.


Prevalence of generalized anxiety disorder and major depression in health-care givers of disabled patients in Majmaah and Shaqra cities, Kingdom of Saudi Arabia.
Anxiety is an emotion characterized by feelings of tension, worried thoughts, and physical changes such as increased blood pressure, etc. The impact of anxiety and depression on health-caregivers of disabled patients are very important because these psychological disorders effects both the health-caregivers and the patients which lead to unexpected bad events. The current study was aimed to find out the prevalence of generalized anxiety disorder and major depression in health-care givers of disabled patients and to observe associations between demographic characteristics and psychological disorders in Majmaah and Shaqra Cities, Kingdom of Saudi Arabia. A cross-sectional community-based study was conducted in rehabilitation centers at Majmaah and Shaqra Cities. The data comprising 100 health-care givers was collected from February to April 2015 through complete enumeration sampling method. A pre-validated "general health quality questionnaire" was used to collect the data. Majority of the participants were male (n = 75, 75%). More than 50% of the participants were holding bachelors' degree. Almost three-fifths of the participants had monthly income between 5000 and 15,000 SAR. The prevalence of somatic disorder was only (2%), anxiety disorder (3%), and depression (1%). However, in psychosocial and stress, the prevalence was (8%), respectively. No significant association was observed between demographic characteristics (nationality, gender, marital status, educational level, occupation, and income) and psychological disorders (somatic, anxiety, psychosocial, depression, and stress) P > 0.05, respectively. The prevalence of psychological disorders was very low. Nationality, gender, marital status, educational level, occupation, and income were not significantly associated with psychological disorders.


Network activation during bimanual movements in humans.
The coordination of movement between the upper limbs is a function highly distributed across the animal kingdom. How the central nervous system generates such bilateral, synchronous movements, and how this differs from the generation of unilateral movements, remain uncertain. Electrophysiologic and functional imaging studies support that the activity of many brain regions during bimanual and unimanual movement is quite similar. Thus, the same brain regions (and indeed the same neurons) respond similarly during unimanual and bimanual movements as measured by electrophysiological responses. How then are different motor behaviors generated? To address this question, we studied unimanual and bimanual movements using fMRI and constructed networks of activation using Structural Equation Modeling (SEM). Our results suggest that (1) the dominant hemisphere appears to initiate activity responsible for bimanual movement; (2) activation during bimanual movement does not reflect the sum of right and left unimanual activation; (3) production of unimanual movement involves a network that is distinct from, and not a mirror of, the network for contralateral unimanual movement; and (4) using SEM, it is possible to obtain robust group networks representative of a population and to identify individual networks which can be used to detect subtle differences both between subjects as well as within a single subject over time. In summary, these results highlight a differential role for the dominant and non-dominant hemispheres during bimanual movements, further elaborating the concept of handedness and dominance. This knowledge increases our understanding of cortical motor physiology in health and after neurological damage.


Neuronal apoptosis inhibitory protein, NAIP, is an inhibitor of procaspase-9.
Ability of the full length NAIP and its BIR3 domain in inhibition of the proteases of the intrinsic apoptosis pathway was investigated. Activity of endogenous executioner caspases was drastically reduced by both recombinant NAIP-BIR3 (NBIR3) and the full length protein. Western blotting experiments showed that the full length NAIP and its BIR3 domain inhibited the cleavage of procaspase-3 by apoptosome activated caspase-9. Moreover, full length NAIP inhibited autocatalytic processing of procaspase-9 in the apoptosome complex indicating that unlike other inhibitor of apoptosis proteins (IAPs) human NAIP is an inhibitor of procaspase-9. Furthermore, inhibition of single-chain caspase-9 (human caspase-9, D315, D330/A point mutations that abrogate the proteolytic processing but not the catalytic activity of caspase-9) by the BIR3 domain indicated that the this domain is the caspase-9 interacting moiety. Consistently, pull-down experiments of single-chain capsase-9 in apoptosome complex by the NBIR3 but not the X-linked inhibitor of apoptosis protein (XIAP)-BIR3 domain confirmed that the protein can associate with procaspase-9 prior to its autoproteolysis upon apoptosome formation. Interaction studies revealed the association of C338W variant of the NBIR3, but not the wild type protein with both SMAC-peptide and the SMAC protein. These data indicate that mutation of C338 to Trp is sufficient to accommodate the interaction of NAIP-BIR3 with SMAC-peptide and protein. Taken together, these results demonstrate that NAIP is evolved to prevent apoptosis right at the initiation stage of apoptosome formation and this inhibition cannot be antagonized by SMAC-type proteins.


The New Role of HNF1A-NAS1/miR-214/INHBA Signaling Axis in Colorectal Cancer.
Colorectal cancer (CRC) is the third most common cancer and one of the leading causes of death worldwide. Seriously threatens human life and health. Previous studies have identified that inhibin βA (INHBA) could induce tumorgenesis and progression of CRC through the regulation of the TGF-β/Smad signal axis. The abnormal expression of INHBA is related to the poor prognosis of patients. The aim of this study was to identify the molecular mechanism of HNF1A-AS1 and miR-214 regulating INHBA and carcinogenesis through bioinformatics combined with experiments. The expression of HNF1A-AS1, miRNA-214-5p, INHBA in pan-cancer and CRC were investigated in the Cancer Genome Atlas (TCGA). The correlation between HNF1A-AS1 and immune-related genes or miRNAs was explored via the Gene Expression Profiling Interactive Analysis (GEPIA) and volcano plots, respectively. The association between HNF1A-AS1 and differentially expressed miRNAs was constructed by TargetScan. The miRDB, miRWalk, and TargetScan databases were utilized to predict the target genes of hsa-miR-214. The expression of INHBA in tissues and cell lines of CRC was examined by RT-qPCR and western blot assay. The INHBA and HNF1A-AS1 expressions were increased in Colon adenocarcinoma (COAD) and Rectum adenocarcinoma (READ) of the TCGA database. Hsa-miR-214 was relatively less expressed in CRC tissues compared with para-cancer tissues. The expression of HNF1A-AS1 was negatively correlated with hsa-miR-214. INHBA was one of the target genes of hsa-miR-214 based on miRDB, miRWalk, and TargetScan databases. The specific binding sites of INHBA-3'UTR and miR-214-5p were identified by starBase. The expression level of INHBA was positively correlated with the T stage of tumor and negatively correlated with overall survival (OS) and disease-free survival (DFS) in CRC patients. The results of RT-qPCR and western blot indicated that the expression of INHBA in tissues and cell lines in CRC was higher than those in para-carcinoma tissues and normal colon cell lines, respectively. These findings suggested that HNF1A-AS1 and miRNA-214-5p were key upstream non-coding RNAs of INHBA. The HNF1A-AS1/miR-214/INHBA signal axis plays a significant role in the tumorgenesis and progression of CRC. By interfering with HNF1A-AS1 and INHBA genes on HT29 and SW480 cells, it was found that HNF1A-AS1 and INHBA genes may be important target genes in CRC.


Cumulative risk effects for the development of behaviour difficulties in children and adolescents with special educational needs and disabilities.
Research has identified multiple risk factors for the development of behaviour difficulties. What have been less explored are the cumulative effects of exposure to multiple risks on behavioural outcomes, with no study specifically investigating these effects within a population of young people with special educational needs and disabilities (SEND). Furthermore, it is unclear whether a threshold or linear risk model better fits the data for this population. The sample included 2660 children and 1628 adolescents with SEND. Risk factors associated with increases in behaviour difficulties over an 18-month period were summed to create a cumulative risk score, with this explanatory variable being added into a multi-level model. A quadratic term was then added to test the threshold model. There was evidence of a cumulative risk effect, suggesting that exposure to higher numbers of risk factors, regardless of their exact nature, resulted in increased behaviour difficulties. The relationship between risk and behaviour difficulties was non-linear, with exposure to increasing risk having a disproportionate and detrimental impact on behaviour difficulties in child and adolescent models. Interventions aimed at reducing behaviour difficulties need to consider the impact of multiple risk variables. Tailoring interventions towards those exposed to large numbers of risks would be advantageous.


Awake prone positioning for hypoxaemic respiratory failure: past, COVID-19 and perspectives.
Prone positioning reduces mortality in the management of intubated patients with moderate-to-severe acute respiratory distress syndrome. It allows improvement in oxygenation by improving ventilation/perfusion ratio mismatching.Because of its positive physiological effects, prone positioning has also been tested in non-intubated, spontaneously breathing patients, or "awake" prone positioning. This review provides an update on awake prone positioning for hypoxaemic respiratory failure, in both coronavirus disease 2019 (COVID-19) and non-COVID-19 patients. In non-COVID-19 acute respiratory failure, studies are limited to a few small nonrandomised studies and involved patients with different diseases. However, results have been appealing with regard to oxygenation improvement, especially when combined with noninvasive ventilation or high-flow nasal cannula.The recent COVID-19 pandemic has led to a major increase in hospitalisations for acute respiratory failure. Awake prone positioning has been used with the aim to prevent intensive care unit admission and mechanical ventilation. Prone positioning in conscious, non-intubated COVID-19 patients is used in emergency departments, medical wards and intensive care units.Several trials reported an improvement in oxygenation and respiratory rate during prone positioning, but impacts on clinical outcomes, particularly on intubation rates and survival, remain unclear. Tolerance of prolonged prone positioning is an issue. Larger controlled, randomised studies are underway to provide results concerning clinical benefit and define optimised prone positioning regimens.


In vivo safety, biodistribution and antitumor effects of uPAR retargeted oncolytic measles virus in syngeneic cancer models.
The urokinase receptor (uPAR) is a clinically relevant target for novel biological therapies. We have previously rescued oncolytic measles viruses fully retargeted against human (MV-h-uPA) or murine (MV-m-uPA) uPAR. Here, we investigated the in vivo effects of systemic administration of MV-m-uPA in immunocompetent cancer models. MV-m-uPA induced in vitro cytotoxicity and replicated in a receptor-dependent manner in murine mammary (4T1) and colon (MC-38 and CT-26) cancer cells. Intravenous administration of MV-m-uPA to 4T1 tumor-bearing mice was not associated with significant clinical or laboratory toxicity. Higher MV-N RNA copy numbers were detected in primary tumors, and viable viral particles were recovered from tumor-bearing tissues only. Non-tumor-bearing organs did not show histological signs of viral-induced toxicity. Serum anti-MV antibodies were detected at day 14 of treatment. Immunohistochemistry and immunofluorescence studies confirmed successful tumor targeting and demonstrated enhanced MV-m-uPA-induced tumor cell apoptosis in treated compared with control mice. Significant antitumor effects and prolonged survival were observed after systemic administration of MV-m-uPA in colon (CT-26) and mammary (4T1) cancer models. The above results show safety and feasibility of uPAR targeting by an oncolytic virus, and confirm significant antitumor effects in highly aggressive syngeneic immunocompetent cancer models.


Effects of tadalafil once daily on maximum urinary flow rate in men with lower urinary tract symptoms suggestive of benign prostatic hyperplasia.
Tadalafil significantly improves lower urinary tract symptoms suggestive of benign prostatic hyperplasia. We post hoc characterized changes in the maximum urinary flow rate using integrated data from 4 international, placebo controlled studies of tadalafil once daily for lower urinary tract symptoms suggestive of benign prostatic hyperplasia. After a 4-week placebo lead-in period 1,500 men were randomized to tadalafil 5 mg or placebo for 12 weeks. Data were analyzed using ANCOVA. Maximum urinary flow rate values were rank transformed for analysis. Baseline maximum urinary flow rate data were available on 1,371 men with a mean age of 63.1 years and end point data were available on 1,197. Tadalafil 5 mg significantly increased maximum urinary flow vs placebo (median 1.1 vs 0.4 ml per second, p = 0.003). At a baseline voided volume of 125 to less than 250 ml the median change in the maximum urinary flow rate was 0.9 and 1.2 ml per second (p = 0.142) in 731 patients, at a baseline of 250 to 450 ml the change was -0.3 and 0.7 ml per second (p = 0.011) in 428, and at a baseline of greater than 450 ml the change was -0.2 and 2.0 ml per second (p = 0.186) in 38 for placebo and tadalafil, respectively. The difference was 0.3, 1.0 and 2.2 ml per second, respectively. At a baseline maximum urinary flow rate of greater than 15 ml per second in 128 patients the median flow rate change was -2.1 and -0.8 ml per second (p = 0.246), at a maximum of 10 to 15 ml per second in 522 the change was 0.2 and 0.8 ml per second (p = 0.044), and at a maximum of less than 10 ml per second in 547 the change was 1.2 and 1.8 ml per second (p = 0.189) for placebo and tadalafil, respectively. Tadalafil improved I-PSS (International Prostate Symptom Score) voiding subscores significantly vs placebo across all baseline maximum urinary flow subgroups (each p <0.001). This integrated analysis revealed a small but statistically significant median maximum urinary flow rate improvement for tadalafil vs placebo. The numerical difference in the maximum urinary flow change from baseline between tadalafil and placebo increased with increased voided volume.


Infection-related Glomerulonephritis and C3 Glomerulonephritis - Similar Yet Dissimilar: A Case Report and Brief Review of Current Literature.
Infection-related glomerulonephritis (IRGN) is an immune complex-mediated glomerulonephritis (GN), often preceded by infection with subsequent recovery of renal function after the resolution of the infection. C3 deposition in the absence of immune complex deposits can be seen in patients with IRGN, but with the emergence of C3 glomerulonephritis (C3GN), the distinction is difficult as the clinical and pathological presentation may be similar. However, their treatment and clinical course vary significantly. A 64-year-old man with a history of hypertension and bioprosthetic aortic valve presented to the Emergency Department with left upper quadrant (LUQ) pain and a purpuric rash on bilateral lower extremities. The patient became septic, and further workup during the hospitalization revealed endocarditis secondary to Streptococcus viridans. On admission, the patient had acute kidney injury (AKI) with a serum creatinine of 3.79 mg/dl, which peaked at 5.72 mg/dl during the hospitalization. Renal biopsy demonstrated segmental necrotizing glomerulonephritis on light microscopy, predominant C3 deposition on immunofluorescence (IF) staining, and mesangial and paramesangial deposits on electron microscopy. This histologic picture can be seen both in IRGN and C3GN. The patient was treated with intravenous ceftriaxone for six weeks for endocarditis and the kidney injury was managed with supportive care. The patient's renal function improved and complement levels normalized, supporting the diagnosis of IRGN retrospectively. IRGN can mimic C3GN, and evaluation for alternate pathways of the complement system may be warranted in patients with atypical presentation of IRGN.


Is there a significant difference in the oral microbiome in vapers vs non-vapers?
This study was a secondary data analysis of a parent study that was initially designed to assess saliva and exhaled breath for passive e-cigarette vapour exposure in children. The initial study collected patients using convenience sampling of the local community between July 2018 and February 2019 and ethical approval was obtained prior to commencing. Eligibility criteria for the parent vapers in the original study included self-reported daily use of e-cigarettes in the presence of children via the use of a questionnaire. Microbiome data for a total of 36 adults were collected and analysed in this study (18 vapers and 18 non-vapers). Vapers have a distinct oral microbiome compared with non-vapers. They tend to have a higher relative abundance of Veilonella, an opportunistic pathogen that can stimulate the growth of other opportunistic pathogens. This species has also been found in carious lesions compared with healthy tooth surface.


Effects of age on the glucoregulatory response following acute glucoprivation induced by 2-deoxyglucose (2DG) in the adrenal medulla of Sprague Dawley rats.
Impairment in glucose homeostasis is one of the factors that may alter the feeding drive, hunger and satiety signals, which essential to maintain a sufficient level of energy for daily activities especially among the elderly. Adrenal medulla is one of the important organs that involves in glucose homeostasis through secretion of catecholamines. The catecholamines biosynthesis pathway utilizes various enzymes and protein kinases. The aims of this study are to investigate the effects of age on the biosynthetic pathway of catecholamines in adrenal medulla by determining the level of blood glucose and blood catecholamines, the gene and protein expression of biosynthetic catecholamine enzymes (TH, DBH and PNMT) as well as protein kinase substrates that involved in the phosphorylation of TH in 2DG-induced rats. Adrenal medulla from male Sprague Dawley rats at the age of 3-months (n=12) and 24-months (n=12) were further divided into two groups: 1) treatment group with 2DG to create glucoprivation condition and 2) the vehicle group which received normal saline as control. The results showed that the level of glucose, adrenaline and noradrenaline were increased in response to acute glucoprivation conditions in both young and old rats. No age-related differences were found in the basal gene expression of the enzymes that involved in the catecholamines biosynthesis pathway. Interestingly the expressions of TH and DBH protein as well as the level of TH phosphorylation at Ser40, PKA, PKC and ERK1/2 substrates were higher in basal condition of the aged rats. However, contradicted findings were obtained in glucoprivic condition, which the protein expressions of DBH, pERK1/2 and substrates for pPKC were increased in young rats. Only substrate for pCDK was highly expressed in the old rats in the glucoprivic condition, while pPKC and pERK1/2 were decreased significantly. The results demonstrate that adrenal medulla of young and old rats are responsive to glucose deficit and capable to restore the blood glucose level by increasing the levels of blood catecholamines. The present findings also suggest that, at least in rats, aging alters the protein expression of the biosynthetic catecholamine enzymes as well as protein kinase substrates that may attenuate the response to glucoprivation.


CRABP2 Is Associated With Thyroid Cancer Recurrence and Promotes Invasion via the Integrin/FAK/AKT Pathway.
Cellular retinoic acid-binding protein 2 (CRABP2) participates in retinoid partitioning between different nuclear receptors. Recently, we identified that CRABP2 is one of the progression-associated genes in thyroid cancer. To explore the prognostic and functional significance of CRABP2, immunohistochemical analysis was performed in thyroid tissues and neoplasms. Overexpression of CRABP2 was observed in malignant thyroid neoplasms but not in benign thyroid lesions. CRABP2 expression was an independent predictive factor for recurrence-free survival in patients with differentiated thyroid cancer. Knockdown of CRABP2 reduced the sensitivity of thyroid cancer cells to retinoic acid. Importantly, CRABP2 expression in thyroid cancer cells was associated with epithelial-mesenchymal transition properties, including anoikis resistance, migration, and invasion capacity. Furthermore, invasion promoted by CRABP2 was mediated at least partly by the integrin/focal adhesion kinase/AKT pathway. In summary, CRABP2 expression is upregulated in thyroid cancer with adverse prognostic implications. The invasion-stimulating effects appear independent of canonical retinoic acid signaling and may serve as a potential therapeutic target.


Sequential changes of bile contents in patients with obstructive jaundice from different etiologies.
The decreasing of serum concentration of bilirubin and the ability of hepatocytes to excrete various biliary contents after release of obstructive jaundice are good indicators of recovery of liver function. We conducted this study to clarify whether different causes of obstructive jaundice have different effects on the biliary excretion and how they are different when obstruction is released. Fifteen patients with obstructive jaundice undergoing percutaneous transhepatic catheter drainage or endoscopic nasobiliary drainage were classified into two groups, depending on the cause of obstruction: common bile duct stones (n = 7) and biliary tract tumors (n = 8). All patients in the gallstone group presented with acute cholangitis while only three patients in the tumor group had a positive bacteria culture in bile. Fasting biles were collected on the day of catheter placement and the 2nd, 4th, 6th, 8th, 10th day thereafter. The sequential changes of biliary concentration of bilirubin, phospholipid, cholesterol, bile salts and serum bilirubin were checked and compared between the two groups. The difference in the improvement of jaundice between the stone and tumor group (p > 0.05) were not significant, nor were the excretion of biliary contents after relief of obstruction. The reduction of serum bilirubin paralleled with the increased excretion of biliary bile salts and bilirubin (gamma = -0.51, p < 0.01 and gamma = -0.4, p < 0.05) in tumor group, but not in the stone group. The decrease of serum bilirubin and the sequential changes of bile contents after relief of obstruction are quite similar in stone and tumor induced obstructive jaundice.


Differing psychosocial risk profiles of college freshmen waterpipe, cigar, and cigarette smokers.
Few studies have examined the psychosocial aspects of tobacco smoking in young adults, particularly among alternative forms such as waterpipe. To address this gap, we examined the association of psychosocial characteristics (i.e., sociodemographics, risk perception, social norms, and pluralistic ignorance) with waterpipe, cigar, and cigarette smoking in college freshmen. Data are from a cross-sectional internet survey conducted during spring semester 2004 at Johns Hopkins University, N=411. Multinomial logistic regression was used to determine the association between psychosocial risk factors and waterpipe, cigar, and cigarette smoking. Results reveal that (1) psychosocial risk profiles of smokers differed by type of smoker and by type of tobacco product smoked, and (2) freshmen perceived the waterpipe as the most attractive product, out of the three products evaluated, to use among their peers. This study provides some of the first data on the association of psychosocial characteristics and various forms of tobacco smoking in young adults. This area of research is of increasing importance as a surge of waterpipe use among college students is becoming evident and interventions to reduce and prevent use are critically needed.


Clustering of Largely Right-Censored Oropharyngeal Head and Neck Cancer Patients for Discriminative Groupings to Improve Outcome Prediction.
Clustering is the task of identifying groups of similar subjects according to certain criteria. The AJCC staging system can be thought as a clustering mechanism that groups patients based on their disease stage. This grouping drives prognosis and influences treatment. The goal of this work is to evaluate the efficacy of machine learning algorithms to cluster the patients into discriminative groups to improve prognosis for overall survival (OS) and relapse free survival (RFS) outcomes. We apply clustering over a retrospectively collected data from 644 head and neck cancer patients including both clinical and radiomic features. In order to incorporate outcome information into the clustering process and deal with the large proportion of censored samples, the feature space was scaled using the regression coefficients fitted using a proxy dependent variable, martingale residuals, instead of follow-up time. Two clusters were identified and evaluated using cross validation. The Kaplan Meier (KM) curves between the two clusters differ significantly for OS and RFS (p-value < 0.0001). Moreover, there was a relative predictive improvement when using the cluster label in addition to the clinical features compared to using only clinical features where AUC increased by 5.7% and 13.0% for OS and RFS, respectively.


Participation of patients 65 years of age or older in cancer clinical trials.
Although 61% of new cases of cancer occur among the elderly, recent studies indicate that the elderly comprise only 25% of participants in cancer clinical trials. Further investigation into the reasons for low elderly participation is warranted. Our objective was to evaluate the participation of the elderly in clinical trials sponsored by the National Cancer Institute (NCI) and assess the impact of protocol exclusion criteria on elderly participation. We conducted a retrospective analysis using NCI data, analyzing patient and trial characteristics for 59,300 patients enrolled onto 495 NCI-sponsored, cooperative group trials, active from 1997 through 2000. Our main outcome measure was the proportion of elderly patients enrolled onto cancer clinical trials compared with the proportion of incident cancer patients who are elderly. Overall, 32% of participants in phase II and III clinical trials were elderly, compared with 61% of patients with incident cancers in the United States who are elderly. The degree of underrepresentation was more pronounced in trials for early-stage cancers than in trials for late-stage cancers (P <.001). Furthermore, protocol exclusion criteria on the basis of organ-system abnormalities and functional status limitations were associated with lower elderly participation. We estimate that if protocol exclusions were relaxed, elderly participation in cancer trials would be 60%. The elderly are underrepresented in cancer clinical trials relative to their disease burden. Older patients are more likely to have medical histories that make them ineligible for clinical trials because of protocol exclusions. Insurance coverage for clinical trials is one step toward improvement of elderly access to clinical trials. Without a change in study design or requirements, this step may not be sufficient.


Coffee consumption and circulating B-vitamins in healthy middle-aged men and women.
Coffee consumption has been associated with several risk factors for coronary heart disease, including increased cholesterol, increased blood pressure, and increased plasma total homocysteine (tHcy). tHcy is determined by several B-vitamins. However, reports about the association between coffee intake and B-vitamin status are few. We measured plasma B-vitamins and tHcy in a cohort of 10,601 healthy, middle-aged Norwegian men and women. Information about lifestyle factors, including coffee consumption, smoking, alcohol use, height, and weight, was obtained by interview. Coffee consumption was dose-dependently associated with reduced plasma B-vitamin concentrations. Compared with coffee abstainers, individuals drinking >or=4 cups/day had 11.7% (P < 0.001), 14.1% (P < 0.001), and 5.5% (P = 0.01) lower plasma concentrations of folate, pyridoxal phosphate, and riboflavin, respectively, and the mean tHcy concentration was 6.8% (P < 0.001) higher. Quantile regression analysis showed essentially no difference in B-vitamin concentrations between coffee consumption categories at low vitamin concentrations but a progressive increase in the difference at higher concentrations. This pattern of differences (effect profile) was found independently of smoking status, alcohol intake, and sex. The decrease in folate explained approximately half of the increase in tHcy. Coffee consumption was associated with reduced circulating B-vitamin concentrations. The observed effect profiles indicated that coffee consumption preferentially affected the upper, but not the lower, part of the B-vitamin concentration distributions. We hypothesize that coffee consumption may increase the loss of surplus B-vitamins by excretion in urine.


Spread through air spaces (STAS) in invasive mucinous adenocarcinoma of the lung: Incidence, prognostic impact, and prediction based on clinicoradiologic factors.
Spread through air spaces (STAS) has recently been demonstrated to exhibit a negative impact on lung adenocarcinoma prognosis. However, most of these studies investigated STAS in nonmucinous adenocarcinoma. Here, we investigated the incidence of STAS in invasive mucinous adenocarcinoma (IMA) of the lung and evaluated whether tumor STAS was a risk factor of disease recurrence in IMA. We also examined clinicoradiologic factors in patients with IMA harboring STAS. We reviewed pathologic specimens and imaging characteristics of primary tumors from 132 consecutive patients who underwent surgical resection for IMA to evaluate STAS. Patients with and without STAS were compared with respect to clinical characteristics as well as computed tomography (CT) imaging using logistic regression. The relationships between all variables including STAS and survival were analyzed. Among a total of 132 patients, full pathologic specimens were available for 119 patients, and STAS was observed in 86 (72.3%). IMA patients with STAS were significantly associated with older age, presence of lobulated and spiculated margins on CT scan (P = 0.009, P = 0.006, and P = 0.027). In multivariate analysis for overall survival (OS), STAS was a borderline independent poor prognostic predictor (P = 0.028). Older age, history of smoking, higher T stage, presence of lymph node metastasis, and consolidative morphologic type remained independent predictors for OS. STAS was associated with reduced OS and was a borderline independent poor prognostic factor in IMA. IMA with STAS was associated with older age and presence of lobulated and spiculated margins on CT scan. SIGNIFICANT FINDINGS OF THE STUDY: Compared with other subtypes, IMA shows a higher incidence of STAS, which is an independent poor prognostic predictor even in IMA. Lobulated and spiculated margins on CT are associated with STAS. Considering that STAS can carry the potential for aerogenous metastasis, predicting STAS using preoperative surrogate CT imaging is desirable to avoid limited resection.


Preparation of Primary Rat Hepatocyte Spheroids Utilizing the Liquid-Overlay Technique.
Herein, we describe a protocol for the preparation and analysis of primary isolated rat hepatocytes in a 3D cell culture format described as spheroids. The hepatocyte cells spontaneously self-aggregate into spheroids without the need for synthetic extracellular matrices or hydrogels. Primary rat hepatocytes (PRHs) are a readily available source of primary differentiated liver cells and therefore conserve many of the required liver-specific functional markers, and elicit the natural in vivo phenotype when compared with common hepatic cells lines. We describe the liquid-overlay technique which provides an ultra-low attachment surface on which PRHs can be cultured as spheroids. © 2019 The Authors. Basic Protocol 1: Preparation of agarose-coated plates Basic Protocol 2: Primary rat hepatocyte isolation procedure Basic Protocol 3: Primary rat hepatocyte spheroid culture Basic Protocol 4: Immunofluorescent analysis of PRH spheroids.


Evaluation of Ixodes scapularis, Amblyomma americanum, and Dermacentor variabilis (Acari: Ixodidae) from Georgia as vectors of a Florida strain of the Lyme disease spirochete, Borrelia burgdorferi.
The ability of three common tick species from Georgia to maintain and transmit the causative agent of Lyme disease, Borrelia burgdorferi, was compared under laboratory conditions. A B. burgdorferi cotton rat isolate (MI-6) from Florida was selected as a strain from the south, and the SH2-82 isolate from New York was used as a positive control. Amblyomma americanum (L.) and Dermacentor variabilis (Say) did not transmit the MI-6 isolate from inoculated hamsters to naive laboratory mice, and nymphal ticks did not maintain this isolate transstadially. Ixodes scapularis Say transmitted both isolates to laboratory mice. There was a significant difference in the percentage of transmission of the two isolates. I. scapularis also transmitted the MI-6 isolate to two of three cotton rats. This study adds support to the premise that I. scapularis is probably the main tick vector of B. burgdorferi in the southeastern United States.


RNA editing of the human serotonin 5-HT2C receptor. alterations in suicide and implications for serotonergic pharmacotherapy.
RNA encoding the human serotonin 5-HT2C receptor (5-HT(2C)R) undergoes adenosine-to-inosine RNA editing events at five positions, resulting in an alteration of amino acids in the second intracellular loop. Several edited 5-HT(2C)Rs possess a reduced G-protein coupling efficiency compared to the completely non-edited isoform. The current studies show that the efficacy of the hallucinogenic drug lysergic acid diethylamide and of antipsychotic drugs is regulated by RNA editing, suggesting that alterations in editing efficiencies or patterns might result in the generation of a 5-HT(2C)R population differentially responsive to serotonergic drugs. An examination of the efficiencies of RNA editing of the 5-HT(2C)R in prefrontal cortex of control individuals vs. subjects diagnosed with schizophrenia or major depressive disorder revealed no significant differences in RNA editing among the three populations. However, subjects who had committed suicide (regardless of diagnosis) exhibited a statistically significant elevation of editing at the A-site, which is predicted to change the amino acid sequence in the second intracellular loop of the 5-HT(2C)R. These findings suggest that alterations in RNA editing may contribute to or complicate therapy in certain psychiatric disorders.


Comparison study of CT and positron emission tomographic data in recent cerebral infarction.
To better understand the pathophysiologic correlates of the computed tomographic (CT) scan changes seen in recent cerebral infarction, 17 patients (20 studies) underwent both x-ray transmission and positron emission CT investigations within 18 days after clinical onset of complicated ischemic stroke in the internal carotid artery territory. The density changes before and after contrast study measured within the CT lesion were correlated to the local cerebral blood flow (CBF), oxygen utilization (CMRO2), and oxygen extraction fraction (OEF) measured with the oxygen-15 steady-state positron technique. Statistically significant linear correlations were found between hypodensity and CBF, hypodensity and CMRO2, and contrast enhancement and CBF, such that the more CBF and CMRO2 were depressed, the more marked was the hypodensity; and the more CBF was elevated, the more marked was the contrast enhancement. Although marked contrast enhancement was associated with decreased OEF (luxury perfusion), it was only rarely associated with increased CBF. Various hypotheses are discussed to explain these findings.


Validation of masticatory function and related factors in maxillectomy patients based on the concept of "oral hypofunction": A retrospective cross-sectional study.
Purpose Although oral functions have been evaluated by various methods in maxillectomy patients, there exists substantial difficulty in the assessment procedures. The purpose of this study was to evaluate masticatory function, maximum occlusal force (MOF), and maximum tongue pressure (MTP) in maxillectomy patients objectively as per the "oral hypofunction" criteria, and examine the factors that correlate with masticatory function.Methods This retrospective cross-sectional study included 50 maxillectomy patients with maxillofacial prostheses (23 males and 27 females, median age, 72 years; interquartile range (IQR), 63.75-77). The records of masticatory function, MOF, and MTP as per the "oral hypofunction" criteria and their profiles (age, number of occlusal supports, and maxillary defect configuration) were reviewed. The number of patients who exceeded a threshold value of "oral hypofunction" and the effects of occlusal support and defect configuration on masticatory function and MOF were calculated. Multiple regression analysis was performed to assess the associations between masticatory function and other variables.Results The median value of masticatory function (114 mg/dL, IQR: 73-167.5) exceeded a threshold value, but not MOF (229.2 N, IQR: 110.2-419.6) and MTP (25.9 kPa, IQR: 21.4-29.0). The number of patients who exceeded each threshold value was 27 (masticatory function), 8 (MOF), and 12 (MTP), respectively. The influence of the number of occlusal supports, but not the defect configuration, was statistically significant. Multiple regression analysis revealed that MOF was independently related to masticatory function (P=0.042).Conclusions Within the limitations of this study, the median value of masticatory function in maxillectomy patients could exceed a threshold and MOF might be an independent predictor of masticatory function.


CO2 induced acute respiratory acidosis and brain tissue intracellular pH: a 31P NMR study in swine.
High concentration carbon dioxide (CO(2)) is used to promote pre-slaughter anaesthesia in swine and poultry, as well as short-lasting surgical anaesthesia and euthanasia in laboratory animals. Questions related to animal welfare have been raised, as CO(2) anaesthesia does not set in momentarily. Carbon dioxide promotes anaesthesia by lowering the intracellular pH in the brain cells, but the dynamics of the changes in response to a high concentration of CO(2) is not known. Based on (31)P NMR spectroscopy, we describe CO(2)-induced changes in intracellular pH in the brains of five pigs inhaling 90% CO(2) in ambient air for a period of 60 s, and compare the results to changes in arterial blood pH, P(CO2), O(2) saturation and HCO(3)(-) concentration. The intracellular pH paralleled the arterial pH and P(CO2) during inhalation of CO(2); and it is suggested that the acute reaction to CO(2) inhalation mainly reflects respiratory acidosis, and not metabolic regulation as for example transmembrane fluxes of H(+)/HCO(3)(-). The intracellular pH decreased to approximately 6.7 within the 60 s inhalation period, and the situation was metabolically reversible after the end of CO(2) inhalation. The fast decrease in intracellular pH supports the conclusion that high concentration CO(2) leads to anaesthesia soon after the start of inhalation.


The cytoskeleton of the electric tissue of Electrophorus electricus, L.
The electric eel Electrophorus electricus is a fresh water teleost showing an electrogenic tissue that produces electric discharges. This electrogenic tissue is distributed in three well-defined electric organs which may be found symmetrically along both sides of the eel. These electric organs develop from muscle and exhibit several biochemical properties and morphological features of the muscle sarcolema. This review examines the contribution of the cytoskeletal meshwork to the maintenance of the polarized organization of the electrocyte, the cell that contains all electric properties of each electric organ. The cytoskeletal filaments display an important role in the establishment and maintenance of the highly specialized membrane model system of the electrocyte. As a muscular tissue, these electric organs expresses actin and desmin. The studies that characterized these cytoskeletal proteins and their implications on the electrophysiology of the electric tissues are revisited.