EGassembler: online bioinformatics service for large-scale processing, clustering and assembling ESTs and genomic DNA fragments.
Expressed sequence tag (EST) sequencing has proven to be an economically feasible alternative for gene discovery in species lacking a draft genome sequence. Ongoing large-scale EST sequencing projects feel the need for bioinformatics tools to facilitate uniform EST handling. This brings about a renewed importance for a universal tool for processing and functional annotation of large sets of ESTs. EGassembler (http://egassembler.hgc.jp/) is a web server, which provides an automated as well as a user-customized analysis tool for cleaning, repeat masking, vector trimming, organelle masking, clustering and assembling of ESTs and genomic fragments. The web server is publicly available and provides the community a unique all-in-one online application web service for large-scale ESTs and genomic DNA clustering and assembling. Running on a Sun Fire 15K supercomputer, a significantly large volume of data can be processed in a short period of time. The results can be used to functionally annotate genes, to facilitate splice alignment analysis, to link the transcripts to genetic and physical maps, design microarray chips, to perform transcriptome analysis and to map to KEGG metabolic pathways. The service provides an excellent bioinformatics tool to research groups in wet-lab as well as an all-in-one-tool for sequence handling to bioinformatics researchers.


Roles of F-box proteins in human digestive system tumors (Review).
F-box proteins (FBPs), the substrate-recognition subunit of E3 ubiquitin (Ub) ligase, are the important components of Ub proteasome system (UPS). FBPs are involved in multiple cellular processes through ubiquitylation and subsequent degradation of their target proteins. Many studies have described the roles of FBPs in human cancers. Digestive system tumors account for a large proportion of all the tumors, and their mortality is very high. This review summarizes for the first time the roles of FBPs in digestive system tumorige-nesis and tumor progression, aiming at finding new routes for the rational design of targeted anticancer therapies in digestive system tumors.


The increased phosphorylation of ribosomal protein S6 in Arbacia punctulata is not a universal event in the activation of sea urchin eggs.
Eggs of the sea urchins Arbacia punctulata (Ap), Lytechinus pictus (Lp), and Strongylocentrotus purpuratus (Sp) were labeled to equilibrium with 32PO3-4. Approximately 65-70% of the label in extractable adenine nucleotides comigrates chromatographically with ATP. Autoradiograms of one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) slab gels show that each species possesses a distinct complement of phosphate-exchangeable phosphoproteins. No changes in the phosphoprotein composition are detected in Lp and Sp eggs as a result of fertilization or development for 2.5 hr (with the possible exception of a 43,000 Mr protein in Lp). In Ap, increases in the phosphorylation of bands at Mr's 30,000, 55,000, and 105,000 are seen during the first 10 min postinsemination. The 30,000 Mr band in Ap eggs has previously been identified as ribosomal protein S6 and the hypothesis presented that its increased phosphorylation may be an important step in the activation of protein synthesis at fertilization (D. G. Ballinger and T. Hunt, 1981, Dev. Biol. 87, 277-285). In Lp and Sp eggs S6 (identified by two-dimensional PAGE) is heavily phosphorylated in the unfertilized state and the extent of labeling does not increase after fertilization. If the increased phosphorylation of S6 seen in Ap is indeed related to translational activation, then these results suggest that different sea urchin species may rely on different mechanisms for the activation of protein synthesis.


A validation study of a new nasogastric polyfunctional catheter.
Pleural and abdominal pressure are clinically estimated by measuring the esophageal and bladder or intragastric pressure (IGP), respectively. A new nasogastric polyfunctional catheter is now commercially available, equipped with two balloons in the lower and distal part; this catheter allows simultaneous esophageal pressure (Pes) and IGP measurements and can be also used to feed the patient. We compared the Pes and IGP measured using this new device with those obtained with a standard balloon catheter taken as gold standard. Twenty-four intubated patients requiring ventilator support (mean age 64.3 ± 16.8 years, body mass index 25.3 ± 3.0 kg/m(2), and PaO(2)/FiO(2) 280.8 ± 123.4 mmHg) were enrolled. Esophageal pressure and IGP were measured with the new nasogastric polyfunctional catheter (Nutrivent, Sidam, Italy) and with a standard balloon catheter (Smart Cath Viasys, USA). The Smart Cath was first inserted in the stomach and then retracted to the esophagus to measure IGP and Pes, respectively. In each patient two paired measurements were averaged. In the Bland-Altman analysis, the bias and agreement bands for Pes, ΔPes (computed as the difference of esophageal pressure between end-inspiration and expiration), and IGP were -0.25 (-2.65 to +2.15), 0.0 (-0.9 to +0.9), and -0.45 (-2.85 to + 1.95) cmH(2)O, respectively. No side effects or complications were recorded. The new polyfunctional catheter showed a clinically acceptable validity in recording esophageal and intragastric pressure. This device should help physicians to better individualize the clinical patient management.


Influence of short-term silicon application on endogenous physiohormonal levels of Oryza sativa L. under wounding stress.
The current study was conducted in order to investigate the short-term effects (6, 12, and 24 h) of silicon (Si) on the endogenous hormonal composition of rice (Oryza sativa L. cv. Dongjin-beyo), with and without wounding stress. Si applied in different concentrations (0.5, 1.0, and 2.0 mM) significantly promoted shoot length, plant biomass, and chlorophyll content of rice plants. Plants treated with different concentrations of sole Si for 6, 12, and 24 h had higher endogenous jasmonic acid contents than control. However, a combined application of wounding stress and Si induced a significantly small quantity of endogenous jasmonic acid as compared with control. On the contrary, endogenous salicylic acid level was significantly higher in sole Si-treated plants, while after wounding stress, a similar trend was observed yet again. After 6, 12, and 24 h of Si applications, with and without wounding stress, ethylene levels were significantly lower in comparison to their respective controls. The findings of the present study perpetrate the beneficial role of Si on the growth and development of rice plant by relieving physical injury and stress. Si also affects endogenous jasmonic acid and ethylene levels, while an inverse correlation exists between jasmonic acid and salicylic acid under wounding stress conditions.


Awakening p53 in vivo by D-peptides-functionalized ultra-small nanoparticles: Overcoming biological barriers to D-peptide drug delivery.
Peptides are a rapidly growing class of therapeutics with many advantages over conventional small molecule drugs. Dextrorotary (D)-peptides, with increased enzymatic stability and prolonged plasma half-life in comparison with natural L-peptides, are considered to have great potential as recognition molecules and therapeutic agents. However, the in vivo efficacy of current therapeutic D-peptides is hindered by their inefficient cellular uptake in diseased tissues. Methods: To overcome physiological and cellular barriers to D-peptides, we designed a gold-based ultra-small nanocarrier coupled with polylysine (PLL) and a receptor-targeted peptide to deliver therapeutic D-peptides. Using a D-peptide p53 activator (DPA) as a proof of concept, we synthesized, functionalized and characterized gold- and DPA-based nanoparticles termed AuNP-DPA. Results: AuNP-DPA were effectively enriched in tumor sites and subsequently internalized by cancer cells, thereby suppressing tumor growth via reactivating p53 signaling. More importantly, through a series of in vivo experiments, AuNP-DPA showed excellent biosafety without the common side effects that hinder p53 therapies in clinic trials. Conclusion: The present study not only sheds light on the development of AuNP-DPA as a novel class of antitumor agents for drugging the p53 pathway in vivo, but also supplies a new strategy to use D-peptides as intracellular PPI inhibitors for cancer-targeted therapy.


Development of a microculture system for stimulation of chicken peripheral blood lymphocytes with concanavalin A.
A microculture system in conjunction with a semiautomatic multiple sample harvester (SAMSH) was used to study the in vitro properties of chicken peripheral lymphocytes. This new procedure enabled doing rapid multiple tests, using relatively few cells, and was highly reproducible. Data were presented to show many variables that are involved in studying the concanavalin A (Con A) response of chicken lymphocytes in a microculture system. Analysis indicated that the conditions for optimal Con A stimulation as measured by incorporation of 3H-TdR include: (a) use of 2 x 10(6) cells per culture in RPMI 1640 culture medium in the absence of any serum, (b) use of 0.4 mug of Con A per culture, (c) incubation at 37 degrees C for 72 hours, and (d) addition of 1 muCi of 3H-TdR to each culture 12 to 24 hours prior to termination. This technique could be used to monitor immunocompetence of the chicken.


Protective roles of bioactive peptides during ischemia-reperfusion injury: From bench to bedside.
Ischemia-reperfusion (I/R) is a well-known pathological condition which may lead to disability and mortality. I/R injury remains an unresolved and complicated situation in a number of clinical conditions, such as cardiac arrest with successful reanimation, as well as ischemic events in brain and heart. Peptides have many attractive advantages which make them suitable candidate drugs in treating I/R injury, such as low toxicity and immunogenicity, good solubility property, distinct tissue distribution pattern, and favorable pharmacokinetic profile. An increasing number of studies indicate that peptides could protect against I/R injury in many different organs and tissues. Peptides also face several therapeutic challenges that limit their clinical application. In this review, we present the mechanisms of action of peptides in reducing I/R injury, as well as further discuss modification strategies to improve the functional properties of bioactive peptides.


Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria.
The inability to transform many clinically important Gram-negative bacteria has hampered genetic studies addressing the mechanism of bacterial pathogenesis. This report describes the development and construction of a delivery system utilizing the broad-host-range transducing bacteriophage P1. The phagemids used in this system contain a P1 pac initiation site to package the vector, a P1 lytic replicon to generate concatemeric DNA, a broad-host-range origin of replication and an antibiotic-resistance determinant to select bacterial clones containing the recircularized phagemid. Phagemid DNA was successfully introduced by infection and stably maintained in members of the families Enterobacteriaceae (Escherichia coli, Shigella flexneri, Shigella dysenteriae, Klebsiella pneumoniae and Citrobacter freundii) and Pseudomonadaceae (Pseudomonas aeruginosa). In addition to laboratory strains, these virions were used successfully to deliver phagemids to a number of strains isolated from patients. This ability to deliver genetic information to wild-type strains raises the potential for use in antimicrobial therapies and DNA vaccine development.


Interaction of volatile organic compounds and underlying liver disease: a new paradigm for risk.
Occupational and environmental exposures to industrial chemicals are known to cause hepatotoxicity and liver injury, in humans and in animal models. Historically, research has focused on severe acute liver injury (e.g. fulminant liver failure) or endstage diseases (e.g. cirrhosis and HCC). However, it has become recently recognized that toxicants can cause more subtle changes to the liver. For example, toxicant-associated steatohepatitis, characterized by hepatic steatosis, and inflammation, was recently recognized in an occupational cohort exposed to vinyl chloride. At high occupational levels, toxicants are sufficient to cause liver damage and disease even in healthy subjects with no comorbidities for liver injury. However, it is still largely unknown how exposure to toxicants initiate and possibly more importantly exacerbate liver disease, when combined with other factors, such as underlying non-alcoholic fatty liver disease caused by poor diet and/or obesity. With better understanding of the mechanism(s) and risk factors that mediate the initiation and progression of toxicant-induced liver disease, rational targeted therapy can be developed to better predict risk, as well as to treat or prevent this disease. The purpose of this review is to summarize established and proposed mechanisms of volatile organic compound-induced liver injury and to highlight key signaling events known or hypothesized to mediate these effects.


Forecasting generation of urban solid waste in developing countries--a case study in Mexico.
Based on a study of the composition of urban solid waste (USW) and of socioeconomic variables in Morelia, Mexico, generation rates were estimated. In addition, the generation of residential solid waste (RSW) and nonresidential solid waste (NRSW) was forecasted by means of a multiple linear regression (MLR) analysis. For residential sources, the independent variables analyzed were monthly wages, persons per dwelling, age, and educational level of the heads of the household. For nonresidential sources, variables analyzed were number of employees, area of facilities, number of working days, and working hours per day. The forecasted values for residential waste were similar to those observed. This approach may be applied to areas in which available data are scarce, and in which there is an urgent need for the planning of adequate management of USW.


Ultrasonographic characteristics of active ocular toxoplasmosis.
To evaluate ophthalmic ultrasonographic findings associated with active ocular toxoplasmosis. Forty-seven eyes with active ocular toxoplasmosis in 47 patients were subjected to ocular ultrasonography using the transpalpebral technique (10-MHz transducer) and fundus photography. Patient medical records were retrospectively reviewed. Ocular ultrasonography revealed vitritis, posterior vitreous detachment, retinal wall thickening, and non-rhegmatogenous retinal detachment in 47 (100%), 36 [76.6%; partial in 12 (25.5%) and total in 23 (48.9%)], 12 (25.5%), and 5 eyes (10.6%). Thirty-five of the 36 eyes with posterior vitreous detachment (97.2%) exhibited posterior hyaloid thickening; moreover, adhesion to the exudative lesion and vitreoschisis were observed in 4 (11.1%) and 12 eyes (25.5%), respectively. Ultrasonography detected the location of the exudative focus in 12 eyes (25.5%). Ultrasonography is helpful for detecting important intraocular findings of acute ocular toxoplasmosis that can be hindered by medial opacity or posterior synechiae.


EEG alpha rhythms and transient chromatic and achromatic pattern visual evoked potentials in children and adults.
Transient chromatic pattern visual evoked potentials (VEPs) have been found to be less repeatable in morphology in children than in adults at low to moderate chromatic contrasts. The purpose of this study is to investigate whether low repeatability of VEP components can be associated with high alpha power, in a comparison of alpha activity in children and adults. Transient chromatic contrast and achromatic resolution VEPs were recorded in children (n = 14, mean 9.6 years) and adults (n = 12, mean 21.8 years) with normal vision and assessed for repeatability. Isoluminant chromatic (magenta-cyan) and luminance-modulated achromatic grating stimuli were presented at and above psychophysical threshold levels, in pattern onset-offset at 2 Hz temporal frequency. EEGs (eyes closed and open) were recorded as single sweeps (1 s long) over three 30 s periods while facing a uniform computer display. An index of VEP detectability by observation was developed based on VEP component repeatability. The index was examined for correlations with alpha-wave parameters. Alpha power was calculated as the sum of the powers of 8-13 Hz frequencies of the EEG sweeps (using the discrete Fourier transform). Alpha power variability was calculated using the standard deviation of the powers of each sweep in a 30 s time period. The children had significantly higher alpha powers than the adults for both the eyes-open and eyes-closed conditions. Alpha power variability was significantly higher for the eyes-open condition only. There was no relationship between alpha power parameters and index of VEP detectability by observation for both the chromatic and achromatic grating stimuli. Poor repeatability of transient pattern VEPs is not associated with high alpha power or its variability in EEG measurements in older children or young adults at Oz.


Improving maternal update rates within the first hour of NICU admission.
Timely communication is essential in attaining maternal satisfaction, developing an excellent physician-patient rapport, and increasing trust. This study reports a significant improvement in maternal communication rates through the quality improvement method. An educational module was developed, and NICU staff was presented with the slides, followed by a performance questionnaire to demonstrate understanding. The first phase was completed by obtaining feedback from mothers through a questionnaire. The first plan-do-study-act (PDSA) cycle, carried out for eight weeks looking at the rates of the maternal update provided within an hour of admission of their neonates to the NICU, was followed by the second PDSA cycle, carried out for ten weeks. The improvement was calculated using conventional statistics and a statistical process control chart. During the first phase of the study, thirty-six percent of the mothers were updated within an hour of admission of their neonates to the NICU. During the first PDSA cycle, we did not notice a special cause variation or process change. A significant shift, eight consecutive points above the mean, was noted on the control chart during PDSA cycle 2. The mean±SD of the weekly update rate increased significantly during PDSA cycle 2 (76.8±11) compared to PDSA cycle 1 (47.5±14), p-value = 0.0002. We improved the maternal update rates through the educational module following the QI improvement model using the PDSA cycles.


Cerebral palsy in the surviving twin associated with infant death of the co-twin.
Monozygotic twins are at greater risk of dying and of serious morbidity than dizygotic twins, and both are at greater risk than singletons. This is only partly explained by the higher proportion of low birthweight infants among twins. To compare, in same sex and different sex twins, birth weight specific neonatal death rates and cerebral palsy prevalence rates in the surviving twin when the co-twin has died in infancy. Analysis of birth and death registration data for same sex and different sex twins for England and Wales 1993-1995 where both were live births. Death certificates of all liveborn twins who died were obtained from the Office for National Statistics. A questionnaire was sent to the general practitioners of all surviving co-twins to determine if the child had any disability. The neonatal death rate in same sex twins was 25.4 and in different sex twins 18.0 per 1000 live births (death rate difference 7.4; 95% confidence interval 4.7 to 10.1; p < 0.001). The higher neonatal death rate in same sex compared with different sex twins is attributable to the higher proportion of same sex twins with low birth weight. Prevalence of cerebral palsy in the low birthweight group (< 1000 g) was marginally higher in same sex (224 per 1000) than different sex (200 per 1000) twin survivors. In the birth weight group 1000-1999 g, same sex twin survivors were at a significantly higher risk of cerebral palsy than those of different sex: 167 v 21 per 1000; difference 145 (95% confidence interval 44 to 231; p < 0.01) per 1000 infant survivors. There are two components to the cause of cerebral palsy in twins. Immaturity per se predisposes to cerebral damage. Also, same sex twins may sustain cerebral damage that is in excess of that due to immaturity.


Age and psoriatic arthritis are important predictors of biologic agent switch in psoriasis.
The demand of biologic switching is increasing for different reasons.We aimed to define reasons for switching biologics and possible predictors for switching risk,and to estimate data on drug survival. 115 patients treated with biologics who switched to a second, third,and/or fourth biologic were eligible for this retrospective study.Patients were divided into 2 groups as switched once,and switched twice or more.Drug survival rates were calculated using the Kaplan-Meier method. All patients switched at least one, 36 patients switched twice, and 9 switched thrice. First-, second-, and third-line biologics were mostly switched due to secondary lack of efficacy for skin disease.Each unit increase in age decreased the risk of having ≥2 switches 4% (p=0.038,OR:0.964,95%CI:0.93-0.998),whereas PsA increased the risk of having ≥2 switches 2.69-fold (p=0.026,OR:2.69,95%CI:1.12-6.44).There was significant difference between biologics in terms of drug survival(p=0.001).Adalimumab had a lower drug survival compared to ustekinumab(p<0.001) and secukinumab(p=0.003) in transition from second-line biologic to third-line biologic. Switching biologics was most commonly due to secondary lack of efficacy for skin disease.Lower ages and the presence of PsA were associated with a higher need for switching in long-term.Ustekinumab and secukinumab are superior to adalimumab in clinical practice in terms of drug survival of second-line biologics.


Copper accumulation and the effect of chelation treatment on cerebral amyloid angiopathy compared to parenchymal amyloid plaques.
Accumulation of fibrillar amyloid β-protein (Aβ) in parenchymal plaques and in blood vessels of the brain, the latter condition known as cerebral amyloid angiopathy (CAA), are hallmark pathologies of Alzheimer's disease (AD) and related disorders. Cerebral amyloid deposits have been reported to accumulate various metals, most notably copper and zinc. Here we show that, in human AD, copper is preferentially accumulated in amyloid-containing brain blood vessels compared to parenchymal amyloid plaques. In light of this observation, we evaluated the effects of reducing copper levels in Tg2576 mice, a transgenic model of AD amyloid pathologies. The copper chelator, tetrathiomolybdate (TTM), was administered to twelve month old Tg2576 mice for a period of five months. Copper chelation treatment significantly reduced both CAA and parenchymal plaque load in Tg2576 mice. Further, copper chelation reduced parenchymal plaque copper content but had no effect on CAA copper levels in this model. These findings indicate that copper is associated with both CAA deposits and parenchymal amyloid plaques in humans, but less in Tg2576 mice. TTM only reduces copper levels in plaques in Tg2576 mice. Reducing copper levels in the brain may beneficially lower amyloid pathologies associated with AD.


Pumping the brakes on RNA velocity by understanding and interpreting RNA velocity estimates.
RNA velocity analysis of single cells offers the potential to predict temporal dynamics from gene expression. In many systems, RNA velocity has been observed to produce a vector field that qualitatively reflects known features of the system. However, the limitations of RNA velocity estimates are still not well understood. We analyze the impact of different steps in the RNA velocity workflow on direction and speed. We consider both high-dimensional velocity estimates and low-dimensional velocity vector fields mapped onto an embedding. We conclude the transition probability method for mapping velocity estimates onto an embedding is effectively interpolating in the embedding space. Our findings reveal a significant dependence of the RNA velocity workflow on smoothing via the k-nearest-neighbors (k-NN) graph of the observed data. This reliance results in considerable estimation errors for both direction and speed in both high- and low-dimensional settings when the k-NN graph fails to accurately represent the true data structure; this is an unknown feature of real data. RNA velocity performs poorly at estimating speed in both low- and high-dimensional spaces, except in very low noise settings. We introduce a novel quality measure that can identify when RNA velocity should not be used. Our findings emphasize the importance of choices in the RNA velocity workflow and highlight critical limitations of data analysis. We advise against over-interpreting expression dynamics using RNA velocity, particularly in terms of speed. Finally, we emphasize that the use of RNA velocity in assessing the correctness of a low-dimensional embedding is circular.


Role of AT(1) receptors and autonomic nervous system in mediating acute pressor responses to ANG II in anesthetized mice.
Hemodynamic responses to angiotensin II and the role of AT(1) and AT(2) receptors and the autonomic nervous system in mediating acute responses to angiotensin II were investigated in anesthetized CD1 mice. Injections of angiotensin II caused dose-related increases in systemic arterial pressure that were antagonized by candesartan. Pressor responses to angiotensin II were not altered by PD-123,319 in doses up to 25 mg/kg iv. At the lowest dose studied (20 microgram/kg iv), the inhibitory effects of candesartan were competitive, whereas at the highest dose (100 microgram/kg iv) the dose-response curve for angiotensin II was shifted to the right in a nonparallel manner with inhibitory effects that could not be surmounted. The inhibitory effects of candesartan were selective and were similar in animals pretreated with enalaprilat (1 mg/kg iv) to reduce endogenous angiotensin II production. Acute pressor responses to angiotensin II were not altered by propranolol (200 microgram/kg iv), phentolamine (200 microgram/kg iv), or atropine (1 mg/kg iv) but were enhanced by hexamethonium (5 mg/kg iv). Increases in total peripheral resistance induced by angiotensin II were inhibited by the AT(1)-receptor antagonist but were not altered by AT(2)-, alpha-, or beta-receptor antagonists. These results suggest that acute pressor responses to angiotensin II are mediated by AT(1) receptors, are buffered by the baroreceptors, and are not modulated by effects on AT(2) receptors and that activation of the sympathetic nervous system plays little if any role in mediating rapid hemodynamic responses to the peptide in anesthetized CD1 mice.


Differential effects of adrenocorticotropic hormone on human and mouse adipose tissue.
Adrenocorticotropic hormone (ACTH) induces lipolysis in a dose-dependent fashion in rodent adipose tissue and adipocytes in vitro. The role of ACTH on lipolysis in human adipose tissue is less clear, however. In this study, we address the hypothesis that ACTH induces lipolysis in human adipose tissue. We used ex vivo organ culture to examine lipolysis in human and mouse adipose tissue. Adipose tissue fragments suspended in culture medium and human ACTH, isoproterenol (positive control), or insulin (negative control) was added in varying concentrations. Lipolysis was measured using glycerol appearance. ACTH receptor mRNA expression was assessed using reverse-transcription polymerase chain reaction (RT-PCR). In mouse adipose tissue, ACTH induced lipolysis in dose-dependent manner; 100 pmol/l ACTH induced 67+/-19% of isoproterenol-stimulated lipolysis and 500 pmol/l ACTH: 86+/-13%. In contrast, human adipose tissue shared no significant response to 100 pmol/l ACTH; ACTH was associated with 9+/-6% and 500 pmol/l of ACTH, 8+/-6% of isoproterenol-stimulated lipolysis. ACTH receptor mRNA was present in mouse adipose tissue, but undetectable in human adipose tissue. These results suggest lipolysis regulation differs between human and mouse adipose tissue in response to ACTH.


Lhermitte-Duclos disease in 3 children: a clinical long-term observation.
We report three boys in whom a diagnosis of Lhermitte-Duclos disease (LDD) was assumed from characteristic neuroimaging findings. LDD was confirmed by an open biopsy in patient 1, while a biopsy in patient 2 was inconclusive. Histologic confirmation in patient 3 was deliberately not attempted. However, a follow-up observation of stable clinical and neuroimaging findings over 2, 5 and 11 years, respectively, support the diagnosis of LDD. Despite extensive expansion of the lesion with brainstem involvement, clinical signs in two boys were minimal, while one patient has cognitive impairment and a complex oculomotor disturbance. So far we found no evidence for an association with Cowden disease (CD). No germline PTEN mutations were detected in these children, but the amount of available biopsy tissue in patients 1 and 2 was insufficient for a complete genetic analysis of tumor tissue. In conclusion, LDD can usually be diagnosed by MRI. In view of the favourable natural history, a conservative "wait and see" strategy is justified, particularly if radical tumor resection is not possible. LDD is often not associated with CD and germline PTEN mutations seem not to be present in isolated LDD.


Alemtuzumab for the treatment of relapsing-remitting multiple sclerosis.
Alemtuzumab, a humanized monoclonal antibody that targets CD52, was recently approved in the EU and Canada for the treatment of patients with active relapsing-remitting multiple sclerosis. Alemtuzumab induces rapid depletion of circulating B- and T-lymphocytes followed by repopulation that leads to a distinctive lymphocyte profile, including an increased proportion of regulatory T-lymphocytes and memory B- and T-lymphocytes. In early open-label studies, alemtuzumab treatment reduced the number of clinical relapses and new MRI lesions in participants with secondary progressive MS. However, most participants had continued worsening of disability, which led to the evaluation of alemtuzumab in patients with early stages of disease in the Genzyme (MA, USA)-sponsored clinical development program in MS. In one Phase II and two Phase III trials, alemtuzumab reduced the number of clinical relapses versus the active comparator, subcutaneous IFN-β-1a, in treatment-naive and treatment-experienced participants with relapsing-remitting multiple sclerosis. Two of these trials showed reduction in risk of confirmed worsening of disability, and all showed reduction in cerebral atrophy. Safety issues include infusion reactions that are mitigated by pretreatment with corticosteroids in addition to symptomatic management with antihistamines; mild to moderate infections; and autoimmune adverse events. In this context, post-marketing risk mitigation strategies will be needed so that potential adverse events can be identified and managed early and effectively.


Phenolic Derivatives with Anti-Acetylcholinesterase Inhibitory Activities from Galeola nudifolia in Vietnam.
A new phenolic derivative, galeomalate A (1), together with five known structurally related compounds (2-6), was isolated from the ethyl acetate extract of Galeola nudifolia collected in Vietnam. The structures were elucidated by various spectroscopic methods, including 1D and 2D NMR, HR-ESI-TOF-MS, and CD data, and chemical conversion of the sugar moiety. All isolated compounds possessed acetylcholinesterase (AChE) inhibitory activities in a dose-dependent manner. Among them, compounds 2 and 3 exhibited the first and second highest inhibitory activity on AChE with IC50 values of 122.13 and 125.49 μM, respectively. Compounds 1 and 4-6 inhibited the AChE activity by mixed modes of action comprising competitive and non-competitive modes, whereas 2 and 3 exerted their inhibitory activities in a competitive manner. Molecular docking analyses suggested that the phenyl-β-D-glucopyranoside unit of 2 and 3 bound to the active site of AChE for the competitive inhibitory activities, while the mixed inhibitory activity of 4 was due to the two binding patterns in the active-site and the active-site entrance of AChE. Furthermore, the docking studies indicated that 1, 5, and 6 would inhibit AChE in a mixed inhibitory manner by adopting three distinct binding patterns of the additional phenyl-β-D-glucopyranoside unit at the active-site entrance.


Increase in rat brain tyrosine hydroxylase activity produced by electroconvulsive shock.
A sustained increase in the turnover of norepinephrine coupled with an increased concentration of that amine has, in a previous study, been observed in the brains of rats exposed to electroconvulsive shock twice daily for one week. The phenomenon has been further examined by studying the effects of a similar regimen of electroconvulsive shock upon tyrosine hydroxylase in the brain, since that enzyme appears to be rate-limiting in norepinephrine synthesis. A small (15%) but significant increase in the whole brain enzyme activity was found in the electroshock-treated animals. A significant increase of tyrosine hydroxylase activity was found in the brainstem (24%) and cortex (20%) of such animals.


AIM2 inflammasome contributes to aldosterone-induced renal injury via endoplasmic reticulum stress.
Inflammatory response and renal fibrosis are the hallmarks of chronic kidney disease (CKD). However, the specific mechanism of aldosterone-induced renal injury in the progress of CKD requires elucidation. Emerging evidence has demonstrated that absent in melanoma 2 (AIM2)-mediated inflammasome activation and endoplasmic reticulum stress (ERS) play a pivotal role in the renal fibrosis. Here, we investigated whether overexpression or deficiency of AIM2 affects ERS and fibrosis in aldosterone-infused renal injury. Interestingly, we found that AIM2 was markedly expressed in the diseased proximal tubules from human and experimental CKD. Mechanically, overactivation of AIM2 aggravated aldosterone-induced ERS and fibrotic changes in vitro while knockdown of AIM2 blunted these effects in vivo and in vitro. By contrast, AIM2 deficiency ameliorated renal structure and function deterioration, decreased proteinuria levels and lowered systolic blood pressure in vivo; silencing of AIM2 blocked inflammasome-mediated signaling pathway, relieved ERS and fibrotic changes in vivo. Furthermore, mineralocorticoid receptor (MR) antagonist eplerenone and ERS inhibitor tauroursodeoxycholic acid (TUDCA) had nephroprotective effects on the basis of AIM2 overactivation in vitro, while they failed to produce a more remarkable renoprotective effect on the treatment of AIM2 silence in vitro. Notably, the combination of TUDCA with AIM2 knockdown significantly reduced proteinuria levels in vivo. Additionally, immunofluorescence assay identified that apoptosis-associated speck-like protein (ASC) recruitment and Gasdermin-D (GSDMD) cleavage respectively occurred in the glomeruli and tubules in vivo. These findings establish a crucial role for AIM2 inflammasome in aldosterone-induced renal injury, which may provide a novel therapeutic target for the pathogenesis of CKD.


Regulation of endothelial-dependent relaxation in human systemic arteries by SKCa and IKCa channels.
Blockade of small-conductance Ca (2)(+)-activated K(+) channels (SK(Ca)) and intermediate conductance Ca(2)(+)-activated K(+) channels (IK(Ca)) can cause inhibition of endothelium-dependent hyperpolarizing factor (EDHF) in many vascular beds from animals, but there is a relative paucity of data in human vessels. Systemic arteries, isolated from women with healthy pregnancies, relax to the endothelial-dependent agonist bradykinin via a nonprostacyclin and non-nitric oxide pathway attributable to EDHF. Therefore, in this study, the authors investigated the effect of pharmacological blockade of SK(Ca) and IK(Ca) on EDHF-mediated relaxation of human omental and myometrial arteries preconstricted with either arginine vasopressin or U46619. Human arteries were isolated from omental and myometrial biopsies taken from healthy women undergoing planned cesarean section at term. Endothelial function was assessed using wire myography. In all vessels examined, nonspecific blockade of IK(Ca) with charybdotoxin attenuated EDHF-attributed relaxation. However, when Tram 34 was used to block IK(Ca), the attenuation of relaxation was evident only with U46619 preconstriction. In arteries from both vascular beds, and with either preconstrictor, a combination of either apamin and charybdotoxin or apamin plus Tram 34 almost ablated EDHF-attributable relaxation. These data support the notion that in human systemic arteries, activation of, primarily, SK(Ca) and IK(Ca)K(+) channel subtypes underlies EDHF-mediated relaxation. These results have important implications for future studies ascertaining the molecular mechanisms of hypertensive disorders (eg, preeclampsia, in which EDHF is thought to be aberrant).


The biology of human natural killer cells.
The human natural killer (NK) cell has long been known to mediate spontaneous, non-major histocompatibility complex-restricted cytotoxicity and antibody-dependent cellular cytotoxicity. It is only fairly recently, however, that the role NK cells play in other functions of the immune system has been realized. The NK cell participates either directly or indirectly in multiple developmental, regulatory and communication networks of the immune system and thus is important in human health and disease. The NK cell has distinct morphologic, phenotypic and ultrastructural characteristics that distinguish it from T and B lymphocytes. Human NK cells are heterogeneous, and functionally different subpopulations of NK cells can be distinguished. NK activity may be regulated by soluble products of hematopoietic as well as non-hematopoietic cells and by a wide variety of exogenous biological response modifiers. Both lymphocytes and monocytes are capable of regulating of NK cell growth and activity. Populations enriched in human NK cells can be obtained utilizing the property of adherence to plastic and subsequent expansion in the presence of IL-2. The adherent lymphokine activated killer (A-LAK) cells represent populations of CD3-CD56+ (up to 98%) IL-2 activated NK cells. They are highly effective in eliminating tumor cells both in vitro and in vivo in animal models of tumor metastasis. A-LAK cells are being used for therapy in the phase I clinical trial in patients with metastatic melanoma and renal cell carcinoma. The studies described emphasize the biologic importance of the NK cell, its therapeutic potential, and a need for more extensive monitoring of NK activity in human disease.


Cytokine gene therapy or infusion as treatment for solid human cancer.
In the induction of tissue-directed immune responses, cytokines tend to be released within the affected tissues. We used two strategies to expose tumor tissues to continuous high levels of cytokines: First, a vaccinia interleukin (IL)2 recombinant was injected directly intratumorally 3-weekly at 10(7) pfus/dose in six patients with the solid tumor malignant mesothelioma (MM). No virus excretion was detectable. At each cycle vaccinia-IL-2 mRNA (SQ [semi-quantitative] reverse transcription polymerase chain reaction) was maximal 24-72 h following injection reduced at 8 days and faded by 21 days. No tumor regression occurred. Second, based on the success of granulocyte macrophage colony-stimulating factor (GM-CSF) in gene transfer experiments, we conducted a study using continuous intratumoral GM-CSF infusion in eight patients with MM using a portable pump at doses of 10 micro/cg/24 h over 8 weeks. Systemic neutrophil agglutination and local catheter-related difficulties occurred. Two patients demonstrated tumor necrosis, one of whom had a marked progressive mononuclear cell infiltration of the tumor associated with a partial response (>50% reduction in tumor area). Murine studies using our MM model in CBA and BALB/C mice have demonstrated that B7-1 and allo-class I transfections induce strong tumor-specific cytotoxic T lymphocyte responses: GM-CSF, IL-12, and IL-2 induced mixed nonspecific plus specific responses, whereas B7-2 and class II transfections were not effective. We conclude that increased intratumoral cytokine concentrations can be generated using both gene transfer and cytokine infusion approaches; however, both have their limitations and, at this stage, have not produced dramatic antitumor effects in humans.


Structural characterization of HC-Pro, a plant virus multifunctional protein.
The helper component proteinase (HC-Pro) is a key protein encoded by plant viruses of the genus Potyvirus. HC-Pro is involved in different steps of the viral cycle, aphid transmission, replication, and virus cell-to-cell and systemic movement and is a suppressor of post-transcriptional gene silencing. Structural knowledge of HC-Pro is required to better understand its multiple functions. To this aim, we purified His-tagged wild-type HC-Pro and a N-terminal deletion mutant (DeltaHC-Pro) from plants infected with recombinant potyviruses. Biochemical analysis of the recombinant proteins confirmed that HC-Pro is a dimer in solution, that the N terminus is not essential for self-interaction, and that a large C-terminal domain is highly resistant to proteolysis. Two-dimensional crystals of the recombinant proteins were successfully grown on Ni2+-chelating lipid monolayers. Comparison of projection maps of negatively stained crystals revealed that HC-Pro is composed of two domains separated by a flexible constriction. Cryo-electron crystallography of DeltaHC-Pro allowed us to calculate a projection map at 9-A resolution. Our data from electron microscopy, biochemical analysis, and secondary structure predictions lead us to suggest a model for structure/function relationships in the HC-Pro protein.


Stromal heparan sulfate differentiates neuroblasts to suppress neuroblastoma growth.
Neuroblastoma prognosis is dependent on both the differentiation state and stromal content of the tumor. Neuroblastoma tumor stroma is thought to suppress neuroblast growth via release of soluble differentiating factors. Here, we identified critical growth-limiting components of the differentiating stroma secretome and designed a potential therapeutic strategy based on their central mechanism of action. We demonstrated that expression of heparan sulfate proteoglycans (HSPGs), including TβRIII, GPC1, GPC3, SDC3, and SDC4, is low in neuroblasts and high in the Schwannian stroma. Evaluation of neuroblastoma patient microarray data revealed an association between TGFBR3, GPC1, and SDC3 expression and improved prognosis. Treatment of neuroblastoma cell lines with soluble HSPGs promoted neuroblast differentiation via FGFR1 and ERK phosphorylation, leading to upregulation of the transcription factor inhibitor of DNA binding 1 (ID1). HSPGs also enhanced FGF2-dependent differentiation, and the anticoagulant heparin had a similar effect, leading to decreased neuroblast proliferation. Dissection of individual sulfation sites identified 2-O, 3-O-desulfated heparin (ODSH) as a differentiating agent, and treatment of orthotopic xenograft models with ODSH suppressed tumor growth and metastasis without anticoagulation. These studies support heparan sulfate signaling intermediates as prognostic and therapeutic neuroblastoma biomarkers and demonstrate that tumor stroma biology can inform the design of targeted molecular therapeutics.


Using a system dynamics model to study the obesity transition by socioeconomic status in Colombia at the country, regional and department levels.
We study the obesity transition by socioeconomic status (SES), gender and age within the Colombian urban population at the country, regional and department levels. The study is informed by cross-sectional data from the 2005 and 2010 ENSIN survey. We used these data to develop a system dynamics model that simulates the dynamics of obesity by body mass index (BMI) categories, gender and SES at the country, regional and department levels from 2005 to 2030. The sample size of the 2005 ENSIN comprised 8515 children younger than 5 years, 32 009 children and adolescents aged 5-17 years and 48 056 adults aged 18-64 years. In 2010, the corresponding numbers were 11 368, 32 524 and 64 425, respectively. The obesity prevalence ratio and prevalence rates for each BMI category. The results show, at the country level, transitions from overweight to obesity were projected to increase sharply among lower SES adults, particularly among women, suggesting that these groups will undergo an obesity transition by 2030. The model projections also indicate that the regions of Colombia are in different stages of the obesity transition. In the case of women, five out of the six regions were expected to undergo an obesity transition by SES over time. For men, only one region was expected to undergo an obesity transition. However, at the department level, trends in the burden of obesity varied. We evidence that the Colombian population could be experiencing an obesity transition where the increase in the GDP could be related to shifts in the burden of obesity from higher to lower SES, especially in women. These patterns support the need for policy planning that considers SES and gender, at the national and subnational levels, as important determinants of overweight and obesity among adults in Colombia.


The DEAH-box RNA helicase RHAU binds an intramolecular RNA G-quadruplex in TERC and associates with telomerase holoenzyme.
Guanine-quadruplexes (G4) consist of non-canonical four-stranded helical arrangements of guanine-rich nucleic acid sequences. The bulky and thermodynamically stable features of G4 structures have been shown in many respects to affect normal nucleic acid metabolism. In vivo conversion of G4 structures to single-stranded nucleic acid requires specialized proteins with G4 destabilizing/unwinding activity. RHAU is a human DEAH-box RNA helicase that exhibits G4-RNA binding and resolving activity. In this study, we employed RIP-chip analysis to identify en masse RNAs associated with RHAU in vivo. Approximately 100 RNAs were found to be associated with RHAU and bioinformatics analysis revealed that the majority contained potential G4-forming sequences. Among the most abundant RNAs selectively enriched with RHAU, we identified the human telomerase RNA template TERC as a true target of RHAU. Remarkably, binding of RHAU to TERC depended on the presence of a stable G4 structure in the 5'-region of TERC, both in vivo and in vitro. RHAU was further found to associate with the telomerase holoenzyme via the 5'-region of TERC. Collectively, these results provide the first evidence that intramolecular G4-RNAs serve as physiologically relevant targets for RHAU. Furthermore, our results suggest the existence of alternatively folded forms of TERC in the fully assembled telomerase holoenyzme.


Identification key for coryneform bacteria derived by numerical taxonomic studies.
Six main groups were formed from a complete linkage dendrogram on 557 bacteria tested for 53 physiological features. The organisms were obtained from culture collections and included representatives of the following genera: Arthrobacter, Brevibacterium, Caseobacter, Cellulomonas, Corynebacterium, Curtobacterium, Micrococcus, Microbacterium, Mycobacterium, Nocardia, Oerskovia and Rhodococcus. The six groups were individually subjected to a numerical taxonomic analysis based on linkage maps, which resulted in a total of 33 subclusters. An identification key to determine the affiliation of the bacteria to the six main clusters and five group-specific schemes is presented. Reference strains are proposed for the 33 subclusters.


Sublingual desensitization in patients with wasp venom allergy: preliminary results.
The aim of this paper is to assess in an open prospective pilot case-control study the tolerability, safety and efficacy of an ultra-rush sublingual immunotherapy (SLIT) protocol with Vespula venom in wasp allergic patients compared to subcutaneous immunotherapy (SCIT). Forty-one wasp allergic patients were treated with sublingual (SLIT group) or subcutaneous (SCIT group) ultrarush immunotherapy with Vespula venom extract. All patients underwent skin tests and serum specific IgE and IgG4 detection before enrollment and after 6, 12 and 24 months of immunotherapy. The SLIT group consisted of 21 (6 females and 15 males) patients who received increasing doses of Vespula venom (Aquagen, ALK-Abellò) until the final dose of 30 drops of extract in 3 hours, containing 100,000 SQ-U/ml. The maintenance dose was of 10 drops of pure venom extract 3 times a week, for a total dose of 100,000 SQ-U weekly (corresponding to 100 microgram of venom extract). The SCIT group consisted of 20 patients (16 males and 4 females) who were treated with subcutaneous ultrarush immunotherapy with Vespula venom extract (Pharmalgen, Alk-Abellò). Patients received 101.1 microgram of Vespula venom in 3 hours and were treated with 100 microgram of wasp venom monthly. During the ultrarush sublingual treatment 2 patients (9.5%) experienced mild side-effects. Specific IgE and specific IgG to wasp venom did not show any significant modification. Four patients were field-stung by a wasp during the treatment (for a total of 6 stings). Two patients (3 stings), with a previous clinical history of a grade III and IV reaction, did not experience any reaction. One patient, with a previous grade II reaction, showed a large local reaction. The fourth patient, with a previous grade III reaction, was re-stung twice (after 12 and 24 months) with two systemic reactions (SR) (mild throat constriction). During the ultrarush SCIT phase, 3 (15%) patients experienced side-effects: 2 of them showed a large local reaction and 1 had headache and stomach ache. Specific IgE showed a significant (P = 0.001) increase after 6 months of treatment and then returned to baseline levels while specific IgG showed a significant (P = 0.001) increase after 6, 12 and 24 months in comparison with baseline. Nine patients were field-stung during the treatment: 8 of them experienced large local reactions; one patient (11%) experienced an SR (dizziness). Our results, even if in a small number of patients, suggest that in patients with Hymenoptera sting allergy SLIT could be efficacious with a good tolerability profile when compared to SCIT. Larger studies are needed to assess efficacy, safety and tolerability profile of wasp venom SLIT.


Modification of vascular function after handgrip exercise training in 73- to 90-yr-old men.
To examine the influence of a unilateral exercise training protocol on brachial artery reactivity (BAR) in 12 men (aged 81 +/- 5 yr). Brachial artery diameters and blood flow parameters were assessed, in both arms, using high-resolution ultrasonography, before and after 5 min of forearm occlusion, before and at the end of each week of a 4-wk training program. Training consisted of a unilateral handgrip training protocol (nondominant arm) at 60% of maximal voluntary handgrip strength, performed for 4 wk, 4 d x wk(-1), 20 min per session, and a cadence of one contraction per 4 s. After training, handgrip strength increased 6.2% (baseline = 32.4 +/- 7.0 kg vs week 4 = 34.4 +/- 6.7 kg) in the trained arm only but failed to reach statistical significance (P = 0.10). No statistical changes were observed for blood pressure or resting HR. In contrast, BAR increased 45% (Pre = 2.9% vs Post = 4.1%, P = 0.05) in the trained arm only. Improvements in BAR were observed after the second week of training, without significant changes in the main vasodilatory trigger, defined as the relevant shear stimulus after forearm occlusion (P > 0.05). These data indicate that a localized short-term exercise program results in significant improvements in vascular function in the trained arm of elderly men compared with the control arm. Furthermore, the findings indicate a statistically significant increase in BAR at the end of the second week of training, despite a similar trigger for dilation versus before training.


Localization of C-protein isoforms in chicken skeletal muscle: ultrastructural detection using monoclonal antibodies.
Monoclonal antibodies (McAbs) specific for the fast (MF-1) and slow (ALD-66) isoforms of C-protein from chicken skeletal muscle have been produced and characterized. Using these antibodies it was possible to demonstrate that skeletal muscles of varying fiber type express different isoforms of this protein and that in the posterior latissimus dorsi muscle both isoforms are co-expressed in the same myofiber (17, 18). Since we had shown that both isoforms were present in all sarcomeres, it was feasible to test whether the two isoforms co-distributed in the same 43-nm repeat within the A-band, thereby establishing a minimum number of C-proteins per repeat in the thick filaments. Here we describe the ultrastructural localization of C-protein in myofibers from three muscle types of the chicken using these same McAbs. We observed that although C-protein was present in a 43-nm repeat along the filaments in all three muscles, there were marked differences in the absolute number and position occupied by the different isoforms. Since McAbs MF-1 and ALD-66 decorated the same 43-nm repeats in the A-bands of the posterior latissimus dorsal muscle, we suggest that at least two C-proteins can co-localize at binding sites 43 nm apart along thick filaments of this muscle.


New therapeutic approach to persistent asthma.
The goal of asthma treatment has moved from achieving symptom relief alone to overall disease control. The latest guidelines propose that first-line treatment of mild persistent asthma requires the introduction of anti-inflammatory agents, preferably inhaled corticosteroids at low dose. In patients in whom control of disease is not achieved with this treatment, it was demonstrated that addition of a long-acting beta 2-agonist is more effective than increasing the dose of inhaled corticosteroid. Addition of a long-acting beta 2-agonist to an inhaled corticosteroid not only improves symptoms and lung function but also reduces the risk of asthma exacerbations, suggesting complementary therapeutic activity. Therefore, use of long-acting bronchodilators with inhaled corticosteroids is a prerequisite for optimal management of the disease. For this reason, the logical development in asthma management would be a combination of these two classes of drug in a single inhaled formulation. Fixed combinations of the long-acting beta 2-agonist salmeterol and the inhaled corticosteroid fluticasone propionate were launched recently. Results from recent clinical trials have demonstrated that the combination of salmeterol and fluticasone propionate improves control of asthma in the majority of patients with moderate-to-severe asthma and enhances compliance by virtue of both drugs being in the one inhaler. The possibility to treat asthma of differing severity is provided by the flexibility to choose between three different doses of fluticasone propionate in the fixed combination devices. Another obvious advantage of the combination is cost savings, as the fixed combination inhaler is cheaper than giving the two drugs separately. The introduction of combination preparations is an important step forward in asthma management, which is expected to be a popular choice with both patients and physicians.


Promoting universal vaccination among workers.
A discussion of vaccines often focuses on protecting children from vaccine-preventable diseases. However, 10 vaccines are available for adults, and occupational health nurses should consider these vaccines when providing health care to adult workers. This article discusses the 10 adult vaccines, indications for using each vaccine, barriers preventing adults from being vaccinated and occupational health nurses from increasing vaccination rates, and strategies nurses can use to promote adult vaccination. Promoting universal vaccination among adults, especially employees, accomplishes two goals. First, universal vaccination promotes a healthy work force, enabling employees to report to work regularly and be more productive while working. Second, vaccinating adults protects the most vulnerable members of the community, young children, those with immunodeficiencies, and those who cannot be vaccinated, via herd immunity.


Searching for nuclear export elements in hepatitis D virus RNA.
To search for the presence of cis elements in hepatitis D virus (HDV) genomic and antigenomic RNA capable of promoting nuclear export. We made use of a well characterized chloramphenicol acetyl-transferase reporter system based on plasmid pDM138. Twenty cDNA fragments corresponding to different HDV genomic and antigenomic RNA sequences were inserted in plasmid pDM138, and used in transfection experiments in Huh7 cells. The relative amounts of HDV RNA in nuclear and cytoplasmic fractions were then determined by real-time polymerase chain reaction and Northern blotting. The secondary structure of the RNA sequences that displayed nuclear export ability was further predicted using a web interface. Finally, the sensitivity to leptomycin B was assessed in order to investigate possible cellular pathways involved in HDV RNA nuclear export. Analysis of genomic RNA sequences did not allow identifying an unequivocal nuclear export element. However, two regions were found to promote the export of reporter mRNAs with efficiency higher than the negative controls albeit lower than the positive control. These regions correspond to nucleotides 266-489 and 584-920, respectively. In addition, when analyzing antigenomic RNA sequences a nuclear export element was found in positions 214-417. Export mediated by the nuclear export element of HDV antigenomic RNA is sensitive to leptomycin B suggesting a possible role of CRM1 in this transport pathway. A cis-acting nuclear export element is present in nucleotides 214-417 of HDV antigenomic RNA.


Genotype and phenotype of COVID-19: Their roles in pathogenesis.
COVID-19 is a novel coronavirus with an outbreak of unusual viral pneumonia in Wuhan, China, and then pandemic. Based on its phylogenetic relationships and genomic structures the COVID-19 belongs to genera Betacoronavirus. Human Betacoronaviruses (SARS-CoV-2, SARS-CoV, and MERS-CoV) have many similarities, but also have differences in their genomic and phenotypic structure that can influence their pathogenesis. COVID-19 is containing single-stranded (positive-sense) RNA associated with a nucleoprotein within a capsid comprised of matrix protein. A typical CoV contains at least six ORFs in its genome. All the structural and accessory proteins are translated from the sgRNAs of CoVs. Four main structural proteins are encoded by ORFs 10, 11 on the one-third of the genome near the 3'-terminus. The genetic and phenotypic structure of COVID-19 in pathogenesis is important. This article highlights the most important of these features compared to other Betacoronaviruses.


The comparative validity and reproducibility of a diet quality index for adults: the Australian Recommended Food Score.
Adult diet quality indices are shown to predict nutritional adequacy of dietary intake as well as all-cause morbidity and mortality. This study describes the reproducibility and validity of a food-based diet quality index, the Australian Recommended Food Score (ARFS). ARFS was developed to reflect alignment with the Australian Dietary Guidelines and is modelled on the US Recommended Food Score. Dietary intakes of 96 adult participants (31 male, 65 female) age 30 to 75 years were assessed in two rounds, five months apart. Diet was assessed using a 120-question semi-quantitative food frequency questionnaire (FFQ). The ARFS diet quality index was derived using a subset of 70 items from the full FFQ. Reproducibility of the ARFS between round one and round two was confirmed by the overall intraclass correlation coefficient of 0.87 (95% CI 0.83, 0.90), which compared favourably to that for the FFQ at 0.85 (95% CI 0.80, 0.89). ARFS was correlated with FFQ nutrient intakes, particularly fiber, vitamin A, beta-carotene and vitamin C (0.53, 95% CI 0.37-0.67), and with mineral intakes, particularly calcium, magnesium and potassium (0.32, 95% CI 0.23-0.40). ARFS is a suitable brief tool to evaluate diet quality in adults and reliably estimates a range of nutrient intakes.


Approaching avoidance. A step essential to the understanding of craving.
Craving is only one component of the mental processes that influence drinking behavior. Alcohol-related cues (ARCs) can set in motion a dynamic competition between inclinations to approach drinking and inclinations to avoid drinking. Craving can thus be integrated into a comprehensive model of decision-making in which ambivalence or conflict is a key element. The relative strength of each component of the ARC reaction can fluctuate over time as well as in response to both subjective states and environmental circumstances. Simultaneously and independently evaluating these opposing responses puts clinicians in a better position to influence the relative weight that the patient assigns to the positive and negative outcomes of alcohol consumption.


Ventricular expansion in wild-type Wistar rats after alcohol exposure by vapor chamber.
Structural magnetic resonance imaging (MRI) reveals widespread brain damage manifest as tissue shrinkage and complementary ventriculomegaly in human alcoholism. For an animal model to parallel the human condition, high alcohol exposure should produce similar radiologically detectable neuropathology. Our previous structural MRI study demonstrated only modest brain dysmorphology of the alcohol-preferring (P) rat with average blood alcohol levels(BALs) of 125 mg/dl achieved with voluntary consumption. Here, we tested the hypothesis that wild-type Wistar rats, exposed to vaporized alcohol ensuring higher BALs than typically achieved with voluntary consumption in rodents, would model MRI findings in the brains of humans with chronic alcoholism. The longitudinal effects of vaporized alcohol exposure on the brains of 10 wild-type Wistar rats compared with 10 sibling controls were investigated with structural MRI, conducted before (MRI 1) and after (MRI 2) 16 of alcohol exposure and after an additional 8 weeks at a higher concentration of alcohol (MRI 3). Two rats in the alcohol group died prior to MRI 2. The remaining vapor-exposed rats(n = 8) achieved BALs of 293 mg/dl by MRI 2 and 445 mg/dl by MRI 3. Whereas the controls gained 17% of their body weight from MRI 1 to MRI 3, the alcohol-exposed group lost 6%.MRI, quantified with atlas-based parcellation, revealed a profile of significant ventricular expansion,after alcohol vapor exposure, in 9 contiguous slices, extending from the dorsolateral to ventrolateral ventricles. In particular, from MRI 1 to MRI 2, this ventricular volume expanded by an average of 6.5% in the controls and by 27.1% in the alcohol-exposed rats but only an additional 1.5% in controls and 2.4% in alcohol-exposed rats from MRI 2 to MRI 3. The midsagittal volume of the full anterior-to-posterior extent of the corpus callosum grew between the first 2 MRIs in both groups followed by regression in the alcohol group by MRI 3. Although group differences were statistically significant, among animals there was substantial variability of the effects of alcohol exposure on brain morphology; some animals showed profound effects, whereas others were essentially unaffected. The ventricular dilatation and callosal shrinkage produced in wild-type rats following involuntary alcohol exposure yielded a modestly successful model of neurodysmorphology phenotypes of human alcoholism. As is the case for the human condition, however, in which some individuals express greater alcoholism-related neuropathology than others, some rats maybe more susceptible than others to extreme alcohol exposure.


RNA polymerase II forms transcription networks in rye and Arabidopsis nuclei and its amount increases with endopolyploidy.
RNA polymerase II (RNAPII) is responsible for the transcription of most eukaryotic genes. In mammalian nuclei, RNAPII is mainly localized in relatively few distinct transcription factories. In this study--applying super-resolution microscopy--it is shown that in plants, inactive (non-phosphorylated) and active (phosphorylated) RNAPII modifications compose distinct 'transcription networks' within the euchromatin. These reticulate structures sometimes attach to each other, but they are absent from heterochromatin and nucleoli. The global RNAPII distribution within nuclei is not influenced by interphase chromatin organization such as Rabl (rye) versus non-Rabl (Arabidopsis thaliana) orientation. Replication of sister chromatids without cell division causes endopolyploidy, a phenomenon widespread in plants and animals. Endopolyploidy raises the number of gene copies per nucleus. Here, it is shown that the amounts of active and inactive RNAPII enzymes in differentiated 2-32C leaf nuclei of A. thaliana proportionally increase with rising endopolyploidy. Thus, increasing the transcriptional activity of cells and tissues seems to be an important function of endopolyploidy.


Assessing the impact of FDG-PET in the management of anal cancer.
To assess the utility of FDG-PET in anal cancer for staging and impact on radiotherapy planning (RTP), response and detection of recurrent disease. Fifty histopathological anal cancer patients were reviewed between 1996 and 2006. The median age was 58 years (range 36-85) with 19 males:31females. Clinical assessment with CT was compared to PET. Impact on management, disease response, recurrence and metastases was evaluated. The non-PET staging was Stage I(8), Stage II(18), Stage III(22), and Stage IV(2)s. The primary was strongly FDG avid in 98% with non-excised tumors compared to CT (58%). PET upstaged 17% with unsuspected pelvic/inguinal nodal disease. Pre-treatment PET identified 11 additional by involved nodal groups in 48 patients causing RTP amendments in 19%. Post-treatment PETs at median 17 weeks (range 9-28) showed complete responses in 20 (80%) and 5 (20%) partial responses (PR). PRs were biopsy positive in 2 and negative in 3. Fifteen had follow-up scans of which all nine PETs detected recurrences were pathologically confirmed. Anal cancer is FDG-PET avid. PET upstages 17% and changes the RTP in 19%. PET can aid in anal cancer staging and identification of residual disease, recurrent/metastatic disease but warrants further prospective studies.


A retrotransposable element from the mosquito Anopheles gambiae .
A family of middle repetitive elements from the African malaria vector Anopheles gambiae is described. Approximately 100 copies of the element, designated T1Ag, are dispersed in the genome. Full-length elements are 4.6 kilobase pairs in length, but truncation of the 5' end is common. Nucleotide sequences of one full-length, two 5'-truncated, and two 5' ends of T1Ag elements were determined and aligned to define a consensus sequence. Sequence analysis revealed two long, overlapping open reading frames followed by a polyadenylation signal, AATAAA, and a tail consisting of tandem repetitions of the motif TGAAA. No direct or inverted long terminal repeats (LTRs) were detected. The first open reading frame, 442 amino acids in length, includes a domain resembling that of nucleic acid-binding proteins. The second open reading frame, 975 amino acids long, resembles the reverse transcriptases of a category of retrotransposable elements without LTRs, variously termed class II retrotransposons, class III elements or non-LTR retrotransposons. Similarity at the sequence and structural levels places T1Ag in this category.


Phaeohyphomycosis in Korea.
Phaeohyphomycosis is a mycotic disease caused by dematiaceous fungi that produce brown yeast-like cells, pseudohyphae, and irregular true hyphae in tissues. Seven Korean cases of subcutaneous phaeohyphomycosis have been reported to date, four males and three females, ranging in age from 9-84 years (mean 50.4 years). Causative organisms of subcutaneous phaeohyphomycosis were 3 of Exophiala jeanselmei, 2 of E. dermatitidis, 1 of Drechslera dematioidea and 1 of Phoma sp. Four cases of abscess and 3 cases of verrucous plaque were present as skin lesions, and were all exposed areas of the body. Patients were treated with itraconazole, ketoconazole, fluconazole or amphotericin B.


Genomics of Aspergillus oryzae: learning from the history of Koji mold and exploration of its future.
At a time when the notion of microorganisms did not exist, our ancestors empirically established methods for the production of various fermentation foods: miso (bean curd seasoning) and shoyu (soy sauce), both of which have been widely used and are essential for Japanese cooking, and sake, a magical alcoholic drink consumed at a variety of ritual occasions, are typical examples. A filamentous fungus, Aspergillus oryzae, is the key organism in the production of all these traditional foods, and its solid-state cultivation (SSC) has been confirmed to be the secret for the high productivity of secretory hydrolases vital for the fermentation process. Indeed, our genome comparison and transcriptome analysis uncovered mechanisms for effective degradation of raw materials in SSC: the extracellular hydrolase genes that have been found only in the A. oryzae genome but not in A. fumigatus are highly induced during SSC but not in liquid cultivation. Also, the temperature reduction process empirically adopted in the traditional soy-sauce fermentation processes has been found to be important to keep strong expression of the A. oryzae-specific extracellular hydrolases. One of the prominent potentials of A. oryzae is that it has been successfully applied to effective degradation of biodegradable plastic. Both cutinase, responsible for the degradation of plastic, and hydrophobin, which recruits cutinase on the hydrophobic surface to enhance degradation, have been discovered in A. oryzae. Genomic analysis in concert with traditional knowledge and technology will continue to be powerful tools in the future exploration of A. oryzae.


Effect of whole-body vibration on BMD: a systematic review and meta-analysis.
Our systematic review and meta-analysis of randomized controlled trials (RCTs) examining whole-body vibration (WBV) effect on bone mineral density (BMD) found significant but small improvements in hip areal BMD (aBMD) in postmenopausal women and in tibia and spine volumetric BMD in children/adolescents, but not in other BMD measurements in postmenopausal women and young adults. Animal experiments report anabolic bone changes in response to WBV, but data in humans are limited. Our objective is to conduct a systematic review and meta-analysis of RCTs examining WBV effect on BMD. Eligible RCTs included randomized or quasi-randomized trials, with follow-up of ≥ 6 months, examining WBV effects on BMD in ambulatory individuals without secondary causes of osteoporosis. The weighted mean differences between WBV and control groups in absolute pre-post change in spine and hip aBMD, and in spine and tibia trabecular volumetric BMD (vBMD) were calculated. eight RCTs in postmenopausal women (five RCTs), young adults (one RCT), and children and adolescents (two RCTs) were included. The regimens were heterogeneous, study durations were relatively short, and available data was mostly per-protocol. In postmenopausal women, WBV was found to significantly increase hip aBMD (0.015 g cm(-2); 95% confidence interval (CI), 0.008-0.022; n = 131) versus controls, but not spine aBMD (n = 181) or tibia trabecular vBMD (n = 29). In young adults, WBV did not increase spine or hip bone mineral content, or tibia trabecular vBMD (n = 53). In children and adolescents, WBV significantly increased spine (6.2 mg cm(-3); 95% CI, 2.5-10.0; n = 65) and tibia (14.2 mg cm(-3); 95% CI, 5.2-23.2; n = 17) trabecular vBMD. We found significant but small improvements in BMD in postmenopausal women and children and adolescents, but not in young adults. WBV is a promising new modality, but before recommendations can be made for clinical practice, large-scale long-term studies are needed to determine optimal magnitude, frequency, and duration.


Spending by California's Department of Developmental Services for Persons with Autism across Demographic and Expenditure Categories.
Few autism spectrum disorder (ASD) studies have estimated non-medical costs for treatment or addressed possible differences in provision of services across gender, race-ethnic, age or demographic or expenditure categories, especially among adults. The California Department of Developmental Services (CDDS) provides services to residents with developmental disabilities. CDDS provided aggregate data on primarily non-medical spending for fiscal year 2012-2013 for persons with ASD with or without intellectual disability (ID) (main sample, n = 42,274), and two sub-samples: ASD only (n = 30,164), and ASD+ID (n = 12,110). Demographic variables included sex, age and race-ethnicity. Spending categories included Employment Support, Community Care Facilities, Day Care, Transportation, and in-home and out-of-home Respite. Per-person spending for males and females were approximately the same: $10,488 and $10,791 for males and females for ages 3-17 and $26,491 and $26,627 for ages 18+. Among race/ethnicity categories, the ranking from highest to lowest among ages 3-17 was white non-Hispanics ($11,480), Asian non-Hispanics ($11,036), "Others" ($11,031), Hispanics ($9,571), and African-American non-Hispanics ($9,482). For ages 18+, the ranking was whites ($31,008), African-Americans ($26,831), "Others" ($25,395), Asians ($22,993), and Hispanics ($18,083). The ASD+ID sub-sample exerted disproportionate influence on findings from the main sample for persons 18+. Combining all ages, the top two expenditure categories for per-person spending were Community Care Facilities ($43,867) and Day Care ($11,244). For most adult age groups, the percentage of recipients participating were highest for Day Care (44.9% - 62.4%) and Transportation (38.6% - 50.9%). Per-person spending for Day Care, Transportation, and Employment Support was relatively low for children but relatively high for adults. White non-Hispanics received the highest per-person spending and Hispanics among the least. Amounts within spending categories varied considerably across age groups. Our estimates may be useful as baseline measures for stakeholders preparing for increasing ASD prevalence, especially among adults.


Molecular phylogeny of Vincetoxicum (Apocynaceae, Asclepiadoideae) from Thailand and integrative taxonomy corroborating a new cryptic species within Vincetoxicum kerrii.
An updated phylogeny of the genus Vincetoxicum s.l. based on DNA sequences of the nuclear internal transcribed spacer (ITS) region and three plastid markers is presented. In total, 21 accessions newly sequenced from Thailand were added to the dataset of the homologous sequences of 75 other Vincetoxicum taxa downloaded from GenBank. In our analysis, the relationships between the well-supported clades largely correspond to those revealed in previous studies. With some exceptions, the phylogenetic positions of the Thai taxa in relation to other conspecifics and congeners generally reflect the geographic distributions of taxa. Moreover, recent extensive sampling throughout Thailand and in-depth investigation have revealed V. kerrii, a slender twiner widespread from South China to Indo-China, to be a species complex. A combination of molecular, morphological, anatomical, ultrastructural and ecological evidence allowed us to reveal a new cryptic species hidden within V. kerrii, described here under the name V. simplex. A comprehensive description, illustrations, photographs, and comparison with the morphologically similar species are provided. Although V. simplex and V. kerrii s.s. resemble one another in various aspects of vegetative and reproductive structures, the latter is phylogenetically closely related to V. irrawadense, which is much less similar morphologically to both V. simplex and V. kerrii s.s. than the latter two are to each other. In addition to the new cryptic species recognized in the present study, a new record for Thailand, V. microstachys, is also reported.


Tau protein and the establishment of an axonal morphology.
Dissociated neuronal cultures from several regions of the nervous system elaborate two populations of neurites which have features of axons and dendrites. The microtubule-associated protein tau appears to segregate to the axon in some of these culture systems, however it does not do so until after the development of morphological polarity. Despite this observation, tau very likely has some role in the development of polarity because in cultured cerebellar macroneurons taken from the rat embryonic day 15 primordial cerebellum, the inhibition of tau expression by antisense techniques resulted in the failure of a single minor neurite to elongate and form an axon-like neurite. Tau antisense given continuously for up to 72 h kept neurons locked in a stage with minor neurites only; however when released from the effects of the antisense they fully recovered. The administration of tau antisense after the development of polarity resulted in the loss of the axon-like neurite, while dendrite-like neurites continued to grow. Together these results suggest that dendritic differentiation in cerebellar macroneurons requires the prior elaboration of an axon-like structure.


In vitro production of Clostridium difficile spores for use in the efficacy evaluation of disinfectants: a precollaborative investigation.
Clostridium difficile is a strict anaerobic spore-forming bacterium, and an increasingly common nosocomial pathogen. The U.S. Environmental Protection Agency (EPA) is responsible for the registration of disinfectants, including products designed to treat environmental surfaces contaminated with spores of C. difficile. Product efficacy data are required for registration; however, there is a lack of methodology for generating high-quality spore suspensions for evaluating product performance. As such, a study was carried out to select a suitable C. difficile strain and to develop a stand-alone method to prepare a spore suspension that meets specific criteria necessary for quantitative testing of disinfectants. The criteria are: (1) a spore titer of > 8 log10/mL, (2) > or = 90% spores to vegetative cells, and (3) resistance of spores (determined by viability) to 2.5 M hydrochloric acid (HCl). Several strains of C. difficile (toxigenic and nontoxigenic) were grown on various media (solid and liquid) for varying lengths of time to determine the best combination of incubation conditions and media to optimize spore production and quality. Once the spore production procedure was optimized, a toxigenic strain of C. difficile [American Type Culture Collection (ATCC) 43598] was selected for use in trials to verify repeatability from one production run to the next. The spore suspension was initiated by spreading vegetative cells of C. difficile (ATCC 43598) on CDC anaerobic 5% sheep blood agar plates and incubating for 7-10 days at 36 +/- 1 degrees C under anaerobic conditions. Spores were harvested when > or = 90% of the cells converted to spores as determined by observation using phase-contrast microscopy. The spores were washed three times with saline-Tween-80, resuspended in cold deionized water, heated to 70 degrees C for 10 min, evaluated microscopically for quality, and enumerated on cycloserine-cefoxitin-fructose agar containing horse blood and taurocholate. The spore suspension was used to inoculate brushed stainless steel carriers (1 cm in diameter) with and without a soil load in accordance with the Standard Quantitative Carrier Disk Test Method (ASTM E-2197-02) to determine carrier load. Once it was determined that > 6 log10 spores/carrier could be recovered, spores were evaluated for resistance to HCI. The sporulation method presented in this report is simple and repeatable and results in spore suspension of high titer (> 8 log10/mL) and quality (> or = 90% spores to vegetative cells) that met acid resistance criteria (spores were resistant to 2.5 M HCI for 10 min). In addition, recovery from brushed stainless steel carriers with and without soil load was > 6 log10 spores/carrier. A 6 log10 performance standard was set forth in the EPA's interim guidance for generating data to support a label claim for effectiveness against C. difficile spores on hard, nonporous surfaces. This precollaborative investigation successfully demonstrated the use of a methodology for in vitro production of C. difficile spores (ATCC 43598) necessary for conducting efficacy tests. A proposal will be submitted to the AOAC INTERNATIONAL Methods Committee on Antimicrobial Efficacy Testing for a collaborative study; see Appendix.


Studies on the antihypertensive, antispasmodic, bronchodilator and hepatoprotective activities of the Carum copticum seed extract.
This study describes the antihypertensive, antispasmodic, bronchodilator and hepatoprotective activities of the aqueous-methanolic extract of Carum copticum Benth. seeds (CSE) to rationalize some of its traditional uses. CSE (3-100 mg/kg) caused a dose-dependent fall in arterial blood pressure in anaesthetized rats. In isolated rabbit aorta and jejunum preparations, CSE (0.1-3.0 mg/ml) caused an inhibitory effect on the K+-induced contractions. The calcium channel blocking (CCB) effect was confirmed when CSE shifted the Ca2+ dose-response curves (DRCs) to right similar to verapamil. In isolated guinea-pig tracheal preparations, it caused inhibition of carbachol and K+-induced bronchoconstriction at 0.1-1.0 mg/ml as well as shifted the dose-response curves (DRCs) of carbachol and histamine to the right with suppression of maximum response suggestive of non-specific bronchodilator effect mediated possibly through CCB. Pretreatment of rats with CSE (500 mg/kg orally for 2 days at 12 h intervals) prevented paracetamol (640 mg/kg) and CCl4 (150 ml/kg)-induced rise in serum alkaline phosphatase (ALP) and aminotransferases (AST and ALT). The same dose of CSE was able to prevent the CCl4-induced prolongation in pentobarbital-induced sleeping time in mice confirming its hepatoprotectivity. These results indicate the presence of calcium antagonist(s) in Carum copticum seeds and thus provides sound mechanistic basis for some of their folkloric uses.


Detection of the floR gene in a diversity of florfenicol resistant Gram-negative bacilli from freshwater salmon farms in Chile.
Florfenicol is an important antibiotic in veterinary medicine that is used extensively in aquaculture, including salmon farming in Chile. We analysed a set of 119 florfenicol-resistant Gram-negative bacilli from seven freshwater Chilean salmon farms for the molecular determinants involved in the florfenicol resistance. Ninety-seven of these strains were glucose non-fermenting bacilli, mainly belonging to the Pseudomonas genus, whereas 22 strains were glucose-fermenters. The floR gene was detected in 26 strains (21.8%) that had been isolated from three of the seven salmon farms. Most of the floR-carrying strains were glucose fermenters (21 strains), and most of the floR-carrying strains were also resistant to streptomycin, chloramphenicol and oxytetracycline. The minimum inhibitory concentrations against florfenicol were assessed in the presence and absence of the efflux pump inhibitor Phe-Arg-beta-naphthylamide (MC-207,110). There was evidence that in the majority of non-fermenting bacteria (82 strains), florfenicol resistance was at least partially mediated by non-specific efflux pump systems. Given the diversity of antibiotic resistance patterns observed in this study in the floR-positive isolates, a single antibiotic has the potential to co-select for a diversity of resistances. For this reason, human health as well as animal health can potentially be impacted by the use of antibiotics in aquaculture. To assess this potential risk, future studies should focus on the ability of different antibiotics used in aquatic environments to co-select for multiple resistances, the molecular basis of this diversity of resistance, and whether the genes conferring resistance can be transferred to other bacteria, including those of human health concern.


Zinc finger transcription factors displaced SREBP proteins as the major Sterol regulators during Saccharomycotina evolution.
In most eukaryotes, including the majority of fungi, expression of sterol biosynthesis genes is regulated by Sterol-Regulatory Element Binding Proteins (SREBPs), which are basic helix-loop-helix transcription activators. However, in yeasts such as Saccharomyces cerevisiae and Candida albicans sterol synthesis is instead regulated by Upc2, an unrelated transcription factor with a Gal4-type zinc finger. The SREBPs in S. cerevisiae (Hms1) and C. albicans (Cph2) have lost a domain, are not major regulators of sterol synthesis, and instead regulate filamentous growth. We report here that rewiring of the sterol regulon, with Upc2 taking over from SREBP, likely occurred in the common ancestor of all Saccharomycotina. Yarrowia lipolytica, a deep-branching species, is the only genome known to contain intact and full-length orthologs of both SREBP (Sre1) and Upc2. Deleting YlUPC2, but not YlSRE1, confers susceptibility to azole drugs. Sterol levels are significantly reduced in the YlUPC2 deletion. RNA-seq analysis shows that hypoxic regulation of sterol synthesis genes in Y. lipolytica is predominantly mediated by Upc2. However, YlSre1 still retains a role in hypoxic regulation; growth of Y. lipolytica in hypoxic conditions is reduced in a Ylupc2 deletion and is abolished in a Ylsre1/Ylupc2 double deletion, and YlSre1 regulates sterol gene expression during hypoxia adaptation. We show that YlSRE1, and to a lesser extent YlUPC2, are required for switching from yeast to filamentous growth in hypoxia. Sre1 appears to have an ancestral role in the regulation of filamentation, which became decoupled from its role in sterol gene regulation by the arrival of Upc2 in the Saccharomycotina.


A Comparison of Parenteral Phenobarbital vs. Parenteral Phenytoin as Second-Line Management for Pediatric Convulsive Status Epilepticus in a Resource-Limited Setting.
Introduction: Pediatric convulsive status epilepticus (CSE) which is refractory to first-line benzodiazepines is a significant clinical challenge, especially within resource-limited countries. Parenteral phenobarbital is widely used in Africa as second-line agent for pediatric CSE, however evidence to support its use is limited. Purpose: This study aimed to compare the use of parenteral phenobarbital against parenteral phenytoin as a second-line agent in the management of pediatric CSE. Methodology: An open-labeled single-center randomized parallel clinical trial was undertaken which included all children (between ages of 1 month and 15 years) who presented with CSE. Children were allocated to receive either parenteral phenobarbital or parenteral phenytoin if they did not respond to first-line benzodiazepines. An intention-to-treat analysis was performed with the investigators blinded to the treatment arms. The primary outcome measure was the success of terminating CSE. Secondary outcomes included the need for admission to the pediatric intensive care unit (PICU) and breakthrough seizures during the admission. In addition, local epidemiological data was collected on the burden of pediatric CSE. Results: Between 2015 and 2018, 193 episodes of CSE from 111 children were enrolled in the study of which 144 met the study requirements. Forty-two percent had a prior history of epilepsy mostly from structural brain pathology (53%). The most common presentation was generalized CSE (65%) caused by acute injuries or infections of the central nervous system (59%), with 19% of children having febrile status epilepticus. Thirty-five percent of children required second-line management. More patients who received parenteral phenobarbital were at a significantly reduced risk of failing second-line treatment compared to those who received parenteral phenytoin (RR = 0.3, p = 0.0003). Phenobarbital also terminated refractory CSE faster (p < 0.0001). Furthermore, patients who received parenteral phenobarbital were less likely to need admission to the PICU. There was no difference between the two groups in the number of breakthrough seizures that occurred during admission. Conclusion: Overall this study supports anecdotal evidence that phenobarbital is a safe and effective second-line treatment for the management of pediatric CSE. These results advocate for parenteral phenobarbital to remain available to health care providers managing pediatric CSE in resource-limited settings. Attachments: CONSORT 2010 checklist Trial registration: NCT03650270 Full trial protocol available: https://clinicaltrials.gov/ct2/show/NCT03650270?recrs=e&type=Intr&cond=Status+Epilepticus&age=0&rank=1.


[A re-do case of recurrent constrictive pericarditis 35 years after pericardiectomy; report of a case].
Reports of recurrent constrictive pericarditis are scarce. We report a re-do case of recurrent constrictive pericarditis 35 years after pericardiectomy. The calcified pericardium was thick and severely adhesive. We performed pericardiectomy on pump, using Harmonic Scalpel and rib scissors for a safe and secure operation. The calcified pericardium looked like a crab's shell. The patient was discharged on 14th post-operative day. Long-term follow-up is necessary for this patient.


The Charnley total hip arthroplasty in patients under age 40.
Considering the high morbidity in young individuals, the authors reviewed 73 Charnley total hip arthroplasties in patients under age 40. The average follow-up period was 6.6 years. A total of 22 hips failed. Thirty-one hip arthroplasties were performed in normally ambulatory patients, with nine failures. Forty-two were performed in patients with limited locomotion, with 13 failures. The main cause of failure was loosening requiring revision (ten hips). There was radiologic evidence of progressive loosening in four other hips. Loss of trochanteric fixation required revision of six hips. Activity and rheumatoid disease were major contributing factors. Alternative methods of treatment should be considered in the young patient. When the operation is indicated, the greater trochanter must not be removed. It is hoped that cementless methods of fixation may produce a lower incidence of failure.


The Spatiotemporal Link of Temporal Expectations: Contextual Temporal Expectation Is Independent of Spatial Attention.
Temporal expectation is the ability to construct predictions regarding the timing of events, based on previously experienced temporal regularities of different types. For example, cue-based expectations are constructed when a cue validly indicates when a target is expected to occur. However, in the absence of such cues, expectations can be constructed based on contextual temporal information, including the onset distribution of the event and recent prior experiences, both providing implicit probabilistic information regarding the timing of the event. It was previously suggested that cue-based temporal expectation is exerted via synchronization of spatially specific neural activity at a predictable time of a target, within receptive fields corresponding to the expected location of the target. Here, we tested whether the same theoretical model holds for contextual temporal effects. Participants (n = 40, 25 females) performed a speeded spatial-cuing detection task with two-thirds valid spatial cues. The hazard-rate function of the target was modulated by varying the foreperiod-the interval between the spatial cue and the target-among trials and was manipulated between groups by changing the interval distribution. Reaction times were analyzed using both frequentist and Bayesian generalized linear mixed models, accounting for hazard and sequential effects. Results showed that the effects of contextual temporal structures on reaction times were independent of spatial attention. This suggests that the spatiotemporal mechanisms, thought to account for cue-based expectation, cannot explain other sources of temporal expectations. We conclude that expectations based on contextual structures have different characteristics than cue-based temporal expectation, suggesting reliance on distinct neural mechanisms.SIGNIFICANCE STATEMENT Temporal expectation is the ability to predict an event onset based on temporal regularities. A neurophysiological model suggested that temporal expectation relies on the synchronization of spatially specific neurons whose receptive fields represent the attended location. This model predicts that temporal expectation would be evident solely within the locus of spatial attention. Existing evidence supported this model for expectation based on associations between a temporal cue and a target, but here we show that it cannot account for temporal expectation that is based on contextual information, that is, the distribution of intervals and recent priors. These findings reveal the existence of different predictive mechanisms for cued and contextual temporal predictions, with the former depending on spatial attention and the latter nonspatially specific.


The impact of the COVID-19 pandemic on cancer diagnosis based on pathology notifications: A comparison across the Nordic countries during 2020.
The severity of the COVID-19 pandemic and subsequent mitigation strategies have varied across the Nordic countries. In a joint Nordic population-based effort, we compared patterns of new cancer cases and notifications between the Nordic countries during 2020. We used pathology notifications to cancer registries in Denmark, the Faroe Islands, Finland, Iceland, Norway and Sweden to determine monthly numbers of pathology notifications of malignant and in situ tumours from January to December 2020 compared to 2019 (2017-2019 for Iceland and the Faroe Islands). We compared new cancer cases per month based on unique individuals with pathology notifications. In April and May 2020, the numbers of new malignant cases declined in all Nordic countries, except the Faroe Islands, compared to previous year(s). The largest reduction was observed in Sweden (May: -31.2%, 95% CI -33.9, -28.3), followed by significant declines in Finland, Denmark and Norway, and a nonsignificant decline in Iceland. In Denmark, Norway, Sweden and Finland the reporting rates during the second half of 2020 rose to almost the same level as in 2019. However, in Sweden and Finland, the increase did not compensate for the spring decline (annual reduction -6.2% and -3.6%, respectively). Overall, similar patterns were observed for in situ tumours. The COVID-19 pandemic led to a decline in rates of new cancer cases in Sweden, Finland, Denmark and Norway, with the most pronounced reduction in Sweden. Possible explanations include the severity of the pandemic, temporary halting of screening activities and changes in healthcare seeking behaviour.


Antithrombin deficiency increases thrombin activity after prolonged cardiopulmonary bypass.
Antithrombin (AT) levels decrease during cardiopulmonary bypass (CPB), particularly when combined with deep hypothermic circulatory arrest (DHCA). Low AT levels might lead to imbalance of pro- and anticoagulant factors promoting systemic thrombotic events. We hypothesized that low levels of AT might lead to increased in vitro thrombin generation when procoagulant factors are added to the patient's plasma after CPB. Blood samples were obtained before heparinization and after separation from CPB from five patients undergoing cardiac surgery with DHCA. AT levels were determined by chromogenic assay and expressed as a percent of normal activity. The balance between procoagulant and anticoagulant elements was manipulated in the patients' plasma by adding normal donor plasma, AT-deficient plasma, or purified AT. The Thrombinoscope system was used to evaluate thrombin generation with and without AT supplementation. AT levels (median, range) were 82.0% (71.0, 109) and 37.0% (34.0, 41.0) of normal before and after separation from CPB, respectively (P < 0.05). Peak thrombin generation (median, range) was 56.6 nM (42.1, 61.0) in plasma after CPB, and it remained at 61.1 nM (54.9, 64.5) when a donor plasma with normal AT (105%) was added. When AT-deficient plasma was added to the patient's plasma, peak thrombin generation (median, range) was increased from 56.6 nM (42.0, 61.0) to 117 nM (95.0, 188) (P < 0.05 versus control). After the addition of purified AT, the peak thrombin generation was reduced to 12.2 nM (9.0, 29.3) (P < 0.05 versus control). Plasma AT activity is severely decreased after CPB with DHCA. Our data suggest that the administration of coagulation factor components without AT repletion may lead to excessive thrombin generation, which clinically, may potentially lead to a hypercoagulable state.