Insertion/deletion-based approach for the detection of Escherichia coli O157:H7 in freshwater environments.
Enterohemorrhagic Escherichia coli O157:H7 is responsible for many outbreaks of gastrointestinal illness and hemolytic uremic syndrome worldwide. Monitoring this pathogen in food and water supplies is an important public health issue. Highly conserved genetic markers, which are characteristic for specific strains, can provide direct identification of target pathogens. In this study, we examined a new detection strategy for pathogenic strains of E. coli O157:H7 serotype based on a conserved signature insertion/deletion (CSI) located in the ybiX gene using TaqMan-probe-based quantitative PCR (qPCR). The qPCR assay was linear from 1.0 × 10(2) to 1.0 × 10(7) genome copies and was specific to O157:H7 when tested against a panel of 15 non-O157:H7 E. coli. The assay also maintained detection sensitivity in the presence of competing E. coli K-12, heterologous nontarget DNA spiked in at a 1000-fold and 800-fold excess of target DNA, respectively, demonstrating the assay's ability to detect E. coli O157:H7 in the presence of high levels of background DNA. This study thus validates the use of strain-specific CSIs as a new class of diagnostic marker for pathogen detection.


In vivo transcription kinetics of a synthetic gene uninvolved in stress-response pathways in stressed Escherichia coli cells.
The fast adaptation of Escherichia coli to stressful environments includes the regulation of gene expression rates, mainly of transcription, by specific and global stress-response mechanisms. To study the effects of mechanisms acting on a global level, we observed with single molecule sensitivity the effects of mild acidic shift and oxidative stress on the in vivo transcription dynamics of a probe gene encoding an RNA target for MS2d-GFP, under the control of a synthetic promoter. After showing that this promoter is uninvolved in fast stress-response pathways, we compared its kinetics of transcript production under stress and in optimal conditions. We find that, following the application of either stress, the mean rates of transcription activation and of subsequent RNA production of the probe gene are reduced, particularly under oxidative stress. Meanwhile, the noise in RNA production decreases under oxidative stress, but not under acidic shift. From distributions of intervals between consecutive RNA productions, we infer that the number and duration of the rate-limiting steps in transcription initiation change, following the application of stress. These changes differ in the two stress conditions and are consistent with the changes in noise in RNA production. Overall, our measurements of the transcription initiation kinetics of the probe gene indicate that, following sub-lethal stresses, there are stress-specific changes in the dynamics of transcription initiation of the probe gene that affect its mean rate and noise of transcript production. Given the non-involvement of the probe gene in stress-response pathways, we suggest that these changes are caused by global response mechanisms of E. coli to stress.


Is activation of the granulocyte by concanavalin-A a reversible process?
The stimulation of granulocyte O2- production by concanavalin-A can be reversed with alpha-methylmannoside. Such cells can be reactivated to generate O2- by adding phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine. Opsonized zymosan, however, is not an effective stimulant to these cells. alpha-Methylmannoside prevents, but does not reverse, depolarization of granulocytes by concanavalin-A. Previously activated cells have a shorter lag time for reactivation by phorbol myristate acetate. Incubation in 2-deoxyglucose of cells previously treated with concanavalin-A and alpha-methylmannoside prevents reactivation. EGTA prevents concanavalin-A-stimulated O2- production only when added prior to the stimulant. EGTA has only a slight effect on reactivation. alpha-Methylmannoside prevents concanavalin-A-stimulated release of lysozyme only when added prior to the stimulant. Prior treatment of cells with concanavalin-A and alpha-methylmannoside inhibits subsequent ingestion of complement-coated particles. We conclude that although the O2--generating system can be reversibly activated with concanavalin-A followed by alpha-methylmannoside, these cells are different from untreated cells. Cells treated in such a way do not respond to all stimuli, remain depolarized, have shortened lag times, no longer require calcium for activation, continue to degranulate, and do not ingest well. Thus, although some changes that accompany the interaction of stimuli with granulocytes are reversible, some are not, and the previously activated cell does not return to a true resting state.


Regulation of neuregulin-mediated acetylcholine receptor synthesis by protein tyrosine phosphatase SHP2.
Synapse-specific expression of the nicotinic acetylcholine receptor (AChR) is believed to be mediated by neuregulin, an epidermal growth factor-like trophic factor released by somatic motoneurons at the neuromuscular junction (NMJ). Neuregulin stimulates ErbB2, ErbB3, and ErbB4, members of the ErbB family of receptor tyrosine kinases. SHP2 is a cytoplasmic protein tyrosine phosphatase containing two Src homology 2 domains near its N terminus, and has been shown to be a positive mediator of mitogenic responses to various growth factors. We found that SHP2 interacted with ErbB2 and ErbB3 after neuregulin stimulation of muscle cells. Expression of SHP2 in C2C12 mouse muscle cells attenuated the neuregulin-induced expression of an AChR epsilon-promoter reporter gene, whereas a catalytically inactive SHP2 mutant or a mutant lacking the N-terminal Src homology 2 (SH2) domain enhanced reporter expression, suggesting that SHP2 negatively regulates the neuregulin signaling pathway. In fibroblast cells that express a mutant SHP2 with a targeted deletion of the N-terminal SH2 domain, neuregulin-mediated activation of the Ras/Raf/extracellular signal-regulated kinase cascade was enhanced. Furthermore, we found that SHP2 immunoreactivity colocalized with the staining of alpha-bungarotoxin, a marker of the NMJ. These results demonstrate a negative role of SHP2 in the neuregulin signal that leads to AChR gene expression at the NMJ.


Birth of rats following nuclear exchange at the 2-cell stage.
We report full-term development of nuclear transfer embryos following nuclear exchange at the 2-cell stage. Nuclei from 2-cell rat embryos were transferred into enucleated 2-cell embryos and developed to term after transfer to recipients (NT2). Pronuclear exchange in zygotes was used for comparison (NT1). Zygotes and 2-cell embryos were harvested from 4-week-old female Sprague-Dawley rats. Nuclear transfer was performed by transferring the pronuclei or karyoplasts into the perivitelline space of recipient embryos followed by electrofusion to reconstruct embryos. Fused couplets were cultured for 4 or 24 h before being transferred into day 1 pseudopregnant recipients (Hooded Wistar) at the 1- or 2-cell stage. In vitro culture was also carried out to check the developmental competence of the embryos. In vitro development to the blastocyst stage was not significantly different between the two groups (NT1, 34.3%; NT2, 45.0%). Two of three recipients from NT1 and two of five recipients from NT2 became pregnant. Six pups (3 from NT1, 3 from NT2) were delivered from the four foster mothers. Three female pups survived; 2 from NT1 and 1 from NT2. At 2 months of age these pups appeared healthy, and were mated with Sprague-Dawley males. One rat derived from NT1 delivered 15 pups (5 males, 10 females) as did the rat from NT2 (7 males, 8 females). Our results show that by using karyoplasts from 2-cell stage embryos as nuclear donors and reconstructing them with enucleated 2-cell embryos, healthy rats can be produced.


An evaluation of the comparative effectiveness of geriatrician-led comprehensive geriatric assessment for improving patient and healthcare system outcomes for older adults: a protocol for a systematic review and network meta-analysis.
Comprehensive geriatric assessment (CGA) is an integrated model of care involving a geriatrician and an interdisciplinary team and can prioritize and manage complex health needs of older adults with multimorbidity. CGAs differ across healthcare settings, ranging from shared care conducted in primary care settings to specialized inpatient units in acute care. Models of care involving geriatricians vary across healthcare settings, and it is unclear which CGA model is most effective. Our objective is to conduct a systematic review and network meta-analysis (NMA) to examine the comparative effectiveness of various geriatrician-led CGAs and to identify which models improve patient and healthcare system level outcomes. An integrated knowledge translation approach will be used and knowledge users (KUs) including patients, caregivers, geriatricians, and healthcare policymakers will be involved throughout the review. Electronic databases including MEDLINE, EMBASE, Cochrane library, and Ageline will be searched from inception to November 2016 to identify relevant studies. Randomized controlled trials of older adults (≥65 years of age) that examine geriatrician-led CGAs compared to any intervention will be included. Primary and secondary outcomes will be selected by KUs to ensure the results are relevant to their decision-making. Two reviewers will independently screen the search results, extract data, and assess risk of bias. Data will be synthesized using an NMA to allow for multiple comparisons using direct (head-to-head) as well as indirect evidence. Interventions will be ranked according to their effectiveness using surface under the cumulative ranking curve (SUCRA). As the proportion of older adults grows worldwide, the demand for specialized geriatric services that help manage complex health needs of older adults with multimorbidity will increase in many countries. Results from this systematic review and NMA will enhance decision-making and the efficient allocation of scarce geriatric resources. Moreover, active involvement of KUs throughout the review process will ensure the results are relevant to different levels of decision-making. PROSPERO CRD42014014008.


TNF mediates a murine model of Addison's crisis.
Addison's crisis is the most serious complication of adrenal insufficiency. To elucidate the mechanism of this disorder following infection, the role of TNF in adrenalectomized murine models of Addison's crisis and Addison's disease (chronic hypoglucocorticoidism) were examined. Adrenalectomy conferred a 40-fold increased sensitivity to the lethal effects of lipopolysacharride (LPS) (P < .001). Enhanced sensitivity to LPS was found to increase with duration of adrenal insufficiency (P < .02). Enhanced lethality to heat-killed Streptococcus pneumonia was also demonstrated (P < 0.02). Necropsy of endotoxin-killed adrenalectomized mice demonstrated similar pathologic findings to those found by others when the control mice were administered a lethal dose of either LPS or TNF. Adrenalectomized TNF receptor Ia and Ib double null mice were demonstrated to be resistant to the lethal effects of LPS (P < 0.02). Pretreatment with anti-TNF, but not control antisera, was found to prevent death in LPS-treated wild-type adrenalectomized mice as well (P < 0.02). Studies into the mechanism by which TNF was precipitating Addison's crisis demonstrated enhanced sensitivity to TNF (3-fold; P < 0.02), and a marked increase in serum TNF concentration (approximately 5-fold; P < 0.001) following LPS challenge. The effect of TNF upon long-term survival in adrenalectomized mice was examined in TNF-receptor Ia- and Ib-deficient mice. Deficiencies in either the TNF-receptor Ia or Ib was noted to confer a survival advantage relative to colony controls following adrenalectomy (P < 0.02). In summary, both LPS-induced Addison's crisis and chronic adrenal insufficiency are disorders of TNF disregulation. Based upon these data, therapeutic strategies targeted at controlling TNF in adrenal insufficiency are suggested.


ECCENTRIC: a fast and unrestrained approach for high-resolution in vivo metabolic imaging at ultra-high field MR.
A novel method for fast and high-resolution metabolic imaging, called ECcentric Circle ENcoding TRajectorIes for Compressed sensing (ECCENTRIC), has been developed and implemented on 7 Tesla human MRI. ECCENTRIC is a non-Cartesian spatial-spectral encoding method optimized for random undersampling of magnetic resonance spectroscopic imaging (MRSI) at ultra-high field. The approach provides flexible and random (k,t) sampling without temporal interleaving to improve spatial response function and spectral quality. ECCENTRIC needs low gradient amplitudes and slew-rates that reduces electrical, mechanical and thermal stress of the scanner hardware, and is robust to timing imperfection and eddy-current delays. Combined with a model-based low-rank reconstruction, this approach enables simultaneous imaging of up to 14 metabolites over the whole-brain at 2-3mm isotropic resolution in 4-10 minutes with high signal-to-noise ratio. In 20 healthy volunteers and 20 glioma patients ECCENTRIC demonstrated unprecedented mapping of fine structural details of metabolism in healthy brains and an extended metabolic fingerprinting of glioma tumors.


The importance of expert review to clarify ambiguous situations for (Q)SAR predictions under ICH M7.
The use of in silico predictions for the assessment of bacterial mutagenicity under the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) M7 guideline is recommended when two complementary (quantitative) structure-activity relationship (Q)SAR models are used. Using two systems may increase the sensitivity and accuracy of predictions but also increases the need to review predictions, particularly in situations where results disagree. During the 4th ICH M7/QSAR Workshop held during the Joint Meeting of the 6th Asian Congress on Environmental Mutagens (ACEM) and the 48th Annual Meeting of the Japanese Environmental Mutagen Society (JEMS) 2019, speakers demonstrated their approaches to expert review using 20 compounds provided ahead of the workshop that were expected to yield ambiguous (Q)SAR results. Dr. Chris Barber presented a selection of the reviews carried out using Derek Nexus and Sarah Nexus provided by Lhasa Limited. On review of these compounds, common situations were recognised and are discussed in this paper along with standardised arguments that may be used for such scenarios in future.


High-dose pyridoxine and magnesium administration in children with autistic disorder: an absence of salutary effects in a double-blind, placebo-controlled study.
Several reports have described salutary effects such as decreased physical aggression and improved social responsiveness being associated with the administration of high doses of pyridoxine and magnesium (HDPM) in open-labeled and controlled studies of patients with autism. Despite this fact, this intervention remains controversial. A 10-week double-blind, placebo-controlled trial was undertaken to examine both the efficacy and safety of HDPM in autism. Twelve patients were enrolled, and 10 patients (mean age 6 years 3 months) were able to complete the study. HDPM at an average dose of 638.9 mg of pyridoxine and 216.3 mg of magnesium oxide was ineffective in ameliorating autistic behaviors as assessed by the Children's Psychiatric Rating Scale (CPRS), the Clinical Global Impression Scale, and the NIMH Global Obsessive Compulsive Scale. Furthermore, no clinically significant side effects were noted during HDPM administration. A trend for a transient change on the CPRS was found that was possibly due to a placebo response. This study raises doubts about the clinical effectiveness of HDPM in autistic disorder.


Skin ulcers caused by ruxolitinib in a patient with chronic cutaneous graft-versus-host disease.
Graft-versus-host disease (GVHD) is a common complication of allogeneic hematopoietic cell transplantation (HCT). In the treatment of chronic GVHD, skin directed therapy, systemic corticosteroids, calcineurin inhibitors (such as cyclosporine (CsA) and tacrolimus), rituximab, mycophenolate mofetil (MMF), extracorporeal photopheresis (ECP) and ruxolitinib are used. We present an 18 year old male with Philadelphia chromosome positive acute B lymphoblastic leukemia, treated with allogeneic HCT from a full matched sibling donor. The patient had grade 2 chronic cutaneous GVHD resistant to corticosteroids, CsA, MMF, and ECP treatment. Three months after initiation of ruxolitinib therapy, the patient developed skin ulcers on his lower extremities. The biopsy revealed that the changes were caused by the drug reactions. We suspected ruxolitinib as the likely cause of these ulcerative lesions after evaluating the adverse drug reaction probability scale. The adverse drug score was 4, therefore, ruxolitinib treatment was discontinued. Ulcerative lesions fully recovered after 4 weeks of follow-up. Ruxolitinib is used in the treatment of chronic GVHD that has been resistant to steroids and other salvage therapies. In our case, ruxolitinib was used as a salvage therapy in a patient who had refractory chronic skin GVHD. Ruxolitinib-related skin lesions with ulcers of lower extremities and whole body erythematous skin lesions were reported previously in patients with myelofibrosis. The pathophysiology of ruxolitinib related skin ulcers is unknown. Skin changes of patients using ruxolitinib should be closely monitored, and newly developing lesions should be suspected of being drug-related and biopsied.


Copy Number Variations in the Survival Motor Neuron Genes: Implications for Spinal Muscular Atrophy and Other Neurodegenerative Diseases.
Proximal spinal muscular atrophy (SMA), a leading genetic cause of infant death worldwide, is an early-onset, autosomal recessive neurodegenerative disease characterized by the loss of spinal α-motor neurons. This loss of α-motor neurons is associated with muscle weakness and atrophy. SMA can be classified into five clinical grades based on age of onset and severity of the disease. Regardless of clinical grade, proximal SMA results from the loss or mutation of SMN1 (survival motor neuron 1) on chromosome 5q13. In humans a large tandem chromosomal duplication has lead to a second copy of the SMN gene locus known as SMN2. SMN2 is distinguishable from SMN1 by a single nucleotide difference that disrupts an exonic splice enhancer in exon 7. As a result, most of SMN2 mRNAs lack exon 7 (SMNΔ7) and produce a protein that is both unstable and less than fully functional. Although only 10-20% of the SMN2 gene product is fully functional, increased genomic copies of SMN2 inversely correlates with disease severity among individuals with SMA. Because SMN2 copy number influences disease severity in SMA, there is prognostic value in accurate measurement of SMN2 copy number from patients being evaluated for SMA. This prognostic value is especially important given that SMN2 copy number is now being used as an inclusion criterion for SMA clinical trials. In addition to SMA, copy number variations (CNVs) in the SMN genes can affect the clinical severity of other neurological disorders including amyotrophic lateral sclerosis (ALS) and progressive muscular atrophy (PMA). This review will discuss how SMN1 and SMN2 CNVs are detected and why accurate measurement of SMN1 and SMN2 copy numbers is relevant for SMA and other neurodegenerative diseases.


Phase II study of G3139, a Bcl-2 antisense oligonucleotide, in combination with dexamethasone and thalidomide in relapsed multiple myeloma patients.
Bcl-2 regulates the mitochondrial apoptosis pathway that promotes chemotherapy resistance. Bcl-2 antisense oligonucleotide, G3139, targets Bcl-2 mRNA. G3139 was administered (3 to 7 mg/kg/d for 7 days) by continuous intravenous infusion. On day 4, patients started thalidomide (100 to 400 mg as tolerated) and dexamethasone (40 mg daily for 4 days) on 21-day cycles for three cycles. Stable and responding patients continued on 35-day cycles for 2 years. Thirty-three patients (median age, 60 years; range, 28 to 76 years) received 220 cycles. Patients received a median of three prior regimens including thalidomide (n = 15) and stem-cell transplantation (n = 31). The regimen was well tolerated; the median number of cycles per patient was eight (range, one to 16+ cycles). Toxicities included reversible increase in creatinine, thrombocytopenia, neutropenia, fatigue, anorexia, constipation, fever, neuropathy, edema, electrolyte disturbances, and hyperglycemia. Fifty-five percent of patients had objective responses, including two complete responses (CRs), four near CRs (positive immunofixation), and 12 partial responses; six patients had minimal responses (MRs). Of patients who received prior thalidomide, seven had objective responses, and three had MRs. The median duration of response was 13 months, and estimated progression-free and overall survival times were 12 and 17.4 months, respectively. Responding patients had significant increase in polyclonal immunoglobulin M (P = .005), indicating innate immune system activation. Western blot analysis of Bcl-2 protein isolated from myeloma cells before and after G3139 demonstrated a decrease of Bcl-2 levels in three of seven patients compared with six of nine patients using reverse transcriptase polymerase chain reaction. G3139, dexamethasone, and thalidomide are well tolerated and result in encouraging clinical responses in relapsed multiple myeloma patients.


Ligand-Induced Variations in Structural and Dynamical Properties Within an Enzyme Superfamily.
Enzyme catalysis is a complex process involving several steps along the reaction coordinates, including substrate recognition and binding, chemical transformation, and product release. Evidence continues to emerge linking the functional and evolutionary role of conformational exchange processes in optimal catalytic activity. Ligand binding changes the conformational landscape of enzymes, inducing long-range conformational rearrangements. Using functionally distinct members of the pancreatic ribonuclease superfamily as a model system, we characterized the structural and conformational changes associated with the binding of two mononucleotide ligands. By combining NMR chemical shift titration experiments with the chemical shift projection analysis (CHESPA) and relaxation dispersion experiments, we show that biologically distinct members of the RNase superfamily display discrete chemical shift perturbations upon ligand binding that are not conserved even in structurally related members. Amino acid networks exhibiting coordinated chemical shift displacements upon binding of the two ligands are unique to each of the RNases analyzed. Our results reveal the contribution of conformational rearrangements to the observed chemical shift perturbations. These observations provide important insights into the contribution of the different ligand binding specificities and effects of conformational exchange on the observed perturbations associated with ligand binding for functionally diverse members of the pancreatic RNase superfamily.


The microbiome as a biomarker of anal precancerous lesions in people with HIV.
Early detection and treatment of human papillomavirus (HPV)-related anal dysplasia in some high-risk groups can help anal cancer prevention, but new tools to improve diagnostic and risk assessment are needed. Here, we aim to discuss the evidence on the role of the microbiome as a potential biomarker for anal high-grade squamous intraepithelial lesions (HSILs) in people with HIV (PWH). This review covers relevant studies on the links between the microbiome and HPV infection, cervical dysplasia/cancer, and anal HPV disease. It focuses on anal samples and precancerous lesions. The review highlights the promising potential of the anal microbiome as a novel biomarker for precancerous lesions in people with HIV, while also discussing limitations and future research needs.


[Dementia and Alzheimer disease. Early detection and treatment].
Early diagnosis and treatment of dementia are intimately connected following recent progress in these topics and limitation of actual treatments to the initial cause of this ailment. Precise diagnosis alone can lead to adequate treatment and a logical procedure is mandatory to define, as precisely as possible, the exact aetiology. The first step will be to establish if the patient is demented, then if it is a curable dementia and finally if the dementia is treatable. If necessary, work up can be completed by a precise typing of the disease. Beside support which is a basic option for these diseases but out of the scope of this paper, the clinician will aim, if possible, to prescribe an aetiological treatment or at least a symptomatic one. This last can be not only non-specific, based on antidepressants, sedatives and neuroleptics, but also specific. This last therapeutic option has tremendously evolved recently with the coming of acetylcholine aimed treatment, particularly cholinesterase inhibitors which are the first medications to be demonstrated as effective for Alzheimer's disease. Even if these drugs have still significant limitations, therapeutic nihilism which was often the rule has to be abandoned and must be an incentive to keep on with research for better diagnostic tests and therapeutic options.


Analysis of nitric oxide-cyclic guanosine monophosphate signaling during metamorphosis of the nudibranch Phestilla sibogae Bergh (Gastropoda: Opisthobranchia).
The gas nitric oxide (NO), and in some cases its downstream second messenger, cyclic guanosine monophosphate (cGMP) function in different taxa to regulate the timing of life-history transitions. Increased taxonomic sampling is required to foster conclusions about the evolution and function of NO/cGMP signaling during life-history transitions. We report on the function and localization of NO and cGMP signaling during metamorphosis of the nudibranch Phestilla sibogae. Pharmacological manipulation of NO or cGMP production in larvae modulated responses to a natural settlement cue from the coral Porites compressa in a manner that suggest inhibitory function for NO/cGMP signaling. However, these treatments were not sufficient to induce metamorphosis in the absence of cue, a result unique to this animal. We show that induction of metamorphosis in response to the settlement cue is associated with a reduction in NO production. We documented the expression of putative NO synthase (NOS) and the production of cGMP during larval development and observed no larval cells in which NOS and cGMP were both detected. The production of cGMP in a bilaterally symmetrical group of cells fated to occupy the distal tip of rhinophores is correlated with competence to respond to the coral settlement cue. These results suggest that endogenous NO and cGMP are involved in modulating responses of P. sibogae to a natural settlement cue. We discuss these results with respect to habitat selection and larval ecology.


Responsible Gambling Telephone Intervention to High-Risk Gamblers by a State-Owned Gambling Operator in Sweden: Study Protocol for a Study on Effectiveness, User Satisfaction, and Acceptability.
Gambling disorder is associated with severe financial, social, and psychological consequences, but treatment-seeking rates have been described to be low. Apart from formal treatment, motivational interventions in nontreatment-seeking high-risk gamblers have been shown to be promising. However, little is known about the effectiveness and acceptability of such motivational interventions carried out by a gambling operator as part of the company's responsible gambling policies. Early experiences of such interventions are limited by the risk that gambling in individuals reached with the intervention may continue with a different gambling operator. The present study aims to evaluate effectiveness and user acceptability of a responsible gambling intervention continuously carried out by the Swedish state-owned gambling operator Svenska Spel Sport & Casino. This intervention for high-risk gamblers, identified either through substantial monetary losses or through a voluntary self-test by the gambler, includes a motivational telephone intervention aiming to encourage the gambler to set deposit limits, practice self-exclusion, or seek help. This protocol paper describes the two-tailed evaluation of this intervention: (1) A retrospective, register-based study of the effectiveness of the motivational intervention on gambling expenditures, deposit limits, and self-exclusions in comparison to control individuals not reached by the intervention, including all clients reached or attempted to be reached during September 2019-April 2020 (total n = 3626), as well as a one-to-one matched comparison of clients reached (n = 1404) and not reached; and (2) a prospective web survey study in individuals reached by the same ongoing telephone intervention practice from November 2020 (target n = 200), measuring clients' attitudes to the intervention, perceived effects of the intervention on gambling, and their self-reported gambling on all operators after the intervention.


Microsomal glutathione S-transferase. Purification, initial characterization and demonstration that it is not identical to the cytosolic glutathione S-transferases A, B and C.
Rat liver microsomal glutathione S-transferase was activated with N-ethylmaleimide, solubilized with Triton X-100, and purified by chromatography on hydroxyapatite and CM-Sepharose. A 36-fold purification resulted in a 36% yield, indicating that the glutathione S-transferase accounts for 2.5-3% of the original microsomal protein. The purified protein moved as a band with an apparent molecular weight of 14 000 on sodium dodecyl sulphate gel electrophoresis and appeared to be nearly homogeneous. The complex formed between the purified microsomal glutathione S-transferase and Triton X-100 has a sedimentation coefficient of 3.2 S, a partial specific volume of 0.844 cm3/g, and a Stokes radius of 5.5 nm. The complex has a molecular weight of 127 000 and contains three or four polypeptide chains and 112-134 detergent molecules. Antibodies directed against soluble glutathione S-transferases A, B and C do not react with the purified microsomal enzyme. This finding, together with differences in molecular weight and substrate specificity, demonstrate that the microsomal glutathione S-transferase is an enzyme distinct from the cytosolic glutathione S-transferases.


Excess deaths during the August 2003 heat wave in Paris, France.
During the August 2003 heat wave in France, almost 15,000 excess deaths were recorded. Paris was severely affected, with an excess death rate of 141%. This study had two aims: to identify individual factors associated with excess deaths during a heat wave in an urban environment and to describe the spatial distribution of deaths within the French capital. The study population included all people who died at home between August 1st and 20th, 2003 (N=961). We identified factors associated with excess deaths by comparing the sociodemographic characteristics of the study population with those of people who died at home during the same period in reference years (2000, 2001, 2002) (N=530). Spatial differences were analysed by calculating comparative mortality rates within Paris during August 2003. Mortality ratio was determined to demonstrate temporal variations in mortality between the heat wave period and reference years. The major factors associated with excess death were: age over 75 years (adjusted OR=1.44 (1.10-1.90), being female (adjusted OR=1.43 (1.11-1.83)), not being married (adjusted OR=1.63 (1.23-2.15)), particularly for men. Being a foreigner appeared to be a protective factor for women. Comparative mortality rates by neighbourhood showed a gradient in excess deaths from North-West to South-East. The mortality ratio was 5.44 (5.10-5.79), with very high rates of excess death in the South (12th, 13th, 14th and 15th "arrondissement"). The August 2003 heat wave in Paris was associated with both an exceptional increase in mortality rates and changes in the characteristics of those dying and spatial distribution of mortality. Understanding the effects of a heat wave on mortality can probably be improved by an analysis of risk at two levels: individual and contextual.


Neurophysiology of photically induced epilepsy in Papio papio.
This chapter critically reviews arguments supporting the role of the frontal cortex in light-sensitive epilepsy of the baboon Papio papio in the light of the most recent neurophysiological research. In particular, it is known that spontaneous or ILS-induced paroxysmal discharges, as well as generalized seizures, originate in the frontorolandic cortical region. In this region during ILS, neurons behave in the same manner as hyperexcitable neurons in focal epileptogenic lesions of animals and man. Aso, section of the corpus callosum causes deterioration or even destruction of the synchronization that exists naturally between the two frontorolandic areas. Lastly experimental focal irritative lesions enhance light sensitivity if they are located in the frontorolandic region and inhibit it if they are located in the occipital cortex. Opposing these arguments are those that support the important role the occipital cortex plays, since its ablation makes excessive light sensitivity in the baboon disappear. Studies of the primary and nonprimary visual messages and pathways have also contributed evidence. In particular, demonstration of the existence of large numbers of direct occipitofrontal connections may help reconcile the two opposing arguments. Other evidence favoring the role of the frontal cortex is furnished by the still fragmentary studies on activation of the motor pathways and by studies in neuropharmacology. The significance and value of this type of epilepsy as an animal model of the generalized reflex epilepsies of man are discussed.


Successful Treatment for Hepatoblastoma in Trisomy 18: A Case Report.
Children with trisomy 18 tend to develop hepatoblastoma. Since the introduction of appropriate management for organ malfunction, individuals with trisomy 18 have come to have a longer life expectancy. However, the predisposition to hepatoblastoma becomes a significant issue for the quality of a case. Here, we present a rare multifocal hepatoblastoma involving predominantly Couinaud segments 5 and 7 in a 10-month-old boy with trisomy 18. Though the first-line cisplatin monotherapy resulted in unsatisfactory tumor shrinkage, the second-line neoadjuvant chemotherapy administrating irinotecan and vincristine gave rise to significant tumor reduction in volume, leading to the completion of partial resection of the liver without the microscopic residual disease. The patient has been free from recurrence for 44 months. Because anatomical right hepatectomy can cause circulatory instability, including acute onset of pulmonary hypertension in trisomy 18 patients, physicians should balance treatment benefits and potential adverse effects. Our successful experience utilizing a combination of efficacious and less cardiotoxic neoadjuvant chemotherapy followed by the partial hepatectomy encourages physicians to treat a patient with trisomy 18 and tackle hepatoblastoma with a genetic background.


Rhinitis alone or rhinitis plus asthma: what makes the difference?
This paper examines the clinical expression of asthma in a group of patients displaying rhinitis according to age, sex, associated symptoms, smoking, familial history of asthma, atopy, type of sensitization to aeroallergens (pollens and/or indoor allergens), total serum immunoglobulin E (IgE), and blood eosinophils. A total of 117 adults with rhinitis were analysed on the basis of symptoms. Among them, 51 also displayed asthma, defined as a history of recurrent episodes of dyspnoea with a reversible airflow obstruction or a positive methacholine challenge. The logistic regression analysis carried out in a stepwise approach, combining several factors, showed that various parameters affected the risk of having asthma associated with rhinitis. A further analysis was made in 74 rhinitis patients comparing 42 subjects without nonallergic airway hyperresponsiveness (NAAH) to 32 patients with asthma and NAAH. Atopy, high total serum IgE levels, elevated blood eosinophil count and maternal asthma were associated with asthma. Furthermore, in atopic patients, pollen sensitization was more closely related to rhinitis alone, whereas sensitization to indoor allergens was a major determinant for the association of asthma with the symptoms of rhinitis. The same risk factors as those found in the clinical part of the study discriminated the patients with rhinitis without NAAH from those with rhinitis, asthma and NAAH. In conclusion, this study gives new insights into the relationships between asthma and rhinitis.


Dual activity lysophosphatidic acid receptor pan-antagonist/autotaxin inhibitor reduces breast cancer cell migration in vitro and causes tumor regression in vivo.
Signal transduction modifiers that modulate the lysophosphatidic acid (LPA) pathway have potential as anticancer agents. Herein, we describe metabolically stabilized LPA analogues that reduce cell migration and invasion and cause regression of orthotopic breast tumors in vivo. Two diastereoisomeric alpha-bromophosphonates (BrP-LPA) were synthesized, and the pharmacology was determined for five LPA G protein-coupled receptors (GPCRs). The syn and anti diastereomers of BrP-LPA are pan-LPA GPCR antagonists and are also nanomolar inhibitors of the lysophospholipase D activity of autotaxin, the dominant biosynthetic source of LPA. Computational models correctly predicted the diastereoselectivity of antagonism for three GPCR isoforms. The anti isomer of BrP-LPA was more effective than syn isomer in reducing migration of MDA-MB-231 cells, and the anti isomer was superior in reducing invasion of these cells. Finally, orthotopic breast cancer xenografts were established in nude mice by injection of MB-231 cells in an in situ cross-linkable extracellular matrix. After 2 weeks, mice were treated with the BrP-LPA alone (10 mg/kg), Taxol alone (10 mg/kg), or Taxol followed by BrP-LPA. All treatments significantly reduced tumor burden, and BrP-LPA was superior to Taxol in reducing blood vessel density in tumors. Moreover, both the anti- and syn-BrP-LPA significantly reduced tumors at 3 mg/kg.


A mechanism for ramified rolling circle amplification.
Amplification of single-stranded DNA circles has wide utility for a variety of applications. The two-primer ramified rolling circle amplification (RAM) reaction provides exponential DNA amplification under isothermal conditions, creating a regular laddered series of double-stranded DNA products. However, the molecular mechanism of the RAM reaction remains unexplained. A RAM reaction model predicts exponential accumulation of a double-stranded DNA product size series, and product-size ratios, that are consistent with observed RAM reaction products. The mechanism involves generation of a series of increasing size intermediate templates; those templates produce RAM products and recursively generate smaller intermediate templates. The model allows prediction of the number of rounds of circular template replication. Real-time RAM reaction data are consistent with the model. Analysis of RAM reaction products shows exponential growth limitation consistent with the model's predictions. The model provides a rationale for the observed products of the RAM reaction, and the molecular yield among those products. Experimental results are consistent with the model.


Improved restoration of thoracic kyphosis using a rod construct with differentiated rigidity in the surgical treatment of adolescent idiopathic scoliosis.
OBJECTIVE The objective of this study was to compare postoperative sagittal alignment among 3 rod constructs with different rigidity profiles. METHODS This was a dual-center retrospective cohort study involving 2 consecutive cohorts in which patients were surgically treated for adolescent idiopathic scoliosis. Lenke Type 5 curves were excluded. Patients were operated on with all-pedicle screw instrumentation using 3 different rod constructs. The first group was operated on using a hybrid construct (HC) consisting of a normal circular rod on the convex side and a beam-like rod (BR) on the concave side. The second group was operated on with a standard construct (SC) using bilateral BRs in the full length of the fusion. The third group was operated on with a modified construct (MC). The modified rods have a beam-like shape in the caudal portion, but in the cranial 2 or 3 fusion levels the rod transitions to a circular shape with a smaller anteroposterior diameter. Radiographs were analyzed preoperatively and at the first postoperative follow-up (range 1-8 weeks). The primary outcome was pre- to postoperative change in thoracic kyphosis (TK), and the secondary outcome was the ability to achieve postoperative TK within the normal range. RESULTS The HC, SC, and MC groups included 23, 70, and 46 patients, respectively. The 3 groups did not differ significantly in preoperative demographic or radiographic parameters. The mean ± standard deviation of the preoperative main curve was 60.7° ± 12.6°, and the mean of curve correction was 62.9% ± 10.4% with no significant difference among groups (p ≥ 0.680). The groups did not differ significantly in coronal balance or proximal or thoracolumbar curve correction (p ≥ 0.189). Mean postoperative TK was 23.1° ± 6.3°, 19.6° ± 7.6°, and 23.4° ± 6.9° in the HC, SC and MC groups, respectively (p = 0.013), and the mean change in TK was -3.5° ± 11.3°, -7.1° ± 11.6°, and 0.1° ± 10.9°, respectively (p = 0.005). The MC group had significantly higher postoperative TK and less loss of TK compared with the SC group (p ≤ 0.018). Postoperative TK ≤ 10° was found in 12 patients (17%) in the SC group, 1 patient (5%) in the HC group, and 1 patient (2%) in the MC group (p = 0.021). There were no differences in proximal alignment, thoracolumbar alignment, or sagittal vertical axis (p ≥ 0.249). Lumbar lordosis was 58.9° ± 11.2° in the HC group versus 52.0° ± 1.3° and 55.0° ± 11.0° and the SC and MC groups, respectively (p = 0.040). CONCLUSIONS In the 3 rod constructs with different rigidity profiles, significantly better restoration of kyphosis was achieved with the use of bilateral modified rods compared with bilateral standard rods. In the MC and HC groups, the rate of severe postoperative hypokyphosis was significantly lower than in the SC group. This is the first study to describe the clinical use of a rod with a reduced proximal diameter and show marked radiographic improvement in sagittal alignment.


Comparing the Reliability of the Glomerular Filtration Rate Estimated with 99m-Technetium Diethylene-Triamine-Pentaacetate versus the Effective Renal Plasma Flow Obtained with 99m-Technetium Ethylene Dicysteine: A Prospective Observational Study.
The glomerular filtration rate (GFR) is important for assessing renal function and must be calculated reliably and reproducibly. This study aimed to compare the reliability and accuracy of GFR estimated with 99m-technetium diethylene-triamine-pentaacetate (99mTc-DTPA) versus that calculated from the effective renal plasma flow (ERPF) (GFR is 20% of ERPF) determined by the 99m-technetium ethylene dicysteine (99mTc-EC) technique. Forty-five patients suffering from cancer requiring platinum compound-based chemotherapy or from chronic renal failure were recruited. The patients were divided into two cohorts: (1) those with normal serum creatinine (SCr) levels (≤2 mg/dL) and (2) deranged SCr levels (>2 mg/dL). For all patients, the relative renal function was estimated by the 99mTc-DTPA and 99mTc-EC methods, 2-4 days apart. A 24-h urine sample for estimating 24-h creatinine clearance (CrCl) was obtained. GFR was also calculated using the Modification of Diet in Renal Disease (MDRD) formula. The GFR estimated via 24-h urine CrCl, 99mTc-DTPA, and ERPF obtained with 99mTc-EC were examined by quantile comparison plots, and all showed evidence of following a non-Gaussian distribution. For SCr values ≤2 mg/dL, the GFR estimated by the MDRD formula consistently shows significantly higher values than the GFR estimated with 99mTc-DTPA or 99mTc-EC. We found a high degree of correlation between the 99mTc-DTPA and 99mTc-EC radionuclide methods of estimating GFR. However, in patients with renal dysfunction, GFR estimated through Gates' method using a gamma camera overestimated the GFR; in these patients, calculating the GFR from the ERPF obtained with 99mTc-EC is more accurate.


Impaired visually guided weight-shifting ability in children with cerebral palsy.
The ability to control voluntary weight shifting is crucial in many functional tasks. To our knowledge, weight shifting ability in response to a visual stimulus has never been evaluated in children with cerebral palsy (CP). The aim of the study was (1) to propose a new method to assess visually guided medio-lateral (M/L) weight shifting ability and (2) to compare weight-shifting ability in children with CP and typically developing (TD) children. Ten children with spastic diplegic CP (Gross Motor Function Classification System level I and II; age 7-12 years) and 10 TD age-matched children were tested. Participants played with the skiing game on the Wii Fit game console. Center of pressure (COP) displacements, trunk and lower-limb movements were recorded during the last virtual slalom. Maximal isometric lower limb strength and postural control during quiet standing were also assessed. Lower-limb muscle strength was reduced in children with CP compared to TD children and postural control during quiet standing was impaired in children with CP. As expected, the skiing game mainly resulted in M/L COP displacements. Children with CP showed lower M/L COP range and velocity as compared to TD children but larger trunk movements. Trunk and lower extremity movements were less in phase in children with CP compared to TD children. Commercially available active video games can be used to assess visually guided weight shifting ability. Children with spastic diplegic CP showed impaired visually guided weight shifting which can be explained by non-optimal coordination of postural movement and reduced muscular strength.


Addressing Learning Needs on the Use of Metagenomics in Antimicrobial Resistance Surveillance.
One Health surveillance of antimicrobial resistance (AMR) depends on a harmonized method for detection of AMR. Metagenomics-based surveillance offers the possibility to compare resistomes within and between different target populations. Its potential to be embedded into policy in the future calls for a timely and integrated knowledge dissemination strategy. We developed a blended training (e-learning and a workshop) on the use of metagenomics in surveillance of pathogens and AMR. The objectives were to highlight the potential of metagenomics in the context of integrated surveillance, to demonstrate its applicability through hands-on training and to raise awareness to bias factors. The target participants included staff of competent authorities responsible for AMR monitoring and academic staff. The training was organized in modules covering the workflow, requirements, benefits and challenges of surveillance by metagenomics. The training had 41 participants. The face-to-face workshop was essential to understand the expectations of the participants about the transition to metagenomics-based surveillance. After revision of the e-learning, we released it as a Massive Open Online Course (MOOC), now available at https://www.coursera.org/learn/metagenomics. This course has run in more than 20 sessions, with more than 3,000 learners enrolled, from more than 120 countries. Blended learning and MOOCs are useful tools to deliver knowledge globally and across disciplines. The released MOOC can be a reference knowledge source for international players in the application of metagenomics in surveillance.


[Lung cancer mortality in Andalucia, 1975-2000].
To describe trends in lung cancer mortality in Andalucia, Spain, during the period from 1975 through 2000. Records of lung cancer deaths for the period from 1975 through 2000 were obtained from the Statistical Institute of Andalucia. The following indicators were calculated: crude rates, age-adjusted rates, truncated rates, and cumulative rates. Mortality rates for men dropped in all subject groups aged over 50 years during the period from 1994 through 2000. In women mortality rates were much lower, although we observed an increase in the 35-39 and 45-49 age groups, which is reflected in the 6.1% rise in the truncated rates (35-64 years) during the period from 1994 through 2000. In older women mortality rates fluctuated more within the different age groups, although on the whole the truncated rate for all those over 65 years old fell by 9.7% in the period between 1995 and 2000. Lung cancer mortality rates among men in Andalusia began to decrease after 1994. In contrast, mortality increased among young women, although their rates are still very low.


Eumycetoma and actinomycetoma--an update on causative agents, epidemiology, pathogenesis, diagnostics and therapy.
Mycetoma is a chronic putrid infection of the cutaneous and subcutaneous tissue concerning predominantly the feet, and more rarely other body parts. Mycetoma can be caused by both fungi (eumycetoma) and bacteria (actinomycetoma). Mode of infection is an inoculation of the causative microorganism via small injuries of the skin. The clinical correlate of both forms of mycetoma is tumescence with abscesses, painless nodules, sinuses and discharge. The latter is commonly serous-purulent and contains grains (filamentous granules) which can be expressed for diagnostic purposes. Distinctive for both eumycetoma and actinomycetoma, are the formation of grains. Grains represent microcolonies of the microorganism in vivo in the vital tissue. The most successful treatment option for eumycetomas offers itraconazole in a dosage of 200 mg twice daily. This triazole antifungal is considered as 'gold standard' for eumycetomas. Alternatively, the cheaper ketoconazole was widely used, however, it was currently stopped by the FDA. Actinomycetomas should be treated by the combination of trimethoprim-sulphamethoxazole (co-trimoxazole 80/400 to 160/800 mg per day) and amikacin 15 mg/kg body weight per day. Mycetomas are neglected infections of the poor. They are more than a medical challenge. In rural areas of Africa, Asia and South America mycetomas lead to socio-economic consequences involving the affected patients, their families and the society in general.


Relationship between soluble HLA-G and HLA-A,-B,-C serum levels, and interferon-gamma production after sublingual immunotherapy in patients with allergic rhinitis.
Allergic rhinitis (AR) is characterized by a T-helper (Th)-2 (Th2) polarized immune response. Soluble human leukocyte antigen (sHLA) molecules play an immunomodulatory role. Specific immunotherapy is the only causal treatment for AR and is able to shift the immune response to Th1 polarization. The aim of this study was to evaluate the relationship between sHLA-G and sHLA-A,-B,-C serum levels and interferon-gamma (IFN-gamma) production in AR patients with pollen allergy before and after a preseasonal course of sublingual immunotherapy (SLIT). A total of 40 AR patients with pollen allergy were enrolled and given a course of preseasonal SLIT for 3 months. Serum sHLA-G and sHLA-A,-B,-C levels were determined by enzyme-linked immunoabsorbent assay, and cell production of IFN-gamma was evaluated by enzyme-linked immunoabsorbent spot assay at baseline and 3 months after the end of the SLIT course. There was a significant relationship between sHLA-G serum level change and IFN-gamma increase as well as between sHLA-A,-B,-C level change and IFN-gamma increase after SLIT. The present study provides the first published evidence that the reduction of sHLA molecules serum levels and the increased IFN-gamma production after SLIT in AR patients with pollen allergy are significantly related phenomena.


Glutamine promotes intestinal SIgA secretion through intestinal microbiota and IL-13.
Glutamine supplementation enhances secretory IgA (SIgA) production in the intestine, but the mechanism is largely unknown. We examined this issue using a mouse model. In mouse model, glutamine supplementation increased both SIgA abundance in intestinal luminal contents and IgA(+) plasma cell numbers in the mouse ileum, and decreased bacterial loads in mouse mesenteric lymph nodes. Glutamine supplementation increased mouse ileal expression of cytokines associated with T cell-dependent and T cell-independent pathways of SIgA induction, including IL-5, IL-6, IL-13, transforming growth factor (TGF-β), a proliferation-inducing ligand (APRIL), B cell-activating factor (BAFF), vasoactive intestinal peptide (VIP) receptor, and retinal dehydrogenases. Injecting an IL-13 antibody during glutamine supplementation reduced J-chain expression in the mouse ileum. Glutamine supplementation increased bacterial invasion into the mouse ileal wall, while disrupting the mouse intestinal microbiota abrogated the influence of glutamine supplementation on SIgA secretion. Glutamine supplementation appears to enhance SIgA secretion in the mouse intestine through the intestinal microbiota and subsequently through T cell-dependent and T cell-independent pathways.


The tandem duplicator phenotype as a distinct genomic configuration in cancer.
Next-generation sequencing studies have revealed genome-wide structural variation patterns in cancer, such as chromothripsis and chromoplexy, that do not engage a single discernable driver mutation, and whose clinical relevance is unclear. We devised a robust genomic metric able to identify cancers with a chromotype called tandem duplicator phenotype (TDP) characterized by frequent and distributed tandem duplications (TDs). Enriched only in triple-negative breast cancer (TNBC) and in ovarian, endometrial, and liver cancers, TDP tumors conjointly exhibit tumor protein p53 (TP53) mutations, disruption of breast cancer 1 (BRCA1), and increased expression of DNA replication genes pointing at rereplication in a defective checkpoint environment as a plausible causal mechanism. The resultant TDs in TDP augment global oncogene expression and disrupt tumor suppressor genes. Importantly, the TDP strongly correlates with cisplatin sensitivity in both TNBC cell lines and primary patient-derived xenografts. We conclude that the TDP is a common cancer chromotype that coordinately alters oncogene/tumor suppressor expression with potential as a marker for chemotherapeutic response.


Genome sequence analysis of dengue virus 1 isolated in Key West, Florida.
Dengue virus (DENV) is transmitted to humans through the bite of mosquitoes. In November 2010, a dengue outbreak was reported in Monroe County in southern Florida (FL), including greater than 20 confirmed human cases. The virus collected from the human cases was verified as DENV serotype 1 (DENV-1) and one isolate was provided for sequence analysis. RNA was extracted from the DENV-1 isolate and was used in reverse transcription polymerase chain reaction (RT-PCR) to amplify PCR fragments to sequence. Nucleic acid primers were designed to generate overlapping PCR fragments that covered the entire genome. The DENV-1 isolate found in Key West (KW), FL was sequenced for whole genome characterization. Sequence assembly, Genbank searches, and recombination analyses were performed to verify the identity of the genome sequences and to determine percent similarity to known DENV-1 sequences. We show that the KW DENV-1 strain is 99% identical to Nicaraguan and Mexican DENV-1 strains. Phylogenetic and recombination analyses suggest that the DENV-1 isolated in KW originated from Nicaragua (NI) and the KW strain may circulate in KW. Also, recombination analysis results detected recombination events in the KW strain compared to DENV-1 strains from Puerto Rico. We evaluate the relative growth of KW strain of DENV-1 compared to other dengue viruses to determine whether the underlying genetics of the strain is associated with a replicative advantage, an important consideration since local transmission of DENV may result because domestic tourism can spread DENVs.


[The calculation of the dose from a 60Co source using 3-dimensional Fourier transform].
The distribution of a dose in tissue-equivalent phantoms was calculated through convolution of scattered photon flux by using three-dimensional Fourier transform. A simplified model of a scattering kernel is presented, wherein multiple scattering radiation is taken into account by a water point source accumulation factor. A 64 x 64 x 64 grid calculation lasts about 2 sec on a routine personal computer (three-dimensional Fourier transform and convolution). The dose were calculated in the cube, 32 x 32 x 32 cm in size and in the cylinder, 12 cm in diameter for 4 x 4-to-20 x 20-cm radiation fields. The results are in good agreement with the reference data and Monte Carlo simulation results.


Is sternotomy always necessary for the treatment of mediastinal ectopic thyroid goiter?
Ectopic thyroid goiter accounts approximately for 1% of all substernal goiters and for 10-15% of all mediastinal masses. Sternotomy is generally accepted as the most adequate approach for the removal of ectopic thyroid goiters of the anterior mediastinum. We report two cases of mediastinal ectopic goiter removal through a cervical incision, without sternotomy. The technique is based on a careful and gentle traction of the mass by means of traction stitches and simultaneously on a blunt digital dissection, in order to exteriorize the lesion in the neck, as much as necessary to ligate its vascular pedicle before completing the removal. When performed with caution and precision, this approach can avoid sternotomy in selected patients with ectopic thyroid goiter. Ectopic thyroid, Sternotomy, Thyroidectomy.


Historical view on ergot alkaloids.
A short survey of the history of ergot, of the original and, for a long time, only source of ergot alkaloids, is given. Once a dreaded poison, ergot has changed its role over the centuries to become a rich treasure house of valuable pharmaceuticals. In the Middle Ages it was the cause of epidemics of ergotism, which cost tens of thousands of people their lives. Ergot was first mentioned by the German physician Lonitzer in 1582 as a remedy used by midwives for quickening childbirth. The isolation of pharmacologically useful alkaloids started in 1906 with the discovery of ergotoxine and its adrenolytic activity by Barger, Carr and Dale. In 1918, Stoll isolated ergotamine, the first chemically pure ergot alkaloid, which found widespread therapeutic use in obstetrics and internal medicine. In 1935 the specific oxytocic principle of ergot, ergonovine, was discovered simultaneously in four separate laboratories. Since then, worldwide investigations on ergot alkaloids resulted in the elucidation of their structures and total syntheses and preparation of valuable therapeutics such as Methergine, Hydergine, Dihydergot, and others.


Bifidobacterium breve MCC-117 Induces Tolerance in Porcine Intestinal Epithelial Cells: Study of the Mechanisms Involved in the Immunoregulatory Effect.
Bifidobacterium breve MCC-117 is able to significantly reduce the expression of inflammatory cytokines in porcine intestinal epithelial (PIE) cells and to improve IL-10 levels in CD4(+)CD25(high) Foxp3(+) lymphocytes in response to heat-stable enterotoxigenic Escherichia coli (ETEC) pathogen-associated molecular patterns (PAMPs), while the immunoregulatory effect of B. adolescentis ATCC15705 was significantly lower than that observed for the MCC-117 strain. Considering the different capacities of the two bifidobacterium strains to activate toll-like receptor (TLR)-2 and their differential immunoregulatory activities in PIE and immune cells, we hypothesized that comparative studies with both strains could provide important information regarding the molecular mechanism(s) involved in the anti-inflammatory activity of bifidobacteria. In this work, we demonstrated that the anti-inflammatory effect of B. breve MCC-117 was achieved by a complex interaction of multiple negative regulators of TLRs as well as inhibition of multiple signaling pathways. We showed that B. breve MCC-117 reduced heat-stable ETEC PAMP-induced NF-κB, p38 MAPK and PI3 K activation and expression of pro-inflammatory cytokines in PIE cells. In addition, we demonstrated that B. breve MCC-117 may activate TLR2 synergistically and cooperatively with one or more other pattern recognition receptors (PRRs), and that interactions may result in a coordinated sum of signals that induce the upregulation of A20, Bcl-3, Tollip and SIGIRR. Upregulation of these negative regulators could have an important physiological impact on maintaining or reestablishing homeostatic TLR signals in PIE cells. Therefore, in the present study, we gained insight into the molecular mechanisms involved in the immunoregulatory effect of B. breve MCC-117.


Federal funding of endoscopic research in the United States: 1972-2002.
Funding for research is critical to the determination of appropriate applications for endoscopy in clinical practice. We sought to evaluate trends in funding provided by the National Institutes of Health (NIH) for endoscopic research from 1972 through 2002. The NIH database of funded biomedical projects for the years 1972 through 2002 was searched using the Computerized Retrieval of Information on Scientific Projects (CRISP) and the keyword "endoscopy." Grants were included if a start date, title, and/or specific aims were available. Grants were classified as primary endoscopy if the title or specific aim had an endoscopy focus. Endoscopy funding was compared with other nonendoscopic funding for the same period. From 1972 through 2002, 133 endoscopy-related grant applications were funded and 98 met inclusion criteria. Funding for endoscopic research increased from one grant (1972-1982) to 4 grants (1983-1992) to 93 grants (1993-2002), a 2325% increase for the 1993-2002 decade alone. Despite this increase, there were substantially fewer funded endoscopy-related applications compared with liver disease (61,804 grants), Helicobacter pylori (866 grants), and cardiac catheterization (1547 grants). Among endoscopic grants, colorectal cancer projects accounted for the largest portion (34%), followed by advanced optical technologies (18%) and Barrett's esophagus (17%). Funding for endoscopic research by the NIH has increased dramatically over the past 30 years, but it still lags behind funding in other fields. Projects focused on colorectal cancer, Barrett's esophagus, and optical technologies were most common among those funded, and the National Cancer Institute was the primary source of funding.


Charge transfer and surface defect healing within ZnO nanoparticle decorated graphene hybrid materials.
To harness the unique properties of graphene and ZnO nanoparticles (NPs) for novel applications, the development of graphene-ZnO nanoparticle hybrid materials has attracted great attention and is the subject of ongoing research. For this contribution, graphene-oxide-ZnO (GO-ZnO) and thiol-functionalized reduced graphene oxide-ZnO (TrGO-ZnO) nanohybrid materials were prepared by novel self-assembly processes. Based on electron paramagnetic resonance (EPR) and photoluminescence (PL) investigations on bare ZnO NPs, GO-ZnO and TrGO-ZnO hybrid materials, we found that several physical phenomena were occurring when ZnO NPs were hybridized with GO and TrGO. The electrons trapped in Zn vacancy defects (VZn(-)) within the core of ZnO NPs vanished by transfer to GO and TrGO in the hybrid materials, thus leading to the disappearance of the core signals in the EPR spectra of ZnO NPs. The thiol groups of TrGO and sulfur can effectively "heal" the oxygen vacancy (VO(+)) related surface defects of ZnO NPs while oxygen-containing functionalities have low healing ability at a synthesis temperature of 100 °C. Photoexcited electron transfer from the conduction band of ZnO NPs to graphene leads to photoluminescence (PL) quenching of near band gap emission (NBE) of both GO-ZnO and TrGO-ZnO. Simultaneously, electron transfer from graphene to defect states of ZnO NPs is the origin of enhanced green defect emission from GO-ZnO. This observation is consistent with the energy level diagram model of hybrid materials.


Editing of the Sendai virus P/C mRNA by G insertion occurs during mRNA synthesis via a virus-encoded activity.
Two forms of the Sendai virus P/C mRNA have been predicted: one an exact copy of the viral genome, and the other with a single G insertion within a run of three G's. We directly cloned the mRNA or portions of it containing the insertion site and screened the resulting colonies with oligonucleotides that could distinguish the presence of three or four G's at this position. We found that 31% of the mRNAs did in fact contain the predicted insertion, whereas the viral genomes contained no heterogeneity at this position. A smaller fraction (7%) of the mRNA contained two to eight G's inserted at this position. The insertions also took place during RNA synthesis in vitro with purified virions but were not detected when the mRNA was expressed in vivo via a vaccinia virus recombinant. When the Sendai virus- and vaccinia virus-derived P/C mRNAs were coexpressed in the same cells under conditions in which each could be distinguished, those from the Sendai genome were altered as before, but those from the vaccinia virus genome remained unaltered. The activity that alters the mRNA is therefore likely to be coded for by the virus and cannot function in trans.


Heritability of submaximal exercise heart rate response to exercise training is accounted for by nine SNPs.
Endurance training-induced changes in hemodynamic traits are heritable. However, few genes associated with heart rate training responses have been identified. The purpose of our study was to perform a genome-wide association study to uncover DNA sequence variants associated with submaximal exercise heart rate training responses in the HERITAGE Family Study. Heart rate was measured during steady-state exercise at 50 W (HR50) on 2 separate days before and after a 20-wk endurance training program in 483 white subjects from 99 families. Illumina HumanCNV370-Quad v3.0 BeadChips were genotyped using the Illumina BeadStation 500GX platform. After quality control procedures, 320,000 single-nucleotide polymorphisms (SNPs) were available for the genome-wide association study analyses, which were performed using the MERLIN software package (single-SNP analyses and conditional heritability tests) and standard regression models (multivariate analyses). The strongest associations for HR50 training response adjusted for age, sex, body mass index, and baseline HR50 were detected with SNPs at the YWHAQ locus on chromosome 2p25 (P = 8.1 × 10(-7)), the RBPMS locus on chromosome 8p12 (P = 3.8 × 10(-6)), and the CREB1 locus on chromosome 2q34 (P = 1.6 × 10(-5)). In addition, 37 other SNPs showed P values <9.9 × 10(-5). After removal of redundant SNPs, the 10 most significant SNPs explained 35.9% of the ΔHR50 variance in a multivariate regression model. Conditional heritability tests showed that nine of these SNPs (all intragenic) accounted for 100% of the ΔHR50 heritability. Our results indicate that SNPs in nine genes related to cardiomyocyte and neuronal functions, as well as cardiac memory formation, fully account for the heritability of the submaximal heart rate training response.


Chitosan/alginate nanogel potentiate berberine uptake and enhance oxidative stress mediated apoptotic cell death in HepG2 cells.
Biopolymer-based nanoscale drug delivery systems have become a promising approach to overcome the limitations associated with conventional chemotherapeutics used for cancer treatment. Herein, we reported to develop a hydrophilic nanogel (NG) composed of Chitosan (Chi) and sodium alginate (Alg) using the ion gelation method for delivering Berberine hydrochloride (BBR), an alkaloid obtained from Berberis aristata roots. The use of different nanocarriers for BBR delivery has been reported previously, but the bioavailability of these carriers was limited due to phagocytic uptake and poor systemic delivery. The developed NG showed enhanced stability and efficient entrapment of BBR ~92 %, resulting in a significant increase in bioavailability. The pH-dependent release behavior demonstrated sustained and effective release of ~86 %, ~74 % and, ~53 % BBR at pH 5.5, 6.6, and 7.4 respectively after 72 h, indicating its potential as a drug carrier. Additionally, the cellular uptake of BBR was significantly higher ~19 % in the BBR-NG (25 μM) than in bulk BBR (100 μM), leading to enhanced ROS generation, mitochondrial depolarisation, and inhibition of cell proliferation and colony formation in HepG2 cells. In summary, the results suggest that the Chi/Alg biopolymer-based nano-formulation could be an effective approach for delivering BBR and enhancing its cellular uptake, efficacy, and cytotoxicity.


The effects of subgingival irrigation with chlorhexidine and stannous fluoride. A preliminary investigation.
This pilot study compared the effectiveness of subgingival irrigation with 0.12% chlorhexidine, 1.64% stannous fluoride, and sterile saline, in addition to scaling and root planing, on levels of Bacteroides porphyromonas and the clinical parameters bleeding tendency, probing depth, and attachment level. A convenience sample of eight patients, exhibiting 32 sites with moderate periodontal disease, was randomly assigned to receive all treatments according to a four-quadrant treatment design. Subgingival irrigation was performed at 0, 1, 2, and 3 weeks following scaling and root planing. Clinical and microbial assessments were measured at baseline, 4, 8, and 12 weeks. Data were analyzed using a two-factor repeated measure analysis of variance, and the Newman-Keuls sequential range test or Friedman test and Kruskal-Wallis test revealed statistically significant (p less than .01) improvements in probing depths, attachment levels, and Bacteroides porphyromonas for all groups at 12 weeks when compared to baseline values. No statistically significant differences occurred between any of the treatment groups at any time period. Based on the findings of this investigation, it has been concluded that four weekly irrigations with 0.12% chlorhexidine, 1.64% stannous fluoride, or saline irrigation did not enhance the beneficial effects of scaling and root planing alone.


The role of local impedance drop in the acute lesion efficacy during pulmonary vein isolation performed with a new contact force sensing catheter-A pilot study.
Our pilot study aimed to evaluate the role of local impedance drop in lesion formation during pulmonary vein isolation with a novel contact force sensing ablation catheter that records local impedance as well and to find a local impedance cut-off value that predicts successful lesion formation. After completing point-by-point radiofrequency pulmonary vein isolation, the success of the applications was evaluated by pacing along the ablation line at 10 mA, 2 ms pulse width. Lesions were considered successful if loss of local capture was achieved. Out of 645 applications, 561 were successful and 84 were unsuccessful. Compared to the unsuccessful ablation points, the successful applications were shorter (p = 0.0429) and had a larger local impedance drop (p<0.0001). There was no difference between successful and unsuccessful applications in terms of mean contact force (p = 0.8571), force-time integral (p = 0.0699) and contact force range (p = 0.0519). The optimal cut-point for the local impedance drop indicating successful lesion formation was 21.80 Ohms on the anterior wall [AUC = 0.80 (0.75-0.86), p<0.0001], and 18.30 Ohms on the posterior wall [AUC = 0.77 (0.72-0.83), p<0.0001]. A local impedance drop larger than 21.80 Ohms on the anterior wall and 18.30 Ohms on the posterior wall was associated with an increased probability of effective lesion creation [OR = 11.21, 95%CI 4.22-29.81, p<0.0001; and OR = 7.91, 95%CI 3.77-16.57, p<0.0001, respectively]. The measurement of the local impedance may predict optimal lesion formation. A local impedance drop > 21.80 Ohms on the anterior wall and > 18.30 Ohms on the posterior wall significantly increases the probability of creating a successful lesion.


Unilateral spatial neglect affected by right-sided stimuli in a three-dimensional virtual environment: A preliminary proof-of-concept study.
Unilateral spatial neglect (USN) is defined as the inability to attend and see on one side, which seriously interferes with daily life. Clinically, patients with left USN commonly demonstrate a striking immediate capture of attention from ipsilesional, right-sided items as soon as a visual scene unfolds (i.e., magnetic attraction [MA]). Therefore, this preliminary study utilized a three-dimensional (3D) virtual environment to evaluate the effects of eliminating stimuli in the rightward space and directing attention to the left on neglect symptoms. Seven patients with USN participated in this study, and two types of visual stimuli were created: the numbers and objects in the 3D virtual environment. To eliminate the visual stimuli on the right side, a moving slit was introduced in the virtual environment. During the experiment, patients were required to orally identify each object and number both in moving and nonmoving slit conditions. A statistical comparison of scores with and without the moving slit in the 3D virtual space indicated significant changes in the object stimuli condition; however, no statistically significant difference was observed in the number stimuli condition. Masking the right side within the 3D virtual space increased the number of objects that can be recognized on the left side by patients with USN. The results may allow interventions in a virtual reality environment that closely resembles the patient's real-life space.Clinical Relevance-Magnetic attraction is a symptom seen in patients in clinical practice, but there is no method of rehabilitation. The proposed moving slit method is expected to be effective because it enables attention guidance in a three-dimensional space.


Stability of compounded thioguanine oral suspensions.
PURPOSE. Updated information on the stability of compounded thioguanine oral suspensions prepared with currently available ingredients, as well as results of testing to determine if the addition of an antioxidant could extend shelf life by inhibiting formation of guanine, are presented. METHODS. Using triturated thioguanine tablets, three compounded suspensions were prepared: (1) a reference formulation containing methylcellulose and simple syrup, (2) an equivalent formulation using Ora-Plus and Ora-Sweet, and (3) an antioxidant-containing formulation prepared by adding ascorbic acid to the equivalent formulation. The compounded batches were stored at room temperature (19-23 °C). The chemical stability of the suspensions was evaluated immediately after compounding and at weekly intervals by a validated liquid chromatography-mass spectrometry (LCMS) assay method; physical stability was evaluated by regular visual checks and weekly pH testing. RESULTS. As demonstrated by serial LCMS testing, mean thioguanine levels in sampled batches of all three suspensions remained above accepted standards and mean guanine formation remained within acceptable limits for up to 63 days. The addition of ascorbic acid appeared to slow guanine formation but did not significantly extend the shelf life of the suspension. CONCLUSION. Compounded oral suspensions of thioguanine 20 mg/mL exhibited acceptable chemical and physical stability for up to nine weeks at 19-23 °C. The addition of ascorbic acid at a concentration of 0.1% to the suspension was not effective in consistently increasing the shelf life of the thioguanine suspensions.


Agronomic, economic, and environmental performance of nitrogen rates and source in Bangladesh's coastal rice agroecosystems.
Farmers in low-elevation coastal zones in South Asia face numerous food security and environmental sustainability challenges. This study evaluated the effects of nitrogen (N) rate and source on the agronomic, economic, and environmental performance of transplanted and rainfed 'aman' (monsoon-season) rice in Bangladesh's non-saline coastal areas. Fifty-one farmers participated in trials distributed across two landscape positions described as 'highlands' (on which field water inundation depth typically remains <30 cm) and 'medium-highlands' (inundation depths 30-90 cm) planted singly with varieties appropriate to each position (BRRI dhan 39 for highlands and the traditional variety Bhushiara for medium-highlands). Researcher designed but farmer-managed dispersed plots were located across three district sub-units (Barisal Sadar, Hizla, Mehendigonj) and compared N source (broadcast prilled urea or deep-placed urea super granules (USG)) at four N rates. Rice grown on medium-highlands did not respond to increasing N rates beyond 28 kg N ha-1, indicating that little fertilization is required to maintain yields and profitability while limiting environmental externalities. In highland locations, clear trade-offs between agronomic and environmental goals were observed. To increase yields and profits for BRRI dhan 39, 50 or 75 kg N ha-1 was often needed, although these rates were associated with declining energy and increasing greenhouse gas (GHG) efficiencies. Compared to prilled urea, USG had no impact on yield, economic, energy and GHG efficiencies in medium-highland locations. USG conversely led to 4.2-5.8% yield improvements at higher N rates on highlands, while also increasing energy efficiency. Given the observed yield, agronomic and economic benefit of USG, our preliminary results that farmers can consider use of USG at 50 kg N ha-1 to produce yields equivalent to 75 kg N ha-1 of prilled urea in highland landscapes, while also reducing environmental externalities. These results suggest that when assessing sustainable intensification (SI) strategies for rice in South Asia's coastal zones, N requirements should be evaluated within specific production contexts (e.g. cultivar type within landscape position) to identify options for increasing yields without negatively influencing environmental and economic indicators. Similar studies in other parts of coastal South Asia could help policy-makers prioritize investments in agriculture with the aim of improving rice productivity while also considering income generation and environmental outcomes.


Analysis of the CoIE1 stability determinant Rcd.
Multimer formation is an important cause of instability for many multicopy plasmids. Plasmid CoIE1 is maintained stably because multimers are converted to monomers by Xer-mediated site-specific recombination at the cer site. However, multimer resolution is not the whole story; inactivation of a promoter (Pcer) within cer causes plasmid instability even though recombination is unaffected. The promoter directs the synthesis of a short transcript (Rcd) which is proposed to delay the division of multimer-containing cells. Mapping of the 5' terminus of Rcd confirms that transcription initiates from Pcer. The 3' terminus shows considerable heterogeneity, consistent with a primary transcript of 95 nt being degraded via intermediates of 79 and 70 nt. Secondary structure predictions for Rcd are presented. Of four mutations which abolish Rcd-mediated growth inhibition, one reduces the activity of Pcer while the other three map to the rcd coding sequence and reduce the steady-state level of the transcript. RNA folding analysis suggests that these three mutant transcripts adopt a common secondary structure in which the major stem-loop differs from that of wild-type Rcd. A survey of 24 cer-like multimer resolution sites revealed six which contain Pcer-like sequences. The putative transcripts from these sites have similar predicted secondary structures to Rcd and contain a highly conserved 15 base sequence. To test the hypothesis that Rcd acts as an anti-sense RNA, interacting with its target gene(s) through the 15 nt sequence, we used DNA hybridization and sequence analysis to find matches to this sequence in the Escherichia coli chromosome. Our failure to find plausible anti-sense targets has led to the suggestion that Rcd may interact directly with a protein target.


Hock lesions in dairy cows in freestall herds: a cross-sectional study of prevalence and risk factors.
Hock lesions (HL) in dairy cows are a common animal welfare problem in modern dairy production with freestall housing systems, but there are no large-scale studies addressing its epidemiology in Sweden. The aims of this cross-sectional study were to investigate the prevalence of HL of different severity in 100 Swedish freestall dairy herds, and to identify cow- and herd-related risk factors. Associations between HL and mastitis as well as culling were also investigated. In total, 3217 cows from 99 herds were included in the statistical analyses. The overall cow prevalence of hair loss on the hock (mild HL) was 68% and the prevalence of ulceration or evident swelling of the hock, with or without hair loss, (severe HL) was 6%. The within-herd prevalence varied among herds, between 23 and 100% for mild HL, and between 0 and 32% for severe ones. Breed (higher risk for Swedish Holstein than for Swedish Red) and days in milk (higher risk at 181-305 days than at 0-90 days) were cow-related risk factors associated with both types of lesions, whereas higher parity and cleaner cows were associated only with increased risk of severe HL. A reduced risk for mild HL was seen in cows housed on mattresses compared to rubber mats, and in cows housed on peat compared to other bedding materials. Also, cows in herds with a high proportion of not yet inseminated heifers older than 17 months had a lower risk of mild HL than cows in herds with a low proportion. Risk for severe HL was lower when cubicles were of recommended width compared to under recommendation, for organic production compared to conventional, and when teat dip or no treatment after milking was used, compared to teat spray. For both mild and severe HL, herringbone milking parlors were associated with higher risk than tandem parlors. We found no significant associations between HL and mastitis or culling. The prevalence of HL is high in Swedish dairy herds, although most lesions are mild. Several cow- and herd-related risk factors were identified and the results can be used to improve recommendations for the prevention of HL in Swedish freestall dairy herds.


Adaptation and limitations of established hemagglutination inhibition assays for the detection of porcine anti-swine influenza virus H1N2 antibodies.
Hemagglutination inhibition (HI) has been a reliable method for determining porcine antibody levels to the well-characterized swine influenza virus (SIV) H1N1 and H3N2 subtypes. However, the recent emergence of the novel H1N2 serotype of SIV and the persistence of 2 other serotypes (H1N1 and H3N2) in the United States swine population represents a significant challenge to diagnostics. Both standardized and modified HI protocols were used in a blinded study to examine a collection of 50 control sera representing a total of 12 swine that were experimentally inoculated with one of the 3 SIV subtypes. Using these control sera data, a statistical basis for analysis was established in an attempt to classify 30 field sera with known case histories or seroprevalance into SIV serotype categories. By this approach 57% of the field sera could be classified into specific categories. The remaining samples that could not be classified reliably were most likely composed of heterogeneous anti-SIV antibody populations. These results indicate that although serological differentiation might be possible in a controlled environment, applications of these methods to field samples are currently problematic. Approaches other than HI will be required to fulfill the current need for SIV diagnostics and surveillance when specific serotype identification is required.


Olfactory inputs activate the medial entorhinal cortex via the hippocampus.
The lateral and medial regions of the entorhinal cortex differ substantially in terms of connectivity and pattern of activation. With regard to olfactory input, a detailed and extensive physiological map of the olfactory projection to the entorhinal cortex is missing, even if anatomic studies suggest that the olfactory afferents are confined to the lateral and rostral entorhinal region. We studied the contribution of the medial and lateral entorhinal areas to olfactory processing by analyzing the responses induced by lateral olfactory tract stimulation in different entorhinal subfields of the in vitro isolated guinea pig brain. The pattern of synaptic activation of the medial and lateral entorhinal regions was reconstructed either by performing simultaneous multisite recordings or by applying current source density analysis on field potential laminar profiles obtained with 16-channel silicon probes. Current source density analysis demonstrated the existence of a direct monosynaptic olfactory input into the superficial 300 microm of the most rostral part of the lateral entorhinal cortex exclusively, whereas disynaptic sinks mediated by associative fibers arising from the piriform cortex were observed at 100-350 microm depth in the entire lateral aspect of the cortex. No local field responses were recorded in the medial entorhinal region unless a large population spike was generated in the hippocampus (dentate gyrus and CA1 region) by a stimulus 3-5x the intensity necessary to obtain a maximal monosynaptic response in the piriform cortex. In these conditions, a late sink was recorded at a depth of 600-1000 microm in the medial entorhinal area (layers III-V) 10.6 +/- 0.9 (SD) msec after a population spike was simultaneously recorded in CA1. Diffuse activation of the medial entorhinal region was also obtained by repetitive low-intensity stimulation of the lateral olfactory tract at 2-8 Hz. Higher or lower stimulation frequencies did not induce hippocampal-medial entorhinal cortex activation. These results suggest that the medial and the lateral entorhinal regions have substantially different roles in processing olfactory sensory inputs.


Visceral analgesia: effects of xylazine, butorphanol, meperidine, and pentazocine in horses.
The visceral analgesic, cardiorespiratory, and behavioral effects induced by xylazine, butorphanol, meperidine, and pentazocine were determined in 9 adult horses with colic. Colic was produced by inflating a balloon in the horses' cecum. Heart rate, respiratory rate, mean arterial blood pressure, and cardiac output increased after cecal balloon inflation. Xylazine and butorphanol decreased the hemodynamic response to cecal balloon inflation. Meperidine and pentazocine had minimal effects on the cardiorespiratory changes induced by cecal balloon inflation. Xylazine produced the most pronounced visceral analgesia. The duration of visceral analgesia was longest with xylazine (approx 90 minutes) followed by butorphanol (approx 60 min) and then by meperidine and pentazocine (approx 30 to 35 min). Accurate assessment of the effects of visceral analgesics is dependent upon the use of objective tests to evaluate pain.


Stanozolol treatment decreases the mitochondrial ROS generation and oxidative stress induced by acute exercise in rat skeletal muscle.
Anabolic androgenic steroids are used in the sport context to enhance muscle mass and strength and to increase muscle fatigue resistance. Since muscle fatigue has been related to oxidative stress caused by an exercise-linked reactive oxygen species (ROS) production, we investigated the potential effects of a treatment with the anabolic androgenic steroid stanozolol against oxidative damage induced on rat skeletal muscle mitochondria by an acute bout of exhaustive exercise. Mitochondrial ROS generation with complex I- and complex II-linked substrates was increased in exercised control rats, whereas it remained unchanged in the steroid-treated animals. Stanozolol treatment markedly reduced the extent of exercise-induced oxidative damage to mitochondrial proteins, as indicated by the lower levels of the specific markers of protein oxidation, glycoxidation, and lipoxidation, and the preservation of the activity of the superoxide-sensitive enzyme aconitase. This effect was not due to an enhancement of antioxidant enzyme activities. Acute exercise provoked changes in mitochondrial membrane fatty acid composition characterized by an increased content in docosahexaenoic acid. In contrast, the postexercise mitochondrial fatty acid composition was not altered in stanozolol-treated rats. Our results suggest that stanozolol protects against acute exercise-induced oxidative stress by reducing mitochondrial ROS production, in association with a preservation of mitochondrial membrane properties.


A quality improvement cycle of the care process in a general surgery hospitalization ward.
There could be important failures in clinical data and plan records that potentially influence the surgical care process. To complete a quality of care improvement cycle on the surgical care process in the General Surgery ward rounds. Structured quality criteria were measured, in order to identify major deficiencies, to implement improvement measures and to reassess the quality of surgical care process. Furthermore, we'll value the viability the implementation of a structured registration system and nutritional assessment method. Comparative quality study (n = 60) before-after the setting of several improvement measures derived from the analysis of the surgical care process. Evaluated criteria were the information received by the patient, adequate recording of the clinical course and plan of care established by the surgeon in the Electronic Health Record, recording of patient's weight and size and nutritional assessment. Informative sessions, subjective, objective, assessment, plan notes on the electronic clinical record and a nutritional assessment test were implemented. There was a significant improvement in all measured criteria (information, data records and nutritional assessment). Simple organizational measures allow a significant improvement in the information process, clinical records and malnutrition risk detection in a surgical ward.


Rate Allocation in Predictive Video Coding Using a Convex Optimization Framework.
Optimal rate allocation is among the most challenging tasks to perform in the context of predictive video coding, because of the dependencies between frames induced by motion compensation. In this paper, using a recursive rate-distortion model that explicitly takes into account these dependencies, we approach the frame-level rate allocation as a convex optimization problem. This technique is integrated into the recent HEVC encoder, and tested on several standard sequences. Experiments indicate that the proposed rate allocation ensures a better performance (in the rate-distortion sense) than the standard HEVC rate control, and with a little loss with respect to an optimal exhaustive research, which is largely compensated by a much shorter execution time.


Accessing healthcare as a person with a rugby-related spinal cord injury in South Africa: the injured player's perspective.
Rugby carries a risk for serious injuries, including acute spinal cord injuries. The lifetime health consequences of these injuries may be far-reaching. In this study, we aimed to describe barriers and facilitators to healthcare and rehabilitation, in individuals with rugby-related spinal cord injuries in South Africa. This study adopted a pragmatic qualitative approach. Stratified purposive sampling was used to select interviews from players from a variety of socio-economic status and geographical areas for inclusion in this study. The final sample consisted of thirty-one (n = 31) participants. Thematic analysis was used to analyze the data. Participants described long-term, accessible, affordable, quality healthcare and rehabilitation as important contributors to their quality of life. Even though public healthcare is accessible from a cost point of view, quality of care, availability of rehabilitation services and factors such as affordability and availability of adequate transport remain a barrier for lower socio-economic groups. This population was unique, as the support from a dedicated organization enabled participants to overcome some barriers, highlighting the challenges of the healthcare system in maintaining the health of people with spinal cord injuries. Every effort should be made to create equitable access to healthcare and rehabilitation for persons with spinal cord injuries in South Africa.


Competition between excited state proton and OH(-) transport via a short water wire: solvent effects open the gate.
We investigate the acid-base proton exchange reaction in a microsolvated bifunctional chromophore by means of quantum chemical calculations. The UV/vis spectroscopy shows that equilibrium of the keto- and enol-forms in the electronic ground state is shifted to the keto conformation in the excited state. A previously unknown mechanism involving a hydroxide ion transport along a short water wire is characterized energetically, which turns out to be competitive with the commonly assumed proton transport. Both mechanisms are shown to have a concerted character, as opposed to a step-wise mechanism. The alternative mechanism of a hydrogen atom transport is critically examined, and evidence for strong solvent dependence is presented. Specifically, we observe electrostatic destabilization of the corresponding πσ* state by the aqueous solvent. As a consequence, no conical intersections are found along the reaction pathway.


Vector Flow Imaging Compared with Conventional Doppler Ultrasound and Thermodilution for Estimation of Blood Flow in the Ascending Aorta.
Transverse oscillation (TO) is a real-time ultrasound vector flow method implemented on a commercial scanner. The TO setup was examined on a flowrig with constant and pulsatile flow. Subsequently, 25 patients undergoing cardiac bypass surgery were scanned intraoperatively with TO on the ascending aorta and compared to transesophageal echocardiography (TEE) and pulmonary artery catheter thermodilution (PACTD). On the flowrig, TO had a precision of 5.5%, 9.4% and 14.7%, a percentage error of 18.2%, 14.6% and 40.7%, and a mean bias of 0.4 cm/s, 36.8 ml/min and 32.4 ml/min for velocity and flow rate (constant and pulsatile) estimation. The correlation coefficients for all flowrig evaluations were 0.99 indicating systematic bias. After bias correction, the percentage error was reduced to 11.5%, 12.6% and 15.9% for velocity and flow rate (constant and pulsatile) estimation. In the in vivo setup, TO, TEE, and PACTD had a precision of 21.9%, 13.7%, and 12.0%. TO compared with TEE and PACTD had a mean bias of 12.6 cm/s and -0.08 l/min, and a percentage error of 23.4%, and 36.7%, respectively. The percentage error was reduced to 22.9% for the TEE comparison, but increased to 43.8% for the PACTD comparison, after correction for the systematic bias found in the flowrig. TO is a reliable and precise method for velocity and flow rate estimation on a flowrig. However, TO with the present setup, is not interchangeable with PACTD for cardiac volume flow estimation, but is a reliable and precise angle-independent ultrasound alternative for velocity estimation of cardiac flow.


Treatment of Executive Function Deficits in autism spectrum disorder with repetitive transcranial magnetic stimulation: A double-blind, sham-controlled, pilot trial.
In youth and young adults with autism spectrum disorder (ASD), executive function (EF) deficits may be a promising treatment target with potential impact on everyday functioning. To conduct a pilot randomized, double-blind, parallel, controlled trial evaluating repetitive transcranial magnetic stimulation (rTMS) for EF deficits in ASD. In Toronto, Ontario (November 2014 to June 2017), a 20-session, 4-week course of 20 Hz rTMS targeting dorsolateral prefrontal cortex (DLPFC) (90%RMT) was compared to sham stimulation in 16-35 year-olds with ASD (28 male/12 female), without intellectual disability, who had impaired everyday EF performance (n = 20 active/n = 20 sham). Outcome measures evaluated protocol feasibility and clinical effects of active vs. sham rTMS on EF performance. The moderating effect of baseline functioning was explored. Of eligible participants, 95% were enrolled and 95% of randomized participants completed the protocol. Adverse events across treatment arms were mild-to-moderate. There was no significant difference between active vs. sham rTMS on EF performance. Baseline adaptive functioning moderated the effect of rTMS, such that participants with lower baseline functioning experienced significant EF improvement in the active vs. sham group. Our pilot RCT demonstrated the feasibility and acceptability of using high frequency rTMS targeting DLPFC in youth and young adults with autism. No evidence for efficacy of active versus sham rTMS on EF performance was found. However, we found promising preliminary evidence of EF performance improvement following active versus sham rTMS in participants with ASD with more severe adaptive functioning deficits. Future work could focus on examining efficacy of rTMS in this higher-need population. Repetitive Transcranial Magnetic Stimulation (rTMS) for Executive Function Deficits in Autism Spectrum Disorder and Effects on Brain Structure: A Pilot Study; https://clinicaltrials.gov/ct2/show/NCT02311751?term = ameis&rank = 1; NCT02311751. The trial was funded by: an American Academy of Child and Adolescent Psychiatry (AACAP) Pilot Research Award, the Innovation Fund from the Alternate Funding Plan of the Academic Health Sciences Centres of Ontario, and an Ontario Mental Health Foundation (OMHF) Project A Grant and New Investigator Fellowship.