How much does it cost to preserve a larynx? An economic study.
Various studies report an increase in costs when induction chemotherapy is included in the treatment of advanced laryngeal cancer, but to our knowledge no studies have yet compared the economic costs of total laryngectomy versus induction chemotherapy in the treatment of advanced laryngeal cancer. We have conducted a retrospective study comparing the costs of treatment and survival in 96 patients with a T3N0-1 glottic carcinoma. Findings showed that the average cost per patient in the group of patients treated by total laryngectomy with or without postoperative radiotherapy was 5,853 Eur, while that for the group of patients who began treatment with induction chemotherapy was 6,452 Eur. The adjusted 5-year survival for patients treated with total laryngectomy with or without postoperative radiotherapy was 80%, and 72% for patients who began treatment with induction chemotherapy. Sixteen of the 35 patients (46%) receiving induction chemotherapy were spared laryngectomy. The use of induction chemotherapy in the treatment of patients with advanced laryngeal carcinomas involved an increase in cost of 600 Eur in relation to treatment with total laryngectomy and postoperative radiotherapy. However, from an economic point of view, we consider induction chemotherapy to be an important consideration in an organ-preservation strategy.


What is a clinically meaningful survival benefit in refractory metastatic colorectal cancer?
Assessment of the clinical benefit of cancer treatments can be highly subjective, influenced by both perspective and context. Such assessments are required in regulatory and policy decision-making, but consistency between jurisdictions is often lacking. Clear and consistent standards for determining when a treatment offers a meaningful benefit, relative to the current standard of care, can help to address issues of equity and transparency in health technology assessment. For metastatic colorectal cancer (mcrc), no standardized Canadian definition of clinically meaningful benefit has yet been proposed. Colorectal Cancer Canada therefore convened a group of medical oncologists expert in colorectal cancer to review the literature about clinical significance. The resulting consensus is intended to apply to any therapeutic agent being considered in the setting of chemotherapy-refractory mcrc. It was agreed that overall survival is the appropriate measure of clinical efficacy in chemorefractory mcrc. As quantitative targets for efficacy, an improvement of 2 months or more in median overall survival or a hazard ratio for survival of 0.75 or lower (or both) are proposed as the threshold for clinically meaningful benefit. That threshold could be influenced by a treatment's effect on quality of life. Treatment toxicity is also relevant to the assessment of clinical benefit in this setting, specifically when significant differences in treatment tolerability are evident.


AXT914 a novel, orally-active parathyroid hormone-releasing drug in two early studies of healthy volunteers and postmenopausal women.
Antagonism of the calcium-sensing receptor in the parathyroid gland leads to parathyroid hormone (PTH) release. Calcilytics are a new class of molecules designed to exploit this mechanism. In order to mimic the known bone-anabolic pharmacokinetic (PK) profile of s.c. administered PTH, such molecules must trigger sharp, transient and robust release of PTH. The results of two early clinical studies with the orally-active calcilytic AXT914, a quinazolin-2ne derivative are reported. These were GCP-compliant, single and multiple dose studies of PK/PD and tolerability in healthy volunteers and postmenopausal women. The first study, examined single ascending doses (4 to 120 mg) and limited multiple doses (60 or 120 mgq.d. for 12 days) of AXT914. The second study was a randomized, double-blind, active- and placebo-controlled, 4-week repeat-dose parallel group study of healthy postmenopausal women (45 and 60 mg AXT914, placebo, 20 μg Forteo/teriparatide/PTH(1-34) fragment). AXT914 was well tolerated at all doses and reproducibly induced the desired PTH-release profiles. Yet, 4 weeks of 45 or 60 mg AXT914 did not result in the expected changes in circulating bone biomarkers seen with teriparatide. However total serum calcium levels increased above baseline in the 45 and 60 mg AXT914 treatment groups (8.0% and 10.7%, respectively), compared to that in the teriparatide and placebo groups (1.3% and 1.0%, respectively). Thus the trial was terminated after a planned interim analysis due to lack of effect on bone formation biomarkers and dose-limiting effects on serum calcium. In conclusion, AXT914 was well tolerated but the observed transient and reproducible PTH-release after repeat oral administration of AXT914 which showed an exposure profile close to that of s c. PTH, did not translate into a bone anabolic response and was associated with a persistent dose-related increase in serum calcium concentrations.


Non-insulin-dependent (type II) diabetes mellitus.
Non-insulin-dependent (type II) diabetes mellitus is an inherited metabolic disorder characterized by hyperglycemia with resistance to ketosis. The onset is usually after age 40 years. Patients are variably symptomatic and frequently obese, hyperlipidemic and hypertensive. Clinical, pathological and biochemical evidence suggests that the disease is caused by a combined defect of insulin secretion and insulin resistance. Goals in the treatment of hyperglycemia, dyslipidemia and hypertension should be appropriate to the patient's age, the status of diabetic complications and the safety of the regimen. Nonpharmacologic management includes meal planning to achieve a suitable weight, such that carbohydrates supply 50% to 60% of the daily energy intake, with limitation of saturated fats, cholesterol and salt when indicated, and physical activity appropriate to the patient's age and cardiovascular status. Follow-up should include regular visits with the physician, access to diabetes education, self-monitoring of the blood or urine glucose level and laboratory-based measurement of the plasma levels of glucose and glycated hemoglobin. If unacceptably high plasma glucose levels (e.g., 8 mmol/L or more before meals) persist the use of orally given hypoglycemic agents (a sulfonylurea agent or metformin or both) is indicated. Temporary insulin therapy may be needed during intercurrent illness, surgery or pregnancy. Long-term insulin therapy is recommended in patients with continuing symptoms or hyperglycemia despite treatment with diet modification and orally given hypoglycemic agents. The risk of pancreatitis may be reduced by treating severe hypertriglyceridemia (fasting serum level greater than 10 mmol/L) and atherosclerotic disease through dietary and, if necessary, pharmacologic management of dyslipidemia. Antihypertensive agents are available that have fewer adverse metabolic effects than thiazides and beta-adrenergic receptor blockers. New drugs are being developed that will enhance effective insulin secretion and action and inhibit the progress of complications.


Conditional Synaptic Vesicle Markers for Drosophila.
The release of neurotransmitters from synaptic vesicles (SVs) at pre-synaptic release sites is the principle means by which information transfer between neurons occurs. Knowledge of the location of SVs within a neuron can thus provide valuable clues about the location of neurotransmitter release within a neuron and the downstream neurons to which a given neuron is connected, important information for understanding how neural circuits generate behavior. Here the development and characterization of four conditional tagged SV markers for Drosophila melanogaster is presented. This characterization includes evaluation of conditionality, specificity for SV localization, and sensitivity of detection in diverse neuron subtypes. These four SV markers are genome-edited variants of the synaptic vesicle-specific protein Rab3. They depend on either the B2 or FLP recombinases for conditionality, and incorporate GFP or mCherry fluorescent proteins, or FLAG or HA epitope tags, for detection.


Defects in the N-linked oligosaccharide biosynthetic pathway in a Trypanosoma brucei glycosylation mutant.
Concanavalin A (ConA) kills the procyclic (insect) form of Trypanosoma brucei by binding to its major surface glycoprotein, procyclin. We previously isolated a mutant cell line, ConA 1-1, that is less agglutinated and more resistant to ConA killing than are wild-type (WT) cells. Subsequently we found that the ConA resistance phenotype in this mutant is due to the fact that the procyclin either has no N-glycan or has an N-glycan with an altered structure. Here we demonstrate that the alteration in procyclin N-glycosylation correlates with two defects in the N-linked oligosaccharide biosynthetic pathway. First, ConA 1-1 has a defect in activity of polyprenol reductase, an enzyme involved in synthesis of dolichol. Metabolic incorporation of [3H]mevalonate showed that ConA 1-1 synthesizes equal amounts of dolichol and polyprenol, whereas WT cells make predominantly dolichol. Second, we found that ConA 1-1 synthesizes and accumulates an oligosaccharide lipid (OSL) precursor that is smaller in size than that from WT cells. The glycan of OSL in WT cells is apparently Man9GlcNAc2, whereas that from ConA 1-1 is Man7GlcNAc2. The smaller OSL glycan in the ConA 1-1 explains how some procyclin polypeptides bear a Man4GlcNAc2 modified with a terminal N-acetyllactosamine group, which is poorly recognized by ConA.


Abattoir survey of bovine tuberculosis in tanta, centre of the Nile delta, with in silico analysis of gene mutations and protein-protein interactions of the involved mycobacteria.
Bovine tuberculosis is a transboundary disease of high economic and public health burden worldwide. In this study, post-mortem examination of 750 cattle and buffalo in Tanta abattoir, Centre of the Nile Delta, revealed visible TB in 4% of animals and a true prevalence of 6.85% (95% CI: 5.3%-8.9%). Mycobacterial culture, histopathology and RT-PCR targeting all members of M. tuberculosis complex were performed, upon which 85%, 80% and 100% of each tested lesions were confirmed as TB, respectively. Mpb70-targeting PCR was conducted on ten RT-PCR positive samples for sequencing and identified nine Mycobacterium (M.) bovis strains and, interestingly, one M. tuberculosis (Mtb) strain from a buffalo. Bioinformatics tools were used for prediction of mutations, nucleotide polymorphisms, lineages, drug resistance and protein-protein interactions (PPI) of the sequenced strains. The Mtb strain was resistant to rifampicin, isoniazid and streptomycin, and to the best of our knowledge, this is the first report of multidrug resistant (MDR)-Mtb originating from buffaloes. Seven M. bovis strains were resistant to ethambutol and ethionamide. Such resistances were associated with KatG, rpoB, rpsL, embB and ethA genes mutations. Other mutations and nucleotide polymorphisms were also predicted, some are reported for the first time and require experimental work for validation. PPI revealed more interactions than what would be expected for a random set of proteins of similar size and had dense interactions between nodes that are biologically connected, as a group. Two M. bovis strains belonged to BOV AFRI lineage (Spoligotypes BOV 1; BOV 2) and eight strains belonged to East-Asian (Beijing) lineage. In conclusion, visible TB was prevalent in the study area, RT-PCR is the best to confirm the disease, MDR-Mtb is associated with buffalo TB, and mycobacteria of different lineages carry many resistance genes to chemotherapeutic agents used in treatment of human TB constituting a major public health risk.


TreeRipper web application: towards a fully automated optical tree recognition software.
Relationships between species, genes and genomes have been printed as trees for over a century. Whilst this may have been the best format for exchanging and sharing phylogenetic hypotheses during the 20th century, the worldwide web now provides faster and automated ways of transferring and sharing phylogenetic knowledge. However, novel software is needed to defrost these published phylogenies for the 21st century. TreeRipper is a simple website for the fully-automated recognition of multifurcating phylogenetic trees (http://linnaeus.zoology.gla.ac.uk/~jhughes/treeripper/). The program accepts a range of input image formats (PNG, JPG/JPEG or GIF). The underlying command line c++ program follows a number of cleaning steps to detect lines, remove node labels, patch-up broken lines and corners and detect line edges. The edge contour is then determined to detect the branch length, tip label positions and the topology of the tree. Optical Character Recognition (OCR) is used to convert the tip labels into text with the freely available tesseract-ocr software. 32% of images meeting the prerequisites for TreeRipper were successfully recognised, the largest tree had 115 leaves. Despite the diversity of ways phylogenies have been illustrated making the design of a fully automated tree recognition software difficult, TreeRipper is a step towards automating the digitization of past phylogenies. We also provide a dataset of 100 tree images and associated tree files for training and/or benchmarking future software. TreeRipper is an open source project licensed under the GNU General Public Licence v3.


Integration and Segregation of Default Mode Network Resting-State Functional Connectivity in Transition-Age Males with High-Functioning Autism Spectrum Disorder: A Proof-of-Concept Study.
The aim of this study is to assess the resting-state functional connectivity (RsFc) profile of the default mode network (DMN) in transition-age males with autism spectrum disorder (ASD). Resting-state blood oxygen level-dependent functional magnetic resonance imaging data were acquired from adolescent and young adult males with high-functioning ASD (n = 15) and from age-, sex-, and intelligence quotient-matched healthy controls (HCs; n = 16). The DMN was examined by assessing the positive and negative RsFc correlations of an average of the literature-based conceptualized major DMN nodes (medial prefrontal cortex [mPFC], posterior cingulate cortex, bilateral angular, and inferior temporal gyrus regions). RsFc data analysis was performed using a seed-driven approach. ASD was characterized by an altered pattern of RsFc in the DMN. The ASD group exhibited a weaker pattern of intra- and extra-DMN-positive and -negative RsFc correlations, respectively. In ASD, the strength of intra-DMN coupling was significantly reduced with the mPFC and the bilateral angular gyrus regions. In addition, the polarity of the extra-DMN correlation with the right hemispheric task-positive regions of fusiform gyrus and supramarginal gyrus was reversed from typically negative to positive in the ASD group. A wide variability was observed in the presentation of the RsFc profile of the DMN in both HC and ASD groups that revealed a distinct pattern of subgrouping using pattern recognition analyses. These findings imply that the functional architecture profile of the DMN is altered in ASD with weaker than expected integration and segregation of DMN RsFc. Future studies with larger sample sizes are warranted.


Comparative cranio-mandibular myology of three species of Batoidea from the Southern Gulf of California, Mexico.
The mandibular apparatus of batoids (skates, electric rays, guitarfishes, stingrays, and sawfishes) is composed of a few skeletal elements to which the muscular bundles, responsible for all movements involved in the feeding mechanism, are inserted. The description of the different mandibular morphologies can help to understand the different feeding guilds in this group. In this study, we examined the cranio-mandibular myology of adult Rostroraja velezi, Narcine entemedor, and Zapteryx exasperata, three species of rays that coexist in the Southern Gulf of California, Mexico. This study described the muscles on the ventral and the dorsal surfaces for each species, identified the origins and insertions of these muscles, as well as the general characteristics of muscle morphology. There were 17 and 18 muscle bundles attached to the feeding apparatus, including five on the dorsal surface. Only the levator rostri, which elevates the rostrum during feeding, showed considerable differences in shape and size among species. The muscles of the adductor complex showed the greatest differences in size among the three species. N. entemedor presented the exclusive muscle X in the lower mandibular area and the extreme reduction of the coracohyoideus in the pharyngeal area derived from the absence of the basihyal cartilage. The information generated in our study supports the morphological specialization of electric rays (N. entemedor) for an almost exclusive suction feeding strategy.


Integration of sensory, spinal, and volitional descending inputs in regulation of human locomotion.
We reported previously that both transcutaneous electrical spinal cord stimulation and direct pressure stimulation of the plantar surfaces of the feet can elicit rhythmic involuntary step-like movements in noninjured subjects with their legs in a gravity-neutral apparatus. The present experiments investigated the convergence of spinal and plantar pressure stimulation and voluntary effort in the activation of locomotor movements in uninjured subjects under full body weight support in a vertical position. For all conditions, leg movements were analyzed using electromyographic (EMG) recordings and optical motion capture of joint kinematics. Spinal cord stimulation elicited rhythmic hip and knee flexion movements accompanied by EMG bursting activity in the hamstrings of 6/6 subjects. Similarly, plantar stimulation induced bursting EMG activity in the ankle flexor and extensor muscles in 5/6 subjects. Moreover, the combination of spinal and plantar stimulation exhibited a synergistic effect in all six subjects, eliciting greater motor responses than either modality alone. While the motor responses to spinal vs. plantar stimulation seems to activate distinct but overlapping spinal neural networks, when engaged simultaneously, the stepping responses were functionally complementary. As observed during induced (involuntary) stepping, the most significant modulation of voluntary stepping occurred in response to the combination of spinal and plantar stimulation. In light of the known automaticity and plasticity of spinal networks in absence of supraspinal input, these findings support the hypothesis that spinal and plantar stimulation may be effective tools for enhancing the recovery of motor control in individuals with neurological injuries and disorders.


Surface self-assembled hybrid nanocomposites with electroactive nanoparticles and enzymes confined in a polymer matrix for controlled electrocatalysis.
A three-dimensional network of highly branched poly(ethyleneimine) (PEI) is designed and synthesized on gold electrode surfaces. A self-assembled monolayer (SAM) of dithiobis(succinimidyl propionate) (DTSP) on a gold electrode was first prepared, which is confirmed by the reductive desorption of Au-S units. The PEI polymer was then covalently immobilized onto the DTSP layer, leaving free primary amine groups acting as a 3D skeleton for high loading of electroactive enzyme-size Prussian blue nanoparticles (PBNPs, 6 nm) via electrostatic trapping. Atomic force microscopy was used to disclose the microscopic structures of the different layers during the surface architecture formation. The resulting surface-bound nanostructured composite shows high electrochemical activity arising from confined PBNPs, and acts as an efficient electrocatalyst towards H2O2 reduction. Facile electron communication is achieved as reflected by a large electron transfer (ET) rate constant (ks) of 200 s-1, and the possible electron propagation mechanisms in the polymer network are discussed. This surface/interfacial nanocomposite can be further used in the accommodation of enzymes for electrochemical bio-catalysis. Glucose oxidase (GOD) was used towards this end, in a proof-of-concept study. This enzyme can be co-trapped in the PEI matrix and is interconnected with PBNPs, leading to highly efficient electrocatalyic oxidation and detection of glucose.


Mass Spectrometry-Based Network Analysis Reveals New Insights Into the Chemodiversity of 28 Species in Aspergillus section Flavi.
Aspergillus section Flavi includes some of the most famous mycotoxin producing filamentous fungi known to mankind. In recent years a number of new species have been included in section Flavi, however these species have been much less studied from a chemical point of view. In this study, we explored one representative strain of a total of 28 fungal species in section Flavi by systematically evaluating the relationship between taxonomy and secondary metabolites with LC-MS/MS analysis for the first time and dereplication through an in-house database and the Global Natural Product Social Molecular Networking (GNPS) platform. This approach allowed rapid identification of two new cyclopiazonic acid producers (A. alliaceus and A. arachidicola) and two new tenuazonic acid producers (A. arachidicola and A. leporis). Moreover, for the first time we report species from section Flavi to produce fumifungin and sphingofungins B-D. Altogether, this study emphasizes that the chemical diversity of species in genus Aspergillus section Flavi is larger than previously recognized, and especially that understudied species are prolific producers of important mycotoxins such as fumi- and sphingofungins not previously reported from this section. Furthermore, our work demonstrates Global Natural Product Social (GNPS) Molecular Networking as a powerful tool for large-scale chemotaxonomic analysis of closely related species in filamentous fungi.


The cardiorenal syndrome in heart failure: an evolving paradigm.
Heart failure constitutes a significant source of morbidity and mortality in the United States, and its incidence and prevalence continue to grow, increasing its burden on the healthcare system. Renal dysfunction in patients with heart failure is common and has been associated with adverse clinical outcomes. This interaction, termed the cardiorenal syndrome, is a complex phenomenon characterized by a pathophysiologic disequilibrium between the heart and the kidney, in which malfunction of 1 organ consequently promotes the impairment of the other. Multiple neurohumoral mechanisms are involved in this cardiorenal interaction, including the deficiency of and/or resistance to compensatory natriuretic peptides, leading to sodium retention, volume overload and organ remodeling. Management of patients with the cardiorenal syndrome can be challenging and should be individualized. Emerging therapies must address the function of both organs to secure better clinical outcomes. To this end, a multidisciplinary approach is recommended to achieve optimal results.


Flashback phenomena after administration of LSD and psilocybin in controlled studies with healthy participants.
LSD and psilocybin are increasingly used in phase I trials and evaluated as therapeutic agents for mental disorders. The phenomenon of reoccurring drug-like experiences after the acute substance effects have worn off was described for both substances and especially attributed to LSD. According to the DSM-V, the persisting and distressing manifestation of these experiences is called hallucinogen-persisting perception disorder (HPPD). Data on both conditions is very limited. This study aims to provide descriptive data on reoccurring drug-like experiences after the administration of LSD and psilocybin in controlled studies with healthy participants. Data from 142 healthy subjects enrolled in six double-blinded, placebo-controlled, randomized cross-over studies were analyzed. In total, 60 subjects received LSD; 27 subjects received LSD, MDMA, and D-amphetamine; 31 subjects received LSD and psilocybin; and 25 subjects received psilocybin and escitalopram. At the end-of-study visit (mean 39.8 days after last study session, SD 37.2), subjects were asked for any reoccurring drug effects since the initial substance effects had worn off. Those reporting reoccurring perception changes more than 24 h after administration were contacted for follow-up (mean follow-up duration: 31.2 months, SD 28.6). Thirteen out of 142 subjects reported reoccurring drug-like experiences (LSD: seven, psilocybin: two, both: four). The reported phenomena were predominantly mild and perceived as neutral to pleasant. Flashbacks were mostly of visual nature, lasted for seconds to minutes, and occurred within a week after the last drug administration. Two subjects reported distressing experiences that subsided spontaneously. One subject reported brief and pleasant visual perception changes which reoccurred for 7 months. None of the subjects reported impairment in their daily lives. None of the cases met DSM-V criteria for HPPD. Reoccurring drug-like experiences after the administration of LSD and psilocybin are a common phenomenon occurring in up to 9.2% of healthy subjects (7.8% for LSD, 8.3% for psilocybin and 14.3% if both substances are administered). Additionally, our work suggests that flashback phenomena are not a clinically relevant problem in controlled studies with healthy participants.


Evaluation of children with neurobrucellosis.
Brucellosis is an endemic disease in many areas throughout the world. Central nervous system involvement is a serious complication of brucellosis with a ratio of 4-11% of all patients. to describe our experience in diagnosis, treatment, and outcome of 25 pediatric patients with neurobrucellosis. This study included a review of medical records of patients who were diagnosed with neurobrucellosis between March 2001 and March 2009. Patients who had both clinical findings consistent with neurobrucellosis and positive microbiologic/serologic examinations of CSF with abnormal CSF findings were enrolled in the study. The study included 25 patients between 1 and 15 years of age (mean 8.8 years), while 15 were males and 10 were females. Most of the patients (52 %) were in the age group of 5-9 years with male predominance. The distribution of cases showed density in June and February. The most commonly presented complaints were headache, fever and sweating while the most commonly observed findings were fever and meningeal irritation signs. All patients had positive cerebrospinal fluid agglutination test for brucellosis. Four different regimens were used based on ceftriaxone, doxycycline, cotrimoxasole, streptomycin, and rifampicin. One patient died, three patients were discharged with sequel, and the remaining patients (84 %) were discharged with full recovery. Clinicians, especially those providing health services in endemic areas like Turkey, should keep in mind that neurobrucellosis can be involved in patients with unexplained symptoms like memory impairment or in patients diagnosed with meningitis (Tab. 5, Fig, 2, Ref. 39).


[Aspects of the radiological evaluation of rheumatoid arthritis].
This paper gives an overview of several aspects of the radiographic evaluation of rheumatoid arthritis (RA). Radiographs give important information about the structural damage caused by RA. On a group level, the natural progression follows a rather linear cause, but on an individual patient level, this can vary widely. Feet are often affected earlier and to a larger extent than hands. Both hands and feet give a good overall impression of the damage in the large joints and are sufficient to monitor the patient. It is recommended to take annual films of hands and feet to monitor patients. Several scoring methods are available for application in clinical trials. Most widely used are the Sharp and Larsen methods with their modifications. For use in clinical practice the SENS method is more feasible. Several drugs are capable of retarding radiographic progression. However, it is difficult to compare results across trials and these difficulties are elaborated.


Sedimentation-equilibrium studies on the heterogeneity of two beta-lactamases.
Solutions of crystalline beta-lactamase I and beta-lactamase II, prepared by Kuwabara (1970), were examined in the ultracentrifuge and their sedimentation coefficients, diffusion coefficients, molecular weights and heterogeneity determined. Each sample was shown to consist of a major component comprising at least 97% of the material and a minor component of much higher molecular weight. The molecular weights of the major components were 27800 for beta-lactamase I and 35600 for beta-lactamase II. Emphasis is placed on a straightforward practical way of analysing the sedimentation-equilibrium results on mixtures of two macromolecular components rather than on a strict theoretical solution. Appendices describe the theory of systems at both chemical and sedimentation equilibrium and the procedure for calculating the combined distribution of two components.


[Prospective study of the presence of atrophic rhinitis and AR-pathogenic Bordetella bronchiseptica and Pasteurella multocida bacteria in piglets of health-certified pig-breeding farms].
Two hundred eight-week-old piglets from 20 pig-breeding farms, on which the clinical findings did not provide any justification for suspecting the presence of atrophic rhinitis (AR), were examined for this disease. Clinical parameters were brachygnathia superior (BS) and snout-distorsion (KD). Piglets with BS (n = 48) were observed on 13 farms and with KD on 8 farms. Morbid anatomical parameters were deviation of the nasal septum (SD) and atrophy of the ventral conchae (AVC). SD was observed in 76 piglets from 17 farms. Piglets with AVC gradation greater than or equal to 1 were recorded on 20 farms and with grade greater than or equal to 3 on 12 farms. The average gradation of AVC over all farms was 1.3 varying from 0.2 to 2.9. Bacteriological studies were done in the piglets for Bordetella bronchiseptica (BB) and AR pathogen Pasteurella multocida (PM+). The score of AVC showed a significant increase on farms on which infections with PM+ and BB was present. It is indicated that pig-herds with an average AVC gradation smaller than 0.7 do have a chance of less than 10% to be infected with PM+. BB was isolated from 52 piglets, from the nose in 49 cases and from the trachea in 27 cases respectively. PM was isolated from 67 piglets. Examination by equine blood agar showed that 15 piglets were positive for PM in the nose, 32 animals being found to be positive for PM by examination using mice. When mice were used in studies of the tonsils, 59 were found to be positive for PM. Of 45 piglets infected with type D, 16 were found te be PM+ following the guinea-pig skin test, the results being regarded as doubtful in two cases. Epidemiological studies for BB and PM+ on 117 breeding-farms in the Province of Overijssel showed that 92% of these farms were infected with BB and 28% with PM+. To be able to qualify breeding-farms as 'not being suspected of AR' concentrated clinical, pathological and bacteriological studies are required.


Academic Leadership for Advancing Diversity, Equity, and Inclusion.
Academic nursing prepares nurses for the future to deliver, improve, and innovate health care. Moving forward on this imperative requires nurse leaders to support diversity, equity, and inclusion efforts thus partnering with individuals, families, communities, and other sectors to advance health equity, reduce health disparities, and improve care. The purpose of this article is to describe the creation, launch, and evaluation of an Office of Diversity, Equity, and Inclusion (DEI) in an academic nursing institution. Lessons learned are shared to assist others in their own journey to establish a DEI structure within an academic setting, especially one with a robust academic-practice partnership. The ideas shared are easily transferable to nonacademic settings.


Functional replacement of the intracellular region of the Notch1 receptor by Epstein-Barr virus nuclear antigen 2.
The intracellular region (RAMIC) of the mouse Notch1 receptor interacts with RBP-J/CBF-1, which binds to the DNA sequence CGTGGGAA and suppresses differentiation by transcriptional activation of genes regulated by RBP-J. Epstein-Barr virus nuclear antigen 2 (EBNA2) is essential for immortalization of human B cells by the virus. EBNA2 is a pleiotropic activator of viral and cellular genes and is targeted to DNA at least in part by interacting with RBP-J. We found that EBNA2 and the Notch1 RAMIC compete for binding to RBP-J, indicating that their interaction sites on RBP-J overlap at least partially. EBNA2 and Notch1 RAMIC transactivated the same set of viral and host promoters, i.e., the EBNA2 response element of the Epstein-Barr virus TP1 and the HES-1 promoter. Furthermore, EBNA2 functionally replaced the Notch1 RAMIC by suppressing differentiation of C2C12 myoblast progenitor cells.


A qualitative study on the oral health of humanitarian migrants in Canada.
There is limited evidence to guide oral health policy and services for the 25,000 refugees and asylum seekers who arrive in Canada yearly. The purpose of this study was to explore and understand the pre-migration use of dental services, oral health knowledge, and the effects of oral disease among newly arrived humanitarian migrants in order to inform policy and practice for the population. Using focused ethnography and the public health model of the dental care process, we conducted face-to-face interviews (50-60 minutes) with a purposive sample of humanitarian migrants who had indicated the need for dental care. We observed mobile dental clinics that provided care to underserved communities in Montreal. Data were analyzed using a thematic and contextual approach that combined inductive and deductive frameworks. Participants included 25 humanitarian migrants from four global geographical regions. Five major thematic categories were explored: problem-based dental consultation, self-assessed oral health status, causes of oral diseases, personal oral hygiene, and good oral health for wellbeing. In their countries of origin, participants consulted a dentist when oral symptoms persisted. They cited excessive sugar consumption and inadequate oral hygiene as causes of oral diseases, and reported significant oral diseases impacts that limited their daily functions and wellbeing once in Canada. Humanitarian migrants were knowledgeable about causes of oral disease and the importance of good oral health, yet poor oral health continued to affect their lives in Canada in important ways.


No increase in alpine snowbed productivity in response to experimental lengthening of the growing season.
Climate change effects on snow cover and thermic regime in alpine tundra might lead to a longer growing season, but could also increase risks to plants from spring frost events. Alpine snowbeds, i.e. alpine tundra from late snowmelt sites, might be particularly susceptible to such climatic changes. Snowbed communities were grown in large monoliths for two consecutive years, under different manipulated snow cover treatments, to test for effects of early (E) and late (L) snowmelt on dominant species growth, plant functional traits, leaf area index (LAI) and aboveground productivity. Spring snow cover was reduced to assess the sensitivity of snowbed alpine species to severe early frost events, and dominant species freezing temperatures were measured. Aboveground biomass, productivity, LAI and dominant species growth did not increase significantly in E compared to L treatments, indicating inability to respond to an extended growing season. Edapho-climatic conditions could not account for these results, suggesting that developmental constraints are important in controlling snowbed plant growth. Impaired productivity was only detected when harsher and more frequent frost events were experimentally induced by early snowmelt. These conditions exposed plants to spring frosts, reaching temperatures consistent with the estimated freezing points of the dominant species ( approximately -10 degrees C). We conclude that weak plasticity in phenological response and potential detrimental effects of early frosts explain why alpine tundra from snowbeds is not expected to benefit from increased growing season length.


Pheochromocytoma: state-of-the-art and future prospects.
This review provides current understanding of the pathophysiology of pheochromocytoma and the wide range of associated clinical manifestations that have led to earlier recognition of the disease. In addition, it reviews optimal screening methods and localization techniques that have enhanced the clinician's ability to make the diagnosis with greater certainty. This article will also discuss alternative antihypertensive regimens and innovative anesthetic and surgical procedures that have made successful management more promising than ever before. Areas requiring further development include additional clinical experience with the measurement of plasma metanephrines that have been shown to have high sensitivity and specificity in the diagnosis of sporadic and familial pheochromocytoma, optimizing cost effectiveness of diagnostic imaging, improving the ability to predict and treat malignant pheochromocytoma, and elucidating not only the surgical approach but, perhaps with rapid advances in molecular genetics, ways of preventing familial pheochromocytoma.


Real-time observation of valence electron motion.
The superposition of quantum states drives motion on the atomic and subatomic scales, with the energy spacing of the states dictating the speed of the motion. In the case of electrons residing in the outer (valence) shells of atoms and molecules which are separated by electronvolt energies, this means that valence electron motion occurs on a subfemtosecond to few-femtosecond timescale (1 fs = 10(-15) s). In the absence of complete measurements, the motion can be characterized in terms of a complex quantity, the density matrix. Here we report an attosecond pump-probe measurement of the density matrix of valence electrons in atomic krypton ions. We generate the ions with a controlled few-cycle laser field and then probe them through the spectrally resolved absorption of an attosecond extreme-ultraviolet pulse, which allows us to observe in real time the subfemtosecond motion of valence electrons over a multifemtosecond time span. We are able to completely characterize the quantum mechanical electron motion and determine its degree of coherence in the specimen of the ensemble. Although the present study uses a simple, prototypical open system, attosecond transient absorption spectroscopy should be applicable to molecules and solid-state materials to reveal the elementary electron motions that control physical, chemical and biological properties and processes.


Stroop interference for food- and body-related words: a meta-analysis.
According to cognitive theories of eating disorders, biased information processing in favour of dysfunctional attitudes about food and body appearance plays a vital role in the development and maintenance of such disorders. Data from 27 studies evaluating Stroop interference for food- and body-related words with negative overtones were included in a meta-analysis in order to investigate whether such processing biases are specific to eating disordered samples. Participants were females characterised as eating disordered, non-eating disordered but nevertheless over-concerned with body appearance and eating, and normal controls. Mean Stroop interference for eating disordered females was of medium effect size (Cohen's d=0.48) and significantly larger than for both non-eating disordered females concerned with body appearance and eating, and normal control females (both d=0.21). Stroop interference for eating disordered females was thus of fairly modest magnitude where it was unclear whether such interference is specific to this sample.


Asymmetric operation of the locomotor central pattern generator in the neonatal mouse spinal cord.
The rhythmic voltage oscillations in motor neurons (MNs) during locomotor movements reflect the operation of the pre-MN central pattern generator (CPG) network. Recordings from MNs can thus be used as a method to deduct the organization of CPGs. Here, we use continuous conductance measurements and decomposition methods to quantitatively assess the weighting and phase tuning of synaptic inputs to different flexor and extensor MNs during locomotor-like activity in the isolated neonatal mice lumbar spinal cord preparation. Whole cell recordings were obtained from 22 flexor and 18 extensor MNs in rostral and caudal lumbar segments. In all flexor and the large majority of extensor MNs the extracted excitatory and inhibitory synaptic conductances alternate but with a predominance of inhibitory conductances, most pronounced in extensors. These conductance changes are consistent with a "push-pull" operation of locomotor CPG. The extracted excitatory and inhibitory synaptic conductances varied between 2 and 56% of the mean total conductance. Analysis of the phase tuning of the extracted synaptic conductances in flexor and extensor MNs in the rostral lumbar cord showed that the flexor-phase-related synaptic conductance changes have sharper locomotor-phase tuning than the extensor-phase-related conductances, suggesting a modular organization of premotor CPG networks consisting of reciprocally coupled, but differently composed, flexor and extensor CPG networks. There was a clear difference between phase tuning in rostral and caudal MNs, suggesting a distinct operation of CPG networks in different lumbar segments. The highly asymmetric features were preserved throughout all ranges of locomotor frequencies investigated and with different combinations of locomotor-inducing drugs. The asymmetric nature of CPG operation and phase tuning of the conductance profiles provide important clues to the organization of the rodent locomotor CPG and are compatible with a multilayered and distributed structure of the network.


HCV and HCC: clinical update and a review of HCC-associated viral mutations in the core gene.
Hepatocellular carcinoma (HCC) is a common cause of cancer-related mortality. The worldwide incidence of HCC and the hepatitis C virus (HCV) has increased over several decades suggesting an etiologic link. Progress has been made recently in several fields related specifically to HCV and HCC. The epidemiology has been better characterized, surveillance and treatment programs have been instituted, and data have emerged detailing the effect of other risk factors for HCC in patients with HCV. Studies of HCV-related HCC suggest differences in oncogenic potential according to genotype and mutations in the viral sequence. These same mutations are associated with interferon treatment failure, insulin resistance, and expression of HCV minicore proteins. These viral mutations provide a focus for investigations into the molecular basis of HCV-related oncogenesis, which may lead to improved diagnostic tests for early (curable) HCC and to interventions to prevent, and/or greatly retard, the oncogenic process.


How much particle surface corrugation is sufficient to improve aerosol performance of powders?
The current study aimed to quantify the different degree of particle surface corrugation and correlate it to the aerosol performance of powders. Powders of different degree of surface corrugation were prepared by spray drying under varying conditions. The solid-state properties of the powders including particle size, morphology, crystal form, true density, and moisture content were characterized. The degree of surface corrugation was quantified by the surface fractal dimension (Ds) obtained by light scattering. The aerosol performance was studied by dispersing the powders using the Rotahaler at 60 L/min into a multi-stage liquid impinger. Fine particle fraction (FPF) was expressed as the wt% of BSA particles of size < or =5 microm in the aerosol. Four powders of increasing degree of particle surface corrugation were prepared, with Ds ranging from 2.06 for the least corrugated to 2.41 for the most corrugated. The powders had a similar size distribution (VMD 3 microm, span 1.4-1.5) and solid-state properties. Increasing the surface corrugation, Ds, slightly from 2.06 to 2.18 enhanced the FPF significantly from 27% to 41%. This was explained by the reduced area of contacts and increased separation distance between the particles. Further increase of corrugation (Ds > or = 2.18) did not improve FPF. Powders with varying degrees of corrugation were successfully obtained by spray drying with their surface roughness quantified by fractal analysis. It was shown that only a relatively small degree of surface corrugation was sufficient to accomplish a considerable improvement in the aerosol performance of the powder.


Portal vein thrombosis in cirrhosis: Controversies and latest developments.
Portal vein thrombosis (PVT) is encountered in liver cirrhosis, particularly in advanced disease. It has been a feared complication of cirrhosis, attributed to significant worsening of liver disease, poorer clinical outcomes and potential inoperability at liver transplantation; also catastrophic events such as acute intestinal ischaemia. Optimal management of PVT has not yet been addressed in any consensus publication. We review current literature on PVT in cirrhosis; its prevalence, pathophysiology, diagnosis, impact on the natural history of cirrhosis and liver transplantation, and management. Studies were identified by a search strategy using MEDLINE and Google Scholar. The incidence of PVT increases with increasing severity of liver disease: less than 1% in well-compensated cirrhosis, 7.4%-16% in advanced cirrhosis. Prevalence in patients undergoing liver transplantation is 5%-16%. PVT frequently regresses instead of uniform thrombus progression. PVT is not associated with increased risk of mortality. Optimal management has not been addressed in any consensus publication. We propose areas for future research to address unresolved clinical questions.


A single-cell view on alga-virus interactions reveals sequential transcriptional programs and infection states.
The discovery of giant viruses infecting eukaryotes from diverse ecosystems has revolutionized our understanding of the evolution of viruses and their impact on protist biology, yet knowledge on their replication strategies and transcriptome regulation remains limited. Here, we profile single-cell transcriptomes of the globally distributed microalga Emiliania huxleyi and its specific giant virus during infection. We detected profound heterogeneity in viral transcript levels among individual cells. Clustering single cells based on viral expression profiles enabled reconstruction of the viral transcriptional trajectory. Reordering cells along this path unfolded highly resolved viral genetic programs composed of genes with distinct promoter elements that orchestrate sequential expression. Exploring host transcriptome dynamics across the viral infection states revealed rapid and selective shutdown of protein-encoding nuclear transcripts, while the plastid and mitochondrial transcriptomes persisted into later stages. Single-cell RNA-seq opens a new avenue to unravel the life cycle of giant viruses and their unique hijacking strategies.


Endoscopic findings and the prevalence of Helicobacter pylori in chronic renal failure patients with dyspepsia.
Peptic ulcer disease (PUD) occurs in up to one fourth of patients with chronic renal failure (CRF). Some of the factors implicated in its causation include hypergastrinaemia, secondary hyperparathyroidism, drugs and, recently, Helicobacter pylori infection. Studies on the latter have been few, with none having been carried out in Kenya. To evaluate the upper gastrointestinal tract endoscopic findings and to determine the prevalence of H. pylori in CRF patients with dyspepsia. STUDY DESIGN AND POPULATION: A prospective study of seventy seven consecutive patients with CRF and dyspepsia compared with consecutive age, sex and socio-economically matched seventy seven controls (no CRF) with dyspepsia. Kenyatta National Hospital (KNH), the major referral and teaching hospital, Nairobi, Kenya. In both the study population and the controls, upper gastrointestinal endoscopy was carried out. H. pylori was tested for using the biopsy urease test and histology. Patients were considered to have H. pylori if they tested positive on both tests. Findings at endoscopy and presence of H. pylori. Inflammatory lesions (gastritis, duodenitis) (42%) and duodenal ulcers (18.4%) were the commonest findings in the two groups combined. The prevalence of H. pylori in the 154 subjects studied was 54.5%. There was no statistically significant difference between the prevalence of H. pylori in CRF patients (53.2%) and the controls (55.8%) (p = 0.746). Patients with endoscopically proven PUD had a very high prevalence of H. pylori (87.3%) regardless of their renal function status. Dyspepsia in patients with or without CRF was due to multiple causes and over 50% were attributable to H. pylori. The prevalence of H. pylon in dyspeptic CRF patients was similar to that in dyspeptic patients with normal renal function.


Preclinical characterization of acyl sulfonimidamides: potential carboxylic acid bioisosteres with tunable properties.
Herein we present the preclinical characterization of novel compounds containing the linear acyl sulfonimidamide functionality. Specifically, we studied the pKa , lipophilicity, in vitro metabolic stability, plasma protein binding, Caco-2 permeability, and aqueous solubility for nine aryl acyl sulfonimidamides. In comparison with widely used carboxylic acid bioisosteres, the acyl sulfonimidamides were found to be less acidic and more lipophilic depending on the substitution pattern in the studied compounds. Importantly, the pKa values (5.9-7.6) were significantly influenced by substituents on the nitrogen atom and the aryl substituents. Moreover, the acyl sulfonimidamides displayed membrane permeabilities ranging from moderate to very high, which correlated with decreased pKa and low to negligible efflux ratios. We foresee that the chiral sulfur center and the two handles for structural diversity of linear acyl sulfonimidamides will offer new opportunities for drug design and for improving the oral bioavailability of acidic drug candidates.


West Nile virus in livestock and wildlife.
WN virus is one of the most ubiquitous arboviruses occurring over a broad geographical range and in a wide diversity of vertebrate host and vector species. The virus appears to be maintained in endemic foci on the African continent and is transported annually to temperate climates to the north in Europe and to the south in South Africa. Reports of clinical disease due to natural WN virus infection in wild or domestic animals were much less common than reports of infection (virus isolation or antibody detection). Until recently, records of morbidity and mortality in wild birds were confined to a small number of cases and infections causing encephalitis, sometimes fatal, in horses were reported infrequently. In the period 1996-2001, there was an increase in outbreaks of illness due to WN virus in animals as well as humans. Within the traditional range of WN virus, encephalitis was reported in horses in Italy in 1998 and in France in 2000. The first report of disease and deaths caused by WN virus infection in domestic birds was reported in Israel in 1997-1999, involving hundreds of young geese. In 1999 WN virus reached North America and caused an outbreak of encephalitis in humans in the New York area at the same time as a number of cases of equine encephalitis and deaths in American crows and a variety of other bird species, both North American natives and exotics. Multi-state surveillance for WN virus has been in place since April 2000 and has resulted in the detection of WN virus in thousands of dead birds from an increasing number of species in North America, and also in several species of mammals. The surveillance system that has developed in North America because of the utility of testing dead birds for the rapid detection of WN virus presence has been a unique integration of public health and wildlife health agencies. It has been suggested that the recent upsurge in clinical WN virus infection in wild and domestic animals as well as in humans may be related to the emergence of one or more new strains of WN virus. Virus isolated in New York in 1999 was found to be identical to that from Israel. It was alarming for WN virus to so easily invade the United States and surprising that it became established so quickly in the temperature climate of New York. Its persistence and rapid expansion in the United States leave a number of unanswered questions. New disease characteristics and patterns have occurred and more are evolving as WN virus further invades the western hemisphere. Additional animal research is needed to answer these questions. Some of the research needs include bird migration as a mechanism of virus dispersal, vector and vertebrate host relationships, virus persistence mechanisms, laboratory diagnosis, viral pathogenesis, risk factor studies, vaccine development, and WN virus impact on wildlife (CDC 2001a). Determination of the primary reservoir host species that are involved in the epidemiology of WN virus and the suitable sentinel species for active surveillance are also important research areas.


Patients with Duchenne and Becker muscular dystrophies are not more asymmetrical than healthy controls on timed performance of upper limb tasks.
This study aimed to investigate possible asymmetries and relationships between performance of dominant and non-dominant upper limbs (UL) in patients with Duchenne and Becker muscular dystrophies (DMD/BMD), to compare UL performance of patients and healthy subjects and to investigate the relationship between timed performance of UL and age, motor function and muscle strength in DMD/BMD patients. Sixteen patients with DMD and 3 with BMD were evaluated with Jebsen-Taylor Test (timed performance), Vignos scale and Dimension 3 of Motor Function Measure (motor function), and Medical Research Council scale (muscle strength) on a single session. ANOVA showed no asymmetry between dominant and non-dominant UL, except in the writing subtest, in patients and in healthy controls. There were relationships between dominant and non-dominant UL performances. Correlations between timed performance, motor function and muscle strength were found, but age was not correlated with these variables. These findings may reduce the assessment time, prevent fatigue and provide more accurate clinical reasoning involving UL in DMD/BMD treatment.


Hepatitis B vaccine containing surface antigen and selected preS1 and preS2 sequences. 1. Safety and immunogenicity in young, healthy adults.
The safety and immunogenicity of a yeast-derived recombinant hepatitis B virus (HBV) vaccine containing surface antigen (S) and selected preS1 and preS2 sequences (S-L*) were compared with those of a vaccine prepared with S alone (Engerix-B). S-L* consisted of composite particles containing S and L* at a ratio of 70/30. L* encompassed amino acid residues 12-52 of preS1 residues 133-145 of preS2, and the entire S domain. A total of 100 healthy, HBV-seronegative, young adults were randomized to receive 20 micrograms/dose of either S-L* or Engerix-B under double-blind conditions according to a 0-, 1-, 2-, 12-month schedule. In vivo humoral and in vitro lymphoproliferative responses to S and preS regions were monitored. Addition of the selected preS sequences to S did not enhance the in vivo humoral anti-HBs response but improved the in vitro stimulating capacity of the antigen (L*) in S-L* primed subjects.


The structural conservation of S100 protein during evolution: analysis by reactivity with a monoclonal antibody.
A hybridoma cell line producing a monoclonal antibody (A4) against bovine S100 protein has been produced by fusing mouse myeloma P3X63/Ag8 cells with spleen cells from a BALB/c mouse immunized with bovine S100 protein. A4 is of the IgG2b subclass and was purified by affinity chromatography on a protein A-Sepharose column. Brain extracts from several mammalian and one avian species reacted both with polyclonal rabbit anti-S100 protein antiserum and with A4 in a radioimmunoassay. Brain extract from dog was a notable exception. It reacted with the rabbit antiserum but not with A4. Therefore A4 reacts with a common epitope that is present on S100 proteins from different vertebrate species but is absent on dog S100 protein.


Advantage of Florisil over charcoal separation in a mechanized testosterone radioimmunoassay.
We have facilitated radiommunoassay of testosterone by mechanizing every pipetting step and separating bound from free hormone by shaking assay tubes with activated magnesium silicate (Florisil) in a multi-tube Vortex mixer. Compared with dextran-charcoal separation, Florisil separation has the advantage that testosterone is only negligibly adsorbed except during active shaking, so that the interval for adding adsorbent does not produce discrepancies in bound hormone estimates between first and last assay tubes in a series. Our method gave CVs of 5% (within assay) and 6% (between assay) for samples containing 4.0 mug of testosterone per liter, considerably better than those obtained with charcoal (respectively 12 and 15%). The assay also had low blank values and demonstrated excellent accuracy.


Computer-assisted laparoscopic pyeloplasty: University of Miami experience with the daVinci Surgical System.
We report our experience with laparoscopic pyeloplasty using the daVinci surgical platform. We routinely performed laparoscopic pyeloplasties prior to acquiring the daVinci system. We prospectively evaluated 26 computer-assisted laparoscopic pyeloplasties (CLP) performed since acquiring the device in March 2003. There were 15 male and 11 female patients with a mean age of 34.5 years, who underwent right-sided procedures in 11 cases and left-sided procedures in 15 cases. Four patients (15%) had secondary ureteropelvic junction obstruction. All procedures were performed through a transperitoneal approach over stents placed preoperatively. The operative time excluded the time needed for stent insertion. Radiographic objective success was defined as adequate cortical drainage (t (1/2) < or =15 minutes) and preserved or improved renal function on MAG-3 diuretic renography. A total of 23 dismembered pyeloplasties and 3 Y-V plasties were performed. In five patients, nephroscopy was performed for stone removal. The mean operative time and blood loss were 245 minutes and 69 mL, respectively. There were no intraoperative complications or open conversions. Three minor postoperative complications were noted. The mean length of hospital stay was 2 days (range 1-5 days). In 19 evaluable patients, at a median follow-up of 6 months (range 2-10 days), 15 (79%) had complete symptom resolution, while 3 (16%) had marked symptom improvement. The overall subjective improvement rate thus was 95%. The objective success rate based on our strict diuretic renography criteria was 100%. The overall clinical success rate was 95% (18/19). Robot-assisted laparoscopic pyeloplasty is a feasible alternative to laparoscopic or open pyeloplasty with excellent short-term subjective and objective success rates.


Posterior Reversible Encephalopathy Syndrome in a Patient with Severe Uremia without Hypertension.
A 28-year-old man was admitted to our hospital with nausea, headache and weakness of the left hand. He had severe uremia without hypertension due to recurrent/chronic pyelonephritis. Brain magnetic resonance imaging showed reversible vasogenic edema in the brainstem and bilateral frontal centrum semiovale. All of his neurological symptoms immediately improved after the introduction of hemodialysis. When a patient with uremia presents with neurological symptoms, posterior reversible encephalopathy syndrome should be considered in the differential diagnosis even if high blood pressure is not observed. Brain magnetic resonance imaging may be helpful in such a case, and an appropriate therapy could be subsequently initiated.


Efficient depletion of host DNA contamination in malaria clinical sequencing.
The cost of whole-genome sequencing (WGS) is decreasing rapidly as next-generation sequencing technology continues to advance, and the prospect of making WGS available for public health applications is becoming a reality. So far, a number of studies have demonstrated the use of WGS as an epidemiological tool for typing and controlling outbreaks of microbial pathogens. Success of these applications is hugely dependent on efficient generation of clean genetic material that is free from host DNA contamination for rapid preparation of sequencing libraries. The presence of large amounts of host DNA severely affects the efficiency of characterizing pathogens using WGS and is therefore a serious impediment to clinical and epidemiological sequencing for health care and public health applications. We have developed a simple enzymatic treatment method that takes advantage of the methylation of human DNA to selectively deplete host contamination from clinical samples prior to sequencing. Using malaria clinical samples with over 80% human host DNA contamination, we show that the enzymatic treatment enriches Plasmodium falciparum DNA up to ∼9-fold and generates high-quality, nonbiased sequence reads covering >98% of 86,158 catalogued typeable single-nucleotide polymorphism loci.


Geographical variability and factors associated with caesarean section delivery in India: a comparative assessment of Bihar and Tamil Nadu.
Caesarean section delivery is a major life-saving obstetric surgical intervention for mothers and babies from pregnancy and childbirth related complications. This paper attempts to investigate the geographical variations and correlating factors of caesarean section delivery in India, particularly focusing on the states of Bihar and Tamil Nadu, accounting for one of the lowest and highest prevalence states of caesarean section delivery respectively. This study is based on secondary data, collected from the fourth round of the National Family Health Survey (NFHS-4), 2015-16. We utilized 190,898 women aged 15-49 years who had a living child during the past 5 years preceding the survey. In this study, caesarean section delivery was the outcome variable. A variety of demographic, socio-economic, and pregnancy- and delivery-related variables were considered as explanatory variables. Descriptive statistics, bivariate percentage distribution, Pearson's Chi-square test, and multivariate binary logistic regression models were employed to draw the inferences from data. Of participants, about 19% of women had undergone caesarean section delivery in the country. The state-wise distribution shows that Telangana (60%) followed by Andhra Pradesh (42%) and Tamil Nadu (36%) represented the topmost states in caesarean delivery, while Bihar (7%), Madhya Pradesh (10%), and Jharkhand (11%) placed at the bottom end. Multivariate logistic models show that the likelihood of caesarean delivery was higher among older women (35-49 years), women with higher levels of education, Muslims, women belonging to the upper quintiles of the household wealth, and those who received antenatal care (ANC), experienced pregnancy loss and delivery complications. Moreover, the odds of caesarean section delivery were remarkably greater for the private health sector than the public health sector in both focused states: Bihar (odds ratio [OR] = 12.84; 95% confidence interval [CI]: 10.90, 15.13) and Tamil Nadu (OR = 2.90; 95% CI: 2.54, 3.31). Findings of this study suggest that improvement in female education, providing economic incentives, and spreading awareness through mass media could raise the caesarean section delivery among women whose vaginal delivery could be unsafe for them as well as for their babies. Moreover, providing adequate ANC and well-equipped public healthcare services would facilitate caesarean delivery among needy women.


Identification of chitin as a structural component of Giardia cysts.
The intestinal parasite Giardia lamblia is a significant cause of diarrheal disease, which is perpetuated by the infective cyst form of the parasite. Although a rational approach to the control of giardiasis would be to inhibit cyst formation, nothing is known of the chemical composition of the cyst wall or of its biosynthesis. In these studies, we have shown that chitin is a major structural component of G. lamblia and G. muris cyst walls. This conclusion is based on the finding that chitinase specifically destroys the cyst wall, as revealed by electron microscopy. The presence of chitin was also shown directly by lectin binding studies. Of 12 lectins with diverse carbohydrate recognition specificity, only the N-acetylglucosamine-specific lectins wheat germ agglutinin, succinylated wheat germ agglutinin, and tomato lectin bound to cyst walls, as shown by fluorescence microscopy and cytochemistry. Wheat germ agglutinin binding was completely abolished by treatment of the cysts with purified chitinase. This effect was specific since it could be prevented by incubating the enzyme with chitin before treatment of the cysts. Treatment of cysts with N-acetyl-beta-glucosaminidase partially inhibited wheat germ agglutinin binding, whereas other glycosidases and proteases had no effect. These findings indicate that chitin is a major structural component of Giardia cyst walls and raise the possibility that inhibitors of chitin synthesis may be of use in preventing encystation and thus controlling spread of the disease.


Targeting acute myeloid leukemia by dual inhibition of PI3K signaling and Cdk9-mediated Mcl-1 transcription.
Resistance to cell death is a hallmark of cancer and renders transformed cells resistant to multiple apoptotic triggers. The Bcl-2 family member, Mcl-1, is a key driver of cell survival in diverse cancers, including acute myeloid leukemia (AML). A screen for compounds that downregulate Mcl-1 identified the kinase inhibitor, PIK-75, which demonstrates marked proapoptotic activity against a panel of cytogenetically diverse primary human AML patient samples. We show that PIK-75 transiently blocks Cdk7/9, leading to transcriptional suppression of MCL-1, rapid loss of Mcl-1 protein, and alleviation of its inhibition of proapoptotic Bak. PIK-75 also targets the p110α isoform of PI3K, which leads to a loss of association between Bcl-xL and Bak. The simultaneous loss of Mcl-1 and Bcl-xL association with Bak leads to rapid apoptosis of AML cells. Concordantly, low Bak expression in AML confers resistance to PIK-75-mediated killing. On the other hand, the induction of apoptosis by PIK-75 did not require the expression of the BH3 proteins Bim, Bid, Bad, Noxa, or Puma. PIK-75 significantly reduced leukemia burden and increased the survival of mice engrafted with human AML without inducing overt toxicity. Future efforts to cotarget PI3K and Cdk9 with drugs such as PIK-75 in AML are warranted.


Biliary cirrhosis and tumors induced by chronic administration of thiobenzamide to rats.
Thiobenzamide (TB), a thiono-containing compound, was administered for 38 weeks to female Sprague-Dawley rats at a dose of 1 g/kg standard diet; the resulting liver pathology was followed up to 8 months after withdrawal of the compound from the diet. TB administration induced the appearance of biliary cirrhosis. In the first weeks of intoxication the progressive distortion of the liver architecture was mainly due to significant proliferation of the bile ductules. Later, the liver assumed a macronodular appearance. In addition to regenerative and degenerative changes of the hepatocytes, preneoplastic lesions were also detected, and some enzymic markers of the mixed-function monooxygenase system were decreased. Cholangiofibrotic areas were evident, and many biliary tubules within them showed mucous metaplasia. At the end of the intoxication period, as well as 4 months after drug suspension, large portions of the liver or entire lobes were substituted by connective tissue surrounding nests of bile ductules and atrophied hepatocellular nodules. Four months later, in the virtual absence of cirrhotic changes, each animal harboured one or more tumors (mainly cholangiomas).


Modelling the ancestral sequence distribution and model frequencies in context-dependent models for primate non-coding sequences.
Recent approaches for context-dependent evolutionary modelling assume that the evolution of a given site depends upon its ancestor and that ancestor's immediate flanking sites. Because such dependency pattern cannot be imposed on the root sequence, we consider the use of different orders of Markov chains to model dependence at the ancestral root sequence. Root distributions which are coupled to the context-dependent model across the underlying phylogenetic tree are deemed more realistic than decoupled Markov chains models, as the evolutionary process is responsible for shaping the composition of the ancestral root sequence. We find strong support, in terms of Bayes Factors, for using a second-order Markov chain at the ancestral root sequence along with a context-dependent model throughout the remainder of the phylogenetic tree in an ancestral repeats dataset, and for using a first-order Markov chain at the ancestral root sequence in a pseudogene dataset. Relaxing the assumption of a single context-independent set of independent model frequencies as presented in previous work, yields a further drastic increase in model fit. We show that the substitution rates associated with the CpG-methylation-deamination process can be modelled through context-dependent model frequencies and that their accuracy depends on the (order of the) Markov chain imposed at the ancestral root sequence. In addition, we provide evidence that this approach (which assumes that root distribution and evolutionary model are decoupled) outperforms an approach inspired by the work of Arndt et al., where the root distribution is coupled to the evolutionary model. We show that the continuous-time approximation of Hwang and Green has stronger support in terms of Bayes Factors, but the parameter estimates show minimal differences. We show that the combination of a dependency scheme at the ancestral root sequence and a context-dependent evolutionary model across the remainder of the tree allows for accurate estimation of the model's parameters. The different assumptions tested in this manuscript clearly show that designing accurate context-dependent models is a complex process, with many different assumptions that require validation. Further, these assumptions are shown to change across different datasets, making the search for an adequate model for a given dataset quite challenging.


The role of venous blood gas in the emergency department: a systematic review and meta-analysis.
The aim of this study is to provide a systematic review of the literature reporting agreement between arterial and venous pH, partial pressure of carbon dioxide (PCO2), bicarbonate (HCO3⁻), base excess and lactate; and to perform a meta-analysis of the differences. Medline and Embase searches using Eduserv Athens from 1950 to present were conducted using the terms 'VBG', 'ABG', 'arterial', 'venous', 'blood', 'gas', 'lactate', 'emergency' and 'department'. References of the published papers were hand searched and full-text versions of those deemed helpful to the question were obtained. Mean difference (MD) and 95% limits of agreement (LOA) were either reported or calculated from the published data. Pooled MDs with 95% confidence intervals (CIs) were calculated for differences between arterial and venous pH, PCO2, bicarbonate and lactate. Thirteen articles relevant to pH, 12 relevant to PCO2, 10 relevant to bicarbonate and three relevant to lactate were found. The pooled MD (venous-arterial) for pH was -0.033 pH units (95% CI -0.039 to 0.027) with narrow 95% LOA, the pooled MD for PCO2 was 4.41 mmHg (95% CI 2.55-6.27) with 95% LOA ranging from -20.4 to 25.8 mmHg, the pooled MD for bicarbonate was 1.03 mmol/l (95% CI 0.56-1.50) with 95% LOA ranging from -7.1 to 10.0 mmol/l and the pooled MD for lactate was 0.25 mmol/l (95% CI 0.15-0.35) with 95% LOA ranging from -2.0 to 2.3 mmol/l. Venous and arterial pH and bicarbonate agree reasonably at all values, but the agreement is highest at normal values. Arteriovenous agreement for PCO2 is poor and PvCO2 cannot be relied upon as an absolute representation of PaCO2. However, normal peripheral PvCO2 has a good negative predictive value for normal arterial PCO2, and a normal PvCO2 can be used as a screen to exclude hypercapnic respiratory disease. There may be a poor agreement between arterial and venous lactate at abnormal values; however, if the venous lactate is normal, it is likely the arterial values of this parameter will also be normal.


Characterization, seasonal occurrence, and diel fluctuation of poly(hydroxyalkanoate) in photosynthetic microbial mats.
In situ poly(hydroxyalkanoate) (PHA) levels and repeating-unit compositions were examined in stratified photosynthetic microbial mats from Great Sippewissett Salt Marsh, Mass., and Ebro Delta, Spain. Unlike what has been observed in pure cultures of phototrophic bacteria, the prevalence of hydroxyvalerate (HV) repeating units relative to hydroxybutyrate (HB) repeating units was striking. In the cyanobacteria-dominated green material of Sippewissett mats, the mole percent ratio of repeating units was generally 1HB:1HV. In the purple sulfur bacteria-dominated pink material the relationship was typically 1HB:2HV. In Sippewissett mats, PHA contributed about 0.5 to 1% of the organic carbon in the green layer and up to 6% in the pink layer. In Ebro Delta mats, PHA of approximately 1HB:2HV-repeating-unit distribution contributed about 2% of the organic carbon of the composite photosynthetic layers (the green and pink layers were not separated). Great Sippewissett Salt Marsh mats were utilized for more extensive investigation of seasonal, diel, and exogenous carbon effects. When the total PHA content was normalized to organic carbon, there was little seasonal variation in PHA levels. However, routine daily variation was evident at all sites and seasons. In every case, PHA levels increased during the night and decreased during the day. This phenomenon was conspicuous in the pink layer, where PHA levels doubled overnight. The daytime declines could be inhibited by artificial shading. Addition of exogenous acetate, lactate, and propionate induced two- to fivefold increases in the total PHA levels when applied in the daylight but had no effect when applied at night. The distinct diel pattern of in situ PHA accumulation at night appears to be related, in some phototrophs, to routine dark energy metabolism and is not influenced by the availability of organic nutrients.


Apparent reduction of endogenous creatinine clearance by salicylate treatment.
A prospective study of nine patients with rheumatoid arthritis and 11 healthy volunteers who started salicylate treatment showed that in 18 of the 20 subjects creatinine clearance values fell the average being 25%. Serum creatinine concentration was increased by an average of 38%, although blood urea levels were unchanged. Since 51Cr-edetic acid clearance tests performed at the same time in 12 of the subjects were not affected, the fall in creatinine clearance was probably not due to impaired glomerular function, and salicylate ingestion may therefore invalidate the creatinine clearance test as an index of glomerular filtration rate. It is clearly important to inquire into the patient's analgesic comsumption when using the creatinine clearance test to assess glomerular function.


Identifying Phytochemicals from Biomedical Literature Utilizing Semantic Knowledge Sources.
Chemicals derived from plants (phytochemicals) are major concepts of interest in the study of medicinal plants. To date, efforts to catalogue and organize phytochemical knowledge have resorted to manual approaches. This study explored the potential to leverage publicly accessible semantic knowledge sources for identifying possible phytochemicals. Within the context of this feasibility study, putative phytochemicals were identified for more than 4,000 plants from the Medical Subject Headings Supplementary Concept Records and the Semantic MEDLINE Database. An examination of phytochemicals identified for five selected plant species using the method developed here reveals that there is a disparity in electronically catalogued phytochemical knowledge compared to information from Dr. Duke's Phytochemical and Ethnobotanical Databases maintained by the United States Department of Agriculture. The results therefore suggest that semantic knowledge sources for biomedicine can be utilized as a source for identifying potential phytochemicals and thus contribute to the overall curation of plant phytochemical knowledge.


Elucidation of the exon-intron structure and size of the human protein kinase C beta gene (PRKCB).
As part of a transcriptional mapping project on human chromosome 16p12, a genomic contig was constructed that spanned the alternatively spliced human protein kinase C beta gene (PRKCB). PRKCB was determined to consist of 18 exons covering approximately 375 kb, with a particularly large intron of over 150 kb between exons 2 and 3. PRKCB is nearly 19 times larger than the highly homologous Drosophila melanogaster protein kinase C gene (dPKC), which has a similar-sized open reading frame but only 13 exons. This increase in size has occurred mostly as a result of expansion of introns, with intron size in the human gene averaging 22 kb compared with 1.5 kb in dPKC. The difference in gene size correlates with the difference in genome size, with the human haploid genome being nearly 18 times larger than the 170 Mb Drosophila haploid genome.


[Testosterone metabolism in the skin of women with hirsutism].
A study was made of the endogenous testosterone (T) level in the blood plasma, the rate of T decomposition and the level of 5 alpha-dehydrotestosterone in the pubic skin of women with hirsutism of different genesis. As to T amounts the investigated form of androgenization can be placed in the following sequence: diencephalic form of ovarian polycystosis greater than congenital adrenogenital syndrome greater than ovarian form of polycystosis (before operation) greater than idiopathic form of hirsutism greater than ovarian form of polycystosis (after operation) greater than healthy women. The rate of T transformation into 5 alpha-dehydrotesosterone was higher but in patients with the idiopathic form of hirsutism whereas the level of 5 alpha-dehydrotestosterone was significantly decreased in all the patients. The highest amount of the metabolite was found in the patients with idiopathic hirsutism, the lowest in the diencephalic form of polycystosis and congenital adrenogenital syndrome. In all the investigated clinical forms of androgenization there is a reduced period of the presence of 5 alpha-dehydrotestosterone in the skin cells. An enhanced process of 5 alpha-dehydrotestosterone transformation into androstenediols is likely to be the most probable cause of this phenomenon. An assumption has been made as to a stimulating effect of androstenediols on the virile pattern hair growth in women.


Outcomes of prolonged mechanical ventilation and tracheostomy in critically ill elderly patients: a historical cohort study.
With an aging global population, the increased proportion of elderly patients in the intensive care unit (ICU) raises important questions regarding optimal management. Currently, data on tracheostomy and its outcomes in the elderly are limited. We aimed to determine the in-hospital survival of elderly ICU patients following tracheostomy, and describe impacts on discharge disposition and functional outcomes. We conducted a historical cohort study at two academic hospitals in Toronto. All patients aged ≥ 70 yr who received a tracheostomy during their ICU stay between January 2010 and June 2016 were included in a retrospective chart review. Data on patient demographics, frailty, tracheostomy indication, and outcomes were collected. The study included 270 patients with a mean (standard deviation) age of 81 (6) yr. The majority were admitted to ICU for respiratory failure (147/270, 54%) and received a tracheostomy for prolonged mechanical ventilation (202/270, 75%). Intensive care unit and hospital mortality were 26% (68/270) and 46% (125/270), respectively. Twenty-five percent (67/270) of patients were decannulated during hospital admission, a median [interquartile range (IQR)] of 41 [25-68] days after tracheostomy. Intensive care unit and hospital length of stay were 31 [17-53] and 81 [46-121] days, respectively. At hospital discharge, 6% (17/270) of patients were discharged home, all were frail (median Clinical Frailty Score of 7) and most were tube-fed (101/270, 70%), unable to speak (81/270, 56%), and nonambulatory (98/270, 68%). In patients aged ≥ 70 yr, tracheostomy during ICU stay marked a transition toward prolonged chronic critical illness. Nearly half of the patients died during the admission, and although a quarter were successfully decannulated, the majority of survivors were left with severe frailty and functional impairment.


Towards a gold standard for quantitative coronary arteriography.
Densitometric quantification of coronary artery stenoses in angiographic images can be problematic for two reasons: (i) the x-rays are inadequately oriented with respect to the vessel segments of interest at image acquisition; (ii) non-linear effects due for instance to beam hardening, scattered radiation and veiling glare may reduce the accuracy. As a consequence, appreciable discrepancies between degrees of stenosis measured in two different projections can occur. To overcome these limitations, we have designed and tested a combined correction that compensates (at subsequent analysis) for the error contributions due to the cited sources. It implies 3D reconstruction of the vessel segments of interest and consequently requires an appropriate biplane coronary angiogram. In experiments performed with a dedicated phantom, application of the correction improved the correlation between measured and true area reduction percentages (without correction: y = 1.04x - 4%, r = 0.97, SEE = 6%, n = 35; with correction: y = 1.02x - 0%, r = 0.99, SEE = 3%, n = 35). Applied to ten area stenoses measured biplane in patients and exhibiting strong interplane discrepancies, the correction had a comparable effect (without correction: y = 0.83x - 11%, r = 0.86, SEE = 9%, n = 10; with correction: y = 0.83x + 2%, r = 0.98, SEE = 4%, n = 10). The new densitometric method could possibly be used as a gold standard in the objective evaluation of geometric methods in patients.


The diagnosis performance of convolutional neural network in the detection of pulmonary nodules: a systematic review and meta-analysis.
Pulmonary nodules are an early imaging indication of lung cancer, and early detection of pulmonary nodules can improve the prognosis of lung cancer. As one of the applications of machine learning, the convolutional neural network (CNN) applied to computed tomography (CT) imaging data improves the accuracy of diagnosis, but the results could be more consistent. To evaluate the diagnostic performance of CNN in assisting in detecting pulmonary nodules in CT images. PubMed, Cochrane Library, Web of Science, Elsevier, CNKI and Wanfang databases were systematically retrieved before 30 April 2023. Two reviewers searched and checked the full text of articles that might meet the criteria. The reference criteria are joint diagnoses by experienced physicians. The pooled sensitivity, specificity and the area under the summary receiver operating characteristic curve (AUC) were calculated by a random-effects model. Meta-regression analysis was performed to explore potential sources of heterogeneity. Twenty-six studies were included in this meta-analysis, involving 2,391,702 regions of interest, comprising segmented images with a few wide pixels. The combined sensitivity and specificity values of the CNN model in detecting pulmonary nodules were 0.93 and 0.95, respectively. The pooled diagnostic odds ratio was 291. The AUC was 0.98. There was heterogeneity in sensitivity and specificity among the studies. The results suggested that data sources, pretreatment methods, reconstruction slice thickness, population source and locality might contribute to the heterogeneity of these eligible studies. The CNN model can be a valuable diagnostic tool with high accuracy in detecting pulmonary nodules.


[Kinetics of uptake of Pherajod by the rabbit eye].
The aqueous humor and lenses of rabbit eyes contain inhibitors of photodynamic destructions. Preparations of vitreous, however, more likely seem to contain stimulatory compounds. The anticataractic drug Pherajod significantly blocks the photodynamic formation of destructive oxygen species. Inhibition by 65% of photodynamic destructions, measured as ethylene release from S-methyl-a-ketobutyric acid in the riboflavin-driven oxygen photoactivation, is observed with aqueous humor preparations isolated 5 min after start of incubation of rabbit eyes in Pherajod solution.


Induction of endometriosis alters the peripheral and endometrial regulatory T cell population in the non-human primate.
Endometriosis is a gynecological condition that is characterized by extreme abdominal pain and also decreased fertility. Regulatory T cells (Tregs) have immunosuppressive activity critical for embryonic implantation and likewise the acceptance of tissue engraftment. Utilizing the induced non-human primate (Papio anubis) model of endometriosis, we hypothesize that endometriosis decreases the peripheral and endomet rial Treg profile, whereas ectopic lesions have increased Treg localization. Peripheral blood and endometrium were obtained throughout the menstrual cycle prior to and after induction of disease. Animals were randomly assigned to control (n = 7) or diseased (n = 16) treatment groups. Endometriosis was induced by i.p. injection of autologous menstrual tissue for 2 consecutive months during menses. Peripheral blood and endometrial tissue were collected at d9-11PO at 1, 3, 6, 9, 12 and 15 months post-induction of disease for fluorescence-activated cell sorting, quantitative RT-PCR and immunohistochemistry. Ectopic lesions were excised at 1 and 6 months post-inoculation and also harvested at necropsy (15 months) and processed for RNA of IHC. Identification of Tregs through analysis of FOXP3 expression was conducted utlilizing several methodologies. Differences were determined by non-parametric statistical analysis between all treatment groups and time points. In control animals, the proportion of peripheral natural Tregs (nTregs) was reduced (P < 0.05) during the mid- and late secretory stages of the menstrual cycle compared with menses. The induction of disease decreased peripheral Treg expression at early time points (P < 0.05) and this remained low throughout the time course, compared with the pre-inoculatory level of an individual. FOXP3 gene expression and Treg populations were also decreased in the eutopic endometrium (P < 0.05) compared with control animals, whereas these parameters were increased in ectopic lesions (P < 0.05), compared with the eutopic endometrium. Our data suggest that a reduction in peripheral Tregs may be a causative factor for endometriosis-associated infertility, while the increase in ectopic Treg expression may aid lesion development. Furthermore, endometriosis appears to disrupt Treg recruitment in both eutopic and ectopic endometrium.


Evaluation of the adsorption potential of biochars prepared from forest and agri-food wastes for the removal of fluoxetine.
Twelve biochars from forest and agri-food wastes (pruning of Quercus ilex, Eucalyptus grandis, Pinus pinaster, Quercus suber, Malus pumila, Prunus spinosa, Cydonia oblonga, Eriobotrya japonica, Juglans regia, Actinidia deliciosa, Citrus sinensis and Vitis vinifera) were investigated as potential low-cost and renewable adsorbents for removal of a commonly used pharmaceutical, fluoxetine. Preliminary adsorption experiments allowed to select the most promising adsorbents, Quercus ilex, Cydonia oblonga, Eucalyptus, Juglans regia and Vitis vinifera pruning material. They were characterized by proximate, elemental and mineral analysis, thermogravimetric analysis, Fourier transform infrared spectroscopy, determination of specific surface area and pH at the point of zero charge. Batch and equilibrium studies were performed, and the influence of pH was evaluated. The equilibrium was reached in less than 15 min in all systems. The maximum adsorption capacity obtained was 6.41 mg/g for the Eucalyptus biochar, which also demonstrated a good behavior in continuous mode (packed column).


Trans-acting untranslated elements of groundnut rosette virus satellite RNA are involved in symptom production.
Isolates of groundnut rosette umbravirus (GRV) contain a satellite RNA (sat-RNA), about 900 nucleotides (nt) in length, different variants of which are responsible for the symptoms of different forms of rosette disease in groundnuts and, in the particular instance of sat-RNA YB3b, for the production of yellow blotch symptoms in Nicotiana benthamiana. Sat-RNA YB3b does not affect the accumulation of GRV genomic or subgenomic RNAs in infected plants. Replication of sat-RNA YB3b and induction of yellow blotch symptoms do not require the production of any sat-RNA-encoded proteins. Experiments with deletion mutants identified three functional untranslated elements in sat-RNA YB3b. One (designated R) comprises nt 47-281, is essential for sat-RNA replication and appears to be cis-acting. The other two (designated A and B) comprise nt 280-470 and 629-849, respectively, are both involved in yellow blotch symptom production and can act in trans. Element A contains the determinant that is unique to sat-RNA YB3b. The process of symptom induction by sat-RNA YB3b apparently involves a novel type of specific interaction of two untranslated RNA elements, which can complement each other, with a host factor or factors.


SJL dystrophic mice express a significant amount of human muscle proteins following systemic delivery of human adipose-derived stromal cells without immunosuppression.
Limb-girdle muscular dystrophies (LGMDs) are a heterogeneous group of disorders characterized by progressive degeneration of skeletal muscle caused by the absence of or defective muscular proteins. The murine model for limb-girdle muscular dystrophy 2B (LGMD2B), the SJL mice, carries a deletion in the dysferlin gene that causes a reduction in the protein levels to 15% of normal. The mice show muscle weakness that begins at 4-6 weeks and is nearly complete by 8 months of age. The possibility of restoring the defective muscle protein and improving muscular performance by cell therapy is a promising approach for the treatment of LGMDs or other forms of progressive muscular dystrophies. Here we have injected human adipose stromal cells (hASCs) into the SJL mice, without immunosuppression, aiming to assess their ability to engraft into recipient dystrophic muscle after systemic delivery; form chimeric human/mouse muscle fibers; express human muscle proteins in the dystrophic host and improve muscular performance. We show for the first time that hASCs are not rejected after systemic injection even without immunosuppression, are able to fuse with the host muscle, express a significant amount of human muscle proteins, and improve motor ability of injected animals. These results may have important applications for future therapy in patients with different forms of muscular dystrophies.


[Conversion of nucleoside triphosphates in an RNA polymerase system].
The behavior of nucleoside triphosphates (NTP) in the RNA-polymerase system from E. coli was studied. The conversion of NTP to the corresponding nucleoside diphosphate (NDP) was observed. This phenomenon was accounted for a contaminating enzyme activity in the RNA-polymerase preparations. The possibility to remove such a contamination was demonstrated, the best technique being the DNA-cellulose affinity chromatography. The purified enzyme does not catalyze the intermediate formation of NDP's from NTP's during RNA synthesis with poly(U) and poly(dG)-poly(dC) as templates.


Cancer pain management among underserved minority outpatients: perceived needs and barriers to optimal control.
Minority patients with cancer are at risk for undertreatment of cancer-related pain. Most studies of patient-related barriers to pain control have surveyed primarily non-Hispanic Caucasian patients. The purpose of the current study was to explore barriers to optimal pain management among African-American and Hispanic patients with cancer through the use of structured patient interviews. Structured interviews allowed the authors to probe for previously unidentified barriers to pain management in these populations. Thirty-one socioeconomically disadvantaged minority patients with cancer (14 African-American patients and 17 Hispanic patients) who had cancer-related pain completed structured interviews that assessed three main content areas: information and communication regarding cancer pain, treatment of cancer pain, and the meaning of cancer pain. The African-American and Hispanic patients reported severe pain and many concerns about pain management. The majority of patients in both ethnic groups expressed a belief in stoicism and concerns about possible addiction to opioid medications and the development of tolerance. The patients described their physicians as the most frequent and trusted source of information about cancer pain. However, patients also reported difficulties with communication and a reluctance to complain of pain. The reported barriers to pain management indicate that socioeconomically disadvantaged African-American and Hispanic patients can benefit from educational interventions on cancer pain that dispel myths about opioids and teach patients to communicate assertively about their pain with their physicians and nurses.


Activities of the semisynthetic glycopeptide LY191145 against vancomycin-resistant enterococci and other gram-positive bacteria.
LY191145 is the prototype of a series of compounds with activities against vancomycin-resistant enterococci derived by modification of the glycopeptide antibiotic LY264826. LY191145 had MICs for vancomycin- and teicoplanin-resistant enterococci of < or = 4 micrograms/ml for 50% of isolates and < or = 16 micrograms/ml for 90% of isolates. Its MICs for vancomycin-resistant, teicoplanin-susceptible enterococci were 1 to 8 micrograms/ml. LY191145 retains the potent activities of its parent compound against staphylococci and streptococci. In vivo studies in a mouse infection model confirmed these activities. This compound indicates the potential of semisynthetic glycopeptides as agents against antibiotic-resistant gram-positive bacteria.


Intracytoplasmic growth and virulence of Listeria monocytogenes auxotrophic mutants.
The intracellular growth of several auxotrophic mutants of Listeria monocytogenes was examined in cell culture, and virulence was evaluated in mice by intravenous injection of log-phase bacteria. L. monocytogenes transposon insertion mutants requiring either uracil, phenylalanine, glycine, proline, or nicotinic acid for growth were fully virulent and grew similarly to the parental strain as shown by their growth rates in cell culture. Those requiring all three aromatic amino acids (phenylalanine, tryptophan, and tyrosine) or adenine were 1.5 log10 less virulent than the wild type. A threonine auxotroph, which showed enhanced growth in the presence of threonine-containing peptides as compared with that in the presence of free threonine, was approximately 1 log10 less virulent than the wild type. When host cells were deprived of specific amino acids required by both the host cell and L. monocytogenes, the bacteria continued to grow intracellularly. These studies suggest that the cytoplasm of eucaryotic cells behaves like rich medium, facilitating the growth of an intracellular bacterial pathogen with complex growth requirements. In addition, results related to amino acid deprivation during intracellular growth and specific extracellular growth requirements of a threonine auxotroph suggest that L. monocytogenes may utilize intracellular peptides as a source of amino acids.


A concept of three light reactions in photosynthesis by green plants.
It has generally been accepted that plant photosynthesis involves two light reactions, one that proceeds best in short-wavelength light and is identified with oxygen evolution (System II) and another that proceeds best in long-wavelength light and is identified with a cycle electron flow (System I). This paper presents a concept of three light reactions in photosynthesis, based on new evidence that System II comprises two rather than one short-wavelength light reaction. These appear to operate in series and to be connected by an electron transport chain peculiar to System II. Parallel to System II is the long-wavelength light reaction of System I.