Convergent molecular evolution of genomic cores in Salmonella enterica and Escherichia coli.
One of the strongest signals of adaptive molecular evolution of proteins is the occurrence of convergent hot spot mutations: repeated changes in the same amino acid positions. We performed a comparative genome-wide analysis of mutation-driven evolution of core (omnipresent) genes in 17 strains of Salmonella enterica subspecies I and 22 strains of Escherichia coli. More than 20% of core genes in both Salmonella and E. coli accumulated hot spot mutations, with a predominance of identical changes having recent evolutionary origin. There is a significant overlap in the functional categories of the adaptively evolving genes in both species, although mostly via separate molecular mechanisms. As a strong evidence of the link between adaptive mutations and virulence in Salmonella, two human-restricted serovars, Typhi and Paratyphi A, shared the highest number of genes with serovar-specific hot spot mutations. Many of the core genes affected by Typhi/Paratyphi A-specific mutations have known virulence functions. For each species, a list of nonrecombinant core genes (and the hot spot mutations therein) under positive selection is provided.


Transport properties of CO2-expanded acetonitrile from molecular dynamics simulations.
Carbon-dioxide-expanded liquids, which are mixtures of organic liquids and compressed CO2, are novel media used in chemical processing. The authors present a molecular simulation study of the transport properties of liquid mixtures formed by acetonitrile and carbon dioxide, in which the CO2 mole fraction is adjusted by changing the pressure, at a constant temperature of 298 K. They report values of translational diffusion coefficients, rotational correlation times, and shear viscosities of the liquids as function of CO2 mole fraction. The simulation results are in good agreement with the available experimental data for the pure components and provide interesting insights into the largely unknown properties of the mixtures, which are being recognized as important novel materials in chemical operations. We find that the calculated quantities exhibit smooth variation with composition that may be represented by simple model equations. The translational and rotational diffusion rates increase with CO2 mole fraction for both the acetonitrile and carbon dioxide components. The shear viscosity decreases with increasing amount of CO2, varying smoothly between the values of pure acetonitrile and pure carbon dioxide. Our results show that adjusting the amount of CO2 in the mixture allows the variation of transport rates by a factor of 3-4 and liquid viscosity by a factor of 8. Thus, the physical properties of the mixture may be tailored to the desired range by changes in the operating conditions of temperature and pressure.


Frequency and risk factors associated with non-alcoholic fatty liver disease in patients with type 2 diabetes mellitus.
To evaluate the frequency of non-alcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes mellitus (DM2) and to describe its risk factors. Blood samples of 78 patients were collected for assessment of glycemic and lipid profile, liver enzymes, TNF-alpha and HOMA-IR. The diagnosis of NAFLD was established by ultrasound. NAFLD was observed in 42% of patients who had greater BMI (p < 0.001), and frequency of hypertension (p < 0.001). Metabolic syndrome was more frequent in those with NAFLD (p = 0.019). The levels of aspartate, alanine aminotransferase, gamma-glutamyl transpeptidase, uric acid, TNF-alpha, insulin and HOMA-IR were significantly higher in patients with NAFLD than those without NAFLD. Almost half of patients with DM2 were found to have NAFLD, and they have more elevated BMI, as well as higher levels of aminotransferases, gamma-GT, uric acid, TNF-alpha, insulin and HOMA-IR than subjects without NAFLD.


Molecular cloning of ion channels in Felis catus that are related to periodic paralyses in man: a contribution to the understanding of the genetic susceptibility to feline neck ventroflexion and paralysis.
Neck ventroflexion in cats has different causes; however, the most common is the hypokalemia associated with flaccid paralysis secondary to chronic renal failure. In humans, the most common causes of acute flaccid paralysis are hypokalemia precipitated by thyrotoxicosis and familial forms linked to mutations in sodium, potassium, and calcium channel genes. Here, we describe the sequencing and analysis of skeletal muscle ion channels in Felis catus that could be related to periodic paralyses in humans, contributing to the understanding of the genetic susceptibility to feline neck ventroflexion and paralysis. We studied genomic DNA from eleven cats, including five animals that were hyperthyroid with hypokalemia, although only one presented with muscle weakness, and six healthy control domestic cats. We identified the ion channel ortholog genes KCNJ2, KCNJ12, KCNJ14, CACNA1S and SCN4A in the Felis catus genome, together with several polymorphic variants. Upon comparative alignment with other genomes, we found that Felis catus provides evidence for a high genomic conservation of ion channel sequences. Although we hypothesized that neck ventroflexion in cats could be associated with a thyrotoxic or familial periodic paralysis channel mutation, we did not identify any previously detected human channel mutation in the hyperthyroid cat presenting hypokalemia. However, based on the small number of affected cats in this study, we cannot yet rule out this molecular mechanism. Notwithstanding, hyperthyroidism should still be considered as a differential diagnosis in hypokalemic feline paralysis.


Functional interaction between serotonin-S2 and dopamine-D2 neurotransmission as revealed by selective antagonism of hyper-reactivity to tryptamine and apomorphine.
The functional significance of the interaction between serotonergic and dopaminergic neurotransmission is still uncertain. To document this interaction further, specific behavioral responses of rats to tryptamine and apomorphine were studied. The sequential injection of these agonists, at time intervals with minimal direct behavioral interference, was used to observe response changes with respect to a single challenge. The antagonists haloperidol, ritanserin and risperidone, with known actions on serotonin-S2 (5-HT2) and dopamine-D2 (D2) receptors were used to evaluate effective antagonism of single and sequential challenges. When tryptamine was preceded by an apomorphine challenge the effective doses of the 5-HT2 antagonists ritanserin and risperidone for 50% inhibition of the seizures increased by a factor of 2.5. The dose-response curve of haloperidol remained virtually unchanged, apparently because of the potent dopamine-D2 antagonism associated with these doses which may block the potentiating effect of apomorphine. When apomorphine was preceded by a tryptamine challenge, the total agitation score of the control animals increased by 59% on the average. Haloperidol was equally effective against the enhanced as against the unenhanced apomorphine response. Ritanserin reduced agitation only by the part corresponding to the tryptamine enhancement. Risperidone's activity against the enhanced agitation started at very low doses and was complete at a dose still about 2.5 times lower than that required against the single apomorphine challenge. Mutual enhancement of tryptamine and apomorphine appears to occur even at a time when the behavioral effects of the first agonist are no longer manifest. The enhanced agitation remains largely dopamine-D2-specific and the enhanced seizures serotonin 5-HT2-specific.(ABSTRACT TRUNCATED AT 250 WORDS)


The design and construction of customised mouthpieces for subaqua diving.
Previous studies have shown that many subaqua divers suffer from temporomandibular joint dysfunction and related problems associated with the use of commercially available diving mouthpieces. This paper highlights factors which play a role in the development of these problems, defines essential features required in mouthpiece design and describes a technique for the construction of a fully customized mouthpiece.


Do monosymptomatic stroke patients with dizziness present a vestibular syndrome without nystagmus? An underestimated entity.
Vestibular symptoms are common in emergency department (ED) patients and have various causes, including stroke. Accurate identification of stroke in patients with vestibular symptoms is crucial for timely management. We conducted a prospective cross-sectional study from 2015 to 2019 to determine stroke prevalence and associated symptoms in ED patients with vestibular symptoms, aiming to improve diagnosis and outcomes. As part of the DETECT project, we screened 1647 ED patients with acute vestibular symptoms. Following a retrospective analysis of 961 head and neck magnetic resonance imaging (MRI) scans, we included 122 confirmed stroke cases and assessed them for vestibular signs and symptoms. Stroke prevalence in dizzy patients was 13% (122/961 MRI scans). Most patients (95%) presented with acute vestibular symptoms with or without nystagmus, whereas 5% had episodic vestibular syndrome (EVS). Nystagmus was present in 50% of stroke patients. Eighty percent had a purely posterior circulation stroke, and nystagmus was absent in 46% of these patients. Seven patients (6%) had lesions in both the anterior and posterior circulation. Vertigo was experienced by 52% regardless of territory. A stroke was identified in 13% of ED patients presenting with acute vestibular symptoms. In 5%, it was EVS. Most strokes were in the posterior circulation territory; vertigo occurred with similar frequency in anterior and posterior circulation stroke, and absence of nystagmus was common in both.


Anabolic androgens restore mating after sexual satiety in male rats.
Androgen receptors and estrogen receptors importantly participate in the neuroendocrine control of masculine mating behavior. Sexual satiety is the long term inhibition of masculine mating behavior after repeated ejaculations and is associated to changes in both androgen receptor and estrogen receptor-alpha expression. Androgen receptor expression is up-regulated by systemic chronic administration of anabolic androgens, 5alpha-dihydrotestosterone or estradiol benzoate. This study was carried out to investigate the effect of these treatments on sexual satiety development and recovery; additionally flutamide or tamoxifen treatments -- alone or together with anabolic androgens -- were also included. Chronic 15-day treatment with 5alpha-dihydrotestosterone (5 mg/kg) or tamoxifen (15 mg/kg) inhibited, whereas estradiol benzoate treatment (5 mg/kg) facilitated, mating behavior during sexual satiety development. The proportion of animals that ejaculated 48 h after sexual satiety was increased after 17-day treatment with a mixture of anabolic androgens containing 2 mg/kg testosterone propionate, 2 mg/kg nandrolone decanoate and 1 mg/kg boldenone undecylenate. This effect was only blocked by the combined administration of flutamide plus tamoxifen. The data suggest that anabolic androgens metabolites synergize to restore mating behavior after sexual satiety.


Genetic interaction of BBS1 mutations with alleles at other BBS loci can result in non-Mendelian Bardet-Biedl syndrome.
Bardet-Biedl syndrome is a genetically and clinically heterogeneous disorder caused by mutations in at least seven loci (BBS1-7), five of which are cloned (BBS1, BBS2, BBS4, BBS6, and BBS7). Genetic and mutational analyses have indicated that, in some families, a combination of three mutant alleles at two loci (triallelic inheritance) is necessary for pathogenesis. To date, four of the five known BBS loci have been implicated in this mode of oligogenic disease transmission. We present a comprehensive analysis of the spectrum, distribution, and involvement in non-Mendelian trait transmission of mutant alleles in BBS1, the most common BBS locus. Analyses of 259 independent families segregating a BBS phenotype indicate that BBS1 participates in complex inheritance and that, in different families, mutations in BBS1 can interact genetically with mutations at each of the other known BBS genes, as well as at unknown loci, to cause the phenotype. Consistent with this model, we identified homozygous M390R alleles, the most frequent BBS1 mutation, in asymptomatic individuals in two families. Moreover, our statistical analyses indicate that the prevalence of the M390R allele in the general population is consistent with an oligogenic rather than a recessive model of disease transmission. The distribution of BBS oligogenic alleles also indicates that all BBS loci might interact genetically with each other, but some genes, especially BBS2 and BBS6, are more likely to participate in triallelic inheritance, suggesting a variable ability of the BBS proteins to interact genetically with each other.


The effect of deoxyribonucleic acid extraction methods from lymphoid tissue on the purity, content, and amplifying ability.
Nowadays, definitive diagnosis of numerous diseases is based on the genetic and molecular findings. Therefore, preparation of fundamental materials for these evaluations is necessary. Deoxyribonucleic acid (DNA) is the first material for the molecular pathology and genetic analysis, and better results need more pure DNA. Furthermore, higher concentration of achieved DNA causes better results and higher amplifying ability for subsequent steps. We aim to evaluate five DNA extraction methods to compare DNA intimacy including purity, concentration, and amplifying ability with each other. The lymphoid tissue DNA was extracted from formalin-fixed, paraffin embedded (FFPE) tissue through five different methods including phenol-chloroform as the reference method, DNA isolation kit (QIAamp DNA FFPE Tissue Kit, Qiagen, Germany), proteinase K and xylol extraction and heat alkaline plus mineral oil extraction as authorship innovative method. Finally, polymerase chain reaction (PCR) and real-time PCR method were assessed to compare each following method consider to DNA purity and its concentration. Among five different applied methods, the highest mean of DNA purity was related to heat alkaline method. Moreover, the highest mean of DNA concentration was related to heat alkaline plus mineral oil. Furthermore, the best result in quantitative PCR was in proteinase K method that had the lowest cycle threshold averages among the other extraction methods. We concluded that our innovative method for DNA extraction (heat alkaline plus mineral oil) achieved high DNA purity and concentration.


The crossover approach to switching antipsychotics: what is the evidence?
Clinicians frequently use a crossover approach in switching antipsychotics, although historically there has been a lack of data addressing the question of switch strategies. To establish if there is now empiric evidence that may guide clinicians in this regard, a MEDLINE search to April 2004 was carried out to identify published, randomized and controlled trials that have addressed this topic. A total of 404 articles were identified in the search, which resulted in the identification of four reports meeting the criteria. The four studies evaluated switching strategies to one of three atypical antipsychotics: aripiprazole, olanzapine (two reports), and ziprasidone. The switching process itself could be subdivided as follows: discontinuation (abrupt vs. gradual); and, replacement (abrupt vs. gradual). Meta-analyses confirmed a lack of difference in outcome, regardless of approach. While a crossover approach does not appear to increase adverse events, the available empiric evidence does not support its clinical superiority on various outcome measures. The existing data therefore argue against the position that a crossover approach in switching antipsychotics represents a 'safer' means of preventing clinical deterioration during the switch.


Validation of a Model for Identification of Patients at Intermediate to High Risk for Complications Associated With Acute Symptomatic Pulmonary Embolism.
For patients with acute symptomatic pulmonary embolism (PE), the Bova score classifies their risk for PE-related complications within 30 days after diagnosis. The original Bova score was derived from 2,874 normotensive patients with acute PE who participated in one of six prospective PE studies. We retrospectively assessed the validity of the Bova risk model in normotensive patients with acute PE diagnosed in an academic urban ED. Two clinician investigators used baseline data for the model's four prognostic variables to stratify patients into the three Bova risk stages (I-III) for 30-day PE-related complications. Intraclass correlation coefficient (ICC) and the κ statistic were used to assess interrater variability. The Bova risk score classified the majority of the cohort of 1,083 patients into the lowest Bova risk stage (stage I, 80%; stage II, 15%; stage III, 5%), The primary end point occurred in 91 of the 1,083 patients (8.4%; 95% CI, 6.7%-10%) during the 30 days after PE diagnosis. Risk stage correlated with the PE-related complication rate (class I, 4.4%; class II, 18%; class III, 42%; ICC, 0.93 [95% CI, 0.92-0.94]; κ statistic, 0.80; P < .001), in-hospital complication rate (class I, 3.7%; class II, 15%; class III, 37%), and 30-day PE-related mortality (class I, 3.1%; class II, 6.8%; class III, 10.5%). The Bova risk score accurately stratifies normotensive patients with acute PE into stages of increasing risk of PE-related complications that occur within 30 days of PE diagnosis.


Membrane proteins of synaptic vesicles and cytoskeletal specializations at the node of Ranvier in electric ray and rat.
Binding sites for antibodies against membrane proteins of synaptic vesicles have been shown to be enhanced at nodes of Ranvier in electromotor axons of the electric ray Torpedo marmorata and sciatic nerve axons of the rat, using indirect immunofluorescence and monoclonal antibodies against the synaptic vesicle transmembrane proteins SV2 and synaptophysin (rat) or SV2 (Torpedo). In the electric lobe of Torpedo, vesicle-membrane constituents occurred at higher density in the proximal axon segments covered by oligodendroglia cells than in the distal axon segments where myelin is formed by Schwann cells. Antibody binding sites were enhanced at nodes forming the borderline of the central and peripheral nervous systems. Filamentous actin was present in the Schwann-cell processes covering both the nodal and the paranodal axon segments as suggested by the pattern of phalloidin labelling. Furthermore, in rat sciatic nerve, Schmidt-Lanterman incisures were intensely labelled by phalloidin. A similar nodal distribution was found for binding sites of antibodies against actin and myosin. Binding of antibodies to tubulin was enhanced at nodes in Torpedo electromotor axons. The apparent nodal accumulation of constituents of synaptic vesicle membranes and the presence of filamentous actin and of myosin are discussed in relation to the substantial constriction of the axoplasm at nodes of Ranvier.


[Psychiatric, medical and environmental factors in patients suffering from environment-related disorders].
A multidisciplinary approach and a multi-modal methodology are needed to assess idiopathic environmental illnesses. 61 patients took part in all diagnostic steps. In the Basel pilot research project on environmental illness, a threefold diagnostic approach was established: patients had a medical and allergological examination, a psychiatric and psychological exploration and an environmental analysis of their homes. There is a clear psychological impact on environmental illness: 46 % of the symptoms could be traced back to psychological factors, and 18 % seemed to be influenced by them. Nevertheless, in 28 % more than one of the three dimensions was seen as important. Values within the self reporting questionnaires show high correspondence. Whereas patients and experts agree in many instances that there are psychological factors, they disagree in attributing clinical relevance to them. This discrepancy is helpful for explaining the difficulties therapists may encounter as to the patients' compliance. Environmental illness should be diagnosed and treated on an interdisciplinary basis including psychosomatic medicine.


Journal Club: the Alvarado score as a method for reducing the number of CT studies when appendiceal ultrasound fails to visualize the appendix in adults.
OBJECTIVE. The purpose of this article is to evaluate the hypothesis that Alvarado scores of 3 or lower identify adult patients who are unlikely to benefit from CT after appendiceal ultrasound fails to show the appendix and is otherwise normal. MATERIALS AND METHODS. We identified 119 consecutive adults for whom the appendix was not seen on otherwise normal appendiceal sonography performed as the first imaging study for suspected appendicitis, who subsequently underwent CT within 48 hours, and whose data permitted retrospective calculation of admission Alvarado scores. Specific benefits of CT were defined as diagnoses of appendicitis or significant alternative findings, and specific benefits were compared between patients with Alvarado scores of 3 or less and 4 or higher. Significant alternative findings on CT were findings other than appendicitis that were treated with medical or surgical therapy during the admission or that were to be addressed during follow-up care. Diagnostic reference standards were discharge diagnoses, pathologic examinations, and clinical follow-up. RESULTS. No patients (0.0%, 0/49) with Alvarado scores 3 or lower had appendicitis, compared with 17.1% (12/70) of patients with Alvarado scores 4 or higher (p = 0.001), and CT showed neither appendicitis nor significant alternative findings in 85.7% (42/49) versus 58.6% (41/70) of these patients, respectively (p = 0.002). The rates of perforated appendicitis, as well as significant alternative CT findings, did not differ significantly. CONCLUSION. Adults with Alvarado scores 3 or lower who have nonvisualized appendixes and otherwise normal appendiceal sonography are at very low risk for appendicitis or significant alternative findings and therefore are not likely to benefit from CT.


Effect of regulatory warnings on antipsychotic prescription rates among elderly patients with dementia: a population-based time-series analysis.
Three warnings of serious adverse events associated with the use of atypical antipsychotic drugs among elderly patients with dementia were sent to health care professionals in Canada. We assessed the impact of these warnings on prescription rates of antipsychotic drugs in this patient population. We used prescription drug claims data from Ontario to calculate prescription rates of atypical and conventional antipsychotic drugs among elderly patients with dementia from May 1, 2000, to Feb. 28, 2007. We performed a time-series analysis to estimate the effect of each warning on rates of antipsychotic drug use. Before the first warning, growth in the use of atypical antipsychotics was responsible for an increasing rate of overall antipsychotic use. Each warning was associated with a small relative decrease in the predicted growth in the use of atypical antipsychotic drugs: a 5.0% decrease after the first warning, 4.9% after the second and 3.2% after the third (each p < 0.05). The overall prescription rate of antipsychotic drugs among patients with dementia increased by 20%, from 1512 per 100 000 elderly patients in September 2002, the month before the first warning, to 1813 per 100 000 in February 2007, 20 months after the last warning. Although the warnings slowed the growth in the use of atypical antipsychotic drugs among patients with dementia, they did not reduce the overall prescription rate of these potentially dangerous drugs. More effective interventions are necessary to improve postmarket drug safety in vulnerable populations.


Visual adaptations to different light environments in Amazonian fishes.
Light measurements were made at seven sites in the Amazon river system near Manaus. It was found that very little light penetrates into the white water of the Solimões because of the large amount of suspended material it contains. In lakes off the Solimões the suspended material settles out, and light penetration is increased. The black waters of the Rio Negro contain dissolved material but little suspended matter, and the penetrating light is mainly of long wavelength. Fishes were also collected from these areas, and the spectral characteristics of their visual pigments, lenses and corneas measured. The visual pigments of the different species vary considerably, fishes from clearer water having pigments absorbing at shorter wavelengths on average than those inhabiting more coloured or turbid waters, and the characins having pigments absorbing at longer wavelengths on average than the cichlids. Yellow lenses and corneas are common among cichlids, but rare in other groups and fishes with such filters were, with one exception, absent from the Solimões where the light levels are especially low. The exception was Colomesus asellus which has an occlusable yellow cornea, which while coloured in the light loses its colour at low levels of illumination. Using the date on light penetration and visual pigments it was calculated that vision would be impossible below 2.3 m in the Solimões, but should be possible at depths up to about 9 m in the clearest waters studied.


In vitro evolution of herpes simplex virus 1 (HSV-1) reveals selection for syncytia and other minor variants in cell culture.
The large dsDNA virus herpes simplex virus 1 (HSV-1) is considered to be genetically stable, yet it can rapidly evolve in response to strong selective pressures such as antiviral treatment. Deep sequencing has revealed that clinical and laboratory isolates of this virus exist as populations that contain a mixture of minor alleles or variants, similar to many RNA viruses. The classic virology approach of plaque purifying virus creates a genetically homogenous population, but it is not clear how closely this represents the mixed virus populations found in nature. We sought to study the evolution of mixed versus highly purified HSV-1 populations in controlled cell culture conditions, to examine the impact of this genetic diversity on evolution. We found that a mixed population of HSV-1 acquired more genetic diversity and underwent a more dramatic phenotypic shift than a plaque-purified population, producing a viral population that was almost entirely syncytial after just ten passages. At the genomic level, adaptation and genetic diversification occurred at the level of minor alleles or variants in the viral population. Certain genetic variants in the mixed viral population appeared to be positively selected in cell culture, and this shift was also observed in clinical samples during their first passages in vitro. In contrast, the plaque-purified viral population did not appear to change substantially in phenotype or overall quantity of minor allele diversity. These data indicate that HSV-1 is capable of evolving rapidly in a given environment, and that this evolution is facilitated by diversity in the viral population.


Human soft tissue analysis using x-ray or gamma-ray techniques.
This topical review is intended to describe the x-ray techniques used for human soft tissue analysis. X-ray techniques have been applied to human soft tissue characterization and interesting results have been presented over the last few decades. The motivation behind such studies is to provide improved patient outcome by using the data obtained to better understand a disease process and improve diagnosis. An overview of theoretical background as well as a complete set of references is presented. For each study, a brief summary of the methodology and results is given. The x-ray techniques include x-ray diffraction, x-ray fluorescence, Compton scattering, Compton to coherent scattering ratio and attenuation measurements. The soft tissues that have been classified using x-rays or gamma rays include brain, breast, colon, fat, kidney, liver, lung, muscle, prostate, skin, thyroid and uterus.


Endemic institutional strongyloidiasis in British Columbia.
The incidental finding in 1984 of Strongyloides stercoralis larvae in a resident of a chronic care institution who had a vague clinical illness prompted a review of the other residents. Five other cases were identified after exhaustive laboratory investigations. Fecal-oral spread was considered the most likely manner of the spread of infection. The possibility of endemic strongyloidiasis in institutions should be considered, even in temperate climates, when there is unexplained persistent illness or high eosinophil counts. Serologic testing is a useful adjunct to fecal examination in such situations.


Ultrastable Graphite-Potassium Anode through Binder Chemistry.
Graphite with abundant reserves has attracted enormous research interest as an anode of potassium-ion batteries (PIBs) owing to its high plateau capacity of 279 mAh g-1 at ≈0.2 V in conventional carbonate electrolytes. Unfortunately, it suffers from fast capacity decay during K+ storage. Herein, an ultrastable graphite-potassium anode is developed through binder chemistry. Polyvinyl alcohol (PVA) is utilized as a water-soluble binder to generate a uniform and robust KF-rich SEI film on the graphite surface, which can not only inhibit the electrolyte decomposition, but also withstand large volume expansion during K+ -insertion. Compared to the PVDF as binder, PVA-based graphite anode can operate for over 2000 cycles (running time of 406 days at C/3) with 97% capacity retention in KPF6 -based electrolytes. The initial Coulombic efficiency (ICE) of graphite anode is as high as 81.6% using PVA as the binder, higher than that of PVDF (40.1%). Benefiting from the strong adhesion ability of PVA, a graphite||fluorophosphate K-ion full battery is further built through 3D printing, which achieves a record-high areal energy of 8.9 mWh cm-2 at a total mass loading of 38 mg cm-2 . These results demonstrate the important role of binder in developing high-performance PIBs.


Factors influencing dietary protein sources in the PREMIER trial population.
Previous research suggests that protein intake, particularly plant protein, may benefit blood pressure control. However, very little has been published regarding protein sources in diets of US adults and factors influencing these choices. The purpose of this report is to describe specific sources of animal and plant proteins in diets of PREMIER clinical trial participants at baseline and how the PREMIER intervention, along with participant demographics, affected protein sources. Adult participants (n=809) who completed the 18-month PREMIER lifestyle intervention trial and had at least one diet recall at each of three study visits were included. Participants were recruited from four clinical centers in the Eastern, Southern, and Northeastern regions of United States. The PREMIER trial, conducted from 1999 to 2002, compared the impact on blood pressure of two structured behavioral interventions focusing on the traditional lifestyle modifications for blood pressure control with or without the Dietary Approaches to Stop Hypertension dietary pattern. Protein sources were assessed by two unannounced 24-hour recalls at each of three study visits. Differences in protein sources were mainly related to participant demographics, with relatively moderate impact of the intervention. The top four protein sources for all the study participants were poultry, dairy, refined grains and beef, each contributing approximately 10% to 17% in descending order to the total protein intake at baseline. Animal and plant protein each comprised approximately 66% and 34%, respectively, to the total daily protein intake at baseline, and such overall contribution pattern remained relatively constant over time. However, sex, race, age, and body weight status all influenced contribution patterns from different food groups significantly. These influences significantly impact choice and are essential elements to consider when designing intervention programs to alter protein contributions from animal vs plant sources.


Substances interfering with direct detection of Mycobacterium tuberculosis in clinical specimens by PCR: effects of bovine serum albumin.
Interfering substances have been reported to inhibit PCR assays for the direct detection of Mycobacterium tuberculosis in clinical specimens. Using an internal control, we determined that 52% of respiratory specimens interfered with our PCR assay. On the basis of these findings, we tried to circumvent the problem by simply diluting prepared sediments. With sediment from a routinely processed sputum known to be inhibitory to PCR, one aliquot was prepared in a routine manner for PCR. Remaining sediment was diluted in phosphate-buffered saline, Middlebrook 7H10 broth, or BACTEC 12B broth; an internal control was added to all reaction mixtures and controls. Internal control was detected only in the sample diluted with BACTEC 12B medium. Components of the BACTEC 12B medium including PANTA reagent (polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin), reconstituting fluid, 0.2% glycerol, 0.05% Tween 80, and 0.05% bovine serum albumin (BSA) were tested in a similar manner. Only 0.05% BSA resulted in amplification of the internal control DNA. Varying concentrations of BSA were added to 11 aliquots of a respiratory sediment known to be inhibitory to the PCR. Internal control was detected in all reaction mixtures containing 0.00038 to 0.1% BSA. To determine the ability of BSA to override inhibition, respiratory specimens were run in triplicate: undiluted, diluted 1:2 with BACTEC 12B medium, or diluted with 0.026% BSA. For 21 of 22 inhibitory specimens, BSA was able to override the presence of interfering substances. These data suggest that the presence of BSA in a PCR assay is critical for the direct detection of M. tuberculosis in respiratory specimens.


Endothelial Toxicity of High Glucose and its by-Products in Diabetic Kidney Disease.
Alterations of renal endothelial cells play a crucial role in the initiation and progression of diabetic kidney disease. High glucose per se, as well as glucose by-products, induce endothelial dysfunction in both large vessels and the microvasculature. Toxic glucose by-products include advanced glycation end products (AGEs), a group of modified proteins and/or lipids that become glycated after exposure to sugars, and glucose metabolites produced via the polyol pathway. These glucose-related endothelio-toxins notably induce an alteration of the glomerular filtration barrier by increasing the permeability of glomerular endothelial cells, altering endothelial glycocalyx, and finally, inducing endothelial cell apoptosis. The glomerular endothelial dysfunction results in albuminuria. In addition, high glucose and by-products impair the endothelial repair capacities by reducing the number and function of endothelial progenitor cells. In this review, we summarize the mechanisms of renal endothelial toxicity of high glucose/glucose by-products, which encompass changes in synthesis of growth factors like TGF-β and VEGF, induction of oxidative stress and inflammation, and reduction of NO bioavailability. We finally present potential therapies to reduce endothelial dysfunction in diabetic kidney disease.


Physiological responses to a cold, wet, and windy environment during prolonged intermittent walking.
The potentially deleterious influence of body cooling on the thermoregulatory and metabolic responses to prolonged walking exercise has not been established. To address this problem, 10 men completed a 6-h intermittent (15 min rest, 45 min exercise) walking protocol in a thermoneutral (+15 degrees C) condition (Neutral) and a cold (+5 degrees C), wet, and windy condition (Cold). The first two exercise periods were conducted at a higher intensity (Higher, 6 km/h and 10% incline) than the subsequent four exercise periods (Lower, 5 km/h and 0% incline). Rectal temperature was lower and heart rate no different in Cold compared with Neutral, whereas the following were higher: oxygen consumption, respiratory exchange ratio, plasma norepinephrine and epinephrine, and blood lactate and glucose. There was no environmental influence on these variables during Higher. In conclusion, heat production during Lower was not sufficient to offset heat loss to the cold environment, and the resulting reduction in rectal temperature and metabolic perturbations may be detrimental if exercise is prolonged.


A subthreshold MOS neuron circuit based on the Volterra system.
We present an analog neuron circuit consisting of a small number of metal-oxide semiconductor (MOS) devices operating in their subthreshold region. The dynamics of the circuit were designed to be equivalent to the well-known Volterra system to facilitate developing the circuit for a particular application. We show that a simple nonlinear transformation of system variables in the Volterra system enables designing a neuron-like oscillator, which can produce sequences in time of identically shaped pulses (spikes) by using current-mode subthreshold MOS circuits. We present experimental results of the fabricated neuron circuits as well as an application in an inhibitory neural network, where the neurons compete with each other in the frequency and time domains.


Actinobacterial acyl coenzyme A synthetases involved in steroid side-chain catabolism.
Bacterial steroid catabolism is an important component of the global carbon cycle and has applications in drug synthesis. Pathways for this catabolism involve multiple acyl coenzyme A (CoA) synthetases, which activate alkanoate substituents for β-oxidation. The functions of these synthetases are poorly understood. We enzymatically characterized four distinct acyl-CoA synthetases from the cholate catabolic pathway of Rhodococcus jostii RHA1 and the cholesterol catabolic pathway of Mycobacterium tuberculosis. Phylogenetic analysis of 70 acyl-CoA synthetases predicted to be involved in steroid metabolism revealed that the characterized synthetases each represent an orthologous class with a distinct function in steroid side-chain degradation. The synthetases were specific for the length of alkanoate substituent. FadD19 from M. tuberculosis H37Rv (FadD19Mtb) transformed 3-oxo-4-cholesten-26-oate (kcat/Km = 0.33 × 10(5) ± 0.03 × 10(5) M(-1) s(-1)) and represents orthologs that activate the C8 side chain of cholesterol. Both CasGRHA1 and FadD17Mtb are steroid-24-oyl-CoA synthetases. CasG and its orthologs activate the C5 side chain of cholate, while FadD17 and its orthologs appear to activate the C5 side chain of one or more cholesterol metabolites. CasIRHA1 is a steroid-22-oyl-CoA synthetase, representing orthologs that activate metabolites with a C3 side chain, which accumulate during cholate catabolism. CasI had similar apparent specificities for substrates with intact or extensively degraded steroid nuclei, exemplified by 3-oxo-23,24-bisnorchol-4-en-22-oate and 1β(2'-propanoate)-3aα-H-4α(3″-propanoate)-7aβ-methylhexahydro-5-indanone (kcat/Km = 2.4 × 10(5) ± 0.1 × 10(5) M(-1) s(-1) and 3.2 × 10(5) ± 0.3 × 10(5) M(-1) s(-1), respectively). Acyl-CoA synthetase classes involved in cholate catabolism were found in both Actinobacteria and Proteobacteria. Overall, this study provides insight into the physiological roles of acyl-CoA synthetases in steroid catabolism and a phylogenetic classification enabling prediction of specific functions of related enzymes.


Bioavailability of sulpiride tablet and capsule in dogs.
The in vitro dissolution rates from four tablet formulations and one capsule formulation were measured in simulated gastric and simulated intestinal fluids. The dissolution t50% in simulated gastric fluid ranged from 1.5 min to 30 min and in simulated intestinal fluid, the values ranged from 4 to 228 min. The relative rates and rank order between products with regard to their dissolution rate were maintained. No correlation was obtained between in vitro dissolution rate and overall bioavailability in dogs as measured by the amounts of unchanged sulpiride excreted in the urine. The total unchanged sulpiride excreted in the 48 hr urine ranged from 30 to 44 % of the dose. The urinary half-lives ranged from 3.9 to 4.9 hr. The results indicate that, in the case of sulpiride, in vitro dissolution rate may not be a good predictor of bioavailability.


Protein kinase C-delta deficiency perturbs bone homeostasis by selective uncoupling of cathepsin K secretion and ruffled border formation in osteoclasts.
Bone homeostasis requires stringent regulation of osteoclasts, which secrete proteolytic enzymes to degrade the bone matrix. Despite recent progress in understanding how bone resorption occurs, the mechanisms regulating osteoclast secretion, and in particular the trafficking route of cathepsin K vesicles, remain elusive. Using a genetic approach, we describe the requirement for protein kinase C-delta (PKCδ) in regulating bone resorption by affecting cathepsin K exocytosis. Importantly, PKCδ deficiency does not perturb formation of the ruffled border or trafficking of lysosomal vesicles containing the vacuolar-ATPase (v-ATPase). Mechanistically, we find that cathepsin K exocytosis is controlled by PKCδ through modulation of the actin bundling protein myristoylated alanine-rich C-kinase substrate (MARCKS). The relevance of our finding is emphasized in vivo because PKCδ-/- mice exhibit increased bone mass and are protected from pathological bone loss in a model of experimental postmenopausal osteoporosis. Collectively, our data provide novel mechanistic insights into the pathways that selectively promote secretion of cathepsin K lysosomes independently of ruffled border formation, providing evidence of the presence of multiple mechanisms that regulate lysosomal exocytosis in osteoclasts.


Protamine sulfate precipitation method depletes abundant plant seed-storage proteins: A case study on legume plants.
Depletion of abundant proteins is one of the effective ways to improve detection and identification of low-abundance proteins. Our previous study showed that protamine sulfate precipitation (PSP) method can deplete abundant ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) from leaf proteins and is suitable for their in-depth proteome investigation. In this study, we provide evidence that the PSP method can also be effectively used for depletion of abundant seed-storage proteins (SSPs) from the total seed proteins of diverse legume plants including soybean, broad bean, pea, wild soybean, and peanut. The 0.05% protamine sulfate (PS) was sufficient to deplete major SSPs from all legumes tested except for peanut where 0.1% PS was required. SDS-PAGE, Western blotting and 2DE analyses of PS-treated soybean and peanut seed proteins showed enriched spots in PS-supernatant than total proteins. Coefficient of variation percentage (%CV) and principal component analysis of 2DE spots support the reproducibility, suitability, and efficacy of the PSP method for quantitative and comparative seed proteome analysis. MALDI-TOF-TOF successfully identified some protein spots from soybean and peanut. Hence, this simple, reproducible, economical PSP method has a broader application in depleting plant abundant proteins including SSPs in addition to RuBisCO, allowing discussion for comprehensive proteome establishment and parallel comparative studies in plants.


Endocrine role of bone: recent and emerging perspectives beyond osteocalcin.
Recent developments in endocrinology, made possible by the combination of mouse genetics, integrative physiology and clinical observations have resulted in rapid and unanticipated advances in the field of skeletal biology. Indeed, the skeleton, classically viewed as a structural scaffold necessary for mobility, and regulator of calcium-phosphorus homoeostasis and maintenance of the haematopoietic niche has now been identified as an important regulator of male fertility and whole-body glucose metabolism, in addition to the classical insulin target tissues. These seminal findings confirm bone to be a true endocrine organ. This review is intended to detail the key events commencing from the elucidation of osteocalcin (OC) in bone metabolism to identification of new and emerging candidates that may regulate energy metabolism independently of OC.


Treatment of leishmaniasis with miltefosine: 2008 status.
Miltefosine is the first recognized oral treatment for leishmaniasis. It was first registered, in 2002, for Indian visceral leishmaniasis, and was reviewed by the present author in 2005. Miltefosine is now being used for the full range of clinical leishmaniasis. The present review addresses non-clinical and clinical advances since 2005. PubMed was accessed for all articles on miltefosine from 2005 to 2008. Miltefosine is effective and can be recommended for visceral disease in India and in Ethiopia, and for cutaneous disease in Colombia and Bolivia. For unusual forms of disease that require long periods of treatment such as diffuse cutaneous leishmaniasis and post-kala-azar dermal leishmaniasis, oral miltefosine is probably the treatment of choice.


Histological phenotypes of enteric smooth muscle disease causing functional intestinal obstruction in childhood.
Functional intestinal obstruction or chronic idiopathic intestinal pseudo-obstruction is due to defects either in the enteric innervation or in intestinal smooth muscle. We have studied full-thickness intestinal biopsies from 27 patients with functional intestinal obstruction due to enteric smooth muscle disease by routine histology and electron microscopy together with histochemical and immunohistochemical techniques to detect changes in the intestinal smooth muscle. Two patients appeared to have an acquired intestinal myopathy as a result of an autoimmune process. In 25 the disorders were congenital, of these seven had segmental abnormalities limited to the rectum and distal colon and 18 had a diffuse disease affecting both the small and large bowel. We identified five apparent histological phenotypes of enteric muscle disease, three of which represent abnormalities in morphogenesis resulting in alterations in intestinal muscle layering and two exemplify intrinsic myocyte defects and/or changes in the extracellular matrix. Careful phenotyping of these patients is important in devising optimal treatment and in understanding the underlying defect as well as the possible genetic mechanisms resulting in these abnormalities. Recognition of autoimmune smooth muscle disease is helpful, since making the diagnosis influences the patient's management.


Computational fluid dynamics analysis of thrombosis potential in left ventricular assist device drainage cannulae.
Cannulation is necessary when blood is removed from the body, for example in hemodialysis, cardiopulmonary bypass, blood oxygenators, and ventricular assist devices. Artificial blood contacting surfaces are prone to thrombosis, especially in the presence of stagnant or recirculating flow. In this work, computational fluid dynamics was used to investigate the blood flow fields in three clinically available cannulae (Medtronic DLP 12, 16, and 24 F), used as drainage for pediatric circulatory support and to calculate parameters that may be indicative of thrombosis potential. The results show that using the 24 F cannula below flow rates of about 0.75 L/min produces hemodynamic conditions, which may increase the risk of blood clotting within the cannula. No reasons are indicated for not using the 12 or 16 F cannulae with flow rates between 0.25 and 3.0 L/min.


Characterization of human colon cancer antigens recognized by autologous antibodies.
The screening of cDNA expression libraries derived from human tumors with autologous antibody (SEREX) has proven to be a powerful method for defining the structure of tumor antigens recognized by the humoral immune system. In the present study, 48 distinct antigens (NY-CO-1-NY-CO-48) reactive with autologous IgG were identified by SEREX analysis in 4 patients with colon cancer. Sequencing analysis showed that 17 of the cDNA clones were previously uncharacterized molecules and 31 represented known gene products. The individual cDNA clones were analyzed in the following manner: a search for mutations or other structural changes; an analysis of mRNA expression in a panel of normal tissues; and a frequency analysis of the antibody response to the expressed product in the sera of colon cancer patients and normal individuals. The initial analysis showed NY-CO-13 to be a mutated version of the p53 tumor suppressor gene. Three of the 48 antigens showed a differential pattern of mRNA expression, with NY-CO-27 (galectin-4) expressed primarily in gastrointestinal tract, and NY-CO-37 and -38 showing a pattern of tissue-specific isoforms. With regard to immunogenicity, 20 of the 48 antigens were detected by allogeneic sera; 14 of these were reactive with sera from both normal donors and cancer patients, and 6 other clones (NY-CO-8, -9, -13, -16, -20 and -38) reacted exclusively with sera from colon cancer patients (ranging from 14% to 27%). Our results on colon cancer illustrate both the complexity and the potential of the SEREX approach for analysis of the humoral immune response against human cancer.


Delineation of the population genetic structure of Culicoides imicola in East and South Africa.
Culicoides imicola Kieffer, 1913 is the main vector of bluetongue virus (BTV) and African horse sickness virus (AHSV) in Sub-Saharan Africa. Understanding the population genetic structure of this midge and the nature of barriers to gene flow will lead to a deeper understanding of bluetongue epidemiology and more effective vector control in this region. A panel of 12 DNA microsatellite markers isolated de novo and mitochondrial DNA were utilized in a study of C. imicola populations from Africa and an outlier population from the Balearic Islands. The DNA microsatellite markers and mitochondrial DNA were also used to examine a population of closely related C. bolitinos Meiswinkel midges. The microsatellite data suggest gene flow between Kenya and south-west Indian Ocean Islands exist while a restricted gene flow between Kenya and South Africa C. imicola populations occurs. Genetic distance correlated with geographic distance by Mantel test. The mitochondrial DNA analysis results imply that the C. imicola populations from Kenya and south-west Indian Ocean Islands (Madagascar and Mauritius) shared haplotypes while C. imicola population from South Africa possessed private haplotypes and the highest nucleotide diversity among the African populations. The Bayesian skyline plot suggested a population growth. The gene flow demonstrated by this study indicates a potential risk of introduction of new BTV serotypes by wind-borne infected Culicoides into the Islands. Genetic similarity between Mauritius and South Africa may be due to translocation as a result of human-induced activities; this could impact negatively on the livestock industry. The microsatellite markers isolated in this study may be utilised to study C. bolitinos, an important vector of BTV and AHSV in Africa and identify sources of future incursions.


Are We There Yet? Exploring the Impact of Translating Cognitive Tests for Dementia Using Mobile Technology in an Aging Population.
This study examines implications of the expanded use of mobile platforms in testing cognitive function, and generates evidence on the impact utilizing mobile platforms for dementia screen. The Saint Louis University Mental State examination (SLUMS) was ported onto a computerized mobile application named the Cambridge University Pen to Digital Equivalence assessment (CUPDE). CUPDE was piloted and compared to the traditional pen and paper version, with a common comparator test for both groups. Sixty healthy participants (aged 50-79) completed both measurements. Differences were tested between overall outcomes, individual items, and relationship with the comparator. Significant differences in the overall scores between the two testing versions as well as within individual items were observed. Even when groups were matched by cognitive function and age, scores on SLUMS original version (M = 19.75, SD = 3) were significantly higher than those on CUPDE (M = 15.88, SD = 3.5), t (15) = 3.02, p < 0.01. Mobile platforms require the development of new normative standards, even when items can be directly translated. Furthermore, these must fit aging populations with significant variance in familiarity with mobile technology. Greater understanding of the interplay and related mechanisms between auditory and visual systems, which are not well understood yet in the context of mobile technologies, is mandatory.


A randomized pilot study of acupuncture as an adjunct therapy in adult asthmatic patients.
This trial aimed to evaluate the feasibility of estimating the effectiveness of acupuncture on asthmatic patients under conventional medical management. PARTICIPATIONS AND METHODS: A prospective randomized, patient/assessor-blinded, sham acupuncture-, and waiting list-controlled pilot trial was conducted. Forty-five eligible asthmatic participants underwent a 1-week run-in period and were then randomized into one of three groups: an active acupuncture group, a sham acupuncture group, and a waiting list group. They were instructed to maintain the use of antiasthmatic medications. Needling was administered three times per week for 4 weeks with a 2-week follow-up in the active and sham acupuncture groups. The primary outcome was daily morning peak expiratory flow (PEF) and the secondary outcomes included forced expiratory volume one second (FEV(1)), quality of life questionnaire for adult Korean asthmatics (QLQAKA), transition dyspnea index (TDI), serum eosinophil count, and total serum immunoglobulin E (IgE). No significant differences in the between- or within-group values of weekly average PEF (recorded daily in the morning) and FEV(1) were found. For QLQAKA and TDI, the active acupuncture group showed a significant improvement over the waiting list group at 2, 4, and 6 weeks after randomization. Acupuncture as an adjunct therapy to conventional medical care does not seem to affect pulmonary function in asthmatic patients. However, 12 sessions of acupuncture treatment during 4 weeks showed a favorable effect on the quality of life in adult asthmatic patients. Further large trials assessing the effectiveness of acupuncture on the quality of life and symptoms in asthmatic patients are needed.


Predictive value of computed tomography in identifying extranodal extension in human papillomavirus-positive versus human papillomavirus-negative head and neck cancer.
Pathologic extranodal extension (pENE) impacts treatment planning and is an important prognostic indicator for patients with head and neck squamous cell carcinoma (HNSCC). Computed tomography (CT) is a commonly used modality for assessment of radiographic ENE (rENE). To determine the predictive value of CT-identified rENE in predicting pENE, we performed a systematic review through a search of 4 databases (PubMed, Scopus, Cochrane, and OVID). Meta-analysis of diagnostic performance based on human papillomavirus (HPV) status was conducted. For HPV-negative HNSCC, pooled sensitivity, specificity, and accuracy were 60.6%, 93.3%, and 82.6%, respectively. Overall positive predictive value (PPV) was 82.7%. For HPV-positive HNSCC, pooled sensitivity, specificity, and accuracy were 77.7%, 72.2%, and 63.8%, respectively. Overall PPV was 68.6%. Significant differences were observed in diagnostic performance parameters between the two cohorts. The radiographic characteristics of HPV-positive and HPV-negative nodal metastases in HNSCC differ and radiographic evaluation of ENE in HPV-positive nodes is challenging. Development of refined imaging characteristics of HPV-positive nodes is needed to improve diagnostic performance.


Cervical cancer screening among homeless women in the Greater Paris Area (France): results of the ENFAMS survey.
Little is known about the prevalence of cervical cancer screening (CCS) and its correlates among homeless women in France. The objectives of this study were to determine the prevalence of women who had never been screened for cervical cancer and to identify the associated factors. This cross-sectional study was based on data collected in the ENFAMS survey, which was conducted in 2013 among 764 sheltered homeless mothers in the Greater Paris Area. Robust Poisson regression models were used to estimate the association between no lifetime CCS and certain sociodemographic and health-related factors (selected from the behavioral model of vulnerable populations). Analyses were carried out separately for women with and without a regular gynaecological follow-up (RGF). The proportion of never-screeners was 33% among the women with an RGF versus 64% among those without an RGF (P<0.001). Among the latter, never having been screened for CCS was associated mainly with socioeconomic conditions, the length of time lived in France, a history of delivery in France and the duration of homelessness. In those with an RGF, the factors were mainly poor health service utilization and language difficulties. This first quantitative study of CCS among homeless women in the Greater Paris Area points to the need for it to be proposed and performed more systematically in primary care. Every contact between this hard-to-reach population and health services should be an opportunity to check their screening status and to ensure that those in need actually undergo a Pap test.


Tales from the trenches: physician assistants' perspectives about precepting students.
A national survey of physician assistants (PAs) found that only 25% of respondents were involved as preceptors for PA students in 2011, and it also identified important barriers and incentives to precept PA students. These findings offer limited information for understanding the nature of this complex phenomenon within its context. The purpose of this study was to further describe key factors that influence the involvement of PAs as preceptors. A qualitative study of four focus groups with clinically practicing PAs (N = 29) was conducted at the American Academy of Physician Assistants' annual conference in May 2012. Semistructured interview questions and follow-up probes were used to elicit participants' perspectives about their own and/or witnessed preceptor experiences. The group discussions were audio-recorded, professionally transcribed, and then analyzed line-by-line by four investigators using conceptual and free coding. The independently coded transcripts were merged, and iterative analysis was used to identify recurrent themes until saturation was reached. Four themes emerged: (1) The preceptor role provides an opportunity to "pay back," or contribute to future colleagues' training while advancing the clinician's own knowledge; (2) Student qualities that incentivize precepting are characterized by motivation and self-directed learning, independent of the level of medical knowledge and/or prior experience; (3) Preceptors value feeling connected with the PA program through initial and ongoing communication; (4) Significant competition for clinical rotation sites for various health professional learners limits the access to sites and preceptors. PAs identified key areas for potential preceptor recruitment and retention interventions.


Extensive distribution of glial cytoplasmic inclusions in an autopsied case of multiple system atrophy with a prolonged 18-year clinical course.
We describe herein an autopsied case of multiple system atrophy (MSA) with prolonged clinical course of 18 years, and evaluate the extent of neurodegeneration and glial cytoplasmic inclusions (GCIs) in the entire brain of this rare case. A 64-year-old woman presented with typical neurological symptoms and imaging features of MSA. Thereafter, she became bedridden, and breathing was assisted through a tracheostomy for 12 years. She died at the age of 82 after 18 years from the initial symptom. Post mortem examination revealed severe neurodegeneration in the inferior olive, pontine nuclei, substantia nigra, locus ceruleus, putamen and cerebellum. Notably, phosphorylated α-synuclein (p-α-syn)-positive GCIs were found in these areas, but their number was very low. In contrast, the density of GCIs was much higher in such regions as the tectum/tegmentum of the brainstem, pyramidal tracts, neocortices and limbic system, which usually contain a small number of GCIs. Another constituent of GCIs, ubiquitin (Ub) and Ub-associated autophagy substrate p62, were also positive in some GCIs, and distribution of Ub/p62 immunoreactivity was proportionate to that of p-α-syn+ GCIs despite the very long duration of the disease. Furthermore, this case had complicated hypoxic encephalopathy, but p-α-syn+ GCIs were also found in the damaged white matter, indicating the contribution of α-syncleinopathy as well as hypoxic effect to the secondary myelin and axonal loss in the white matter. Together, this rare case suggests the contribution of the disease duration to the prevalence of GCIs, and the possible involvement of the limbic system in extensive-stage disease.


Musical rhythms in heart period dynamics: a cross-cultural and interdisciplinary approach to cardiac rhythms.
The purpose of this study was to expand classic heart period analysis methods by techniques from ethnomusicology that explicitly take complex musical rhythm principles into consideration. The methods used are based on the theory of African music, the theory of symbolic dynamics, and combinatorial theory. Heart period tachograms from 192 24-h electrocardiograms of 96 healthy subjects were transformed into binary symbol sequences that were interpretable as elementary rhythmic (percussive) patterns, the time lines in African music. Using a hierarchical rhythm pattern scheme closely related to the Derler Rhythm Classification (from jazz theory), we calculated the predominance and stability of pattern classes. The results show that during sleep certain classes, specific to individuals, occurred in a cyclically recurrent manner and many times more often than expected. Simultaneously, other classes disappeared more or less completely. Moreover, the most frequent classes obviously originate from phase-locking processes in autonomic regulation (e.g., between respiratory and cardiac cycles). In conclusion, the new interdisciplinary method presented here demonstrates that heart period patterns, in particular those occurring during night sleep, can be interpreted as musical rhythms. This method may be of great potential use in music therapy research.


Depression over Time in Persons with Stroke: A Network Analysis Approach.
Network analysis has been used to elucidate the relationships among depressive symptoms, but this approach has not been typically used in persons with stroke. Using a sample of 835 persons with stroke from Stroke Recovery in Underserved Populations 2005-2006 dataset, this study used network analysis to (1) examine changes in relationships between depressive symptoms over time, and (2) test whether baseline network characteristics were prognostic for depression persistence. Network analysis was performed on depressive symptoms collected at discharge from inpatient rehabilitation and at 3-months and 12-months post-discharge. The depressive symptom network at discharge was less connected than at both post-discharge follow-ups. Trouble focusing and feeling good as others were the most predictable symptoms at post-discharge, even though they were less connected to other depressive symptoms. Among participants with elevated baseline depression severity, those whose depression persisted 12 months later had more strongly connected networks at discharge than those who recovered 12 months later. This study was unable to determine the directionality of edges. The depression scale was administered differently across time points. These results suggest that baseline network connectivity can predict the course of post-stroke depression, similar to non-stroke populations. More broadly, the study highlights the importance of examining relationships between individual depressive symptoms rather than only sum-scores.


Efficient multiscale regularization with applications to the computation of optical flow.
A new approach to regularization methods for image processing is introduced and developed using as a vehicle the problem of computing dense optical flow fields in an image sequence. The solution of the new problem formulation is computed with an efficient multiscale algorithm. Experiments on several image sequences demonstrate the substantial computational savings that can be achieved due to the fact that the algorithm is noniterative and in fact has a per pixel computational complexity that is independent of image size. The new approach also has a number of other important advantages. Specifically, multiresolution flow field estimates are available, allowing great flexibility in dealing with the tradeoff between resolution and accuracy. Multiscale error covariance information is also available, which is of considerable use in assessing the accuracy of the estimates. In particular, these error statistics can be used as the basis for a rational procedure for determining the spatially-varying optimal reconstruction resolution. Furthermore, if there are compelling reasons to insist upon a standard smoothness constraint, the new algorithm provides an excellent initialization for the iterative algorithms associated with the smoothness constraint problem formulation. Finally, the usefulness of the approach should extend to a wide variety of ill-posed inverse problems in which variational techniques seeking a "smooth" solution are generally used.


Associations between types of combat violence and the acquired capability for suicide.
Research suggests that combat exposure might increase risk for suicide. The interpersonal-psychological theory of suicide (IPTS) proposes that exposure to painful and provocative experiences such as combat contribute to fearlessness about death and increased pain tolerance, which serve to enhance the individual's capability to attempt suicide. Violent and aggressive combat experiences, in particular, should demonstrate relatively stronger associations to this capability. The current study tests this proposition in a sample of deployed active duty combatants. Results indicate that all types of combat exposure independently contribute to capability for suicide. Consistent with the IPTS, when considering all types of combat simultaneously, combat characterized by violence and high levels of injury and death are associated with relatively stronger associations to this capability.


Resolving spatial inconsistencies in chromosome conformation measurements.
Chromosome structure is closely related to its function and Chromosome Conformation Capture (3C) is a widely used technique for exploring spatial properties of chromosomes. 3C interaction frequencies are usually associated with spatial distances. However, the raw data from 3C experiments is an aggregation of interactions from many cells, and the spatial distances of any given interaction are uncertain. We introduce a new method for filtering 3C interactions that selects subsets of interactions that obey metric constraints of various strictness. We demonstrate that, although the problem is computationally hard, near-optimal results are often attainable in practice using well-designed heuristics and approximation algorithms. Further, we show that, compared with a standard technique, this metric filtering approach leads to (a) subgraphs with higher statistical significance, (b) lower embedding error, (c) lower sensitivity to initial conditions of the embedding algorithm, and (d) structures with better agreement with light microscopy measurements. Our filtering scheme is applicable for a strict frequency-to-distance mapping and a more relaxed mapping from frequency to a range of distances. Our filtering method for 3C data considers both metric consistency and statistical confidence simultaneously resulting in lower-error embeddings that are biologically more plausible.


Sacral insufficiency fracture.
Insufficiency fractures of the sacrum are not uncommon and usually occur in osteoporotic bone with minimal or unremembered trauma. However, they appear to be relatively under-diagnosed and this pictorial review aims to highlight the condition, discuss the expected imaging features and some of the potential imaging pitfalls. Owing to its relationship with osteoporosis, the majority occur in elderly females and are frequently bilateral, often presenting as low back pain. Plain radiographs are generally normal and both clinician and radiologist need to consider the possibility of sacral insufficiency fracture to allow prompt accurate diagnosis and correct treatment. Lumbar spine MRI is among the first investigations performed and can enable the correct diagnosis to be made. Occasionally the MR appearances can mimic tumour or osteomyelitis. The "H" sign on an isotope bone scan is considered diagnostic in the right clinical setting, but this sign is often not present. CT is useful to confirm the diagnosis and exclude tumour or infection.


Vital capacity and one-second forced expiratory volume in Australian male factory workers.
The vital capacity (VC) and one-second forced expiratory volume (FEV1) of 482 men were measured and corrected for temperature but not pressure. The relationship between both VC and FEV1 and height and age was examined and found to be linear. In using these relationships for significance testing the 95% confidence limit is 1.65 standard deviations below the predicted value. Our results are expressed in graphical form to facilitate interpolation and will thus be of use to the clinical practitioner in Australia. Neither weight nor smoking habit was found to influence the relationship of VC or FEV1 with age or height.


Epistatic interaction between haplotypes of the ghrelin ligand and receptor genes influence susceptibility to myocardial infarction and coronary artery disease.
Data from both experimental models and humans provide evidence that ghrelin and its receptor, the growth hormone secretagogue receptor (ghrelin receptor, GHSR), possess a variety of cardiovascular effects. Thus, we hypothesized that genetic variants within the ghrelin system (ligand ghrelin and its receptor GHSR) are associated with susceptibility to myocardial infarction (MI) and coronary artery disease (CAD). Seven single nucleotide polymorphisms (SNPs) covering the GHSR region as well as eight SNPs across the ghrelin gene (GHRL) region were genotyped in index MI patients (864 Caucasians, 'index MI cases') from the German MI family study and in matched controls without evidence of CAD (864 Caucasians, 'controls', MONICA Augsburg). In addition, siblings of these MI patients with documented severe CAD (826 'affected sibs') were matched likewise with controls (n = 826 Caucasian 'controls') and used for verification. The effect of interactions between genetic variants of both genes of the ghrelin system was explored by conditional classification tree models. We found association of several GHSR SNPs with MI [best SNP odds ratio (OR) 1.7 (1.2-2.5); P = 0.002] using a recessive model. Moreover, we identified a common GHSR haplotype which significantly increases the risk for MI [multivariate adjusted OR for homozygous carriers 1.6 (1.1-2.5) and CAD OR 1.6 (1.1-2.5)]. In contrast, no relationship between genetic variants and the disease could be revealed for GHRL. However, the increase in MI/CAD frequency related to the susceptible GHSR haplotype was abolished when it coincided with a common GHRL haplotype. Multivariate adjustments as well as permutation-based methods conveyed the same results. These data are the first to demonstrate an association of SNPs and haplotypes within important genes of the ghrelin system and the susceptibility to MI, whereas association with MI/CAD could be identified for genetic variants across GHSR, no relationship could be revealed for GHRL itself. However, we found an effect of GHRL dependent upon the presence of a common, MI and CAD susceptible haplotype of GHSR. Thus, our data suggest that specific haplotypes of the ghrelin ligand and its receptor act epistatically to affect susceptibility or tolerance to MI and/or CAD.


From coercion to collaboration: two weeks in the life of a therapeutic community.
The purpose of this paper is to describe an aspect of the functioning of a therapeutic community conceptualized in terms of a sociotherapy framework as adumbrated, for example, by Edelson (1970), Rapoport (1960), and White et al. (1964). This approach, as distinct from group therapy, is concerned with the social system and social conditions that form the fabric of any given behavior. In the hospital context it is the therapeutic community that provides the fabric in which interpersonal and intergroup tensions can be addressed and clarified. We will illustrate this approach by detailing a discussion between therapy and nursing staff members who consult to, and participate in, the various group meetings of the Therapeutic Community Program (TCP), which together provide the framework of this particular community. Specifically, the discussion was about the evolving nature of the engagement between the community and a borderline patient, who attempted to alter the structure and functioning of an aspect of the community program in order to meet her own needs and avoid inner conflict. The discussion will focus on the way in which the community members adhered to existing structures and functioning, and were able to utilize staff consultation to this end, despite the patient's strong pressure on them to alter it. In describing this particular case we hope to illustrate how a well-functioning therapeutic community can "hold" one of its members in a manner that allows change to take place, and also serves as an important adjunct and facilitator to individual therapy.


Expression analysis of cytokines IL-5, IL-6, IL-8, IL-17 and VEGF in breast cancer patients.
To investigate the relationship between changes in peripheral blood vascular endothelial growth factor (VEGF), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-17 (IL-17) concentrations in breast cancer patients and their significance and clinical value in breast cancer staging and invasive metastasis. From September 2021 to April 2022, 60 breast cancer patients from Chongqing Medical University Hospital No. 2022 were enrolled in the breast breast cancer surgery group, while 30 patients with benign breast disease were enrolled in the control group during the same period. Venous blood samples were collected at admission and 1 week after surgery to determine the expression of these factors in serum. Statistical methods such as Wilcoxon test and Spearman correlation analysis were used to analyze the relationship between the above factors and the clinicopathological characteristics of the patients. By analyzing data from patients with benign and malignant breast tumors, an association was found with serum levels of IL-6, IL-17 and VEGF. Their respective areas under the operating characteristic curve were 0.649, 0.734 and 0.656 (P < 0.05). There were significant differences in the cytokine expression levels of IL-17 and VEGF in different molecular typing (P values were 0.008 and 0.040, respectively). The expression levels of IL-17 and VEGF were higher in HER-2 receptor-positive and triple-negative patients than in hormone receptor-positive patients (P < 0.05). Also, no significant correlation was found between the various cytokines mentioned in the article and breast cancer lymph node metastasis and Tumor Node Metastasis stage (TNM stage). In addition, in the breast cancer surgery group, postoperative VEGF levels were lower (P < 0.05) and IL-6 levels were higher (P < 0.05) compared to preoperative levels. Serum IL-6, IL-17, and VEGF are strongly associated with breast cancer development and can be used as a reference indicators for breast cancer diagnosis. In addition, post-operative VEGF levels decreases and IL-6 levels increases compared to pre-operative levels, which can also be used as an a postoperative follow-up indicator. In contrast, IL-5 and IL-8 have not found to be significantly associated with breast cancer patients in this study, which requires further study.


Glass thermometer injuries: it is not just about the mercury.
Glass mercury thermometers were once used as the criterion standard for measuring core body temperature. Once broken, however, there is the dual hazard of broken glass and exposure to mercury. Previous studies have focused only on the mercury hazard rather than injuries related to thermometer glass. To identify injury patterns from glass thermometers, including glass-related injury and mercury exposure. We performed a retrospective cohort study of glass thermometer-related injuries in children evaluated in a pediatric emergency department between October 1995 and October 2007. Case identification was performed using a computer-assisted screening tool followed by a manual chart review. Collected data included age, sex, injury characteristics, physical examination findings, radiologic imaging, interventions, and disposition. To analyze injury rates during these years, we used a multiplicative Poisson model allowing for varying exposures. Thirty-three patients were identified among 627,592 who presented to the emergency department during the study period. Approximately 1 to 6 patients presented every year, including 3 patients in 2007. Decline in injury incidence is less than 9% per year (P = 0.041). Median patient age was 2.4 years (interquartile range, 0.4-3.8 years); 12 patients (36%) were female. Twenty-two patients (66%) underwent radiologic imaging to identify potential foreign body (21 underwent radiography and 1 underwent computed tomographic scanning). There were 15 mouth injuries (45%), 13 anal injuries (39%), and 1 ear injury. Glass was identified by imaging in 5 patients (15%) and by sigmoidoscopic evaluation in 1 patient (3%). Mercury exposure was identified in 14 patients (42%). Persistent use of glass mercury thermometers has resulted in pediatric injury especially in children younger than 4 years. We reported the different mechanisms of injury with the hope of eliminating its use and reinforcing the use of alternative thermometers.


Developmental Differences in Infants' Attention to Social and Nonsocial Threats.
Research has demonstrated that humans detect threatening stimuli more rapidly than nonthreatening stimuli. Although the literature presumes that biases for threat should be normative, present early in development, evident across multiple forms of threat, and stable across individuals, developmental work in this area is limited. Here, we examine the developmental differences in infants' (4- to 24-month-olds) attention to social (angry faces) and nonsocial (snakes) threats using a new age-appropriate dot-probe task. In Experiment 1, infants' first fixations were more often to snakes than to frogs, and they were faster to fixate probes that appeared in place of snakes vs. frogs. There were no significant age differences, suggesting that a perceptual bias for snakes is present early in life and stable across infancy. In Experiment 2, infants fixated probes more quickly after viewing any trials that contained an angry face compared to trials that contained a happy face. Further, there were age-related changes in infants' responses to face stimuli, with a general increase in looking time to faces before the probe and an increase in latency to fixate the probe after seeing angry faces. Together, this work suggests that different developmental mechanisms may be responsible for attentional biases for social vs. nonsocial threats.


Aneurysmal bone cyst in the horse.
An aneurysmal bone cyst was diagnosed in the distal metaphysis of Mt3 in a 9-month-old-Quarter Horse colt. The lesion developed between the fifth and ninth month of life and clinically appeared as a non-painful swelling which did not cause a lameness. Radiographs of the lesion showed expansion of the cortex and incomplete septa of new bone extending from the cortex into surrounding soft tissue. Surgical intervention revealed a blood filled cavity. The animal died during the immediate post-surgical period. Histopathologic evaluation of the lesion was performed.


Indole based cyclooxygenase inhibitors: synthesis, biological evaluation, docking and NMR screening.
The close structural similarity between the two cyclooxygenase (COXs) isoforms and the absence of selective inhibitors without side effects continues to stimulate the development of novel approaches towards selective anti-inflammatory drugs. In the present study a small library of new indolic compounds involving two different substitutions patterns at the indole scaffold was synthesized. In order to establish a relation between the spatial distribution of known functional groups related with inhibitory activity, two substitution patterns were explored: one with substituents at N-1, C-3, C-5 positions and another at C-2, C-3 and C5 positions. Accordingly, indole positions C-5, C-3 and N-1 were substituted with: sulfonamide or methylsulfone at C-5, p-halo-benzyl group at C-3, and an alkyl chain with a trifluoromethyl group at N-1. Alternatively, a p-halo-benzyl group was introduced at C-2, leaving the indolic nitrogen free. Inhibitory studies were performed and the activity results obtained against both COXs isoforms were rationalized based on docking and NMR studies. Docking studies show that dialkyation at C-2 and C-3 favors a binding with an orientation similar to that of the known selective inhibitor SC-558. From the tested compounds, this substitution pattern is correlated with the highest inhibitory activity and selectivity: 70% COX-2 inhibition at 50 μM, and low COX-1 inhibition (18 ± 9%). Additionally, Saturation Transfer Difference NMR experiments reveal different interaction patterns with both COXs isoforms that may be related with different orientations of the sulfonamide group in the binding pocket. Despite the moderated inhibitory activities found, this study represents an innovative approach towards COXs inhibitory activity rationalization and to the design of anti-inflammatory drugs.


Straight back syndrome as a clue to diagnosing asymptomatic congenital valvular heart disease and limiting the risk of weightlifting.
Although both are initially asymptomatic, mitral valve prolapse/myxomatous mitral valve disease (MVP/MMVD) and bicuspid aortic valve (BAV), with its associated aortic disease, are currently the two most common congenital valvular heart diseases. Severe mitral regurgitation due to rupture of chordae tendineae (CTR) prompts surgery for MVP/MMVD. Surgery for BAV is performed for severe aortic stenosis and/or regurgitation, often with management of root and/or ascending aortic enlargement. There may be an association between straight back syndrome (SBS) and MVP/MMVD, which may be a key to earlier diagnosis. Other associations link weightlifting with ascending aortic enlargement and with CTR, where the common theme is blood pressure elevation. As the number of people with fitness center memberships continues to increase, this potentially exposes more undiagnosed individuals with MVP/MMVD or BAV to risk from weightlifting. Challenges include making the public aware of this risk and preparing the osteopathic physician to recognize patients at risk through a structured history-taking and targeted cardiovascular examination.


Nanoparticle-mediated drug delivery system for cardiovascular disease.
Administration of drugs and other therapeutic agents has been the central strategy of contemporary medicine for cardiovascular disease. The use of a drug delivery system (DDS) is always demanded to enhance the efficacy and safety of therapeutic agents, and improve the signal-to-noise ratio of imaging agents. Nano-scale materials modify in vivo drug kinetics, depending on (patho)physiological mechanisms such as vascular permeability and incorporation by the mononuclear phagocyte system, which constitute 'passive-targeting' properties of nano-DDS. By contrast, an 'active-targeting' strategy employs a specific targeting structure on nano-DDS, which binds to the target molecule that is specific for a certain disease process, such as tumor specific antigens and the induction of adhesion molecules. In this review, we summarize recent studies that applied nano-DDS for the diagnosis and treatment of cardiovascular disease, especially focusing on atherosclerosis and myocardial ischemia-reperfusion (IR) injury. Pathophysiological changes in atherosclerosis and myocardial IR injury are successfully targeted by nano-DDS and preclinical studies in animals showed positive effects of nano-DDS enhancing efficacy and reducing adverse effects. The development of nano-DDS in clinical medicine is keenly being awaited.


[Review on the progress of tobacco epidemic monitoring methods].
Tobacco use is a major preventable contributor to premature death and disease globally. Tobacco epidemic monitoring plays a vital role in tobacco control, considered an essential component of tobacco control in the WHO Framework Convention on Tobacco Control and MPOWER package of tobacco control. In this paper, we reviewed the methods of tobacco monitoring worldwide, mainly including face-to-face surveys, telephone interviews, online questionnaire surveys, data sharing, model estimation, smoking cessation support, and the combination of multiple methods. Meanwhile, we analyzed the strengths and weaknesses of those monitoring methods. Tobacco monitoring methods vary in countries and regions. The review might provide a reference for tobacco epidemic monitoring in China.


Variations in power of opinion leaders in online communication networks.
Online social media has completely transformed how we communicate with each other. While online discussion platforms are available in the form of applications and websites, an emergent outcome of this transformation is the phenomenon of 'opinion leaders'. A number of previous studies have been presented to identify opinion leaders in online discussion networks. In particular, Feng (2016 Comput. Hum. Behav. 54, 43-53. (doi:10.1016/j.chb.2015.07.052)) has identified five different types of central users besides outlining their communication patterns in an online communication network. However, the presented work focuses on a limited time span. The question remains as to whether similar communication patterns exist that will stand the test of time over longer periods. Here, we present a critical analysis of the Feng framework both for short-term as well as for longer periods. Additionally, for validation, we take another case study presented by Udanor et al. (2016 Program 50, 481-507. (doi:10.1108/PROG-02-2016-0011)) to further understand these dynamics. Results indicate that not all Feng-based central users may be identifiable in the longer term. Conversation starter and influencers were noted as opinion leaders in the network. These users play an important role as information sources in long-term discussions. Whereas network builder and active engager help in connecting otherwise sparse communities. Furthermore, we discuss the changing positions of opinion leaders and their power to keep isolates interested in an online discussion network.


Simple model for the nonlinear optical response of gases in the transparency region.
We present a simple model for the nonlinear optical response of atomic gases for pulses with center wavelengths in the transparency region and peak fields for which ionization is not prevalent. By comparing with simulations based on the Schrödinger equation for a hydrogen atom we demonstrate that the model accurately captures the dispersion of the nonlinear polarization as well as noninstantaneous effects for a variety of photon energies and also a two-color pulse. Our approach should be of utility in simulating near- and mid-infrared pulse propagation in dielectric media for which extreme nonlinear effects can arise.


The Players: Cells Involved in Glomerular Disease.
Glomerular diseases are common and important. They can arise from systemic inflammatory or metabolic diseases that affect the kidney. Alternately, they are caused primarily by local glomerular abnormalities, including genetic diseases. Both intrinsic glomerular cells and leukocytes are critical to the healthy glomerulus and to glomerular dysregulation in disease. Mesangial cells, endothelial cells, podocytes, and parietal epithelial cells within the glomerulus all play unique and specialized roles. Although a specific disease often primarily affects a particular cell type, the close proximity, and interdependent functions and interactions between cells mean that even diseases affecting one cell type usually indirectly influence others. In addition to those cells intrinsic to the glomerulus, leukocytes patrol the glomerulus in health and mediate injury in disease. Distinct leukocyte types and subsets are present, with some being involved in different ways in an individual glomerular disease. Cells of the innate and adaptive immune systems are important, directing systemic immune and inflammatory responses, locally mediating injury, and potentially dampening inflammation and facilitating repair. The advent of new genetic and molecular techniques, and new disease models means that we better understand both the basic biology of the glomerulus and the pathogenesis of glomerular disease. This understanding should lead to better diagnostic techniques, biomarkers, and predictors of prognosis, disease severity, and relapse. With this knowledge comes the promise of better therapies in the future, directed toward halting pathways of injury and fibrosis, or interrupting the underlying pathophysiology of the individual diseases that lead to significant and progressive glomerular disease.


Size-structured risk assessments govern Daphnia migration.
One of the more fascinating phenomena in nature is animal mass migrations and in oceans and freshwaters, diel variations in depth distribution of zooplankton are a phenomenon that has intrigued scientists for more than a century. In our study, we show that zooplankton are able to assess the threat level of ultraviolet radiation and adjust their depth distribution to this level at a very fine tuned scale. Moreover, predation risk induces a size-structured depth separation, such that small individuals, which we show are less vulnerable to predation than larger, make a risk assessment and continue feeding in surface waters during day, offering a competitive release from down-migrating larger animals. Hence, we mechanistically show that such simple organisms as invertebrate zooplankton are able to make individual, size-specific decisions regarding how to compromise between threats from both predators and UV radiation, and adjust their diel migratory patterns accordingly.


Secretory overproduction of Arthrobacter simplex 3-ketosteroid delta 1-dehydrogenase by Streptomyces lividans with a multi-copy shuttle vector.
The gene for 3-ketosteroid delta 1-dehydrogenase (ksdD) of Arthrobacter simplex was expressed in Streptomyces lividans and the secreted enzyme was overproduced by using a multi-copy shuttle vector composed of pIJ702 and pUC19. Deletional analysis of the recombinant plasmid showed that the entire coding sequence of the ksdD gene was located within a 7-kb segment of the chromosomal DNA obtained from the enzyme-producing strain of A. simplex. When S. lividans carrying the recombinant plasmid was grown in an appropriate medium, the cells produced about 100-fold more 3-ketosteroid delta 1-dehydrogenase than the original strain. Although the percentage of enzyme secreted was changed during cultivation, a maximum 55% of the enzyme was secreted into the cultured medium of S. lividans, while A. simplex did not produce the enzyme extracellularly. Secretory overproduction of 3-ketosteroid delta 1-dehydrogenase in S. lividans was also identified by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and on native gel, and the enzyme reaction was confirmed by reverse-phase HPLC using 4-androstene-3,17-dione as a substrate.


5-Fluorouracil and cisplatin in the treatment of advanced oral cancer.
The benefit of the effect of chemotherapy in patients with advanced head and neck squamous cell tumors have been demonstrated by recent meta-analyses of randomized studies. However, the role of chemotherapy-especially in advanced oral cancer-is not fully clear, because of the very small amount of phase II literature available. From January 1994 to December 2000, a total of 44 pts aged 33-75 years (mean age 60 years) with advanced and histologically proved squamous cell carcinoma's of the oral cavity received at least one chemotherapy course. Seven patients had stage III and 37 stage IV disease. The chemotherapy was the initial therapy in a group of 21 patients. In a second group of 23 patients the chemotherapy was delivered after relapse of their disease. The pre-chemotherapy treatment of the second group was radiotherapy in 11, surgery in 4, combination of radiotherapy and surgery in 8 patients. The chemotherapy regimen consisted of cisplatin 100 mg/m(2) in 3-h infusion, day 1 and 5-FU 1000 mg/m(2) in 24-h infusion, days 1-5. Treatment was repeated every 21 days. A total of 154 treatment courses (3.5 per patient, ranged 1-10) were administered. Myelotoxicity, nausea and vomiting were the major treatment complications. The overall response rate to the induction chemotherapy was 52.3%, with 19% complete (CR), and 33.3% partial response's (PR) and to the chemotherapy for recurrent/metastatic disease 30.4% with 8.7% CR, and 21.7% PR. No difference was found in the median survival of the two subgroups (12 months). The median survival of the responders was 15 months (95% CI 11.3-18.7 months), and of the non-responders 9 months (95% CI 5.6-12.4 months) (P = 0.0067). Chemotherapy with cisplatin and 5-FU combination is effective in pts with advanced squamous cell oral cancer and appears to improve the survival of patients who have a good response.


TMEM16F and dynamins control expansive plasma membrane reservoirs.
Cells can expand their plasma membrane laterally by unfolding membrane undulations and by exocytosis. Here, we describe a third mechanism involving invaginations held shut by the membrane adapter, dynamin. Compartments open when Ca activates the lipid scramblase, TMEM16F, anionic phospholipids escape from the cytoplasmic monolayer in exchange for neutral lipids, and dynamins relax. Deletion of TMEM16F or dynamins blocks expansion, with loss of dynamin expression generating a maximally expanded basal plasma membrane state. Re-expression of dynamin2 or its GTPase-inactivated mutant, but not a lipid binding mutant, regenerates reserve compartments and rescues expansion. Dynamin2-GFP fusion proteins form punctae that rapidly dissipate from these compartments during TMEM16F activation. Newly exposed compartments extend deeply into the cytoplasm, lack numerous organellar markers, and remain closure-competent for many seconds. Without Ca, compartments open slowly when dynamins are sequestered by cytoplasmic dynamin antibodies or when scrambling is mimicked by neutralizing anionic phospholipids and supplementing neutral lipids. Activation of Ca-permeable mechanosensitive channels via cell swelling or channel agonists opens the compartments in parallel with phospholipid scrambling. Thus, dynamins and TMEM16F control large plasma membrane reserves that open in response to lateral membrane stress and Ca influx.


Barcoding and traditional health practitioner perspectives are informative to monitor and conserve frogs and reptiles traded for traditional medicine in urban South Africa.
Previous literature suggests that Indigenous cultural practices, specifically traditional medicine, are commonplace among urban communities contrary to the general conception that such practices are restricted to rural societies. We reviewed previous literature for records of herptiles (frog and reptile species) sold by traditional health practitioners in urban South Africa, then used visual confirmation surveys, DNA barcoding and folk taxonomy to identify the herptile species that were on sale. Additionally, we interviewed 11 IsiZulu and SePedi speaking traditional health practitioners to document details of the collection and pricing of herptile specimens along with the practitioners' views of current conservation measures for traditional medicine markets. The 34 herptile species recorded in previous literature on traditional medicine markets included endangered and non-native species. Spectrophotometry measurements of the DNA we extracted from the tissue of herptiles used in traditional medicine were an unreliable predictor of whether those extractions would be suitable for further experimental work. From our initial set of 111 tissue samples, 81 sequencing reactions were successful and 55 of those sequences had species-level matches to COI reference sequences on the NCBI GenBank and/or BOLD databases. Molecular identification revealed that traditional health practitioners correctly labelled 77% of the samples that we successfully identified with DNA barcoding in this study. Our mixed methodology approach is useful for conservation planning as it updates knowledge of animal use in Indigenous remedies and can accurately identify species of high conservation priority. Furthermore, this study highlights the possibility of collaborative conservation planning with traditional health practitioners.