RU486-treated rats show endocrine and morphological responses to therapies analogous to responses of women with polycystic ovary syndrome treated with similar therapies.
Administration of 4 mg of the antiprogestagen RU486 to 4-day-cyclic rats over 8 consecutive days starting on the day of estrus (Day 1) induced and anovulatory cystic ovarian condition with endocrine and morphological features similar to those exhibited in polycystic ovarian disease (PCO). To determine whether the RU486-treated rat responds in an analogous fashion to therapies similar to those that have been used to treat human PCO, RU486-treated rats were injection on Days 5 and 7 with 1) 1 mg of an LHRH antagonist (LHRHa), 2) 5 IU of human FSH (hFSH), 3) 2 mg of the antiandrogen flutamide (FLU), 4) 1 mg of the antiestrogen tamoxifen (TMX), or 5) 1 mg of the dopamine agonist bromocriptine (BRC). Controls were intact cyclic rats decapitated on estrus and rats injected with RU486 and the corresponding vehicles (saline or 70% ethanol) used with LHRHa, hFSH, FLU, TMX, and BRC injections. RU486-treated rats were decapitated on Day 9, and the serum concentrations of LH, FSH, prolactin (PRL), testosterone (T), and estradiol-17 beta (E2) were determined. Pituitary and ovary weight, number of follicular cysts, size of the corpora lutea, and rates of follicular growth and atresia were also noted. Finally, the ovulatory response to ovine LH (oLH) in rats treated with RU486 and injected with various doses of hFSH (5, 10, or 20 IU) was evaluated. While administration of LHRHa and of TMX decreased the serum concentrations of LH, T and E2 and the LH/FSH and T/E2, ratios, and injections of BRC and of FLU increased the serum concentration of LH and T, the administration of hFSH (10 IU) to RU486-treated rats increased only the serum levels of E2. All treatments decreased, though in different degrees, both the number of cysts and the rate of follicular atresia, and stimulated follicular growth. The positive effects on follicular growth and atresia were significantly higher in those rats injected with hFSH. Moreover, RU486-treated rats injected with different doses of hFSH ovulated in a dose-dependent manner in response to oLH. Rates deprived of the actions of progesterone through the administration of the antiprogestagen RU486 had 1) endocrine and morphologic alterations comparable to those observed in women with PCO, 2) analogous responses to therapies similar to those that have been used to treat human PCO, and 3) an ovulatory response to combined treatment with FSH and LH. These results establish the fundamental adequacy of using the RU486-treated rat as a PCO model.


Escherichia coli thioredoxin: a subunit of bacteriophage T7 DNA polymerase.
T7 DNA polymerase (DNA nucleotidyltransferase; deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 22.214.171.124) is composed of an 84,000 dalton protein specified by the gene 5 of the phage and a 12,000 dalton protein (TsnC protein) specified by the tsnC gene of E. coli [Modrich, P. & Richardson, C. C. (1975) J. Biol. Chem. 250 5515-5522]. Both proteins are necessary for T7 DNA polymerase activity and for the replication of T7 DNA. The TsnC protein is identical to thioredoxin of E. coli by the following criteria: (1) Homogeneous preparations of both proteins have TsnC and thioredoxin activity. (2) Both proteins show similar stability to heat. (3) They have identical mobilities, corresponding to a molecular weight of 12,000, on polyacrylamide gels containing sodium dodecyl sulfate. (4) Their amino-acid compositions are indistinguishabe. (5) Antibody prepared against thioredoxin inhibits TsnC activity. (6) TsnC protein isolated from purified T7 DNA polymerase has thioredoxin activity. In addition, preparations of T7 DNA polymerase itself exhibit thioredoxin activity and are partially inhibited by antibody to thioredoxin.


Fossilized melanosomes and the colour of Cretaceous dinosaurs and birds.
Spectacular fossils from the Early Cretaceous Jehol Group of northeastern China have greatly expanded our knowledge of the diversity and palaeobiology of dinosaurs and early birds, and contributed to our understanding of the origin of birds, of flight, and of feathers. Pennaceous (vaned) feathers and integumentary filaments are preserved in birds and non-avian theropod dinosaurs, but little is known of their microstructure. Here we report that melanosomes (colour-bearing organelles) are not only preserved in the pennaceous feathers of early birds, but also in an identical manner in integumentary filaments of non-avian dinosaurs, thus refuting recent claims that the filaments are partially decayed dermal collagen fibres. Examples of both eumelanosomes and phaeomelanosomes have been identified, and they are often preserved in life position within the structure of partially degraded feathers and filaments. Furthermore, the data here provide empirical evidence for reconstructing the colours and colour patterning of these extinct birds and theropod dinosaurs: for example, the dark-coloured stripes on the tail of the theropod dinosaur Sinosauropteryx can reasonably be inferred to have exhibited chestnut to reddish-brown tones.


A systematic review of the evidence base for telehospice.
Abstract The use of telehealth technologies to overcome the geographic distances in the delivery of hospice care has been termed telehospice. Although telehospice research has been conducted over the last 10 years, little is known about the comprehensive findings within the field. The purpose of this systematic article was to focus on available research and answer the question, What is the state of the evidence related to telehospice services? The article was limited to studies that had been published in the English language and indexed between January 1, 2000 and March 23, 2010. Indexed databases included PubMed and PsycINFO and contained specified key words. Only research published in peer review journals and reporting empirical data, rather than opinion or editorials, were included. A two-part scoring framework was modified and applied to assess the methodological rigor and pertinence of each study. Scoring criteria allowed the evaluation of both quantitative and qualitative methodologies. Twenty-six studies were identified with the search strategy. Although limited in number and in strength, studies have evaluated the use of a variety of technologies, attitudes toward use by providers and consumers, clinical outcomes, barriers, readiness, and cost. A small evidence base for telehospice has emerged over the last 10 years. Although the evidence is of medium strength, its pertinence is strong. The evidence base could be strengthened with randomized trials and additional clinical-outcome-focused research in larger randomized samples and in qualitative studies with better-described samples.


[Acute gastric and duodenal ulcers in burns].
The authors made a clinical trial of acute gastric and duodenal ulcers arising in burnt patients. Treatment results and autopsy material have been analysed for 1410 and 165 burnt patients, respectively. Of 1410 patients, 8(0.6%) patients developed acute gastric and duodenal ulcers. Of 165 dead of burn disease, 23(14%) died of gastroduodenal hemorrhage from ulcer lesions. The causes of acute ulcer emergence were studied and factors of risk for burnt patients to develop acute ulcer and gastroduodenal erosions were determined. This allowed the authors to propose measures to prevent onset of acute ulcer in burnt patients.


Corneal xerophthalmia and keratomalacia.
Detailed examinations were conducted on 162 consecutive children with nutritional keratopathy. Surface changes ranged from mild haziness through generalized xerosis and formation of thickened keratinized plaques. Diffuse stromal edema occured early in the disease. Stromal loss took two forms: relatively small, sharply demarcated, eccentric, noninfiltrated cylindrical ulcers of varying depth; and localized or generalized, usually full-thickness necrosis. All forms of involvement were sometimes present in the same individual or even the same eye. Traumatic separation of a keratinized plaque, or decompensation of a dellen, accounted for some instances of stromal baring. In other cases, stromal melting appeared to progress below an intact epithelium. Focal areas of necrosis healed rapidly, as adherent leukomas; larger lesions sloughed, forming extensive descemetoceles. With therapy, the vast majority of children retained central corneal clarity.


Comparison of Effects of Two Aging Methods on the Physicochemical Traits of Pork Loin.
The objective of this study was to compare effects of two different aging methods on physical, chemical, and microbial traits of pork loin: Dry and wet-aged meat was hung in the cooler at 8±1°C and 85±2.1% humidity for 14 days, while wet-aged meat was immersed in a 3.5% salt solution of brine in vacuum pouches. On day 7, pH and moisture content were higher in dry-aged loins than in wet-aged, while drip loss and total plate counts (p<0.05) were lower on day 14. As aging continued, the pH and drip loss of dry-aged loins decreased, while their total plate counts and water holding capacity (WHC) increased (p<0.05). After 7 and 14 days of aging, redness in dry-aged loins was higher than that in wet -aged muscles (p<0.05). On day 14 of aging, hardness, chewiness, and adhesiveness were lower in dry-aged pork loin as compared to those in wet-aged samples (p<0.05). Consequently, the results suggested that dry and wet aging methods differently affects meat quality traits of pork loin.


Phlebotomy Staffing.
Laboratory directors are tasked with staffing laboratories in a manner that provides adequate services and maintains economic sustainability. To determine the national normative rates of phlebotomy staffing and the types of laboratory operational characteristics that may be associated with the magnitude of those staffing levels. Study participants provided data on inpatient and outpatient phlebotomy sites, including the numbers of patients receiving phlebotomy services, phlebotomy staff, and billable tests. From these data, we calculated performance indicators including the numbers of phlebotomies/phlebotomy full-time equivalent staff, outpatient phlebotomy visits/full-time equivalent staff, and average outpatient phlebotomy wait times. Participants also completed a survey of their laboratory phlebotomy practices. This study was conducted during the third quarter of 2017. Forty-two institutions participated in this study, providing eligible results for 40 selected inpatient sites and 70 selected outpatient sites. The ratios for all performance indicators spanned between 3.3- and 142-fold. The median average outpatient phlebotomy wait time was 8 minutes. None of the performance indicators were associated with the practice variables that we chose to test. The distribution of phlebotomy staffing performance indicators among the laboratories participating in this study varied widely, even among those groups performing similar volumes of tests.


Global exponential stability of competitive neural networks with different time scales.
The dynamics of cortical cognitive maps developed by self-organization must include the aspects of long and short-term memory. The behavior of such a neural network is characterized by an equation of neural activity as a fast phenomenon and an equation of synaptic modification as a slow part of the neural system. We present a new method of analyzing the dynamics of a biological relevant system with different time scales based on the theory of flow invariance. We are able to show the conditions under which the solutions of such a system are bounded being less restrictive than with the K-monotone theory, singular perturbation theory, or those based on supervised synaptic learning. We prove the existence and the uniqueness of the equilibrium. A strict Lyapunov function for the flow of a competitive neural system with different time scales is given and based on it we are able to prove the global exponential stability of the equilibrium point.


A randomized trial of amantadine in Huntington disease.
Huntington chorea, like levodopa-induced dyskinesias, may be responsive to amantadine hydrochloride treatment. To measure the effect of amantadine treatment on Huntington chorea and to test a hypothesis that the adventitious movements are associated with reduced central proprioception that can be corrected by amantadine treatment. A randomized placebo-controlled cross-over trial with 2 weeks of treatment. A tertiary referral center. Twenty-four subjects with Huntington disease took amantadine hydrochloride, 100 mg 3 times daily for 2 weeks, and placebo for 2 weeks. Chorea of the face, trunk, and limbs while seated was videotaped at baseline and after each study phase. Segments were viewed in random order by blinded reviewers and scored. Proprioception was determined using arm restraints in which the right and left elbows were set at slightly different angles, and the errors in selecting the more extended elbow over 40 trials were recorded. The chorea score was not correlated with a proprioception deficit. Neither chorea nor proprioception were significantly affected by amantadine therapy. The chorea score was 9.6 (3.1) points at baseline and 9.7 (3.7) points when the patient was receiving amantadine therapy. The 95% confidence interval for the difference between placebo effect and amantadine effect was -1.43 to 1.0 points. Despite this, 19 subjects felt improved during the amantadine phase compared with 6 subjects improved in the placebo phase (P =.006) and the quality of life was better (P<.001). Elbow proprioception was not shown to be related to Huntington chorea. Amantadine hydrochloride treatment at doses of 300 mg/d had no effect, on average, for Huntington chorea, although most patients felt subjectively better during the short course of amantadine treatment.


Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals.
We describe an attractive cloning system for the seed-specific expression of recombinant proteins using three non-food/feed crops. A vector designed for direct subcloning by Gateway® recombination was developed and tested in Arabidopsis, tobacco and petunia plants for the production of a chimeric form (GAD67/65) of the 65 kDa isoform of glutamic acid decarboxylase (GAD65). GAD65 is one of the major human autoantigens involved in type 1 diabetes (T1D). The murine anti-inflammatory cytokine interleukin-10 (IL-10) was expressed with the described system in Arabidopsis and tobacco, whereas proinsulin, another T1D major autoantigen, was expressed in Arabidopsis. The cost-effective production of these proteins in plants could allow the development of T1D prevention strategies based on the induction of immunological tolerance. The best yields were achieved in Arabidopsis seeds, where GAD67/65 reached 7.7% of total soluble protein (TSP), the highest levels ever reported for this protein in plants. IL-10 and proinsulin reached 0.70% and 0.007% of TSP, respectively, consistent with levels previously reported in other plants or tissues. This versatile cloning vector could be suitable for the high-throughput evaluation of expression levels and stability of many valuable and difficult to produce proteins.


An fMRI study of cortical representation of mechanical allodynia in patients with neuropathic pain.
To investigate cerebral activity associated with allodynia in patients with neuropathic pain. The brain responses of 27 patients with peripheral (5), spinal (3), brainstem (4), thalamic (5), lenticular (5), or cortical (5) lesions were studied with fMRI as innocuous mechanical stimuli were addressed to either the allodynic territory or the homologous contralateral region. When applied to the normal side, brush and cold rubbing stimuli did not evoke pain and activated a somatosensory "control" network including contralateral primary (SI) and secondary (SII) somatosensory cortices and insular regions. The same stimuli became severely painful when applied to the allodynic side and activated regions in the contralateral hemisphere that mirrored the "control" network, with, however, lesser activation of the SII and insular cortices. Increased activation volumes were found in contralateral SI and primary motor cortex (MI). Whereas ipsilateral responses appeared very small and restricted after control stimuli, they represented the most salient effect of allodynia and were observed mainly in the ipsilateral parietal operculum (SII), SI, and insula. Allodynic stimuli also recruited additional responses in motor/premotor areas (MI, supplementary motor area), in regions involved in spatial attention (posterior parietal cortices), and in regions linking attention and motor control (mid-anterior cingulate cortex). On a background of deafferentation in the hemisphere contralateral to stimuli, enhanced or additional responses to innocuous stimuli in the ipsilateral hemisphere may contribute to the shift of perception from innocuous toward painful and ill-defined sensations.


Association of heat shock protein 70 with enterovirus capsid precursor P1 in infected human cells.
Members of the human heat shock (HSP) family of related proteins are involved in the intracellular folding, transport, and assembly of proteins and protein complexes. We have observed that human heat shock protein 70 (HSP70) is associated with the capsid precursor P1 of poliovirus and coxsackievirus B1 in infected HeLa cells. Antiserum generated against HSP70 coimmunoprecipitated the poliovirus protein P1, an intermediate in capsid assembly. Similarly, alpha-virion serum coimmunoprecipitated HSP70 from virus-infected cell extracts, but not from mock-infected cell extracts. The HSP70-P1 complex was stable in high-salt medium but was sensitive to incubation with 2 mM ATP, which is a characteristic of other known functional complexes between HSP70 and cellular proteins. The P1 in the complex was predominantly newly synthesized, and the half-life of complexed P1 was nearly twice as long as that of total P1. The HSP70-P1 complex was found to sediment at 3S to 6S, suggesting that it may be part of, or a precursor to, the "5S promoter particles" thought to be an assembly intermediate of picornaviruses. The finding that HSP70 was associated with the capsid precursors of at least two enteroviruses may suggest a functional role of these complexes in the viral life cycles.


Medical practice without antianxiety drugs.
The benzodiazepines and hydroxyzine were removed from a statewide Medicaid pharmacopeia. Barbiturate sedatives, hypnotic and antipsychotic medication, and tricyclic antidepressants remained available, but no substitution was made for about two-thirds of the deleted ataractic medications. The authors conclide that most physicans regard ataractics as unique and do not use them interchangeably with the other drugs.


Superiority of ibutilide (a new class III agent) over DL-sotalol in converting atrial flutter and atrial fibrillation. The Ibutilide/Sotalol Comparator Study Group.
To compare the efficacy and safety of a single dose of ibutilide, a new class III antiarrhythmic drug, with that of DL-sotalol in terminating chronic atrial fibrillation or flutter in haemodynamically stable patients. Double blind, randomised study. 43 European hospitals. 308 patients (mean age 60 years, 70% men, 48% with heart disease) with sustained atrial fibrillation (n = 251) or atrial flutter (n = 57) (duration three hours to 45 days) were randomised to three groups to receive a 10 minute infusion of 1 mg ibutilide (n = 99), 2 mg ibutilide (n = 106), or 1.5 mg/kg DL-sotalol (n = 103). Infusion was discontinued at termination of the arrhythmia. Successful conversion of atrial fibrillation or flutter, defined as termination of arrhythmia within one hour of treatment. Both drugs were more effective against atrial flutter than against atrial fibrillation. Ibutilide was superior to DL-sotalol for treating atrial flutter (70% and 56% v 19%), while the high dose of ibutilide was more effective for treating atrial fibrillation than DL-sotalol (44% v 11%) and the lower dose of ibutilide (44% v 20%, p < 0.01). The mean (SD) time to arrhythmia termination was 13 (7) minutes with 2 mg ibutilide, 19 (15) minutes with 1 mg ibutilide, and 25 (17) minutes with DL-sotalol. In all patients, the duration of arrhythmia before treatment was a predictor of arrhythmia termination, although this was less obvious in the group that received 2 mg ibutilide. This dose converted almost 48% of atrial fibrillation that was present for more than 30 days. Concomitant use of digitalis or nifedipine and prolongation of the QTc interval were not predictive of arrhythmia termination. Bradycardia (6.5%) and hypotension (3.7%) were more common side effects with DL-sotalol. Of 211 patients given ibutilide, two (0.9%) who received the higher dose developed polymorphic ventricular tachycardia, one of whom required direct current cardioversion. Ibutilide (given in 1 or 2 mg doses over 10 minutes) is highly effective for rapidly terminating persistent atrial fibrillation or atrial flutter. This new class III drug, under monitored conditions, is a potential alternative to currently available cardioversion options.


Antigenic structure of the Murray Valley encephalitis virus E glycoprotein.
To complement our battery of St Louis encephalitis (SLE) virus monoclonal antibodies (MAbs), we isolated and characterized MAbs reactive with another member of the SLE virus serocomplex, Murray Valley encephalitis (MVE) virus. From 40 fusion products, we isolated 10 stable hybridomas. The combination of SLE and MVE virus MAbs defined eight epitopes on the MVE envelope (E) glycoprotein. Six of these epitopes (E-1a, E-1c, E-1d, E-3, E-4a, E-4b) were identical to those previously demonstrated on SLE virus. Two new epitopes (E-5 and E-6) were also identified. As with SLE virus, the MVE E-1c epitope elicited the most potent virus-neutralizing and protective MAb. Unlike SLE virus, however, one of the cross-reactive epitopes (E-5) elicited neutralizing antibody and protected animals from MVE virus challenge. These results indicate that, while the antigenic domains on viruses within the SLE virus serocomplex are quite similar, epitopes involved in virus neutralization or protection from virus challenge may vary and can be topologically distinct.


Meta-analysis of a possible signal of increased mortality associated with cefepime use.
On the basis of meta-analyses, concern has been raised regarding a possible signal of increased mortality associated with the use of cefepime versus other beta-lactam antibiotics. To further investigate this possible signal, we accessed findings and data from published and unpublished cefepime clinical trials. We performed meta-analyses using trial- and patient-level data from comparative trials. Trial-level analyses were performed using summary data from all patients in the trials, and patient-level analyses were performed on trials for which patient-level data were available. Thirty-day, all-cause mortality was analyzed using the Mantel-Haenszel adjusted risk difference (ARD) method. The trial-level meta-analysis was based on 88 trials (9467 cefepime patients and 8288 comparator patients). The 30-day, all-cause mortality rates were 6.21% (588/9467) for the cefepime patients and 6.00% (497/8288) for comparator patients (ARD per 1000 population, 5.38; 95% confidence interval [CI], -1.53 to 12.28). In the patient-level analysis (35 trials, 5058 cefepime patients, and 3976 comparator patients), 30-day, all-cause mortality rates were 5.63% (285/5058) for cefepime patients and 5.68% (226/3976) for comparator patients (ARD per 1000 population, 4.83; 95% CI, -4.72 to 14.38). A sensitivity analysis based solely on the 24 febrile neutropenia trials did not show a statistically significant increase in mortality with cefepime use (ARD per 1000 population, 9.67; 95% CI, -2.87 to 22.21). In both trial-level and patient-level meta-analyses, we did not identify a statistically significant increase in mortality among cefepime-treated patients, compared with those treated with other antibacterials.


Concussion Guidelines Step 2: Evidence for Subtype Classification.
Concussion is a heterogeneous mild traumatic brain injury (mTBI) characterized by a variety of symptoms, clinical presentations, and recovery trajectories. By thematically classifying the most common concussive clinical presentations into concussion subtypes (cognitive, ocular-motor, headache/migraine, vestibular, and anxiety/mood) and associated conditions (cervical strain and sleep disturbance), we derive useful definitions amenable to future targeted treatments. To use evidence-based methodology to characterize the 5 concussion subtypes and 2 associated conditions and report their prevalence in acute concussion patients as compared to baseline or controls within 3 d of injury. A multidisciplinary expert workgroup was established to define the most common concussion subtypes and their associated conditions and select clinical questions related to prevalence and recovery. A literature search was conducted from January 1, 1990 to November 1, 2017. Two experts abstracted study characteristics and results independently for each article selected for inclusion. A third expert adjudicated disagreements. Separate meta-analyses were conducted to do the following: 1) examine the prevalence of each subtype/associated condition in concussion patients using a proportion, 2) assess subtype/associated conditions in concussion compared to baseline/uninjured controls using a prevalence ratio, and 3) compare the differences in symptom scores between concussion subtypes and uninjured/baseline controls using a standardized mean difference (SMD). The most prevalent concussion subtypes for pediatric and adult populations were headache/migraine (0.52; 95% CI = 0.37, 0.67) and cognitive (0.40; 95% CI = 0.25, 0.55), respectively. In pediatric patients, the prevalence of the vestibular subtype was also high (0.50; 95% CI = 0.40, 0.60). Adult patients were 4.4, 2.9, and 1.7 times more likely to demonstrate cognitive, vestibular, and anxiety/mood subtypes, respectively, as compared with their controls (P < .05). Children and adults with concussion showed significantly more cognitive symptoms than their respective controls (SMD = 0.66 and 0.24; P < .001). Furthermore, ocular-motor in adult patients (SMD = 0.72; P < .001) and vestibular symptoms in both pediatric and adult patients (SMD = 0.18 and 0.36; P < .05) were significantly worse in concussion patients than in controls. Five concussion subtypes with varying prevalence within 3 d following injury are commonly seen clinically and identifiable upon systematic literature review. Sleep disturbance, a concussion-associated condition, is also common. There was insufficient information available for analysis of cervical strain. A comprehensive acute concussion assessment defines and characterizes the injury and, therefore, should incorporate evaluations of all 5 subtypes and associated conditions.


Spatial encoding in dorsomedial prefrontal cortex and hippocampus is related during deliberation.
Deliberation is thought to involve the internal simulation of the outcomes of candidate actions, the valuation of those outcomes, and the selection of the actions with the highest expected value. While it is known that deliberation involves prefrontal cortical areas, specifically the dorsomedial prefrontal cortex (dmPFC), as well as the hippocampus (HPC) and other brain regions, how these areas process prospective information and select actions is not well understood. We recorded simultaneously from ensembles in dmPFC and CA1 of dorsal HPC in rats during performance of a spatial contingency switching task, and examined the relationships between spatial and reward encoding in these two areas during deliberation at the choice point. We found that CA1 and dmPFC represented either goal locations or the current position simultaneously, but that when goal locations were encoded, HPC and dmPFC did not always represent the same goal location. Ensemble activity in dmPFC predicted when HPC would represent goal locations, but on a broad timescale on the order of seconds. Also, reward encoding in dmPFC increased during hippocampal theta cycles where CA1 ensembles represented the goal location. These results suggest that dmPFC and HPC share prospective information during deliberation, that dmPFC may influence whether HPC represents prospective information, and that information recalled about goal locations by HPC may be integrated into dmPFC reward representations on fast timescales.


Halogen-bonded iodonium ion catalysis: a route to α-hydroxy ketones via domino oxidations of secondary alcohols and aliphatic C-H bonds with high selectivity and control.
A domino synthesis of α-hydroxy ketones has been developed from benzylic secondary alcohols employing catalytic iodonium ions stabilized by DMSO. The reaction proceeds through an unprecedented sequential oxidation of alcohols to ketone and its α-hydroxylation in a controlled manner. The spectroscopic evidence establishes the possibility of formation of a stable halogen-bonded adduct between DMSO and iodonium ions.


Impact of Changes in Clinical Microbiology Laboratory Location and Ownership on the Practice of Infectious Diseases.
The number of onsite clinical microbiology laboratories in hospitals is decreasing, likely related to the business model for laboratory consolidation and labor shortages, and this impacts a variety of clinical practices, including that of banking isolates for clinical or epidemiologic purposes. To determine the impact of these trends, infectious disease (ID) physicians were surveyed regarding their perceptions of offsite services. Clinical microbiology practices for retention of clinical isolates for future use were also determined. Surveys were sent to members of the Infectious Diseases Society of America's (IDSA) Emerging Infections Network (EIN). The EIN is a sentinel network of ID physicians who care for adult and/or pediatric patients in North America and who are members of IDSA. The response rate was 763 (45%) of 1,680 potential respondents. Five hundred forty (81%) respondents reported interacting with the clinical microbiology laboratory. Eighty-six percent of respondents thought an onsite laboratory very important for timely diagnostic reporting and ongoing communication with the clinical microbiologist. Thirty-five percent practiced in institutions where the core microbiology laboratory has been moved offsite, and an additional 7% (n = 38) reported that movement of core laboratory functions offsite was being considered. The respondents reported that only 24% of laboratories banked all isolates, with the majority saving isolates for less than 30 days. Based on these results, the trend toward centralized core laboratories negatively impacts the practice of ID physicians, potentially delays effective implementation of prompt and targeted care for patients with serious infections, and similarly adversely impacts infection control epidemiologic investigations.


Two forms of the nucleoside diphosphate kinase of Pseudomonas aeruginosa 8830: altered specificity of nucleoside triphosphate synthesis by the cell membrane-associated form of the truncated enzyme.
Nucleoside diphosphate kinase (EC 184.108.40.206) (Ndk) is a ubiquitous enzyme functioning in the intracellular distribution of terminal phosphate bond energy among the various nucleotides used in synthetic and regulatory functions in cells. We have previously reported that in Pseudomonas aeruginosa, this important enzyme is transcriptionally regulated by the gene algR2 and posttranslationally regulated by a phosphoprotein phosphatase for the phosphorylated form of Ndk. We report here that an intracellular protease cleaves the 16-kDa form of Ndk to a 12-kDa form that undergoes autophosphorylation with an efficiency almost identical to that of the 16-kDa form. The 12-kDa form was found to be predominantly associated with the P. aeruginosa cell membrane fraction, whereas the 16-kDa form was predominantly cytoplasmic. In the membrane-associated state, the 12-kDa form of Ndk was found to synthesize GTP in preference to other nucleoside triphosphates. The specificity toward GTP synthesis could be abolished by the addition of Tween 20 or Triton X-100. The activity itself could be abolished by the addition of anti-Ndk antibody to the assay mixture. The formation of the 12-kDa form of Ndk and its association with the cell membrane were found to be related to the growth stage of P. aeruginosa, with less than 1% of the 12-kDa Ndk detectable in the membrane fraction at early log phase in comparison with the levels present at late stationary phase.


Cadmium-induced changes in antioxidant enzymes in suspension culture of soybean cells.
Cadmium (Cd), similarly to other heavy metals, inhibits plant growth. We have recently showed that Cd(2+) either stimulates (1-4 microM) or inhibits (>or= 6 microM) growth of soybean (Glycine max L.) cells in suspension culture (Sobkowiak & Deckert, 2003, Plant Physiol Biochem. 41: 767-72). Here, soybean cell suspension cultures were treated with various concentrations of Cd(2+) (1-10 microM) and the following enzymes were analyzed by native electrophoresis: superoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APOX). We found a significant correlation between the cadmium-induced changes of soybean cell culture growth and the isoenzyme pattern of the antioxidant enzymes. The results suggest that inhibition of growth and modification of antioxidant defense reactions appear in soybean cells when Cd(2+) concentration in culture medium increases only slightly, from 4 to 6 microM.


Mutants of the membrane-binding region of Semliki Forest virus E2 protein. II. Topology and membrane binding.
The p62/E2 protein of Semliki Forest virus (SFV) is a typical transmembrane glycoprotein, with an amino-terminal lumenal domain, a transmembrane (hydrophobic) domain, and a carboxy-terminal cytoplasmic domain (or tail). Our hypothesis has been that the membrane-binding polypeptide region (membrane anchor) of this protein consists of both the transmembrane domain and the adjacent positively charged peptide, Arg-Ser-Lys, which is part of the cytoplasmic domain. We have investigated three anchor mutants of the p62 protein with respect to both their disposition and their stability in cell membranes. The construction of the three mutants has been described (Cutler, D.F., and H. Garoff, J. Cell Biol., 102:889-901). They are as follows: A1, changing the basic charge cluster from Arg-Ser-Lys(+2) to Gly-Ser-Glu(-1); A2, replacing an Ala in the middle of the hydrophobic stretch with a Glu; A3, changing the charge cluster from Arg-Ser-Lys(+2) to Gly-Ser-Met(0). All three mutants retain the transmembrane configuration of the wild-type p62. In a cell homogenate they have a cytoplasmic domain that is accessible to protease. In living cells an anti-peptide antibody specific for the cytoplasmic tail of p62 reacts with the tails of both wild-type and mutant p62s following its introduction into the cytoplasm. All three mutant proteins have Triton X-114 binding properties similar to the wild-type p62. However, when the membranes of cells expressing the three mutants or the wild-type p62 protein are washed with sodium carbonate, pH 11.5, three to four times as much mutant protein as wild-type p62 is released from the membranes. Thus the stability in cell membranes of the three mutant p62 proteins is significantly reduced.


Potentiation of natural killer cells to overcome cancer resistance to NK cell-based therapy and to enhance antibody-based immunotherapy.
Natural killer (NK) cells are cellular components of the innate immune system that can recognize and suppress the proliferation of cancer cells. NK cells can eliminate cancer cells through direct lysis, by secreting perforin and granzymes, or through antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC involves the binding of the Fc gamma receptor IIIa (CD16), present on NK cells, to the constant region of an antibody already bound to cancer cells. Cancer cells use several mechanisms to evade antitumor activity of NK cells, including the accumulation of inhibitory cytokines, recruitment and expansion of immune suppressor cells such as myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs), modulation of ligands for NK cells receptors. Several strategies have been developed to enhance the antitumor activity of NK cells with the goal of overcoming cancer cells resistance to NK cells. The three main strategies to engineer and boost NK cells cytotoxicity include boosting NK cells with modulatory cytokines, adoptive NK cell therapy, and the employment of engineered NK cells to enhance antibody-based immunotherapy. Although the first two strategies improved the efficacy of NK cell-based therapy, there are still some limitations, including immune-related adverse events, induction of immune-suppressive cells and further cancer resistance to NK cell killing. One strategy to overcome these issues is the combination of monoclonal antibodies (mAbs) that mediate ADCC and engineered NK cells with potentiated anti-cancer activity. The advantage of using mAbs with ADCC activity is that they can activate NK cells, but also favor the accumulation of immune effector cells to the tumor microenvironment (TME). Several clinical trials reported that combining engineered NK cells with mAbs with ADCC activity can result in a superior clinical response compared to mAbs alone. Next generation of clinical trials, employing engineered NK cells with mAbs with higher affinity for CD16 expressed on NK cells, will provide more effective and higher-quality treatments to cancer patients.


Measuring parathyroid hormone (PTH) in patients with oxidative stress--do we need a fourth generation parathyroid hormone assay?
Oxidation of PTH at methionine residues results in loss of biological activity. PTH may be oxidized in patients with renal disease. The aim of this study was to develop an assay considering oxidation of PTH. Oxidized hPTH was analyzed by high resolution nano-liquid chromatography coupled to ESI-FTT tandem mass spectrometry (nanoLC-ESI-FT-MS/MS) directly and after proteolytic cleavage. The oxidized hPTH(1-84) sample shows TIC-peaks at 18-20 min and several mass peaks due to mass shifts caused by oxidations. No significant signal for oxidized hPTH(1-84) species after removal of oxidized PTH molecules by a specific column with monoclonal antibodies (MAB) raised against the oxidized hPTH was detectable. By using this column in samples from 18 patients on dialysis we could demonstrate that measured PTH concentrations were substantially lower when considering oxidized forms of PTH. The relationship between PTH concentrations determined directly and those concentrations measured after removal of the oxidized PTH forms varies substantially. In some patients only 7% of traditionally measured PTH was free of oxidation, whereas in other patients 34% of the traditionally measured PTH was real intact PTH. In conclusion, a huge but not constant proportion of PTH molecules are oxidized in patients requiring dialysis. Since oxidized PTH is biologically inactive, the currently used methods to detect PTH in daily clinical practice may not adequately reflect PTH-related bone and cardiovascular abnormalities in patients on dialysis.


Clinical effectiveness of percutaneous adhesiolysis and predictive factors of treatment efficacy in patients with lumbosacral spinal stenosis.
Patients with lumbosacral spinal stenosis (LSS) do not always obtain satisfactory pain relief from transforaminal epidural steroid injection (TFESI) because perineural/epidural adhesions prevent the spread of injectate into the epidural space. Percutaneous adhesiolysis (PA) can eliminate the deleterious effects of adhesion. This study was to evaluate the effectiveness of PA among patients with LSS refractory to TFESI and to ascertain the prognostic factors determining PA efficacy. Retrospective study. Spine hospital. Sixty-five patients with LSS refractory to TFESI who underwent PA with NaviCath® were reviewed. We recorded Numeric Rating Scale for back pain (NRS back) and leg pain (NRS leg), and Oswestry Disability Index (ODI), at pretreatment, 2 weeks, and 3 months after treatment. Successful pain relief and functional improvement were described as a 50% and 40% or more reduction in NRS and ODI, respectively. Clinical data and radiological findings were obtained to assess the possible predictive factors for PA efficacy. Among the 65 patients, 45 (69.2%), 40 (61.5%), and 39 (60.0%) patients showed successful outcomes in NRS back, NRS leg, and ODI at 2 weeks, respectively. Among 63 patients who were followed up at 3 months, 34 (54.0%), 32 (50.8%), and 30 (47.6%) patients showed successful results, respectively. Spondylolisthesis, previous lumbar surgery, and foraminal stenosis were associated with a significantly higher proportion of unsuccessful result in NRS and ODI (%). PA may be a useful treatment in patients with LSS refractory to TFESI and reduce the surgical requirement. Previous surgery, spondylolisthesis, and foraminal stenosis may be associated with poor prognosis.


Mechanistic insights into tRNA cleavage by a contact-dependent growth inhibitor protein and translation factors.
Contact-dependent growth inhibition is a mechanism of interbacterial competition mediated by delivery of the C-terminal toxin domain of CdiA protein (CdiA-CT) into neighboring bacteria. The CdiA-CT of enterohemorrhagic Escherichia coli EC869 (CdiA-CTEC869) cleaves the 3'-acceptor regions of specific tRNAs in a reaction that requires the translation factors Tu/Ts and GTP. Here, we show that CdiA-CTEC869 has an intrinsic ability to recognize a specific sequence in substrate tRNAs, and Tu:Ts complex promotes tRNA cleavage by CdiA-CTEC869. Uncharged and aminoacylated tRNAs (aa-tRNAs) were cleaved by CdiA-CTEC869 to the same extent in the presence of Tu/Ts, and the CdiA-CTEC869:Tu:Ts:tRNA(aa-tRNA) complex formed in the presence of GTP. CdiA-CTEC869 interacts with domain II of Tu, thereby preventing the 3'-moiety of tRNA to bind to Tu as in canonical Tu:GTP:aa-tRNA complexes. Superimposition of the Tu:GTP:aa-tRNA structure onto the CdiA-CTEC869:Tu structure suggests that the 3'-portion of tRNA relocates into the CdiA-CTEC869 active site, located on the opposite side to the CdiA-CTEC869 :Tu interface, for tRNA cleavage. Thus, CdiA-CTEC869 is recruited to Tu:GTP:Ts, and CdiA-CT:Tu:GTP:Ts recognizes substrate tRNAs and cleaves them. Tu:GTP:Ts serves as a reaction scaffold that increases the affinity of CdiA-CTEC869 for substrate tRNAs and induces a structural change of tRNAs for efficient cleavage by CdiA-CTEC869.


The association of smoking and the cost of military training.
To determine if premature discharge from the US Air Force was associated with the smoking status of recruits. A total of 29 044 US Air Force personnel recruited from August 1995 to August 1996 were administered baseline behavioural risk assessment surveys during basic military training. They were tracked over a 12 month period to determine those who were prematurely discharged. Excess training costs as a result of premature discharge. In this 12 month period, 14.0% of those entering the US Air Force were discharged at a one year follow up. In both univariate and multivariate models, the best single predictor of early discharge was smoking status. Overall, 11.8% of non-smokers versus 19.4% of smokers were prematurely discharged (relative risk 1.795). Using US Department of Defense data on the cost of military training, recruits who smoke in the US Air Force are associated with $18 million per year in excess training costs. Applied to all service branches, smoking status, which represents a constellation of underlying behaviours and attitudes that can contribute to early discharge, is associated with over $130 million per year in excess training costs.


Colon Perfusion Patterns During Colorectal Resection Using Visible Light Spectroscopy.
The impact of blood supply to the anastomosis on development of anastomotic leakage is still a matter of debate. Considering that bowel perfusion may be affected by manipulation during surgery, perfusion assessment of the anastomosis alone may be of limited value. We propose perfusion assessment at different time points during surgery to explore the dynamics of bowel perfusion during colorectal resection and its impact on outcome. In this prospective cohort study, patients undergoing elective colorectal resection were eligible. Colon perfusion was evaluated using visible light spectroscopy. Main outcome was the difference in colon perfusion, quantified by measuring tissue oxygen saturation (StO2) in the colonic serosa, before and after anastomosis during surgery. We included 58 patients between July 2013 and November 2015. Colon perfusion increased by an average of 5.9% StO2 during surgery (95% confidence interval 3.1, 8.8; P < 0.001). The number of patients with abnormal perfusion (defined as StO2 < 65%) decreased from 50% at the beginning to 24% by the end of surgery. Six patients (10%) developed anastomotic leaks (AL), of which five patients had abnormal perfusion at the beginning of surgery, whereas four patients had normal StO2 at the anastomosis. Colon perfusion significantly increased during colorectal surgery. Considering that one quarter of patients had suboptimal anastomotic perfusion without developing AL, impaired blood flow at the anastomosis alone does not seem to be critical. Further investigations including more patients are necessary to evaluate the impact of perioperative parameters on colon perfusion, anastomotic healing and surgical outcome.


Tumor localization using diffuse optical tomography and linearly constrained minimum variance beamforming.
We present a tumor localization method for diffuse optical tomography using linearly constrained minimum variance (LCMV) beamforming. Beamforming is a spatial filtering technique where signals from certain directions can be enhanced while noise and interference from other directions are suppressed. In our method, we tessellate the domain into small voxels and regard each voxel as a possible position of abnormality (e.g., tumor).We then design a spatial filter based on the linearly constrained minimum variance criterion and apply it to each voxel in the domain. The abnormality is localized by observing the peak in the filter output signals. We test our method using simulated 3D examples. We assume a cubic transmission geometry and consider different cases where the abnormality is an absorber, a scatterer, and both. We also give examples showing the resolution of our method and its performance under different perturbation levels and noise levels. Simulation results show that LCMV beamforming can localize the abnormality well with good computational efficiency. It can be used alone for tumor localization and also as an effective preprocessing tool for improving the image reconstruction performances of other inverse methods.


Environmental determinants of atopic eczema phenotypes in relation to asthma and atopic sensitization.
There is still uncertainty about the determinants of atopic eczema (AE). To explain the heterogeneity of the disease, different phenotypes of AE have been suggested. The cross-sectional PARSIFAL study included 14 893 school-age children of farmers or children attending Steiner schools and their respective reference groups. A detailed questionnaire was completed, and house dust was collected for the measurement of endotoxin and glucans. Atopic sensitization was defined by allergen-specific IgE levels in the serum. In multivariate analyses, helping with haying was the only variable related to a farming environment having a consistent inverse association with both current symptoms and a doctor's diagnosis of AE [aOR = 0.65 (95% CI: 0.46-0.93) and 0.73 (0.51-1.05)], respectively. Severe lower respiratory tract infections (LRTI) in the first 2 years of life and usage of antibiotics ever were found to be positively related only to asthma-associated AE, whereas the effect of LRTI on AE without asthma had an opposite effect. Levels of beta(1-->3)-glucans in mattress dust were inversely related to a doctor's diagnosis of asthma-associated AE [aOR = 0.75 (0.57-0.98)], and endotoxin levels to current symptoms of asthma-associated AE [aOR = 0.73 (0.57-0.94)]. The analyses of the PARSIFAL study revealed two different phenotypes of AE, depending on the association with asthma and wheezing ever. With regard to the hygiene hypothesis, help with haying, exposure to beta(1-->3)-glucans and endotoxin were found to be inversely associated with the AE phenotype associated with asthma and wheezing.


Thermal-Conductivity-Enhancing Copper-Plated Expanded Graphite/Paraffin Composite for Highly Stable Phase-Change Materials.
Paraffin (PA)/expanded graphite (EG) is an important composite phase change material with low cost, high heat storage, good thermal conductivity and cycling stability. Its thermal conductivity needs to be further improved for application in the thermal management system of power lithium-ion batteries. In this paper, copper plated expanded graphite (CPEG) with 3D porous structure was prepared by electroless copper plating method, which was used as thermal conductivity enhancing material to replace part of EG in PA/EG composite materials. For the optimized phase change material composed of 80 %PA-14 %EG-6 %CPEG, the copper content is very low (0.768 wt %), but its thermal conductivity can be significantly improved without loss of latent heat and thermal cycling stability. Its thermal conductivity is increased from 11 times to 16.5 times that of paraffin while compared with the copper-free composite material (80 %PA-20 %EG). The PA/EG/CPEG composite material exhibits good temperature control effect on power lithium-ion batteries.


Pioglitazone, a PPARγ agonist, reduces nicotine craving in humans, with marginal effects on abuse potential.
Possibly through their actions upon glia, peroxisome proliferator-activated receptor agonists (PPAR) have been shown to alter the abuse potential of addictive drugs in several preclinical models. The current study extends this research into the human laboratory as the first clinical study into the effects of the PPAR gamma agonist, pioglitazone, on the abuse potential of nicotine. Heavy smokers were recruited for this 3-week study. Upon admission, participants were randomized to either active (45mg, n=14) or placebo (0mg, n=13) PIO maintenance conditions for the duration of the study. After 5-7days of stabilization on a 7mg nicotine patch, participants began laboratory testing. On the 1st-4th test days, participants could self-administer cigarettes or receive money by making verbal choices for either option. On the 5th day, participants were administered 10 puffs of their usual brand of cigarette in the morning and later chose between smoking and money by making finger presses on a computer mouse in a progressive ratio self-administration task. Later on the 5th day participants also underwent a smoking cue exposure session. The 8th-11th test days were identical to the 1st-4th test days with the exception that during one of the test weeks de-nicotinized cigarettes were available, and during the other nicotinized cigarettes were available. Nicotinized cigarettes were always administered on the 5th and 12th days. On some measures PIO increased indicators of abuse potential, though this effect was typically not statistically significant. However, PIO did significantly reduce measures of craving.


[Ultrastructure of L forms. IV. L forms of nonagglutinating vibrios].
A study was made of the ultrastructure of stable L-forms of Nag vibrios aged 24 hours. Cells of all types of the L-forms had cytoplasmic membranes, and a three-layered structure, which was found not everywhere. Externally of the cytoplasmic membrane, in some areas of the individual cells there were revealed a plastic layer of cell wall and a basal membrane. However, in difference to bacterial forms of the vibryos, rigidity of the cell wall was disturbed, and the links between the cell wall and the cytoplasmic membrane were indetectable. There were regularly revealed lamellar of myelin-like membranous structures in the cytoplasm, which did not occur in bacterial forms, and also lamellar mesosomes. The latter were found in the sites of cell division. Viability of small bodies as the minimal reproductive forms of the L-cultures is confirmed by the presence in them of a nucleoid and of the binary division.


Sleep assessments for a mild traumatic brain injury trial in a military population.
Baseline sleep characteristics were explored for 71 U.S. military service members with mild traumatic brain injury (mTBI) enrolled in a post-concussive syndrome clinical trial. The Pittsburgh Sleep Quality Index (PSQI), sleep diary, several disorder-specific questionnaires, actigraphy and polysomnographic nap were collected. Almost all (97%) reported ongoing sleep problems. The mean global PSQI score was 13.5 (SD=3.8) and 87% met insomnia criteria. Sleep maintenance efficiency was 79.1% for PSQI, 82.7% for sleep diary and 90.5% for actigraphy; total sleep time was 288, 302 and 400 minutes, respectively. There was no correlation between actigraphy and subjective questionnaires. Overall, 70% met hypersomnia conditions, 70% were at high risk for obstructive sleep apnea (OSA), 32% were symptomatic for restless legs syndrome, and 6% reported cataplexy. Nearly half (44%) reported coexisting insomnia, hypersomnia and high OSA risk. Participants with post-traumatic stress disorder (PTSD) had higher PSQI scores and increased OSA risk. Older participants and those with higher aggression, anxiety or depression also had increased OSA risk. The results confirm poor sleep quality in mTBI with insomnia, hypersomnia, and OSA risk higher than previously reported, and imply sleep disorders in mTBI may be underdiagnosed or exacerbated by comorbid PTSD.


Meaning of context in recapturing self-care after stroke or spinal cord injury.
This study identifies the meaning of context in recapturing self-care after a stroke or spinal cord injury (SCI). Recapturingdenotes the process of engaging in self-care to regain the ability to participate in self-care activities. Five people who had had a stroke and 6 people with SCI were interviewed 1-3 months after onset. The interviews were open ended and transcribed verbatim. They were analyzed by using the empirical, phenomenological, psychological method, which identified 6 main characteristics describing the role of context in recapturing self-care: (1) support from others, (2) an air of expectation, (3) extended time, (4) new daily structure, (5) therapeutic relationship as enabling possibility, and (6) gradual change in challenge. These findings showed that rehabilitation professionals play an important role in creating a context that contributes to recapturing self-care by allowing extended time, enabling patients to see possibilities, and creating expectations for them to do things on their own.


Studies on the sex-specific lethals of Drosophila melanogaster. II. Further studies on a male-specific lethal gene, maleless.
Effects of a second chromosome male-specific lethal gene, maleless (mle), of Drosophila melanogaster were further studied. It was shown that, although no maternal effect was seen with respect to the male-specific lethality, the lethal stage was influenced by whether parental females were homozygous or heterozygous for mle. Thus, in the former mle/mle males died mostly in the late third instar larval stage, while in the latter practically all males survived to the pupal stage. In the dying mle/mle male pupae complete differentiation of adult external head and thorax structures was often observed but that of abdominal structures was incomplete forming only a few segments in most cases. Imaginal discs from third instar mle/mle male larvae which were produced by mle/mle mothers and were destined to die as larvae were able to differentiate into adult structures upon transplantation into normal third instar larval hosts. A somewhat elaborated version of the previously presented hypothesis (FUKUNAGA, TANAKA and OISHI 1975) was discussed as to the possible presence of a class of sex-specific lethals which are not related to the process of primary sex differentiation.


Numerical analysis of blood flow through a stenosed artery using a coupled, multiscale simulation method.
A global system model of the systemic circulation is combined with a local finite element solution to simulate blood flow in a stenosed coronary artery. Local fluid dynamic issues arise in connection with the detailed flow patterns within the stenosed coronary artery while the global system model is used to simulate the response of the rest of the circulation to the local perturbation. A PISO type finite element technique is employed to compute the local blood flow. The Navier-Stokes equations are solved with the assumption of viscous incompressible flow across the stenosed coronary artery. A detailed lumped parameter model simulates the characteristics of the coronary circulation and is imbedded in a coarse-grained lumped parameter model of the entire cardiovascular system. These two methods are coupled in that the lumped parameter calculations provide the time-dependent boundary conditions for the local finite element calculation. In turn, the local fluid dynamical computation provides estimates for the pressure drop across the stenosis, which is subsequently used to refine the lumped parameter calculation. Results are obtained for an axisymmetric coronary artery model with a stenosis of 90% area reduction over one cardiac cycle. Numerical results show that the flow rate and resistance are strongly coupled. Compared with the flow rate distribution computed from the global simulation with constant resistance, the coupled solution predicts a flow rate with only slight changes. The high flow rate during diastole increases the stenosis pressure drop and resistance. In turn, this increased resistance of the stenosis slightly reduces the flow rate computed in the lumped parameter simulation.


Automated ultrasound edge-tracking software comparable to established semi-automated reference software for carotid intima-media thickness analysis.
Many commercial ultrasound systems are now including automated analysis packages for the determination of carotid intima-media thickness (cIMT); however, details regarding their algorithms and methodology are not published. Few studies have compared their accuracy and reliability with previously established automated software, and those that have were in asymptomatic adults. Therefore, this study compared cIMT measures from a fully automated ultrasound edge-tracking software (EchoPAC PC, Version 110.0.2; GE Medical Systems, Horten, Norway) to an established semi-automated reference software (Artery Measurement System (AMS) II, Version 1.141; Gothenburg, Sweden) in 30 healthy preschool children (ages 3-5 years) and 27 adults with coronary artery disease (CAD; ages 48-81 years). For both groups, Bland-Altman plots revealed good agreement with a negligible mean cIMT difference of -0·03 mm. Software differences were statistically, but not clinically, significant for preschool images (P = 0·001) and were not significant for CAD images (P = 0·09). Intra- and interoperator repeatability was high and comparable between software for preschool images (ICC, 0·90-0·96; CV, 1·3-2·5%), but slightly higher with the automated ultrasound than the semi-automated reference software for CAD images (ICC, 0·98-0·99; CV, 1·4-2·0% versus ICC, 0·84-0·89; CV, 5·6-6·8%). These findings suggest that the automated ultrasound software produces valid cIMT values in healthy preschool children and adults with CAD. Automated ultrasound software may be useful for ensuring consistency among multisite research initiatives or large cohort studies involving repeated cIMT measures, particularly in adults with documented CAD.


Rice cyclophilin OsCYP18-2 is translocated to the nucleus by an interaction with SKIP and enhances drought tolerance in rice and Arabidopsis.
Cyclophilin 18-2 (CYP18-2) genes, homologues of human peptidyl-prolyl isomerase-like 1 (PPiL1), are conserved across multicellular organisms and Schizosaccharomyces pombe. Although PPiL1 is known to interact with ski-interacting protein (SKIP), a transcriptional co-regulator and spliceosomal component, there have been no functional analyses of PPiL1 homologues in plants. Rice cyclophilin 18-2 (OsCYP18-2) bound directly to amino acids 56-95 of OsSKIP and its binding was independent of cyclosporin A, a cyclophilin-binding drug. Moreover, OsCYP18-2 exhibited PPIase activity regardless of its interaction with OsSKIP. Therefore, the binding site for OsCYP18-2's interaction with SKIP was distinct from the PPIase active site. OsCYP18-2's interaction with SKIP full-length protein enabled OsCYP18-2's translocation from the cytoplasm into the nucleus and AtSKIP interacted in planta with both AtCYP18-2 and OsCYP18-2. Drought and salt stress induced similar expression of OsCYP18-2 and OsSKIP. Overexpression of OsCYP18-2 in transgenic rice and Arabidopsis thaliana plants enhanced drought tolerance and altered expression and pre-mRNA splicing patterns of stress-related genes in Arabidopsis under drought conditions. Furthermore, OsCYP18-2 caused transcriptional activation with/without OsSKIP in the GAL4 system of yeast; thus the OsSKIP-OsCYP18-2 interaction has an important role in the transcriptional and post-transcriptional regulation of stress-related genes and increases tolerance to drought stress.


Tropical forests and the changing earth system.
Tropical forests are global epicentres of biodiversity and important modulators of the rate of climate change. Recent research on deforestation rates and ecological changes within intact forests, both areas of recent research and debate, are reviewed, and the implications for biodiversity (species loss) and climate change (via the global carbon cycle) addressed. Recent impacts have most likely been: (i) a large source of carbon to the atmosphere, and major loss of species, from deforestation and (ii) a large carbon sink within remaining intact forest, accompanied by accelerating forest dynamism and widespread biodiversity changes. Finally, I look to the future, suggesting that the current carbon sink in intact forests is unlikely to continue, and that the tropical forest biome may even become a large net source of carbon, via one or more of four plausible routes: changing photosynthesis and respiration rates, biodiversity changes in intact forest, widespread forest collapse via drought, and widespread forest collapse via fire. Each of these scenarios risks potentially dangerous positive feedbacks with the climate system that could dramatically accelerate and intensify climate change. Given that continued land-use change alone is already thought to be causing the sixth mass extinction event in Earth's history, should such feedbacks occur, the resulting biodiversity and societal consequences would be even more severe.


Disorders of consciousness: differential diagnosis and neuropathologic features.
Disorders of consciousness present intriguing challenges to the neurologist and neurorehabilitation specialist. Assessment is constrained by the lack of reliable methods of assessing consciousness, and there are no treatment interventions known to influence the course of recovery from these conditions. In addition, the relationship between the clinical features associated with these disorders and their corresponding pathophysiologic substrate is also unclear. Our understanding of disorders of consciousness has not kept pace with the advances in neurosurgical management that have decreased mortality following severe injury. There is still considerable confusion regarding differential diagnosis and prognostication concerning states of severely altered consciousness. The purpose of this article is to discuss the content and neural basis of consciousness and to review the terminology most often used to describe altered states of consciousness. The neurobehavioral criteria for differentiating among specific syndromes associated with severe alterations in consciousness are presented. Representative case studies are utilized to illustrate the characteristic clinical profiles of coma, vegetative state, persistent and permanent vegetative state, minimally conscious state, akinetic mutism, and locked-in syndrome. Areas of ambiguity and controversy are emphasized and future directions for research are suggested.


Multimodality Neuromonitoring in Adult Traumatic Brain Injury: A Narrative Review.
Neuromonitoring plays an important role in the management of traumatic brain injury. Simultaneous assessment of cerebral hemodynamics, oxygenation, and metabolism allows an individualized approach to patient management in which therapeutic interventions intended to prevent or minimize secondary brain injury are guided by monitored changes in physiologic variables rather than generic thresholds. This narrative review describes various neuromonitoring techniques that can be used to guide the management of patients with traumatic brain injury and examines the latest evidence and expert consensus guidelines for neuromonitoring.


Guanosine-induced decrease in side population of lung cancer cells: lack of correlation with ABCG2 expression.
Cancers contain a 'side population' (SP), a subset of cells that is greatly enriched in stem cells and which contains malignant progenitors. SP cells are characterised by high efflux capability for Hoechst 33342 dye and for anti-cancer therapeutic agents through transporters; ABCG2 (ATP-binding cassette transporter G2) is currently most closely associated with the SP phenotype. Guanosine is an important intercellular signalling molecule; it stimulates stem cell proliferation in vivo and affects cholesterol efflux in vitro through activation of ABCG transporter (ABCG1), raising the possibility that it might also affect ABCG2 and hence the SP. We examined the effects of guanosine on the SP of A549 lung cancer cells. Fluorescence-activated cell sorting (FACS) revealed that exposure to 10 microM guanosine significantly decreased the proportion of SP cells after 48 hours but not after 6 hours. In contrast, Western blot analysis showed that 10 microM guanosine significantly decreased ABCG2 expression after 6 hours, but not after 48 hours. These data demonstrate that guanosine affects both the proportion of SP cells and ABCG2 transporters, but the lack of correlation between ABCG2 expression and the SP phenotype indicates that transporters other than ABCG2 are involved in maintaining the SP phenotype in A549 lung cancer cells.


Deamidation in human gamma S-crystallin from cataractous lenses is influenced by surface exposure.
A major component of human nuclear cataracts is water-insoluble, high molecular weight protein. A significant component of this protein is disulfide bonded gamma S-crystallin that can be reduced to monomers by dithiothreitol. Analysis of this reduced gamma S-crystallin showed that deamidation of glutamine and asparagine residues is a principal modification. Deamidation is one of the modifications of lens crystallins associated with aging and cataractogenesis. One proposed hypothesis of cataractogenesis is that it develops in response to altered surface charges that cause conformational changes, which, in turn, permit formation of disulfide bonds and crystallin insolubility. This report, showing deamidation among the disulfide bonded gamma S-crystallins from cataractous lenses, supports this hypothesis.


The standard of care for lymphedema: current concepts and physiological considerations.
The standard of care for lymphedema encompasses risk reduction, early detection, Phase-I Complete Decongestive Therapy in a clinical setting, and Phase-II Self Management at home. This review discusses these lymphedema treatment components, including relevant physiological aspects that form the basis of treatment and an understanding of which are critical to successful outcomes. Phase-I therapy includes manual lymphatic drainage (MLD), short-stretch compression bandaging, decongestive exercise, skin care, and sometimes intermittent pneumatic compression (IPC). Home management includes many of the same elements, with adaptations for home treatment. Significant to the success of Phase-I and Phase-II care is effective bandaging and lymphatic drainage treatment via manual (MLD) or automated (IPC) processes. Truncal clearance plays a key role in facilitating lymphatic drainage from affected limbs by achieving effective pressure gradients, reducing lymphatic network resistance, and stimulating lymphatic contractility. The transition from Phase-I to Phase-2 presents challenges to patients and clinicians. Poor patient compliance with time-consuming technique dependent-home care regimens contribute to losing gains achieved in the clinic. Ineffective treatment results in complications, including increased pain, reduced range of motion and mobility, increased risk of infection, and other physical and psychological sequelae. The use of an advanced programmable IPC device for lymphedema treatment may support successful home management by addressing some of the home treatment barriers. Device selection requires careful consideration of the mode of action and device functionality. An advanced programmable device that provides truncal and proximal clearance and low applied pressure represents the current choice for such in-home devices.


Apparent increase in the thickness of superconducting particles at low temperatures measured by electron holography.
We predict that superconducting particles will show an apparent increase in thickness at low temperatures when measured by electron holography. This will result not from a real thickness increase, rather from an increase in the mean inner potential sensed by the electron wave traveling through the particle, originating in expansion of the electronic wavefunction of the superconducting electrons and resulting negative charge expulsion from the interior to the surface of the superconductor, giving rise to an increase in the phase shift of the electron wavefront going through the sample relative to the wavefront going through vacuum. The temperature dependence of the observed phase shifts will yield valuable new information on the physics of the superconducting state of metals.


Khellinone derivatives as blockers of the voltage-gated potassium channel Kv1.3: synthesis and immunosuppressive activity.
The voltage-gated potassium channel Kv1.3 constitutes a promising new target for the treatment of T-cell-mediated autoimmune diseases such as multiple sclerosis. In this study, we report the discovery of two new classes of Kv1.3 blockers based on the naturally occurring compound khellinone, 5-acetyl-4,7-dimethoxy-6-hydroxybenzofuran: (1) khellinone dimers linked via the alkylation of the 6-hydroxy groups and (2) chalcone derivatives of khellinone formed by Claisen-Schmidt condensation of the 5-acetyl group with aryl aldehydes. In particular, the chalcone 3-(4,7-dimethoxy-6-hydroxybenzofuran-5-yl)-1-phenyl-3-oxopropene (16) and several of its derivatives inhibited Kv1.3 with K(d) values of 300-800 nM and a Hill coefficient of 2, displayed moderate selectivity over other Kv1-family K(+) channels, suppressed T-lymphocyte proliferation at submicromolar concentrations, and showed no signs of acute toxicity in mice. Because of their relatively low molecular weight and lipophilicity and their high affinity to Kv1.3, aryl-substituted khellinone derivatives represent attractive lead compounds for the development of more potent and selective Kv1.3 blocking immunosuppressants.


Recipes and mechanisms of cellular reprogramming: a case study on budding yeast Saccharomyces cerevisiae.
Generation of induced pluripotent stem cells (iPSCs) and converting one cell type to another (transdifferentiation) by manipulating the expression of a small number of genes highlight the progress of cellular reprogramming, which holds great promise for regenerative medicine. A key challenge is to find the recipes of perturbing genes to achieve successful reprogramming such that the reprogrammed cells function in the same way as the natural cells. We present here a systems biology approach that allows systematic search for effective reprogramming recipes and monitoring the reprogramming progress to uncover the underlying mechanisms. Using budding yeast as a model system, we have curated a genetic network regulating cell cycle and sporulation. Phenotypic consequences of perturbations can be predicted from the network without any prior knowledge, which makes it possible to computationally reprogram cell fate. As the heterogeneity of natural cells is important in many biological processes, we find that the extent of this heterogeneity restored by the reprogrammed cells varies significantly upon reprogramming recipes. The heterogeneity difference between the reprogrammed and natural cells may have functional consequences. Our study reveals that cellular reprogramming can be achieved by many different perturbations and the reprogrammability of a cell depends on the heterogeneity of the original cell state. We provide a general framework that can help discover new recipes for cellular reprogramming in human.


The outcome of the functioning filter after subsequent cataract extraction.
To evaluate and compare the outcome of functioning filtration surgery followed by cataract surgery with posterior intraocular lens implantation by both phacoemulsification and extracapsular cataract extraction (ECCE) techniques in glaucomatous eyes. We retrospectively evaluated the clinical course of 77 eyes (68 patients) that after successful trabeculectomy, underwent cataract surgery by either phacoemulsification or ECCE techniques. We determined the frequency of partial and absolute failure following cataract surgery by either phacoemulsification or ECCE in eyes with functioning trabeculectomies. Partial failure of intraocular pressure (IOP), control after cataract extraction was defined as the need for an increased number of antiglaucoma medications or argon laser trabeculoplasty to maintain IOP < or =21mm Hg. Complete failure of IOP control after cataract surgery was defined as an IOP >21 mm Hg on at least two consecutive measurements one or more weeks apart or the performance of additional filtration surgery. Failure rates were calculated using the Kaplan-Meier actuarial method. Failure rates between phacoemulsification and ECCE subgroups were compared using the log rank test. The probability of partial failure by the third postoperative year after cataract surgery was 39.5% in the phacoemulsification subgroup and 37.3% in the ECCE subgroup. This small difference is not statistically significant (P = 0.48). The probability of complete failure by the fourth postoperative year after cataract surgery was 12.0% in the phacoemulsification subgroup and 12.5% in the ECCE subgroup. This difference is also not statistically significant (P = 0.77). At the 6-month follow-up visit, visual acuity of both groups improved one or more lines in 87.0% of patients, and worsened one or more lines in 3.9% of patients. Sixty-one percent achieved visual acuity of 20/40 or better. The most frequent complication was posterior capsular opacification requiring laser capsulotomy that occurred in 31.2% of patients. Cataract extraction by either phacoemulsification or ECCE following trabeculectomy surgery may be associated with a partial loss of the previously functioning filter and the need for more antiglaucoma medications to control IOP.


Underlying Skills of Oral and Silent Reading Fluency in Chinese: Perspective of Visual Rapid Processing.
Reading fluency is a critical skill to improve the quality of our daily life and working efficiency. The majority of previous studies focused on oral reading fluency rather than silent reading fluency, which is a much more dominant reading mode that is used in middle and high school and for leisure reading. It is still unclear whether the oral and silent reading fluency involved the same underlying skills. To address this issue, the present study examined the relationship between the visual rapid processing and Chinese reading fluency in different modes. Fifty-eight undergraduate students took part in the experiment. The phantom contour paradigm and the visual 1-back task were adopted to measure the visual rapid temporal and simultaneous processing respectively. These two tasks reflected the temporal and spatial dimensions of visual rapid processing separately. We recorded the temporal threshold in the phantom contour task, as well as reaction time and accuracy in the visual 1-back task. Reading fluency was measured in both single-character and sentence levels. Fluent reading of single characters was assessed with a paper-and-pencil lexical decision task, and a sentence verification task was developed to examine reading fluency on a sentence level. The reading fluency test in each level was conducted twice (i.e., oral reading and silent reading). Reading speed and accuracy were recorded. The correlation analysis showed that the temporal threshold in the phantom contour task did not correlate with the scores of the reading fluency tests. Although, the reaction time in visual 1-back task correlated with the reading speed of both oral and silent reading fluency, the comparison of the correlation coefficients revealed a closer relationship between the visual rapid simultaneous processing and silent reading. Furthermore, the visual rapid simultaneous processing exhibited a significant contribution to reading fluency in silent mode but not in oral reading mode. These findings suggest that the underlying mechanism between oral and silent reading fluency is different at the beginning of the basic visual coding. The current results also might reveal a potential modulation of the language characteristics of Chinese on the relationship between visual rapid processing and reading fluency.


An electromyographic assessment pilot study on the reliability of the forearm muscles during multi-planar maximum voluntary contraction grip and wrist articulation in young males.
Electromyographic systems are widely used in scientific and clinical practice. The reproducibility and reliability of these measures are crucial when conducting scientific research and collecting experimental data. To test the reliability of surface electromyography signals from both the Flexor Digitorum Superficialis (FDS) and Extensor Carpi Radialis Brevis (ECRB) muscles of both the left and right arms during an individual, static multi-planar maximum voluntary contraction handgrip task using the Myon 320 system (Myon AG, Switzerland). Eight right-handed male participants performed two maximal handgrip tests in five separate wrist positions using both hands. Muscle activity was recorded from both forearms. Reliability was measured using the Standard Error of Measurement (SEM), Coefficient of Variation (CV) and Intra-class correlation coefficients. Wrist joint position correlations within and between the FDS and ECRB muscle activities were also analysed. Absolute reliability was shown across all positions for both hands with CV and SEM recorded at below 10%. The output measures indicate that the Myon 320 system (Myon AG, Switzerland) produces good to fair reliability when assessing forearm muscle activity. Correlations in the left FDS muscles were negative. Correlations between the left ECRB and left FDS muscles were variable but positive between the right ECRB and right FDS muscles. The data sets retrieved from all participants were reliably evaluated. Wrist position correlations within and between the FDS and ECRB muscles may have been influenced by hand dominance. The findings demonstrate that the methods and systems outlined in this study can be used reliably in future research.


Plain and metrizamide CT of lumbar disk disease: comparison with myelography.
Computed tomography with and without administration of intrathecal contrast material was evaluated in 100 patients with suspected lumbar disk disease. Metrizamide computed tomography was performed in 75 patients and plain computed tomography was performed in 25. Metrizamide computed tomography was more accurate than plain computed tomography. It also disclosed many lesions not shown by metrizamide myelography at the lumbosacral level. Very few lesions were revealed by myelography that were not seen by metrizamide computed tomography. The small amount of intrathecal metrizamide needed for scanning has practically no side effects. Experience in 12 patients indicates that the procedure may be performed safely on an outpatient basis. This study suggests that computed tomography should be given serious consideration as the primary definitive radiographic examination of suspected lumbar disk disease.


Colloidal polarization of yolk/shell particles by reconfiguration of inner cores responsive to an external magnetic field.
Yolk/shell particles, which were hollow silica particles containing a movable magnetic silica core (MSC), were prepared by removing a middle polystyrene layer from multilayered particles of MSC/polystyrene/silica shell with heat treatment followed by a slight etching with a basic solution. An ac electric field was applied to the suspension of the yolk/shell particles to form pearl chains (1D structure) of yolk/shell particles. Observation with an optical microscope showed that the MSCs in the silica compartment of the pearl chains had a zigzag structure under the electric field. An external magnetic field applied to the suspension could form a novel structure of doublet MSC in the shell compartment of the quasi-pearl chain structure. Application of a magnetic field was also performed for 2D hexagonally close-packed assemblies of the yolk/shell particles, which could two-dimensionally form a doublet structure of MSCs as if they were polarized in the compartment. Switching on/off the magnetic field successfully controlled the positional ordering of cores in the consolidated silica shell.


Racial and ethnic differences in health care access and health outcomes for adults with type 2 diabetes.
To evaluate health care access and utilization and health status and outcomes for type 2 diabetic patients according to race and ethnicity and to determine whether health status is influenced by health care access and utilization. National samples of Caucasians, African-Americans, and Mexican-Americans were studied in the third National Health and Nutrition Examination Survey. Information on medical history and treatment of diabetes, health care access and utilization, and health status and outcomes was obtained by structured questionnaires and by clinical and laboratory assessments. Almost all patients in each race and ethnic group had one primary source of ambulatory medical care (92-97%), saw one physician at this source (83-92%), and had at least semiannual physician visits (83-90%). Almost all patients > or = 65 years of age had health insurance (99-100%), and for those patients < 65 years of age, Caucasians (91%) and African-Americans (89%) had higher rates of coverage than Mexican-Americans (66%). Rates of treatment with insulin or oral agents (71-78%), eye examination in the previous year (61-70%), blood pressure check in the previous 6 months (83-89%), and the proportion of hypertension that was diagnosed (84-91%) were similar for each race and ethnic group. Lower proportions of African-Americans and Mexican-Americans self-monitored their blood glucose (insulin-treated, 27 vs. 44% of Caucasians), had their cholesterol checked (62-68 vs. 81%), and had their dyslipidemia diagnosed (45 vs. 58%). African-American and Mexican-American patients had a somewhat higher proportion than Caucasian patients, with HbA1c > or = 7% (58-66 vs. 55%), blood pressure > or = 140/90 mmHg among those with diagnosed hypertension (60-65 vs. 55%), and clinical proteinuria (11-14 vs. 5%). In contrast, they had better levels of total cholesterol (> or = 240 mg/dl) (28 -30 vs. 34%) and HDL cholesterol (> or = 45 mg/dl) (46 -59 vs. 38%), and African-American and Mexican-American men were less overweight than Caucasian men (BMI > or = 30) (34-37 vs. 44%), although the opposite was true for women. LDL cholesterol levels and the proportion of patients who smoked cigarettes or were hospitalized in the past year were similar among all three groups. In logistic regression analysis, there was little evidence that levels of blood glucose, blood pressure, lipids, or albuminuria were associated with access to or utilization of health care or with socioeconomic status. There are some differences by race and ethnicity in health care access and utilization and in health status and outcomes for adults with type 2 diabetes. However, the magnitude of these differences pale in comparison with the suboptimal health status of all three race and ethnic groups relative to established treatment goals. Health status does not appear to be influenced by access to health care.


A community-based, environmental chronic disease prevention intervention to improve healthy eating psychosocial factors and behaviors in indigenous populations in the Canadian Arctic.
Diet-related chronic diseases are highly prevalent among indigenous populations in the Canadian Arctic. A community-based, multi-institutional nutritional and lifestyle intervention-Healthy Foods North-was implemented to improve food-related psychosocial factors and behaviors among Inuit and Inuvialuit in four intervention communities (with two comparison communities) in Nunavut and the Northwest Territories, Canada, in 2008. The 12-month program was developed from theory (social cognitive theory and social ecological models), formative research, and a community participatory process. It included an environmental component to increase healthy food availability in local stores and activities consisting of community-wide and point-of-purchase interactive educational taste tests and cooking demonstrations, media (e.g., radio ads, posters, shelf labels), and events held in multiple venues, including recreation centers and schools. The intervention was evaluated using pre- and postassessments with 246 adults from intervention and 133 from comparison communities (311 women, 68 men; mean age 42.4 years; 78.3% retention rate). Outcomes included psychosocial constructs (healthy eating knowledge, self-efficacy, and behavioral intentions), frequency of healthy and unhealthy food acquisition, healthiness of commonly used food preparation methods, and body mass index (kg/m(2)). After adjustment for demographic, socioeconomic status, and body mass index variables, respondents living in intervention communities showed significant improvements in food-related self-efficacy (β = 0.15, p = .003) and intentions (β = 0.16, p = .001) compared with comparison communities. More improvements from the intervention were seen in overweight, obese, and high socioeconomic status respondents. A community-based, multilevel intervention is an effective strategy to improve psychosocial factors for healthy nutritional behavior change to reduce chronic disease in indigenous Arctic populations.


Inorganic phosphate inhibits sympathetic neurotransmission in canine saphenous veins.
Inorganic phosphate has been proposed as the initiator of metabolic vasodilatation in active skeletal muscle. The present study was primarily designed to determine if this substance has an inhibitory effect on adrenergic neurotransmission. Rings of canine saphenous veins were suspended for isometric tension recording in organ chambers. A comparison was made of the ability of inorganic phosphate (3 to 14 mM) to relax rings contracted to the same degree by electrical stimulation, exogenous norepinephrine, and prostaglandin F2 alpha. The relaxation during electrical stimulation was significantly greater at all concentrations of phosphate. In strips of saphenous veins previously incubated with [3H]norepinephrine, the depression of the contractile response caused by phosphate during electrical stimulation was accompanied by a significant reduction in the overflow of labeled neurotransmitter. Thus inorganic phosphate inhibits sympathetic neurotransmission and hence may have a key role in the sympatholysis in the active skeletal muscles during exercise. By contrast, in this preparation, it has a modest direct relaxing action on the vascular smooth muscle.


The Ligand Gated Ion Channel Database.
The ligand gated ion channels (LGICs) are ionotropic receptors to neurotransmitters. Their physiological effect is carried out by the opening of an ionic channel upon binding of a particular neurotransmitter. These LGICs constitute superfamilies of receptors formed by homologous subunits. A database has been developed to handle the growing wealth of cloned subunits. This database contains nucleic acid sequences, protein sequences, as well as multiple sequence alignments and phylogenetic studies. This database is accessible via the worldwide web (http://www.pasteur.fr/units/neubiomol/LGIC.h tml), where it is continuously updated. A downloadable version is also available [currently v0.1 (98.06)].


Teachers' Perspectives on Preventing Suicide in Children and Adolescents in Schools: A Qualitative Study.
Given the important role teachers play as gatekeepers in school suicide prevention, this study explored teachers' perspectives on what should be done to improve current suicide prevention efforts. The study, in Queensland, Australia, was part of a large-scale survey examining teachers' knowledge, attitudes and experience of suicidality. One hundred and fifteen teachers responded to an online survey question regarding their views on the requirements for school suicide prevention. Qualitative analysis identified five themes from teachers' responses: awareness and stigma reduction, support services for students, education and training, bullying and the role of social media. The results of this study provide some profound insights into teachers' perspectives on suicide and highlight the critical need for improved suicide prevention efforts in schools.


Performance evaluation of reverse osmosis technology for selected antibiotics removal from synthetic pharmaceutical wastewater.
This study addresses the possibility for low pressure reverse osmosis membrane (RE 2521, CSM) process to serve as an alternative to remove selected antibiotics (ampicillin and amoxicillin) from synthetic wastewater by changing operating conditions such as pH = 3, 6.5 and 10; Pressure = 9, 11 and13 (bar); antibiotic concentration = 10, 255 and 500(mg/L), and temperature = 20, 30 and 40°C. The experiment was designed based on Box-benken, which is a Response Surface methodology design (RSM), using Design Expert software. The concentration of antibiotics was measured by applying a UV-spectrophotometer (Cecil), at the wavelength of 254 nm. Results showed a range of rejection percentage from 73.52% to 99.36% and 75.1% to 98.8%, for amoxicillin and ampicillin, respectively. Considering the solute rejections and the membrane porosity show that the prevailing rejection mechanism of the examined antibiotics by the membrane was the size exclusion effect. The permeate flux for both of the antibiotics was 12-18.73 L/m2.h. Although the permeate flux and antibiotic rejection are influenced by operating pressure, pH, and temperature individually, the interaction between operating parameters did not have noticeable effects. According to the results obtained in this study, the application of RO membrane is recommended for the selected antibiotics to be removed to a considerable degree (up to 95%).


Antiviral Nucleotide Incorporation by Recombinant Human Mitochondrial RNA Polymerase Is Predictive of Increased In Vivo Mitochondrial Toxicity Risk.
Nucleoside or nucleotide inhibitors are a highly successful class of antivirals due to selectivity, potency, broad coverage, and high barrier to resistance. Nucleosides are the backbone of combination treatments for HIV, hepatitis B virus, and, since the FDA approval of sofosbuvir in 2013, also for hepatitis C virus (HCV). However, many promising nucleotide inhibitors have advanced to clinical trials only to be terminated due to unexpected toxicity. Here we describe the in vitro pharmacology of compound 1, a monophosphate prodrug of a 2'-ethynyluridine developed for the treatment of HCV. Compound 1 inhibits multiple HCV genotypes in vitro (50% effective concentration [EC50], 0.05 to 0.1 μM) with a selectivity index of >300 (50% cytotoxic concentration [CC50], 30 μM in MT-4 cells). The active triphosphate metabolite of compound 1, compound 2, does not inhibit human α, β, or γ DNA polymerases but was a substrate for incorporation by the human mitochondrial RNA polymerase (POLRMT). In dog, the oral administration of compound 1 resulted in elevated serum liver enzymes and microscopic changes in the liver. Transmission electron microscopy showed significant mitochondrial swelling and lipid accumulation in hepatocytes. Gene expression analysis revealed dose-proportional gene signature changes linked to loss of hepatic function and increased mitochondrial dysfunction. The potential of in vivo toxicity through mitochondrial polymerase incorporation by nucleoside analogs has been previously shown. This study shows that even moderate levels of nucleotide analog incorporation by POLRMT increase the risk of in vivo mitochondrial dysfunction. Based on these results, further development of compound 1 as an anti-HCV compound was terminated.


Fate and availability of glyphosate and AMPA in agricultural soil.
The fate of glyphosate and its degradation product aminomethylphosphonic acid (AMPA) was studied in soil. Labeled glyphosate was used to be able to distinguish the measured quantities of glyphosate and AMPA from the background values since the soil was sampled in a field where glyphosate had been used formerly. After addition of labeled glyphosate, the disappearance of glyphosate and the formation and disappearance of AMPA were monitored. The resulting curves were fitted according to a new EU guideline. The best fit of the glyphosate degradation data was obtained using a first-order multi compartment (FOMC) model. DT(50) values of 9 days (glyphosate) and 32 days (AMPA) indicated relatively rapid degradation. After an aging period of 6 months, the leaching risk of each residue was determined by treating the soil with pure water or a phosphate solution (pH 6), to simulate rain over a non-fertilized or fertilized field, respectively. Significantly larger (p < 0.05) amounts of aged glyphosate and AMPA were extracted from the soil when phosphate solution was used as an extraction agent, compared with pure water. This indicates that the risk of leaching of aged glyphosate and AMPA residues from soil is greater in fertilized soil. The blank soil, to which 252 g glyphosate/ha was applied 21 months before this study, contained 0.81 ng glyphosate/g dry soil and 10.46 ng AMPA/g dry soil at the start of the study. Blank soil samples were used as controls without glyphosate addition. After incubation of the blank soil samples for 6 months, a significantly larger amount of AMPA was extracted from the soil treated with phosphate solution than from that treated with pure water. To determine the degree of uptake of aged glyphosate residues by crops growing in the soil, (14)C-labeled glyphosate was applied to soil 6.5 months prior to sowing rape and barley seeds. After 41 days, 0.006 +/- 0.002% and 0.005 +/- 0.001% of the applied radioactivity was measured in rape and barley, respectively.


A comparison of 48-week treatment efficacy between clevudine and entecavir in treatment-naïve patients with chronic hepatitis B.
Clevudine and entecavir are currently available in Korea as antiviral drugs against chronic hepatitis B (CHB). We aimed to compare the efficacy of clevudine and entecavir therapy. Treatment-naïve CHB patients who received 30 mg of clevudine or 0.5 mg of entecavir a day were analyzed. Mean reduction of hepatitis B virus (HBV) DNA levels, complete virological response (cVR, undetectable HBV DNA by real-time PCR), biochemical response (recovery to normal ALT level), and hepatitis B e antigen (HBeAg) seroconversion rate at the 48th week of treatment were assessed. A number of 59 patients in clevudine group and 61 patients in entecavir group were included. Mean HBV DNA reductions from baseline were similar in the clevudine and entecavir groups, -6.4 versus -6.8 log(10) copies/mL in HBeAg-positive (p = 0.417) and -6.9 versus -7.0 log(10) copies/mL in HBeAg-negative patients (p = 0.640). The proportion of patients who achieved cVR was not different between the two groups, 53 versus 55% in HBeAg-positive (p = 1.000) and 100 versus 95% in HBeAg-negative patients (p = 0.452). Biochemical response rates and HBeAg seroconversion rates were also similar in both the groups. Two (3.4%) patients in clevudine group showed virologic breakthrough with rtM204I mutation using direct sequencing analysis. Clinical myopathy occurred in two (3.4%) patients in clevudine group. Mean reduction of viral loads was similar between clevudine and entecavir groups during 48 weeks. However, virologic breakthrough and significant myopathy were noted only in clevudine-treated patients. Therefore, more attention should be paid to patients receiving clevudine.