The clinical development process for a novel preventive vaccine: An overview.
Each novel vaccine candidate needs to be evaluated for safety, immunogenicity, and protective efficacy in humans before it is licensed for use. After initial safety evaluation in healthy adults, each vaccine candidate follows a unique development path. This article on clinical development gives an overview on the development path based on the expectations of various guidelines issued by the World Health Organization (WHO), the European Medicines Agency (EMA), and the United States Food and Drug Administration (USFDA). The manuscript describes the objectives, study populations, study designs, study site, and outcome(s) of each phase (Phase I-III) of a clinical trial. Examples from the clinical development of a malaria vaccine candidate, a rotavirus vaccine, and two vaccines approved for human papillomavirus (HPV) have also been discussed. The article also tabulates relevant guidelines, which can be referred to while drafting the development path of a novel vaccine candidate.


Effects of Clinical Exposure to Osteopathic Manipulative Medicine on Confidence Levels of Medical Students.
Training in osteopathic manipulative medicine (OMM) is a unique component of the osteopathic medical school curriculum. Indicators of successful OMM programming include student comfort in explaining and performing OMM as well as confidence in using OMM on future patients. Research on the amount of clinical exposure sufficient to achieve this goal is limited. To gauge the impact of clinical OMM exposure on medical students' self-assessed understanding of OMM, their ability to discuss, explain, and perform OMM, and their plan to use OMM in their future practice. Fourth-year osteopathic medical students were e-mailed surveys before (baseline), during, and after 4 weeks (postrotation) of an elective OMM rotation. Answers were scaled from 0 to 10, with 0 being not at all comfortable/confident and 10 being the most comfortable/confident. Thirty-five students participated in the survey. A significant mean (SD) increase was found between the baseline and postrotation scores for students' understanding of OMM principles (1.43 [0.51]; P<.001), comfort discussing OMM principles with patients (1.27 [0.88]; P<.001), comfort with explaining OMM to someone unfamiliar with it (1.32 [0.82]; P<.001), comfort with performing an osteopathic structural examination (2.23 [1.44]; P<.001), and confidence incorporating OMM into future practice (1.86 [0.47]; P<.001). Increased clinical exposure to OMM increased the confidence of osteopathic medical students in all dimensions surveyed. This observation can help guide the development of undergraduate osteopathic clinical programming as well as standards for entry of allopathic residents into ACGME programs with osteopathic designation.


Predictive ability of fasting plasma glucose for a diabetic 2-h postload glucose value in oral glucose tolerance test: spectrum effect.
The performance of diagnostic tests may vary according to patient characteristics. The aim of this study is to find out the factors, if any, that may affect the performance of fasting plasma glucose (FPG) to predict a diabetic 2-h postload glucose level (> or =200 mg/dl) in oral glucose tolerance test (OGTT). One hundred ninety-six patients with known risk factors for diabetes mellitus to whom OGTT was applied were included. Factors that may have an effect on the performance of FPG in prediction of a diabetic value in OGTT were determined by using logistic regression and likelihood ratios (LRs). The cutoff of FPG predicting a 2-h postload glucose of > or =200 mg/dl was calculated by receiver operating characteristic curve as 110 mg/dl (sensitivity, 76.7%; specificity, 75.9%). Waist-to-hip ratio (WHR) and body mass index (BMI) influenced sensitivity, whereas age, family history, and presence of hyperlipidemia affected specificity of FPG. Significant factors for positive LR were age and hyperlipidemia, whereas sex, smoking, hyperlipidemia, physical inactivity, WHR, and BMI influenced negative LR. Fasting plasma glucose performance as a diagnostic test can be affected by many factors that are clearly stated as risk factors for diabetes mellitus. These data emphasize how the interpretation of a diagnostic test varies as the patient characteristics vary; the criteria that we confidently rely on may not be that reliable, changing between just two different patients.


Preacinetobactin not acinetobactin is essential for iron uptake by the BauA transporter of the pathogen Acinetobacter baumannii.
New strategies are urgently required to develop antibiotics. The siderophore uptake system has attracted considerable attention, but rational design of siderophore antibiotic conjugates requires knowledge of recognition by the cognate outer-membrane transporter. Acinetobacter baumannii is a serious pathogen, which utilizes (pre)acinetobactin to scavenge iron from the host. We report the structure of the (pre)acinetobactin transporter BauA bound to the siderophore, identifying the structural determinants of recognition. Detailed biophysical analysis confirms that BauA recognises preacinetobactin. We show that acinetobactin is not recognised by the protein, thus preacinetobactin is essential for iron uptake. The structure shows and NMR confirms that under physiological conditions, a molecule of acinetobactin will bind to two free coordination sites on the iron preacinetobactin complex. The ability to recognise a heterotrimeric iron-preacinetobactin-acinetobactin complex may rationalize contradictory reports in the literature. These results open new avenues for the design of novel antibiotic conjugates (trojan horse) antibiotics.


A synthetic peptide induces long-term protection from lethal infection with herpes simplex virus 2.
Immunization against viral pathogens is generally directed toward the induction of virus neutralizing antibody (VNA) and the maintenance of the potential for a second-set (IgG) response. Indeed, an elevated level of specific antibody is considered a reliable clinical indicator that a state of immunity exists in the host. However, in the case of herpes simplex virus (HSV), the presence of circulating VNA does not necessarily correlate with protection. Thus, it has been found that secondary infections occur in individuals even with high neutralizing titers to HSV, suggesting that antibody to the virus may be useless or even deleterious. In consideration of these facts, we were interested in inducing a T cell response to HSV. We had already shown that synthetic peptides corresponding to the NH3-terminal region of the glycoprotein D (gD) molecule of HSV could induce a strong T cell response when injected into mice, but did not, by themselves, confer protection. In this report, we examined the ability of peptides, covalently coupled to palmitic acid and incorporated into liposomes, to induce virus-specific T cell responses that confer protection against a lethal challenge of HSV-2. We have demonstrated that long-term protective immunity is achieved with a single immunization in the absence of neutralizing antibody when antigen is presented in this form. Furthermore, T cells but not serum from such immune mice can adoptively transfer this protection.


Efficacy and safety study of docetaxel as salvage chemotherapy in metastatic gastric cancer failing fluoropyrimidine and platinum combination chemotherapy.
Fluoropyrimidine (F) and platinum (P) combination chemotherapy has been widely used for the first line treatment of advanced gastric cancer (AGC). Docetaxel (D) has shown promising activity in this disease. The present study retrospectively investigated the efficacy of D monotherapy as salvage chemotherapy for AGC that is failing F and P combination chemotherapy. A total of 34 patients, fitting the eligibility criteria, were included in this study. D was administered at a dose of 75 mg/m(2) IV every 3 weeks, with dexamethasone prophylaxis. Twenty-nine patients had measurable lesions. The median treatment-free interval was 38.5 days, and 91.2% of patients had progressed within 4 months of withdrawal of the first line chemotherapy. A total of 133 cycles of D were administered, with a median of 3.5 (1 approximately 8) cycles. From an intention-to-treat analysis, 6 patients achieved partial responses (PR), with a response rate of 20.7% (95% CI, 6.0~35.4). The duration of objective PRs in these six were 2.3+, 2.5+, 2.9, 3.0+, 6.2 and 6.8 months, respectively. Six patients showed a stable disease, but 15 showed progression. The median time to progression was 4.2 months (95% CI, 2.8 approximately 5.5), with a median overall survival since the start of D monotherapy of 8.4 months (95% CI, 5.5 approximately 11.3). Grade 3/4 neutropenia and febrile neutropenia occurred in 12.9% of patients and 3.1% of cycles. The incidence of grade 3 or worse non-hematological toxicities were as follows; peripheral sensory neuropathy 9.7%, asthenia 3.2% and allergic reaction 2.7%. Docetaxel, 75 mg/m(2), is active in AGC as second-line chemotherapy after failure of prior exposure to the F and P combination chemotherapy, with a favorable toxicity profile.


High-resolution myogenic lineage mapping by single-cell mass cytometry.
Muscle regeneration is a dynamic process during which cell state and identity change over time. A major roadblock has been a lack of tools to resolve a myogenic progression in vivo. Here we capitalize on a transformative technology, single-cell mass cytometry (CyTOF), to identify in vivo skeletal muscle stem cell and previously unrecognized progenitor populations that precede differentiation. We discovered two cell surface markers, CD9 and CD104, whose combined expression enabled in vivo identification and prospective isolation of stem and progenitor cells. Data analysis using the X-shift algorithm paired with single-cell force-directed layout visualization defined a molecular signature of the activated stem cell state (CD44+/CD98+/MyoD+) and delineated a myogenic trajectory during recovery from acute muscle injury. Our studies uncover the dynamics of skeletal muscle regeneration in vivo and pave the way for the elucidation of the regulatory networks that underlie cell-state transitions in muscle diseases and ageing.


[The role of radiological methods in complex diagnostics of pancreonecrosis].
The results of treatment of 77 patients with necrotic forms of acute pancreatitis are critically revised. The diagnostic algorithm of pancreonecrosis, consisting of ultrasound, computed scanning, magnetic resonance cholangiopancreatography and endoscopic retrograde cholangiopancreatography proved to be useful and effective. The Balthazar computed tomography severity index allows to prognose the course of the disease more accurately.


Mathematical model of beta-lactam penetration into a biofilm of Pseudomonas aeruginosa while undergoing simultaneous inactivation by released beta-lactamases.
We present a mathematical model that describes penetration of an antibacterial agent into a bacterial biofilm and, in particular, the penetration of a beta-lactam compound into a biofilm of Pseudomonas aeruginosa. As well as dealing with this penetration, and the consequent bacterial lysis, the model considered diffusion of the released beta-lactamases in the extracellular space and the consequent inactivation there of further incoming antibiotic; it also allowed for any chosen fraction of the total beta-lactamase to be permanently accessible to exogenous substrate. The modelling scheme was validated against analytical solutions under appropriately simplified conditions. Using published experimental data we show here that lysis of cells in the surface layers of a film could have an important protective effect on the viability of underlying bacteria, especially in thick biofilms.


Macroevolutionary interpretations of symmetry and synchroneity in the fossil record.
Quantitative analyses of global diversity in the marine fossil record over Phanerozoic time reveal an historically ordered pattern of sequential dominance and increasing diversity. Explanatory models applied to this empirical pattern lead to irreconcilable differences of interpretation. The issue may be resolved by determining the expected distributions and limits of temporal covariation among clades generated by a random branching process. Results also challenge the claim that asymmetries in intra-clade diversity variation provide a directional arrow for the history of life.


Low molecular weight heparin (KABI 2165) as thromboprophylaxis in elective visceral surgery. A randomized, double-blind study versus unfractionated heparin.
In two randomized double-blind studies perioperative bleeding complications and thromboembolic events were assessed in 189 patients (pts) undergoing elective visceral surgery after subcutaneous administration of a low molecular weight (LMW) heparin fragment (KABI fragment 2165) or unfractionated (UF) heparin. The first study comparing 1 X 7'500 anti-factor Xa IU LMW heparin daily with 2 X 5'000 IU UF heparin was interrupted because of excessive bleeding complications (LMW heparin: 11/23 pts, UF heparin: 2/20 pts, p less than 0.01). In the second study (146 pts) the dose of LMW heparin was reduced to 1 X 2'500 anti-factor Xa IU. Bleeding complications (LMW heparin: 14.9%, UF heparin: 15.3%) and thromboembolic events (LMW heparin: 2.86%, UF heparin: 2.94%) were equal among the two groups. 2'500 anti-factor Xa IU/day of this LMW heparin fragment, corresponding to 15 mg/day, is the lowest dose of a LMW heparin used in a randomized clinical trial and was found to be a safe and efficient regimen in perioperative thrombosis prophylaxis. An advantage of LMW heparin over UF heparin is its once daily administration.


Small cell carcinoma of the prostate. Part I. A clinicopathologic study of 20 cases.
Clinical information and histological slides of 20 cases of small cell carcinoma of the prostate seen at The University of Texas M. D. Anderson Hospital and Tumor Institute at Houston over a 23-year period were reviewed. Patient's ages ranged from 30 to 89 years (median, 67 years). In nine cases, pure adenocarcinoma of the prostate preceded recognition of the small cell component by 7 months to 8 years (median, 18 months); five of these were initially at Stage A. There was a small cell component at presentation in 11 cases (10, Stage D). Small cell carcinoma was merging with the adenocarcinoma in 11 cases and represented 30% to 90% of total tumor volume. Eleven of 20 patients died of their disease. Those presenting initially with a pure adenocarcinoma survived between 7 months and 9 years (median, 24 months). After the recognition of the small cell carcinoma component, regardless of a prior history of adenocarcinoma, death followed within 1.5 years (median, 5 months). This study suggests a biologic difference in behavior in prostatic carcinoma containing a small cell carcinoma component. The small cell component may manifest early or late in the disease.


Spectral entropy as an electroencephalographic measure of anesthetic drug effect: a comparison with bispectral index and processed midlatency auditory evoked response.
The authors compared the behavior of two calculations of electroencephalographic spectral entropy, state entropy (SE) and response entropy (RE), with the A-Line ARX Index (AAI) and the Bispectral Index (BIS) and as measures of anesthetic drug effect. They compared the measures for baseline variability, burst suppression, and prediction probability. They also developed pharmacodynamic models relating SE, RE, AAI, and BIS to the calculated propofol effect-site concentration (Ceprop). With institutional review board approval, the authors studied 10 patients. All patients received 50 mg/min propofol until either burst suppression greater than 80% or mean arterial pressure less than 50 mmHg was observed. SE, RE, AAI, and BIS were continuously recorded. Ceprop was calculated from the propofol infusion profile. Baseline variability, prediction of burst suppression, prediction probability, and Spearman rank correlation were calculated for SE, RE, AAI, and BIS. The relations between Ceprop and the electroencephalographic measures of drug effect were estimated using nonlinear mixed effect modeling. Baseline variability was lowest when using SE and RE. Burst suppression was most accurately detected by spectral entropy. Prediction probability and individualized Spearman rank correlation were highest for BIS and lowest for SE. Nonlinear mixed effect modeling generated reasonable models relating all four measures to Ceprop. Compared with BIS and AAI, both SE and RE seem to be useful electroencephalographic measures of anesthetic drug effect, with low baseline variability and accurate burst suppression prediction. The ability of the measures to predict Ceprop was best for BIS.


NMR Metabolite Profiles of the Bivalve Mollusc Mytilus galloprovincialis Before and After Immune Stimulation With Vibrio splendidus.
The hemolymph metabolome of Mytilus galloprovincialis injected with live Vibrio splendidus bacteria was analyzed by 1H-NMR spectrometry. Changes in spectral hemolymph profiles were already detected after mussel acclimation (3 days at 18 or 25 °C). A significant decrease of succinic acid was accompanied by an increase of most free amino acids, mytilitol, and, to a smaller degree, osmolytes. These metabolic changes are consistent with effective osmoregulation, and the restart of aerobic respiration after the functional anaerobiosis occurred during transport. The injection of Vibrio splendidus in mussels acclimated at 18°C caused a significant decrease of several amino acids, sugars, and unassigned chemical species, more pronounced at 24 than at 12 h postinjection. Correlation heatmaps indicated dynamic metabolic adjustments and the relevance of protein turnover in maintaining the homeostasis during the response to stressful stimuli. This study confirms NMR-based metabolomics as a feasible analytical approach complementary to other omics techniques in the investigation of the functional mussel responses to environmental challenges.


Retinotomy Closure Following Subretinal Stem Cell Transplant With a 30-Gauge Needle.
The authors report two cases of posterior retinotomy closure following subretinal stem cell transplantation for age-related macular degeneration with a 30-gauge needle - a larger bore needle than those used in prior studies. Partial retinotomy closure was seen on optical coherence tomography within 24 hours in one patient, whereas complete closure occurred by the 5-month and 1-year follow-up visits. A trace retinal hemorrhage occurred in one case, with resolution by 12 weeks. These findings demonstrate the likelihood of uncomplicated, spontaneous retinotomy closure following subretinal stem cell transplantation with a 30-gauge needle. [Ophthalmic Surg Lasers Imaging Retina. 2016;47:869-873.].


Differential contribution of two organelles of endosymbiotic origin to iron-sulfur cluster synthesis and overall fitness in Toxoplasma.
Iron-sulfur (Fe-S) clusters are one of the most ancient and ubiquitous prosthetic groups, and they are required by a variety of proteins involved in important metabolic processes. Apicomplexan parasites have inherited different plastidic and mitochondrial Fe-S clusters biosynthesis pathways through endosymbiosis. We have investigated the relative contributions of these pathways to the fitness of Toxoplasma gondii, an apicomplexan parasite causing disease in humans, by generating specific mutants. Phenotypic analysis and quantitative proteomics allowed us to highlight notable differences in these mutants. Both Fe-S cluster synthesis pathways are necessary for optimal parasite growth in vitro, but their disruption leads to markedly different fates: impairment of the plastidic pathway leads to a loss of the organelle and to parasite death, while disruption of the mitochondrial pathway trigger differentiation into a stress resistance stage. This highlights that otherwise similar biochemical pathways hosted by different sub-cellular compartments can have very different contributions to the biology of the parasites, which is something to consider when exploring novel strategies for therapeutic intervention.


[Effect of leaves of Ginkgo biloba on hair regrowth in C3H strain mice].
Effects of 70% ethanolic extract from leaves of Ginkgo biloba (GBE) on the hair regrowth in normal and high butter diet-pretreated C3H strain mice which posterior hair we shaved were investigated. GBE showed a promoting effect on the hair regrowth. GBE had the inhibitory effects on blood platelet aggregation, thrombin activity and fibrinolysis. GBE inhibited the increase of serum the triglyceride level in high cholesterol diet-treated rats. These results suggested that GBE promotes the hair regrowth and could be used as a hair tonic.


The effect of β-caryophyllene on nonalcoholic steatohepatitis.
The pathogenesis of nonalcoholic steatohepatitis (NASH) is not fully understood, but many studies have suggested that oxidative stress plays a key role. The methionine- and choline-deficient diet (MCD) administration model can reproduce histopathological features of human NASH and is widely used for investigating NASH. C57BL/6J mice have been used in many studies, but strain differences in pathogenesis have not been sufficiently investigated. We administred MCD to two mouse strains and then compared difference between strains and investigated the effects of β-caryophyllene (BCP), which possesses an antioxidant effect, on development and progression of NASH. ICR and C57BL/6J mice were administred a control diet, MCD, MCD containing 0.02% BCP, or MCD containing 0.2% BCP. After 4 or 8 weeks, mice were sacrificed. In both strains, MCD administration induced hepatic steatosis and inflammation. These lesions were more severe in C57BL/6J mice than ICR mice, and liver fibrosis was observed at 8 weeks in C57BL/6J mice. These changes were attenuated by BCP coadministration. The mRNA expression of monocyte chemotactic and activating factor (MCP)-1 and fibrosis-related factors increased in C57BL/6J mice, and these increases were reduced by BCP coadministration. The mRNA expression of antioxidant-related factors decreased in both strains, and these decreases were attenuated by BCP coadministration. Based on these results, the C57BL/6J mouse was a more suitable model for MCD-induced NASH than the ICR mouse. In addition, it was suggested that antioxidant effect of BCP might suppressed the damage of hepatocytes caused by oxidative stress and following inflammation and fibrosis.


Canadian Cardiovascular Society guidelines on the use of cardiac resynchronization therapy: evidence and patient selection.
Recent landmark trials provided the impetus to update the recommendations for cardiac resynchronization therapy (CRT). This article provides guidance on the prescription of CRT within the confines of published data. A future article will explore the implementation of these guidelines. These guidelines are intended to serve as a framework for the prescription of CRT within the Canadian health care system and beyond. They were developed through a critical evaluation of the existing literature, and expert consensus. The panel unanimously adopted each recommendation. The 8 recommendations relate to ensuring the adequacy of medical therapy before the initiation of CRT, the use of symptom severity to select candidates for CRT, differing recommendations based on the presence or absence of sinus rhythm, the presence of left bundle branch block vs other conduction patterns, and QRS duration. The use of CRT in the setting of chronic right ventricular pacing, left ventricular lead placement, and the routine assessment of dyssynchrony to guide the prescription of CRT are also included. The strength of evidence was weighed, taking full consideration of any risks of bias, as well as any imprecision, inconsistency, and indirectness of the available data. The strength of each recommendation and the quality of evidence were adjudicated. Trade-offs between desirable and undesirable consequences of alternative management strategies were considered, as were values, preferences, and resource availability. These guidelines were externally reviewed by experts, modified based on those reviews, and will be updated as new knowledge is acquired.


Establishment and characterization of a recurrent malignant peripheral nerve sheath tumor cell line: RsNF.
Malignant peripheral nerve sheath tumor (MPNST) is a highly aggressive and recurrent soft tissue sarcoma. It most commonly occurs secondary to neurofibromatosis type I, and it has a 5-year survival rate of only 8-13%. To better study the tumor heterogeneity of MPNST and to develop diverse treatment options, more tumor-derived cell lines are needed to obtain richer biological information. Here, we established a primary cell line of relapsed MPNST RsNF cells derived from a patient diagnosed with NF1 and detected the presence of NF1 mutations and SUZ12 somatic mutations through whole-exome sequencing(WES). Through tumor molecular marker targeted sequencing and single-cell transcriptome sequencing, it was found that chromosome 7 copy number variation (CNV) was gained in this cell line, and ZNF804B, EGFR, etc., were overexpressed on chromosome 7. Therefore, RsNF cells can be used as a useful tool in NF1-associated MPNST genomic amplification studies and to develop new therapeutic strategies.


Changes in oxygenated hemoglobin link freezing of gait to frontal activation in patients with Parkinson disease: an fNIRS study of transient motor-cognitive failures.
Recent studies have suggested that deficits in executive function contribute to freezing of gait (FOG), an episodic disturbance common among patients with Parkinson's disease (PD). To date, most findings provide only indirect evidence of this relationship. Here, we evaluated a more direct link between FOG and frontal lobe dysfunction. Functional, near infrared spectroscopy measured frontal activation, i.e., oxygenated hemoglobin (HbO2) levels in Brodmann area 10 before and during FOG. Eleven patients with PD and eleven healthy older adults were studied. Changes in frontal lobe activation before and during FOG that occurred during turns were determined. Altogether, 49 FOG episodes were observed-28 occurred during turns that were anticipated (i.e., the patient knew in advance that the turn was coming), 21 during unanticipated turns that were performed "abruptly", according to the examiner's request. During anticipated turns, HbO2 increased by 0.22 ± 0.08 µM (p = 0.004) before FOG and by an additional 0.19 ± 0.13 µM (p = 0.072) during FOG. In contrast, during unanticipated turns, HbO2 did not increase before or during FOG. HbO2 decreased by 0.32 ± 0.08 µM (p = 0.004) during turns without FOG; in healthy controls HbO2 did not change during turns. These findings support the existence of an association between FOG episodes and changes in frontal lobe HbO2. Increased activation in Brodmann area 10 before FOG, specifically during anticipated turns, highlights the connections between motor planning, information processing, and FOG. These results support the idea that alterations in executive control play a role in this debilitating motor disturbance.


X-ray crystallography and biological metal centers: is seeing believing?
Metalloenzyme crystal structures have a major impact on our understanding of biological metal centers. They are often the starting point for mechanistic and computational studies and inspire synthetic modeling chemistry. The strengths and limitations of X-ray crystallography in determining properties of biological metal centers and their corresponding ligand spheres are explored through examples, including ribonucleotide reductase R2 and particulate methane monooxygenase. Protein crystal structures locate metal ions within a protein fold and reveal the identities and coordination geometries of amino acid ligands. Data collection strategies that exploit the anomalous scattering effect of metal ions can establish metal ion identity. The quality of crystallographic data, particularly the resolution, determines the level of detail that can be extracted from a protein crystal structure. Complementary spectroscopic techniques can provide crucial information regarding the redox state of the metal center as well as the presence, type, and protonation state of exogenous ligands. The final result of the crystallographic characterization of a metalloenzyme is a model based on crystallographic data, supported by information from biophysical and modeling studies, influenced by sample handling, and interpreted carefully by the crystallographer.


COVID-19 and OCD: Potential impact of exposure and response prevention therapy.
This brief clinical review critically assesses the use of exposure and response prevention therapy (ERP) for patients with obsessive-compulsive disorder (OCD) in light of the COVID-19 pandemic. We discuss the ethical and practical considerations that clinicians employed in past infectious disease outbreaks, as well as general safety measures routinely practiced in the conduct of exposure therapy. During this time, concerns regarding the feasibility of ERP have emerged, especially with strict guidelines on social distancing and on following other preventative behaviors. While ERP may have to be modified to follow public health guidelines, this review outlines a) how ERP has been adapted in the context of other infectious triggers; b) the potential impacts on OCD patients of attenuated ERP, and c) minimizing concerns related to litigation. A case report is provided detailing ERP personalized given COVID-19 related considerations. In all, we advise against modifying therapies in ways that may jeopardize the efficacy of patient care or progress.


Precision Zeeman-Stark spectroscopy of the metastable a1[3Sigma+] state of PbO.
The metastable a(1)[(3)Sigma(+)] state of PbO has been suggested as a suitable system in which to search for the electric dipole moment of the electron. We report here the development of experimental techniques allowing high-sensitivity measurements of Zeeman and Stark effects in this system, similar to those required for such a search. We observe Zeeman quantum beats in fluorescence from a vapor cell of PbO, with shot-noise limited extraction of the quantum beat frequencies, high counting rates, and long coherence times. We argue that improvement in sensitivity to the electron electric dipole moment by at least 2 orders of magnitude appears possible using these techniques.


Coronary microcirculation and cardiovascular pathology.
The recent arrival of new techniques for exploring the coronary microcirculation has facilitated assessment of both the incidence and consequences of disorders of this network in a large number of cardiovascular diseases. The microcirculation is affected in numerous cardiomyopathies in the presence of different cardiovascular risk factors and also following cardiac transplantation. Dysfunction of the microcirculation may correspond to a reduction in the surface of the maximum section of coronary arterioles, which involves multiple mechanisms, although this phenomenon does not appear to play a role in ischaemic heart disease. Reduced coronary flow is most frequently related to vascular rarefaction of multifactorial origin, including greater or lesser degrees of intimal proliferation, perivascular fibrosis, hypertrophy of the media and extrinsic compression.


Therapeutic effect of adenosine on experimentally induced acute ulcerative colitis model in rats.
To examine the therapeutic effect of external adenosine on an acetic acid-induced acute ulcerative colitis model in rats. Thirty male mature rats were divided into three groups as control, acute colitis (AC) and AC+adenosine group (AC+AD). AC was induced by rectal administration of 4% acetic acid (AA). 5mg/kg/day adenosine was performed i.p for 4 weeks to AC+AD group. Rectum and colon were excised for microscopic and histopathological histopathologic evaluations, and immunohistochemical analysis of nuclear factor kappa B (NF-kB). Blood samples were collected for biochemical detection of TNF-α, Pentraxin-3 and malondialdehyde (MDA) levels. AC group had generalized hyperemia and hemorrhage with increased macroscopic and histopathological scores compared with control (P <0.0001) while adenosine treatment decreased these scores significantly (P <0.001), with reduced distribution of disrupted epithelium, leukocyte infiltrates, and focal hemorrhage. AC group showed significantly increased immunoexpression of NF-kB in rectum, plasma and tissue levels of TNF-α, plasma Pentraxin-3 and MDA levels (P <0.0001) while adenosine reduced these levels (P < 0.05). Adenosine appears to promote healing of colon and rectum exposed to AA-induced AC, suggesting a boosting effect of adenosine on the intestinal immune system to cure ulcerative colitis.


Increase in sleep related GH and Prl secretion after chronic arginine aspartate administration in man.
Arginine aspartate was administered orally (250 mg/dg/day) for one week to 5 healthy male human volunteers aged 20-35. After this period, the 24 h patterns of growth hormone (GH) and prolactin (Prl) secretion were determined by radioimmunoassay on blood samples withdrawn every 20 min and correlated with the polygraphic recordings of electroencephalogram, electromyogram and electrooculogram. the results were compared to data obtained in identical conditions with the same subjects but after a week of placebo administration. In all 5 subjects the slow wave sleep related GH peak was about 60% higher after a week of arginine aspartate administration than in the control period, with individual changes of +24, +25, +42, +47 and +162%. The nocturnal mean plasma Prl of each subject was higher after arginine aspartate than before. The nocturnal rise of plasma Prl increased from a mean value of +21.5% during the placebo period to +95% at the end of the arginine aspartate treatment. These hormonal modifications were not accompanied by any detectable alteration of sleep organization and specially of the slow wave sleep pattern, suggesting a direct neuroendocrine action of the drug.


[Antibiotic drug minocycline: a potential therapeutic drug for methamphetamine-related disorders].
Accumulating evidence suggests a role of inflammation in the pathophysiology of a number of neuropsychiatric diseases. The second generation antibiotic drug minocycline has potent neuroprotective and anti-inflammatory effects. We reported that minocycline could attenuate behavioral abnormalities and dopaminergic neurotoxicity in mice after administration of methamphetamine or 3,4-methylenedioxymethamphetamine (MDMA). Furthermore, we reported that minocycline was effective in the animal models of schizophrenia. Moreover, a double-blind, placebo-control, cross-over study showed that minocycline was effective in the rewarding effects in healthy human subjects. In this article, we would like to discuss minocycline as a potential therapeutic drug for methamphetamine-related disorders.


Musculoskeletal Disorders: Prevalence and Associated Factors among District Hospital Nurses in Haiphong, Vietnam.
Musculoskeletal disorders (MSDs) are a major occupational health problematic among healthcare workers, and the prevalence is especially high among nurses. In high income countries, the prevention of MSDs is an occupational health priority. But in Vietnam, there is no data available among health professionals. To determine the prevalence and associated factors of musculoskeletal disorders among district hospital nurses in Haiphong city. A cross-sectional study was conducted on 1179 nurses working in 15 district hospitals using the Standardized Nordic Questionnaire. A very high prevalence of MSDs in the past 12 months (74.7%) and during the last 7 days (41.1%), with the two most common sites being the low back (44.4%) and neck (44.1%), was found; 37.8% complained that MSDs symptoms limit their work. When analyzing factors related to MSDs, the results showed that women were 2.1 times more likely to develop MSDs than men; people with a previous history of MSDs were more likely to develop MSDs symptoms in the past 12 months than those with no history (OR = 7.1); nurses with symptoms of psychological distress and frequent absenteeism in the workplace had a higher prevalence of MSDs compared to the rest (p<0.001). Due to the high prevalence of MSDs among nurses in district hospitals in Haiphong, preventive actions are needed to improve the working conditions and to raise the awareness of nurses about MSDs prevention.


A Comprehensive Qualitative Review of Studies Evaluating the Impact of Local US Laws Restricting the Sale of Flavored and Menthol Tobacco Products.
To assess the quality of evidence on the effectiveness of local US laws restricting the sale of flavored tobacco products. We conducted a systematic search and qualitative scoping review of English-language papers published through May 2020 that evaluated flavored tobacco sales policies implemented by US jurisdictions during 2010-2019. We constructed a conceptual model for flavored and menthol tobacco sales restriction outcomes, assigned GRADE quality of evidence ratings to policy outcomes evaluated through the included studies, and summarized factors that might explain weak or inconsistent findings. We found moderate to high quality of evidence associating policy implementation with reduced availability, marketing, and sales of policy-restricted products, and decreased youth and adult tobacco use of these products; however, policy exclusions and exemptions, implementation challenges, tobacco industry actions (e.g., marketing of concept-named flavored products; exploiting policy exemptions for certain store types), and consumer responses (e.g., cross-border or illicit purchasing) might undermine or mitigate intended policy effects. Flavored and menthol tobacco product sales restrictions implemented and evaluated in US jurisdictions appear to have achieved some of their intended outcomes; however, deficiencies in study designs, methods, and metrics could contribute to equivocal findings on quality of evidence associating policy implementation and outcomes. Gaps in the evidence are beginning to be filled with research using more rigorous study designs, improved measurement and analytic methods, and longer-term follow-up. In the absence of comprehensive federal action, US jurisdictions have the obligation to restrict flavored and menthol product sales to protect vulnerable populations from tobacco-related harms. The considerable expenditure of financial resources, political will, and time dedicated to policy adoption and implementation argue for evaluation studies designed to maximize the quality of evidence. This review offers generalizable insights into evaluation findings that can inform efforts to enhance tobacco control policy implementation and impact in the US and globally.


[Inflammatory fibrosarcoma: a clinical morphological study of six cases].
The report describes the clinical and morphological characteristics of inflammatory fibrosarcoma (6). Tumor occurred in the mediastinum, small pelvis and liver in patients aged 12-64 yrs. It featured infiltrative growth. Primary tumor relapsed in 2 cases, distant metastases - 4. Four patients died from tumor progression within 7-14 months after diagnosis. Tumors consisted of neoplastic fibroblasts and lymphofibroblasts with polymorphous nuclei and high mitotic activity.


Scroll-wave dynamics in human cardiac tissue: lessons from a mathematical model with inhomogeneities and fiber architecture.
Cardiac arrhythmias, such as ventricular tachycardia (VT) and ventricular fibrillation (VF), are among the leading causes of death in the industrialized world. These are associated with the formation of spiral and scroll waves of electrical activation in cardiac tissue; single spiral and scroll waves are believed to be associated with VT whereas their turbulent analogs are associated with VF. Thus, the study of these waves is an important biophysical problem. We present a systematic study of the combined effects of muscle-fiber rotation and inhomogeneities on scroll-wave dynamics in the TNNP (ten Tusscher Noble Noble Panfilov) model for human cardiac tissue. In particular, we use the three-dimensional TNNP model with fiber rotation and consider both conduction and ionic inhomogeneities. We find that, in addition to displaying a sensitive dependence on the positions, sizes, and types of inhomogeneities, scroll-wave dynamics also depends delicately upon the degree of fiber rotation. We find that the tendency of scroll waves to anchor to cylindrical conduction inhomogeneities increases with the radius of the inhomogeneity. Furthermore, the filament of the scroll wave can exhibit drift or meandering, transmural bending, twisting, and break-up. If the scroll-wave filament exhibits weak meandering, then there is a fine balance between the anchoring of this wave at the inhomogeneity and a disruption of wave-pinning by fiber rotation. If this filament displays strong meandering, then again the anchoring is suppressed by fiber rotation; also, the scroll wave can be eliminated from most of the layers only to be regenerated by a seed wave. Ionic inhomogeneities can also lead to an anchoring of the scroll wave; scroll waves can now enter the region inside an ionic inhomogeneity and can display a coexistence of spatiotemporal chaos and quasi-periodic behavior in different parts of the simulation domain. We discuss the experimental implications of our study.


Macrophage migration inhibitory factor.
Macrophage migration inhibitory factor (MIF) is a ubiquitous protein that is found in virtually all cells. Its precise function in the majority of cells is not known, but studies performed over the last decade indicate that it is a critical upstream regulator of the innate and acquired immune response. MIF is released under a variety of circumstances, regulates cytokine secretion and the expression of receptors that are involved in innate immunity, inhibits p53 function, and activates components of the mitogen-activated protein kinase and Jun-activation domain-binding protein-1 (Jab-1) pathways. Compelling in vitro and in vivo evidence has focused attention on this protein as a new therapeutic target for inflammatory and autoimmune diseases. Unique structural features, including an intrinsic catalytic activity, offer attractive opportunities for the discovery and design of therapeutic MIF inhibitors.


The effect of interventions targeting gut microbiota on depressive symptoms: a systematic review and meta-analysis.
Despite their popularity, the efficacy of interventions targeting gut microbiota to improve depressive symptoms is unknown. Our objective is to summarize the effect of microbiome-targeting interventions on depressive symptoms. We conducted a systematic review and meta-analysis. We searched MEDLINE, Embase, PsycINFO, Database of Abstracts of Reviews of Effects, Cochrane Database of Systematic Reviews and the Cochrane Controlled Register of Trials from inception to Mar. 5, 2021. We included studies that evaluated probiotic, prebiotic, synbiotic, paraprobiotic or fecal microbiota transplant interventions in an adult population (age ≥ 18 yr) with an inactive or placebo comparator (defined by the absence of active intervention). Studies must have measured depressive symptoms with a validated scale, and used a randomized controlled trial study design. We conducted a random effects meta-analysis of change scores, using standardized mean difference as the measure of effect. Sixty-two studies formed the final data set, with 50 included in the meta-analysis. Probiotic, prebiotic, and synbiotic interventions on depressive symptoms showed statistically significant benefits. In the single studies evaluating each of fecal microbiota transplant and paraprobiotic interventions, neither showed a statistically significant benefit. Despite promising findings of benefit of probiotic, prebiotic and synbiotic interventions for depressive symptoms in study populations, there is not yet strong enough evidence to favour inclusion of these interventions in treatment guidelines for depression. Critical questions about species administered, dosage and timing relative to other antidepressant medications remain to be answered. PROSPERO no. 143178.


[Itchy rash caused by the oak processionary caterpillar].
BACKGROUND As a result of climate change, we will have to deal more and more often with the consequences of the oak processionary caterpillar. CASE DESCRIPTION In June, we saw a 6-year-old girl with a severely itchy rash all over her body. The rash had developed a day after school and consisted of erythematous papules on an erythematous base. Several other children at the school were suffering from a similar skin rash. The girl also seemed to have a bad cold. We diagnosed 'oak processionary caterpillar dermatitis'. It turned out there was indeed an oak processionary caterpillar nest at her school. The nest was removed. We prescribed cetirizine oral solution, followed by triamcinolone cream, after which the itch and skin rash disappeared. CONCLUSION The oak processionary caterpillar can cause a lot of symptoms in the period from April to July, including skin rash and eye and upper airway symptoms. The symptoms are usually self-limiting and automatically disappear within 2 weeks. In case of severe itching, menthol-based creams, topical corticosteroids and antihistamines may be prescribed.


Adenosine kinase and ribokinase--the RK family of proteins.
Ribokinase (RK) and adenosine kinase (AK) catalyze the phosphorylation of ribose and adenosine to ribose-5-phosphate and AMP, respectively. Belonging to the RK family of proteins, these enzymes share a number of unique structural and functional elements. Extensive work has been carried out on many aspects of these enzymes in recent years, and we summarize the wealth of information currently available on them. The topics covered include descriptions of the primary and three-dimensional structures of AK and RK, their phylogenetic relationships, biochemical aspects of these enzymes including their reaction mechanisms and ionic requirements, and also work on certain inhibitors of these enzymes. The cellular metabolism and transport of ribose and adenosine are also briefly discussed, as well as the beneficial effects of ribose and adenosine in physiology and how these effects can be harnessed for pharmacological purposes.


Sperm cryopreservation in the spermcasting Australian flat oyster Ostrea angasi by a programmable freezing method.
Cryopreservation offers long-term storage of gametes without constraint from seasonal gamete maturation, provides opportunities to improve the efficiency of breeding and genetic programs, and protects endangered species from extinction due to epidemic diseases and natural disasters. In this study, a protocol for cryopreserving sperm of the spermcasting Australian flat oyster Ostrea angasi was developed by optimizing key factors influencing the quality of cryopreserved sperm. Dimethyl sulfoxide (DMSO) was non-toxic to sperm within the concentration and duration assessed in the toxicity experiment whereas 10% methanol or a higher concentration was toxic to sperm from the exposure duration of 30 min onwards. DMSO produced higher post-thaw sperm motility among the treatments with a single cryoprotectant. The inclusion of trehalose or glucose with DMSO further increased the post-thaw sperm motility (%) and plasma membrane integrity (PMI). Sperm equilibrated for 30 min showed higher post-thaw motility and PMI than those for 10 or 50 min. Higher post-thaw sperm motility and PMI were achieved at the freezing rate of -3 °C/min than at -7 °C/min. Sperm packaged in 0.5 ml straws had a higher post-thaw motility and PMI than those packaged in 0.25 ml straws. In this study, 44.4% post-thaw sperm motility and 49.2% PMI were achieved when sperm were equilibrated in 10% DMSO +0.45 M trehalose for 30 min, packaged in 0.5 ml straws, frozen at -3 °C/min from 4 °C to -80 °C, and thawed at 40 °C for 8 s. The availability of viable cryopreserved sperm would open an option for future breeding and genetic improvement programs for the spermcasting Australian flat oyster.


Hypoxia promotes the growth and metastasis of ovarian cancer cells by suppressing ferroptosis via upregulating SLC2A12.
Ovarian cancer has been a worldwide health burden for women and its progression is highly hypoxia-independent. Here, we investigated the exact mechanisms by which hypoxia contributes to the malignant progression of ovarian cancer. MTT, transwell, colony formation, and scratch wound healing assays were carried out for cellular functions. The underlying mechanism by which hypoxia functions was explored by RNA-seq, enrichment analysis, western blotting, qRT-PCR, flow cytometry, ChIP, luciferase reporter, and ELISA. Finally, animal experiments including the xenograft model and tumor metastasis model were constructed to validate the role of SLC2A12 in vivo. Hypoxia treatment promoted the cell proliferation, mobility, and colony growth abilities of the two ovarian cancer cell lines HO-8910 and A2780. RNA-seq and enrichment analysis showed that SLC2A12 was hyper-expressed under hypoxia condition and it may be related to glutathione and lipid metabolism. Besides, the expression of SLC2A12 was negatively correlated with overall survival. Hypoxia suppressed ferroptosis by SLC2A12 because silencing SLC2A12 declined the cell viability of HO-8910 and A2780 cells under hypoxia conditions, while the ferroptosis inhibitor ferrostatin-1 (Fer-1) breached that result and upregulated the expression of glutathione peroxidase 4 (GPX4). Moreover, hypoxia increased the expression of hypoxia inducible factor 1 A (HIF-1A), and the accumulated HIF-1A binds to hypoxia inducible factor 1 B (HIF1B) to form HIF-1 complex, then promoted the binding of hypoxic response elements (HRE) to SLC2A12 promoter by HIF-1/HRE signal. Subsequently, SLC2A12 regulated glutathione metabolism and in turn inhibited ferroptosis. The animal experiments indicated that silencing SLC2A12 could significantly inhibit tumor growth and metastasis in vivo. Hypoxia promoted ovarian cancer progression by upregulating SLC2A12 and then regulating glutathione metabolism to inhibit ferroptosis.


Mouse alopecia areata models: an array of data on mechanisms and genetics.
Laboratory mice have become the premier animal model for most human and domestic animal diseases, and they have long been the model of choice for studying mammalian genetics, especially since the advent of genetic engineering. Many remarkable discoveries have been made through intense study of these wonderful small mammals, and undoubtably many more will be made. It is no surprise that one mouse model for alopecia areata (AA) has been found possibly many more will be, some of which exhibit rare phenotypes found in subpopulations of humans with the disease, such as nail deformities, thyroid disease, inflammatory bowel disease, and autoimmune polyendocrine syndrome type 1. Intense investigation by many groups into the first model, the adult onset form of AA (using the C3H/HeJ inbred strain), found similarities as well as differences with commonly held ideas concerning human AA. Regardless of some of the controversies, much insight has been gained from studying these and other rodent and domestic animal models which has opened up new ideas and discussions of AA and its treatment.


Synthetic Biology for Specialty Chemicals.
In this review, we address recent advances in the field of synthetic biology and describe how those tools have been applied to produce a wide variety of chemicals in microorganisms. Here we classify the expansion of the synthetic biology toolbox into three different categories based on their primary function in strain engineering-for design, for construction, and for optimization. Next, focusing on recent years, we look at how chemicals have been produced using these new synthetic biology tools. Advances in producing fuels are briefly described, followed by a more thorough treatment of commodity chemicals, specialty chemicals, pharmaceuticals, and nutraceuticals. Throughout this review, an emphasis is placed on how synthetic biology tools are applied to strain engineering. Finally, we discuss organism and host strain diversity and provide a future outlook in the field.


A stress-responsive Escherichia coli protein, CysQ is a highly effective solubility enhancer for aggregation-prone heterologous proteins.
When used as an N-terminal fusion expression partner, the Escherichia coli stress-responsive protein, CysQ dramatically increased the cytoplasmic solubility of various aggregation-prone heterologous proteins: Pseudomonas putida cutinase (CUT), human granulocyte colony-stimulating factor (hG-CSF), human ferritin light chain (hFTN-L), arginine deiminase (ADI), human interleukin-2 (IL2), human activation induced cytidine deaminase (AID), and deletion mutant of human glutamate decarboxylase (GAD448-585). As compared with well-known fusion tags such as glutathione-S-transferase (GST) and maltose-binding protein (MBP), the performance of CysQ as solubility enhancer was evidently better than GST and was similar to or better than MBP for the seven heterologous proteins above. This is likely due to the intrinsic ability of CysQ to form its native conformation, probably promoting the binding of molecular chaperones during the folding of CysQ-fusion protein. When used as a substrate, p-nitrophenyl butyrate (PNB) was successfully hydrolyzed to p-nitrophenol by CysQ-CUT fusion mutant. Even after CysQ was removed, the solubility of hFTN-L and hG-CSF, the secondary structure of hG-CSF, and self-assembly activity of hFTN-L were successfully maintained. Conclusively, it seems that CysQ is a highly effective solubility enhancer and fusion expression partner for the production of a variety of bio-active recombinant proteins.


Becoming sandwiched in later life: Consequences for individuals' well-being and variation across welfare regimes.
The experience of being sandwiched between support obligations towards both aging parents and adult offspring is likely to become more common and more relevant. We aim at assessing the effect of demographic and social sandwiching on the psychological health and subjective well-being of individuals experiencing these transitions, and to what extent, these effects vary across welfare regimes. Data are from 63,585 individuals aged 50-75 participating in the Survey of Health, Ageing and Retirement in Europe (SHARE). We estimate within- and between-individual effects using hybrid regression models to predict depressive symptoms (EURO-D) and subjective well-being (CASP). Among demographically sandwiched women, transitioning into social sandwiching and into supporting only parents were associated with a moderate but statistically significant increase in EURO-D and decline in CASP scores. The same association is not observed for male respondents. The pattern of variation among women living in countries characterized by different welfare regimes suggests that social sandwiching is less detrimental in the Nordic regimes than in other welfare contexts. Results from the between-individuals part of the model indicate that there is a selection into social sandwiching of more healthy individuals into support roles. However, the within-individuals part of the model indicates that the transition into social sandwiching has a detrimental effect on women's (but not men's) psychological health and well-being. The explanations for this gendered effect of social sandwiching may be found in the "invisible" support provided by women and the gendered division of specific care tasks.


Recent N-atom containing compounds from indo-pacific invertebrates.
A large variety of unique N-atom containing compounds (alkaloids) without terrestrial counterparts, have been isolated from marine invertebrates, mainly sponges and ascidians. Many of these compounds display interesting biological activities. In this report we present studies on nitrogenous compounds, isolated by our group during the last few years, from Indo-Pacific sponges, one ascidian and one gorgonian. The major part of the review deals with metabolites from the Madagascar sponge Fascaplysinopsis sp., namely, four groups of secondary metabolites, the salarins, tulearins, taumycins and tausalarins.


Dialect identification versus evaluation of risk in language screening.
This article proposes that any dialect-neutral screening test should consist of two parts: one part using contrastive items to screen for Language Variation Status (Mainstream American English [MAE] or a degree of variation from MAE), and a second part using noncontrastive items to screen for degree of risk for language disorder (low, medium, or high). The two scores are interpreted together in forming an overall clinical profile of a child. The implications for further diagnostic testing of both MAE and non-MAE speakers who exhibit risk for language disorder are discussed. Most importantly, for the African American child, viewing the results of the two types of proposed screening items together clears the "variation due to speech and language dialect" issue out of the way so that if further evaluation is needed, it is done for valid reasons, not superficial ones.


Stature and birth rank. A study of schoolchildren in St Helena.
More than 1200 St Helenian schoolchildren were studied in 1973 and 1978 to determine how social factors affected growth. The association of short stature with large family size was confirmed and shown to be concentrated in the earlier members of the sibship. This was independent of birth interval. Association between birth interval and stature was most pronounced for the spacing following the index child, less for the preceding interval, and least for the family average interval. These results are similar to those of British children, for whom St Helenians form a reasonable model. It is argued that these results are not readily explained by genetic factors nor are they explained by differences of birthweight, but they are compatible with deprivation of parental care and love, for which older children have to compete with other sibings. Deprivation sufficient to retard growth is not confined to extreme circumstances or to broad social groups, but occurs selectively within the heart of the normal family.


Toxoplasma gondii induces prostaglandin E2 synthesis in macrophages via signal pathways for calcium-dependent arachidonic acid production and PKC-dependent induction of cyclooxygenase-2.
In this study, the intracellular signaling pathway of PGE2 synthesis in macrophages (RAW264.7) induced by Toxoplasma gondii was investigated. The T. gondii-induced PGE2 production in macrophages increased in a time-dependent manner, as PGE2 induction began at 4 h, peaked at 12 h, and then plateaued at a high level. COX-2 mRNA in macrophages was detectable as early as 4 h after treatment; the maximal expression was observed at 8 h. The earliest induction of COX-2 protein occurred at 4 h and peaked at 16 h; meanwhile, COX-1 mRNA level and protein production remained unchanged throughout. Indomethacin and nimesulide inhibited tachyzoite-induced PGE2 production and COX-2 mRNA expression in macrophages but they had no significant effect on COX-2 protein expression. EGTA, TFP and BAPTA/AM inhibited both arachidonic acid (AA) and PGE2 production without effecting COX-2 protein expression, but verapamil inhibited neither AA nor PGE2 production. H7 was found to inhibit PGE2 production, and COX-2 mRNA expression and protein expression by tachyzoite or LPS stimulated macrophages in a dose-dependent manner. Our results demonstrate that T. gondii induces PGE2 biosynthesis in RAW264.7 macrophages by regulating AA production through a calcium-dependent pathway and induction of COX-2 expression by a PKC-dependent pathway.


Alpha-tocopherol in the ventricular cerebrospinal fluid of Parkinson's disease patients: dose-response study and correlations with plasma levels.
To determine if ventricular cerebrospinal fluid (vCSF) alpha-tocopherol levels in Parkinson's disease (PD) patients can be increased by oral alpha-tocopherol supplementation and whether vCSF levels are linearly related to plasma alpha-tocopherol levels. In spite of its putative neuroprotective properties, alpha-tocopherol has failed to alter PD clinical progression. However, the ability of supplemental alpha-tocopherol to affect brain or vCSF levels has never been assessed in humans nor has a dose response curve for alpha-tocopherol in vCSF been established. Five PD patients with Ommaya catheters received oral dl-alpha-tocopherol over 5 months. Each patient ingested alpha-tocopherol daily with monthly dosage increases (400, 800, 1,600, 3,200, 4,000 IU/day). Plasma and vCSF samples were obtained at baseline and at the end of each month. Alpha-tocopherol levels were determined in triplicate by high-pressure liquid chromatography with fluorometric and electrochemical detection. At baseline, endogenous alpha-tocopherol was detected in plasma and vCSF, with a greater than one-hundred-fold difference between the fluid compartments (mean plasma level 18.76 microM/l (SD +/- 4.69) versus mean CSF level 0.114 microM/l (SD +/- 0.084). A clear dose-response curve occurred in plasma, with statistically significant increases over baseline developing even with 400 IU/d. With higher doses, a significant increase continued without evidence of saturation. However, there was no significant increase in vCSF alpha-tocopherol levels at any dose, including the supraclinical (4,000 IU/d). There was no correlation between plasma and vCSF alpha-tocopherol levels. Oral alpha-tocopherol supplementation, even at supraclinical doses, fails to increase vCSF alpha-tocopherol levels. This lack of change may be due to limited passage across the blood-brain barrier or very rapid alpha-tocopherol metabolism. All prior negative studies on efficacy of alpha-tocopherol in PD may need reevaluation in light of these pharmacologic data.


Oral care training in the basic education of care professionals.
To investigate the quantity and quality of oral care training in the basic education of future long-term care (LTC) professionals in Norway. The level of oral hygiene has often proved inadequate in LTC facilities. It has been maintained that this could be due to insufficient knowledge of oral care among care professionals. A self-administered questionnaire was sent to all 270 schools in Norway which offered basic education of LTC personnel in 2004/05. Information on theoretical and practical oral care training, scope of oral care in teaching material and curriculum, educational background of the teaching staff and schools opinion regarding adequacy of their training programme was collected. Of the 203 respondents (75% response rate), 188 (participants) included oral care in their educational programme. Approximately two-thirds of the participating schools provided 3 h or more of oral care training and many of the important themes were presented in the textbooks that were recommended. Moreover, the practical exercises performed in practice placement supplemented the knowledge. The results could not confirm that LTC professional's basic education concerning oral care was inadequate. There may therefore be other explanations for the poor oral hygiene in many LTC facilities.


Iterative Development of a Home Review Program to Promote Preschoolers' Vocabulary Skills: Social Validity and Learning Outcomes.
Purpose This article describes the iterative development of a home review program designed to augment vocabulary instruction for young children (ages 4 and 5 years) occurring at school through the use of a home review component. Method A pilot study followed by two experiments used adapted alternating treatment designs to compare the learning of academic words taught at school to words taught at school and reviewed at home. At school, children in small groups were taught academic words embedded in prerecorded storybooks for 6 weeks. Children were given materials such as stickers with review prompts (e.g., "Tell me what brave means") to take home for half the words. Across iterations of the home intervention, the home review component was enhanced by promoting parent engagement and buy-in through in-person training, video modeling, and daily text message reminders. Visual analyses of single-subject graphs, multilevel modeling, and social validity measures were used to evaluate the additive effects and feasibility of the home review component. Results Social validity results informed each iteration of the home program. The effects of the home program across sites were mixed, with only one site showing consistently strong effects. Superior learning was evident in the school + home review condition for families that reviewed words frequently at home. Although the home review program was effective in improving the vocabulary skills of many children, some families had considerable difficulty practicing vocabulary words. Conclusion These studies highlight the importance of using social validity measures to inform iterative development of home interventions that promote feasible strategies for enhancing the home language environment. Further research is needed to identify strategies that stimulate facilitators and overcome barriers to implementation, especially in high-stress homes, to enrich the home language environments of more families.


Offspring hospitalization for substance use and changes in parental mental health: A Finnish register-based study.
Prior research indicates that parental psychiatric disorders increase their offspring's risk of substance use problems. Though the association is likely bidirectional, the effects of an adult child's substance use on parental mental health remain understudied. We examined parents' psychotropic medication use trajectories by parental sex and educational attainment before and after a child's alcohol- or narcotics-attributable hospitalization. We identified Finnish residents, born 1979-1988, with a first hospitalization for substance use during emerging adulthood (ages 18-29, n = 12,851). Their biological mothers (n = 12,283) and/or fathers (n = 10,765) were followed for the two years before and after the hospitalization. Psychotropic medication use was measured in three-month periods centered around the time of child's hospitalization, and the probability of psychotropic medication use at each time point was assessed using generalized estimating equations logit models. Among mothers, the prevalence of psychotropic medication use increased during the year before, peaked during the 0-3 months after hospitalization, and remained at a similarly elevated level until the end of follow-up. The prevalence among fathers increased gradually and linearly across follow-up, with minimal changes evident either directly before or after the hospitalization. Parents' educational attainment did not modify these trajectories. Our results highlight the importance of considering linked lives when quantifying substance use-attributable harms and underscore the need for future research examining the intergenerational spillover effects of substance use in both directions, particularly in mother-child dyads.


Modeling of a rotary motor driven by an anisotropic piezoelectric composite laminate.
This paper proposes an analytical model of a rotary motor driven by an anisotropic piezoelectric composite laminate. The driving element of the motor is a three-layer laminated plate. A piezoelectric layer is sandwiched between two anti-symmetric composite laminae. Because of the material anisotropy and the anti-symmetric configuration, torsional vibration can be induced through the inplane strain actuated by the piezoelectric layer. The advantages of the motor are its magnetic field immunity, simple structure, easy maintenance, low cost, and good low-speed performance. In this paper, the motor is considered to be a coupled dynamic system. The analytical model includes the longitudinal and torsional vibrations of the laminate and the rotating motion of the rotor under action of contact forces. The analytical model can predict the overall characteristics of the motor, including the modal frequency and the response of motion of the laminate, the rotating speed of the rotor, the input power, the output power, and the efficiency of the motor. The effects of the initial compressive force, the applied voltage, the moment of rotor inertia, and the frictional coefficient of the contact interface on the characteristics of the motor are simulated and discussed. A selection of the numerical results from the analytical model is confirmed by experimental data.


The liver angiotensin receptor involved in the activation of glycogen phosphorylase.
Specific angiotensin binding to rat hepatocytes and purified liver plasma membranes was measured by using biologically active [(3)H]angiotensin (sp. radioactivity 14Ci/mmol). The kinetic parameters for angiotensin binding to hepatocytes are: K(+1) (association rate constant). 100mum(-1).min(-1); K(-1) (dissociation rate constant), 2min(-1); K(d) (dissociation constant). 30nm; maximal binding capacity, 0.42pmol/10(6) cells or 260000 sites/cell. Angiotensin binding to membranes is profoundly affected by GTP (0.1mm) and NaCl (100mm); these regulatory compounds greatly enhance both the rate of association and of dissociation and also the extent of dissociation. K(d) amounts to 10nm in the presence of GTP+NaCl and to 1.5nm in their absence; maximal binding capacity is 0.70pmol/mg of protein, both with or without GTP+NaCl. The relative affinities of 11 angiotensin structural analogues were deduced from competition experiments for [(3)H]angiotensin binding to hepatocytes and to membranes (in the latter case, GTP + NaCl were not included, in order to study the higher affinity state of the receptor). These are highly correlated with their biological activity (activation of glycogen phosphorylase in hepatocytes). Binding to membranes occurs in the same concentration range as the biological effect. On the other hand, the existence of numerous spare receptors is suggested by the observation that binding of the agonists to hepatocytes requires 25-fold higher concentrations than those needed for their biological activity. These data clearly suggest that the detected binding sites correspond to the physiological receptors involved in the glycogenolytic action of angiotensin on rat liver.


Tracking alcohol consumption over time.
Researchers are tracking long-term changes in alcohol consumption and related behaviors or outcomes in order to detect trends in the entire population or certain subgroups, test models of alcohol-related outcomes, and understand the consequences of interventions. Such analyses must consider the complexity of typical lifetime consumption patterns. Major approaches to measuring alcohol consumption over time include aggregate measures of consumption (e.g., sales data), cross-sectional surveys, and longitudinal or panel surveys. When analyzing the data, researchers must try to ensure the comparability of measurements over time. The stability of various measures and the potential for combining different types of data are also important considerations when tracking alcohol consumption over time. If these requirements are met, the regular collection of data on aspects of alcohol consumption will greatly increase researchers' understanding of the forces influencing a population's alcohol consumption and its consequences.


Infant statistical-learning ability is related to real-time language processing.
Infants are adept at learning statistical regularities in artificial language materials, suggesting that the ability to learn statistical structure may support language development. Indeed, infants who perform better on statistical learning tasks tend to be more advanced in parental reports of infants' language skills. Work with adults suggests that one way statistical learning ability affects language proficiency is by facilitating real-time language processing. Here we tested whether 15-month-olds' ability to learn sequential statistical structure in artificial language materials is related to their ability to encode and interpret native-language speech. Specifically, we tested their ability to learn sequential structure among syllables (Experiment 1) and words (Experiment 2), as well as their ability to encode familiar English words in sentences. The results suggest that infants' ability to learn sequential structure among syllables is related to their lexical-processing efficiency, providing continuity with findings from children and adults, though effects were modest.


Regulation of cell survival by sphingosine-1-phosphate receptor S1P1 via reciprocal ERK-dependent suppression of Bim and PI-3-kinase/protein kinase C-mediated upregulation of Mcl-1.
Although the ability of bioactive lipid sphingosine-1-phosphate (S1P) to positively regulate anti-apoptotic/pro-survival responses by binding to S1P1 is well known, the molecular mechanisms remain unclear. Here we demonstrate that expression of S1P1 renders CCL39 lung fibroblasts resistant to apoptosis following growth factor withdrawal. Resistance to apoptosis was associated with attenuated accumulation of pro-apoptotic BH3-only protein Bim. However, although blockade of extracellular signal-regulated kinase (ERK) activation could reverse S1P1-mediated suppression of Bim accumulation, inhibition of caspase-3 cleavage was unaffected. Instead S1P1-mediated inhibition of caspase-3 cleavage was reversed by inhibition of phosphatidylinositol-3-kinase (PI3K) and protein kinase C (PKC), which had no effect on S1P1 regulation of Bim. However, S1P1 suppression of caspase-3 was associated with increased expression of anti-apoptotic protein Mcl-1, the expression of which was also reduced by inhibition of PI3K and PKC. A role for the induction of Mcl-1 in regulating endogenous S1P receptor-dependent pro-survival responses in human umbilical vein endothelial cells was confirmed using S1P receptor agonist FTY720-phosphate (FTY720P). FTY720P induced a transient accumulation of Mcl-1 that was associated with a delayed onset of caspase-3 cleavage following growth factor withdrawal, whereas Mcl-1 knockdown was sufficient to enhance caspase-3 cleavage even in the presence of FTY720P. Consistent with a pro-survival role of S1P1 in disease, analysis of tissue microarrays from ER(+) breast cancer patients revealed a significant correlation between S1P1 expression and tumour cell survival. In these tumours, S1P1 expression and cancer cell survival were correlated with increased activation of ERK, but not the PI3K/PKB pathway. In summary, pro-survival/anti-apoptotic signalling from S1P1 is intimately linked to its ability to promote the accumulation of pro-survival protein Mcl-1 and downregulation of pro-apoptotic BH3-only protein Bim via distinct signalling pathways. However, the functional importance of each pathway is dependent on the specific cellular context.


Effects of insulin, glucagon, dexamethasone, cyclic GMP and spermine on the stability of phosphatidate phosphohydrolase activity in cultured rat hepatocytes.
Hepatocytes were preincubated with 10mM-glucagon and 100 microM-corticosterone to increase phosphatidate phosphohydrolase activity. Addition of 10 nM-glucagon or 100 microM-8-bromo cyclic GMP to a second incubation mixture that contained cycloheximide increased the half-life of the phosphohydrolase activity. Dexamethasone (100 nM) had no significant effect, but insulin (500 pM) or spermine (1 mM) decreased the half-life. None of these compounds altered the general rate of degradation of proteins labelled with [3H]leucine. There appears to be a specific control of the half-life of phosphatidate phosphohydrolase activity, which could contribute to its long-term regulation in the liver.


Medical necessity of routine admission of children with mild traumatic brain injury to the intensive care unit.
OBJECTIVE Approximately 475,000 children are treated for traumatic brain injury (TBI) in the US each year; most are classified as mild TBI (Glasgow Coma Scale [GCS] Score 13-15). Patients with positive findings on head CT, defined as either intracranial hemorrhage or skull fracture, regardless of severity, are often transferred to tertiary care centers for intensive care unit (ICU) monitoring. This practice creates a significant burden on the health care system. The purpose of this investigation was to derive a clinical decision rule (CDR) to determine which children can safely avoid ICU care. METHODS The authors retrospectively reviewed patients with mild TBI who were ≤ 16 years old and who presented to a Level 1 trauma center between 2008 and 2013. Data were abstracted from institutional TBI and trauma registries. Independent covariates included age, GCS score, pupillary response, CT characteristics, and Injury Severity Score. A composite outcome measure, ICU-level care, was defined as cardiopulmonary instability, transfusion, intubation, placement of intracranial pressure monitor or other invasive monitoring, and/or need for surgical intervention. Stepwise logistic regression defined significant predictors for model inclusion with p < 0.10. The authors derived the CDR with binary recursive partitioning (using a misclassification cost of 20:1). RESULTS A total of 284 patients with mild TBI were included in the analysis; 40 (14.1%) had ICU-level care. The CDR consisted of 5 final predictor variables: midline shift > 5 mm, intraventricular hemorrhage, nonisolated head injury, postresuscitation GCS score of < 15, and cisterns absent. The CDR correctly identified 37 of 40 patients requiring ICU-level care (sensitivity 92.5%; 95% CI 78.5-98.0) and 154 of 244 patients who did not require an ICU-level intervention (specificity 63.1%; 95% CI 56.7-69.1). This results in a negative predictive value of 98.1% (95% CI 94.1-99.5). CONCLUSIONS The authors derived a clinical tool that defines a subset of pediatric patients with mild TBI at low risk for ICU-level care. Although prospective evaluation is needed, the potential for improved resource allocation is significant.


Evaluating the impacts of marine debris on cetaceans.
Global in its distribution and pervading all levels of the water column, marine debris poses a serious threat to marine habitats and wildlife. For cetaceans, ingestion or entanglement in debris can cause chronic and acute injuries and increase pollutant loads, resulting in morbidity and mortality. However, knowledge of the severity of effects lags behind that for other species groups. This literature review examines the impacts of marine debris on cetaceans reported to date. It finds that ingestion of debris has been documented in 48 (56% of) cetacean species, with rates of ingestion as high as 31% in some populations. Debris-induced mortality rates of 0-22% of stranded animals were documented, suggesting that debris could be a significant conservation threat to some populations. We identify key data that need to be collected and published to improve understanding of the threat that marine debris poses to cetaceans.


[Estimation of in vitro Monclavit-1 virucide activity against influenza virus A (H5N1)].
The virucide activity of Monclavit-1 against the influenza virus A/Chicken/Kurgan/2/Russia/05 (H5N1) was investigated in the MDCK cell culture. It was shown that at a temperature of 14.0 +/- 1.0 degrees C Monclavit-1 in a 20-fold dilution within an hour inactivated the cytopathic activity of the virus and the specific hemagglutinin generation by 100 and 87.5% respectively. In a 40-fold dilution it inactivated at the average by 50% both the cytopathic activity and the specific hemagglutinin generation. In an 80-fold dilution it inactivated the cytopathic activity at the average by 20%. In a 160-fold dilution it did not inactivate the pathogen. At a temperature of 25.0 +/- 1.0 degrees C Monclavit-1 inactivated the influenza virus A/Chicken/Kurgan/2/Russia/05 (H5N1) by 90.0% only in the highest concentration.


Neuropep-1 ameliorates learning and memory deficits in an Alzheimer's disease mouse model, increases brain-derived neurotrophic factor expression in the brain, and causes reduction of amyloid beta plaques.
Alzheimer's disease (AD) is a neurodegenerative disease characterized by amyloid beta (Aβ) deposits, hyperphosphorylated tau deposition, and cognitive dysfunction. Abnormalities in the expression of brain-derived neurotrophic factor (BDNF), which plays an important role in learning and memory formation, have been reported in the brains of AD patients. A BDNF modulating peptide (Neuropep-1) was previously identified by positional-scanning synthetic peptide combinatorial library. Here we examine the neuroprotective effects of Neuropep-1 on several in vitro neurotoxic insults, and triple-transgenic AD mouse model (3xTg-AD). Neuropep-1 protects cultured neurons against oligomeric Aβ1-42, 1-methyl-4-phenylpyridinium, and glutamate-induced neuronal cell death. Neuropep-1 injection also significantly rescues the spatial learning and memory deficits of 3xTg-AD mice compared with vehicle-treated control group. Neuropep-1 treatment markedly increases hippocampal and cortical BDNF levels. Furthermore, we found that Neuropep-1-injected 3xTg-AD mice exhibit dramatically reduced Aβ plaque deposition and Aβ levels without affecting tau pathology. Neuropep-1 treatment does not alter the expression or activity of full-length amyloid precursor protein, α-, β-, or γ-secretase, but levels of insulin degrading enzyme, an Aβ degrading enzyme, were increased. These findings suggest Neuropep-1 may be a therapeutic candidate for the treatment of AD.


Acquired childhood aphasia with convulsive disorder (Landau-Kleffner syndrome). A case report.
The Landau-Kleffner syndrome is a rare form of acquired childhood aphasia associated with convulsive disorder. These children are typically misdiagnosed. A 15-year-old girl whose aphasia was not diagnosed until the age of 13 is described. Pitfalls in diagnosis are discussed, particularly with regard to interpretation of the results of audiometric testing. Attention is drawn to the existence of this rare form of aphasia and to the lack of appropriate educational facilities for aphasic children in general.


Are Gulf War veterans experiencing illness due to exposure to smoke from Kuwaiti oil well fires? Examination of Department of Defense hospitalization data.
There has been much concern among the public and veterans that specific environmental exposures incurred during the Gulf War were the cause of subsequent illness among Gulf War veterans. In this historical cohort study, the authors compared the postwar morbidity of US military personnel exposed to smoke from the 1991 Kuwaiti oil well fires with that of unexposed personnel. Complete exposure and demographic data were available for 405,142 active-duty Gulf War veterans who did not remain in the region after the war. The authors used data from all Department of Defense hospitals for the period August 1, 1991-July 31, 1999 to estimate rates of hospitalization due to any cause, hospitalization due to a diagnosis in one of 15 major categories, and hospitalization due to one of nine diagnoses likely to be manifestations of smoke exposure. Exposures to particulate matter from oil-well-fire smoke were based on the integration of meteorologic data, diffusion modeling, and troop location data. The authors constructed seven exposure groups combining duration and amount of exposure. In Cox modeling, three of the 25 models showed an increased adjusted risk of hospitalization. However, there was no evidence of a dose-response relation. Despite some limitations, these data do not support the hypothesis that Gulf War veterans have an increased risk of postwar morbidity from exposure to Kuwaiti oil-well-fire smoke.


Transforming growth factor β recruits persistent MAPK signaling to regulate long-term memory consolidation in Aplysia californica.
In this study, we explore the mechanistic relationship between growth factor signaling and kinase activity that supports the protein synthesis-dependent phase of long-term memory (LTM) consolidation for sensitization ofAplysia Specifically, we examine LTM for tail shock-induced sensitization of the tail-elicited siphon withdrawal (T-SW) reflex, a form of memory that requires both (i) extracellular signal-regulated kinase (ERK1/2; MAPK) activity within identified sensory neurons (SNs) that mediate the T-SW and (ii) the activation of transforming growth factor β (TGFβ) signaling. We now report that repeated tail shocks that induce intermediate-term (ITM) and LTM for sensitization, also induce a sustained post-training phase of MAPK activity in SNs (lasting at least 1 h). We identified two mechanistically distinct phases of post-training MAPK: (i) an immediate phase that does not require ongoing protein synthesis or TGFβ signaling, and (ii) a sustained phase that requires both protein synthesis and extracellular TGFβ signaling. We find that LTM consolidation requires sustained MAPK, and is disrupted by inhibitors of protein synthesis and TGFβ signaling during the consolidation window. These results provide strong evidence that TGFβ signaling sustains MAPK activity as an essential mechanistic step for LTM consolidation.


Determination of available amino acids and energy in alfalfa meal, feather meal, and poultry by-product meal by various methods.
Several experiments were conducted to evaluate various methodologies for determining availability of amino acids (AA) and energy in alfalfa meal (AM), feather meal (FM), and poultry by-product meal (PBPM). Digestibility of AA was determined with 48-h precision-fed cockerel assays using conventional (CONV) and cecectomized (CEC) cockerels. Amino acid bioavailability was assessed with chick growth assays. True digestibility values of most AA were significantly lower for CEC than for CONV cockerels. In the chick growth assays, partitioning weight gains to reflect only growth attributable to supplemental crystalline AA or test feedstuff AA consumption often yielded higher bioavailability values than when total weight gains were used for FM and PBPM but lower ones for AM. The magnitude of differences between AA digestibility (cockerels) and AA bioavailability (chicks) values varied among feedstuffs; the largest differences were observed for FM. The TMEn values of AM and PBPM were lower for CEC birds than for CONV birds, whereas TMEn of FM was similar for both types of birds.


Measuring quality of life in Chinese cancer patients: a new version of the Functional Living Index for Cancer (Chinese).
Since its translation into Chinese, the Functional Living Index for Cancer (FLIC) has not been widely received due to some of its difficulties. We modified its visual analogue scale (VAS) to an ordered categorical scale and changed some of the wording in the instrument. This study examined the measurement properties of the modified FLIC. The modified version of FLIC and the Functional Assessment of Cancer Therapy (FACT-G Chinese version 4) were filled in by 140 patients recruited from the National Cancer Centre Singapore. The patients' FLIC scores were compared with their clinical characteristics to establish known-group validity. Convergent and divergent validity of FLIC were examined by correlation analysis with FACT-G and its sub-scales. Cronbach's alpha and relative efficiency were also examined. FLIC and most of its sub-scales could indicate a clear and statistically significant difference of quality of life (QOL) according to patients' performance status and treatment status. FLIC strongly correlated with FACT-G. The Physical, Psychological, and Symptoms sub-scales of FLIC converged to and diverged from FACT-G sub-scales as conceptually expected. Cronbach's alpha indicated a satisfactory level of reliability. FLIC appeared to be more efficient than FACT-G, meaning that a smaller sample size will be required for FLIC than for FACT-G to achieve the same research purpose. The modified version of FLIC was found to have achieved satisfactory measurement properties. This is a user-friendly alternative to the original FLIC.


A genetic screen identifies BEND3 as a regulator of bivalent gene expression and global DNA methylation.
Epigenetic mechanisms are essential to establish and safeguard cellular identities in mammals. They dynamically regulate the expression of genes, transposable elements and higher-order chromatin structures. Consequently, these chromatin marks are indispensable for mammalian development and alterations often lead to disease, such as cancer. Bivalent promoters are especially important during differentiation and development. Here we used a genetic screen to identify new regulators of a bivalent repressed gene. We identify BEND3 as a regulator of hundreds of bivalent promoters, some of which it represses, and some of which it activates. We show that BEND3 is recruited to a CpG-containg consensus site that is present in multiple copies in many bivalent promoters. Besides having direct effect on the promoters it binds, the loss of BEND3 leads to genome-wide gains of DNA methylation, which are especially marked at regions normally protected by the TET enzymes. DNA hydroxymethylation is reduced in Bend3 mutant cells, possibly as consequence of altered gene expression leading to diminished alpha-ketoglutarate production, thus lowering TET activity. Our results clarify the direct and indirect roles of an important chromatin regulator, BEND3, and, more broadly, they shed light on the regulation of bivalent promoters.


Systematic reviews of topical fluorides for dental caries: a review of reporting practice.
This paper aims to assess systematic reviews on the caries-preventive effect of topical fluorides, identifying key content and reporting quality issues to be considered by researchers planning a review in this area. Published systematic reviews and meta-analyses of any topical fluoride intervention for caries control were included. Relevant databases were searched (December 2009), along with reference lists of included publications. Thirty-eight reports were identified and assessed. A majority of these focused on the child/adolescent population, fluoride toothpastes, no treatment/placebo comparisons, and had caries increment as the main outcome. Complete reporting of eligibility criteria (PICOS) was uncommon, except in Cochrane reviews. Less than half reported searching multiple sources and only one third reported a search strategy. Duplicate study selection and data extraction was reported in 27 (71%) and 16 (42%) reviews, respectively; quality assessment of included studies was not reported in one third of the reviews. Meta-analysis was reported in 20 (52%) reviews, with six not reporting the methods of synthesis used, 17 formally assessing heterogeneity, and 12 reporting analyses for its exploration. This study shows that some content features have been covered more often than others in existing fluoride reviews, while some relevant features are yet to be addressed. Also, reporting of several methodological aspects are below an acceptable level, except for Cochrane reviews. Current reporting guidelines for systematic reviews of interventions (e.g. PRISMA) and sources of high-quality existing reviews (e.g. The Cochrane Library) should be closely followed to enhance the validity and relevance of future topical fluoride reviews.


Comparison of synergism between colistin, fosfomycin and tigecycline against extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates or with carbapenem resistance.
To investigate the synergistic and bactericidal effects of antimicrobial combinations of any two of colistin, fosfomycin and tigecycline against the nine extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae (KP) clinical isolates, including 4 carbapenem-susceptible strains and five imipenem and/or meropenem-resistant strains. In vitro synergism and bactericidal activity of combination of colistin, fosfomycin and tigecycline were evaluated by time-kill studies in standard inoculum of bacterial densities of a suspension containing 5 × 105 CFU/mL by using 1/2× MIC for each alone, and both 1/2× and 1/4× MIC for any two drugs. The settings of low MIC dosing were allowed to rapidly survey the most active drug combination. The most active combination group was colistin plus tigecycline, showing synergy in 8 isolates and bactericidal activities in 6 isolates by using concentrations of 1/2× MIC and 1/4× MIC, respectively. The least active combination was tigecycline plus fosfomycin, which showed synergy in only 4 isolates and no bactericidal activities by using concentrations of 1/2× MIC and 1/4× MIC, respectively. The combination of tigecycline and colistin may be considered as a last-resort approach to the ESBL-producing KP infections, especially those isolates with carbapenem resistance.


Evaluation of volatile constituents of Cochlospermum angolense.
The chemical composition of the essential oils obtained from the leaves and roots of Cochlospermum angolense (Welw) growing wild in Angola was analyzed for the first time by capillary gas chromatography (GC) and gas chromatography/mass spectrometry (GC-MS). The investigation led to the identification of 67 and 130 compounds from the leaves and roots, respectively. Both oils were strongly characterized by the presence of sesquiterpenoids (68.8% in the leaves and 53.2% in the roots), while monoterpenoids were present in minor percentages (9.8% in the leaves and 26.2% in the root). The main constituents of the leaves were germacrene D (9.4%), alpha-cadinol (7.4%) and 10-epi-cubenol (6.2%), while the most abundant compounds in the root essential oil were the sesquiterpenes beta-caryophyllene (19.7%) and isoborneol (6.6%). The analysis by HS-SPME of the roots, leaves, fruits and seeds were also reported for the first time. Different volatile profiles were detected.


Engineered defective interfering RNAs of Sindbis virus express bacterial chloramphenicol acetyltransferase in avian cells.
We are investigating the feasibility of using the positive-strand RNA virus Sindbis virus and its defective interfering (DI) particles as vectors for introducing foreign genes into cells. In previous work we showed by deletion mapping of a cloned cDNA derived from one of the DI RNAs that only nucleotides at the 3' and 5' termini of the RNA are essential for the DI RNA to be amplified after it is transfected into cells in the presence of helper virus. As a first step in developing a vector we replaced 75% of the internal nucleotides of this DI cDNA with foreign sequences including the bacterial chloramphenicol acetyltransferase (CAT; EC 22.214.171.124) gene. DI RNAs transcribed from this cDNA were replicated and packaged by helper Sindbis virus and became a major viral RNA species in infected cells by the third passage after transfection. They were also translated to produce enzymatically active CAT. CAT activity was measured at passage 3 but could also be detected in transfected cells. DI RNAs containing the CAT gene were translated in vivo and in vitro to produce two polypeptides immunoprecipitable by anti-CAT antibodies. One polypeptide was identical in size to the authentic CAT polypeptide; the other was the size expected for a protein initiated at an upstream, viral-specific AUG in frame with the CAT AUG. These studies establish that DI genomes of Sindbis virus can tolerate the insertion and direct the expression of at least one foreign gene.


Dose response to inhaled salbutamol in chronic obstructive airways disease.
High dose inhaled salbutamol is increasingly used in the management of chronic obstructive airways disease. To determine the range of doses to achieve optimal bronchodilatation and the proportion of patients requiring high dose therapy we have studied 23 patients with chronic obstructive airways disease. Cumulative dose responses were measured to six incremental doses of salbutamol (0.2 to 1.2 mg) delivered by metered dose inhaler. Results were analysed by polynomial regression to calculate the smallest dose required to produce 90% maximal bronchodilatation in each patient. While 5/23 (22%) required greater than 1 mg the majority, 14/23 (61%), achieved 90% maximal bronchodilation with salbutamol 0.6 mg or less. The 8 patients with severe airflow limitation (FEV1 less than or equal to 1 litre) showed a similar pattern of response. We conclude that in chronic obstructive airways disease there are wide individual variations in the dose of inhaled salbutamol producing 90% maximal bronchodilatation with only a minority requiring high dose therapy.


Tc17 cells mediate vaccine immunity against lethal fungal pneumonia in immune deficient hosts lacking CD4+ T cells.
Vaccines may help reduce the growing incidence of fungal infections in immune-suppressed patients. We have found that, even in the absence of CD4(+) T-cell help, vaccine-induced CD8(+) T cells persist and confer resistance against Blastomyces dermatitidis and Histoplasma capsulatum. Type 1 cytokines contribute to that resistance, but they also are dispensable. Although the role of T helper 17 cells in immunity to fungi is debated, IL-17 producing CD8(+) T cells (Tc17 cells) have not been investigated. Here, we show that Tc17 cells are indispensable in antifungal vaccine immunity in hosts lacking CD4(+) T cells. Tc17 cells are induced upon vaccination, recruited to the lung on pulmonary infection, and act non-redundantly in mediating protection in a manner that requires neutrophils. Tc17 cells did not influence type I immunity, nor did the lack of IL-12 signaling augment Tc17 cells, indicating a distinct lineage and function. IL-6 was required for Tc17 differentiation and immunity, but IL-1R1 and Dectin-1 signaling was unexpectedly dispensable. Tc17 cells expressed surface CXCR3 and CCR6, but only the latter was essential in recruitment to the lung. Although IL-17 producing T cells are believed to be short-lived, effector Tc17 cells expressed low levels of KLRG1 and high levels of the transcription factor TCF-1, predicting their long-term survival and stem-cell like behavior. Our work has implications for designing vaccines against fungal infections in immune suppressed patients.