[The resistance of the bacterial flora in cancer patients to drug therapy].
Agents etiologically relevant to hospital infection have been studied in an oncologic clinic for the period of 20 years since 1969. Pronounced changes in the profile of basic infections have been observed. In 1969, Staphylococcus accounted for 46% of cases of infection and E. coli was isolated in 22% whereas in 1987 and 1988, the respective figures for staphylococcus were 18 and 16% only while for E. coli--8.4 and 8.6%, respectively. At the same time, the occurrence of all types of streptococcal infections has risen from 17 to 26-27%. As regards gram-negative bacilli such as Klebsiella spp., Ps. aeruginosa, Enterobacter, Acinetobacter and Serrata spp., their share has increased and spectrum has become wider. The level and spectrum of drug resistance have changed, too. The level of plasmid genes accounting for resistance and pathogenicity in bacterial genomes, particularly, in gram-negative bacilli, have increased. Formation of multidrug resistant strains was shown to depend both on intensity of drug treatment in the clinic and presence and activity of R plasmids, particularly, with a wide spectrum of hosts.


Conformational analysis of a model synthetic prodiginine.
A conformational analysis of a synthetic model prodiginine was carried out. In solution this compound showed a strong preference for the β conformation, in which all the heterocycles are mutually cis. This conformation provided an ideal alignment of the three N-H groups for interacting with anions when the molecule is protonated. A different conformation was also detected in d(6)-DMSO for the mesylate salt, assigned to the α conformation, in which the C ring is engaged in an intramolecular hydrogen bond with the OMe group. The formation of a homodimer was observed in concentrated CDCl(3) solutions of the neutral free base form of this prodiginine derivative. DFT calculations and the solid state structures of the hydrochloric and methanesulfonic acid salts were in good agreement with the results observed in solution. A complete study of the relative energies of different tautomers, isomers, and supramolecular complexes supported the preference for the β conformation both in water and in the gas phase.


Behavioral and socio-emotional functioning in children with selective mutism: a comparison with anxious and typically developing children across multiple informants.
We examined differences among 158 children, 44 with selective mutism (SM; M = 8.2 years, SD = 3.4 years), 65 with mixed anxiety (MA; M = 8.9 years, SD = 3.2 years), and 49 community controls (M = 7.7 years, SD = 2.6 years) on primary caregiver, teacher, and child reports of behavioral and socio-emotional functioning. Children with SM were rated lower than controls on a range of social skills, but the SM and MA groups did not significantly differ on many of the social skills and anxiety measures. However, children with SM were rated higher than children with MA and controls on social anxiety. Findings suggest that SM may be conceptualized as an anxiety disorder, with primary deficits in social functioning and social anxiety. This interpretation supports a more specific classification of SM as an anxiety disorder for future diagnostic manuals than is currently described in the literature. The present findings also have implications for clinical practice, whereby social skills training merits inclusion in intervention for children with anxiety disorders as well as children with SM.


Bacterial respiration of arsenic and selenium.
Oxyanions of arsenic and selenium can be used in microbial anaerobic respiration as terminal electron acceptors. The detection of arsenate and selenate respiring bacteria in numerous pristine and contaminated environments and their rapid appearance in enrichment culture suggest that they are widespread and metabolically active in nature. Although the bacterial species that have been isolated and characterized are still few in number, they are scattered throughout the bacterial domain and include Gram-positive bacteria, beta, gamma and epsilon Proteobacteria and the sole member of a deeply branching lineage of the bacteria, Chrysiogenes arsenatus. The oxidation of a number of organic substrates (i.e. acetate, lactate, pyruvate, glycerol, ethanol) or hydrogen can be coupled to the reduction of arsenate and selenate, but the actual donor used varies from species to species. Both periplasmic and membrane-associated arsenate and selenate reductases have been characterized. Although the number of subunits and molecular masses differs, they all contain molybdenum. The extent of the environmental impact on the transformation and mobilization of arsenic and selenium by microbial dissimilatory processes is only now being fully appreciated.


Disruption of the blood-brain barrier following ALA-mediated photodynamic therapy.
Photodynamic therapy (PDT) is a local antineoplastic treatment with the potential for tumor cell specificity. PDT using either hematoporphyrin derivatives or 5-aminolevulinic acid (ALA) has been reported to induce brain edema indicating disruption of the blood-brain barrier (BBB). We have evaluated the ability of ALA-mediated PDT to open the BBB in rats. This will permit access of chemotherapeutic agents to brain tumor cells remaining in the resection cavity wall, but limit their penetration into normal brain remote from the site of illumination. ALA-PDT was performed on non-tumor bearing inbred Fischer rats at increasing fluence levels. Contrast T(1)-weighted high field (3 T) magnetic resonance imaging (MRI) scans were used to monitor the degree of BBB disruption which could be inferred from the intensity and volume of the contrast agent visualized. PDT at increasing fluence levels between 9 and 26 J demonstrated an increasing contrast flow rate. A similar increased contrast volume was observed with increasing fluence rates. The BBB was found to be disrupted 2 hours following PDT and 80-100% restored 72 hours later at the lowest fluence level. No effect on the BBB was observed if 26 J of light was given in the absence of ALA. ALA-PDT was highly effective in opening the BBB in a localized region of the brain. The degradation of the BBB was temporary in nature at fluence levels of 9 J, opening rapidly following treatment and significantly restored during the next 72 hours. No signs of tissue damage were seen on histological sections at this fluence level. However, higher fluences did demonstrate permanent tissue changes localized in the immediate vicinity of the light source.


Glucose stimulates translocation of the homeodomain transcription factor PDX1 from the cytoplasm to the nucleus in pancreatic beta-cells.
One of the mechanisms whereby glucose stimulates insulin gene transcription in pancreatic beta-cells involves activation of the homeodomain transcription factor PDX1 (pancreatic/duodenal homeobox-1) via a stress-activated pathway involving stress-activated protein kinase 2 (SAPK2, also termed RK/p38, CSBP, and Mxi2). In the present study we show, by Western blotting and electrophoretic mobility shift assay, that in human islets of Langerhans incubated in low glucose (3 mM) PDX1 exists as an inactive 31-kDa protein localized exclusively in the cytoplasm. Transfer of the islets to high (16 mM) glucose results in rapid (within 10 min) conversion of PDX1 to an active 46-kDa form that was present predominantly in the nucleus. Activation of PDX1 appears to involve phosphorylation, as shown by incorporation of 32Pi into the 46-kDa form of the protein. These effects of glucose could be mimicked by chemical stress (sodium arsenite), or by overexpression of SAPK2 in the beta-cell line MIN6. Overexpression of SAPK2 also stimulated PDX1-dependent transcription of a -50 to -250 region of the human insulin gene promoter linked to a firefly luciferase reporter gene. The effects of glucose were inhibited by the SAPK2 inhibitor SB 203580, and by wortmannin and LY 294002, which inhibit phosphatidylinositol 3-kinase, although the effects of stress (arsenite) were inhibited only by SB 203580. These results demonstrate that glucose regulates the insulin gene promoter through activation and nuclear translocation of PDX1 via the SAPK2 pathway.


Medical care costs: trends and outlook.
Data on health care expenditures continue to show favorable developments in terms of restraining spending. The growth in spending has eased significantly, particularly for hospital care and physicians' services. Spending in the health care sector has remained relatively flat in relation to the nation's economy, reflecting significant changes to the delivery and financing of medical care. Although coming under some pressure, managed care, especially the health maintenance organization (HMO), continues to exert a dominant influence in changing the landscape of medical care in the United States. Through Medicare and Medicaid, government is expanding its role in medical care. However, indications are that spending growth has begun to trend upward. Estimates by the Health Care Financing Administration show that the health care sector's utilization of the nation's resources will once again move to an upward track.


Factors that are associated with the risk of acquiring Plasmodium knowlesi malaria in Sabah, Malaysia: a case-control study protocol.
Plasmodium knowlesi has long been present in Malaysia, and is now an emerging cause of zoonotic human malaria. Cases have been confirmed throughout South-East Asia where the ranges of its natural macaque hosts and Anopheles leucosphyrus group vectors overlap. The majority of cases are from Eastern Malaysia, with increasing total public health notifications despite a concurrent reduction in Plasmodium falciparum and P. vivax malaria. The public health implications are concerning given P. knowlesi has the highest risk of severe and fatal disease of all Plasmodium spp in Malaysia. Current patterns of risk and disease vary based on vector type and competence, with individual exposure risks related to forest and forest-edge activities still poorly defined. Clustering of cases has not yet been systematically evaluated despite reports of peri-domestic transmission and known vector competence for human-to-human transmission. A population-based case-control study will be conducted over a 2-year period at two adjacent districts in north-west Sabah, Malaysia. Confirmed malaria cases presenting to the district hospital sites meeting relevant inclusion criteria will be requested to enrol. Three community controls matched to the same village as the case will be selected randomly. Study procedures will include blood sampling and administration of household and individual questionnaires to evaluate potential exposure risks associated with acquisition of P. knowlesi malaria. Secondary outcomes will include differences in exposure variables between P. knowlesi and other Plasmodium spp, risk of severe P. knowlesi malaria, and evaluation of P. knowlesi case clustering. Primary analysis will be per protocol, with adjusted ORs for exposure risks between cases and controls calculated using conditional multiple logistic regression models. This study has been approved by the human research ethics committees of Malaysia, the Menzies School of Health Research, Australia, and the London School of Hygiene and Tropical Medicine, UK.


Priorities for comparative effectiveness reviews in cardiovascular disease.
Comparative effectiveness reviews offer a systematic method to critically appraise existing research and to identify unaddressed clinical areas in cardiovascular disease where significant morbidity, mortality, and variation in the use of resources persist. To delineate and help select areas where comparative effectiveness reviews are needed, the Effective Health Care Program of the Agency for Healthcare Research and Quality involved stakeholders in prioritization of the research agenda. We involved a diverse panel of stakeholders representing a broad range of clinical, policy, and patient perspectives. To assist in prioritization of topics for evidence synthesis, we created a framework evaluating 12 cardiovascular disease subcategories that reflect American College of Cardiology/American Heart Association disease-based guidelines. We performed an environmental scan for each disease subcategory to populate this framework with existing knowledge, levels of evidence, and degrees of public interest. Through a formalized process, 4 disease subcategories were prioritized: chronic coronary artery disease, ventricular arrhythmias, heart failure, and cerebrovascular disease. Within these subcategories, 11 topics that address the comparative safety and effectiveness of existing treatments and evaluate emerging treatments were nominated by the stakeholder panel to proceed for feasibility assessment before developing comparative effectiveness reviews. Using a systematic process deriving consensus from multiple stakeholders across cardiovascular disease states, we generated a prioritized list of evidence synthesis topics to inform decision makers. The topics vetted through this process seek to determine the comparative safety and effectiveness of a range of treatments, both established and emerging, and are immediately relevant for prevalent disease states.


Parthenolide and DMAPT exert cytotoxic effects on breast cancer stem-like cells by inducing oxidative stress, mitochondrial dysfunction and necrosis.
Triple-negative breast cancers (TNBCs) are aggressive forms of breast carcinoma associated with a high rate of recidivism. In this paper, we report the production of mammospheres from three lines of TNBC cells and demonstrate that both parthenolide (PN) and its soluble analog dimethylaminoparthenolide (DMAPT) suppressed this production and induced cytotoxic effects in breast cancer stem-like cells, derived from dissociation of mammospheres. In particular, the drugs exerted a remarkable inhibitory effect on viability of stem-like cells. Such an effect was suppressed by N-acetylcysteine, suggesting a role of reactive oxygen species (ROS) generation in the cytotoxic effect. Instead z-VAD, a general inhibitor of caspase activity, was ineffective. Analysis of ROS generation, performed using fluorescent probes, showed that both the drugs stimulated in the first hours of treatment a very high production of hydrogen peroxide. This event was, at least in part, a consequence of activation of NADPH oxidases (NOXs), as it was reduced by apocynin and diphenylene iodinium, two inhibitors of NOXs. Moreover, both the drugs caused downregulation of Nrf2 (nuclear factor erythroid 2-related factor 2), which is a critical regulator of the intracellular antioxidant response. Prolonging the treatment with PN or DMAPT we observed between 12 and 24 h that the levels of both superoxide anion and hROS increased in concomitance with the downregulation of manganese superoxide dismutase and catalase. In addition, during this phase dissipation of mitochondrial membrane potential occurred together with necrosis of stem-like cells. Finally, our results suggested that the effect on ROS generation found in the first hours of treatment was, in part, responsible for the cytotoxic events observed in the successive phase. In conclusion, PN and DMAPT markedly inhibited viability of stem-like cells derived from three lines of TNBCs by inducing ROS generation, mitochondrial dysfunction and cell necrosis.


Association between decline in brain dopamine activity with age and cognitive and motor impairment in healthy individuals.
Although it is documented that brain dopamine activity declines with age, the functional significance of this is not known. This study assessed the relation between measures of brain dopamine activity and indexes of motor and cognitive function in healthy individuals. Thirty healthy volunteers aged 24-86 years were studied with positron emission tomography and [11C]raclopride to assess dopamine D2 receptors. All subjects underwent a neuropsychological test battery that included tasks found to be sensitive to dopamine alterations in patients with neurodegenerative disease and control tasks. Transfer of [11C]raclopride from plasma to brain in the striatum and cerebellum was not affected by age. In contrast, D2 receptor availability in the caudate and putamen declined with age. Correlations between D2 receptors and neuropsychological test performance were strongest for the motor task (Finger Tapping Test) and were also significant for most tasks involving frontal brain regions, including measures of abstraction and mental flexibility (Wisconsin Card Sorting Test) and attention and response inhibition (Stroop Color-Word Test, interference score). These relationships remained significant after control for age effects. Age-related decreases in brain dopamine activity are associated with a decline in motor function and may also contribute to impaired performance on tasks that involve frontal brain regions. Interventions that enhance dopamine activity may improve performance and quality of life for the elderly. The fact that correlations remained significant after age effects were partialed out suggests that dopamine activity may influence motor and cognitive performance irrespective of age.


Improved serodiagnosis of cystic echinococcosis using the new recombinant 2B2t antigen.
A standardized test for the serodiagnosis of human cystic echinococcosis (CE) is still needed, because of the low specificity and sensitivity of the currently available commercial tools and the lack of proper evaluation of the existing recombinant antigens. In a previous work, we defined the new ELISA-B2t diagnostic tool for the detection of specific IgGs in CE patients, which showed high sensitivity and specificity, and was useful in monitoring the clinical evolution of surgically treated CE patients. Nevertheless, this recombinant antigen gave rise to false-negative results in a percentage of CE patients. Therefore, in an attempt to improve its sensitivity, we constructed B2t-derived recombinant antigens with two, four and eight tandem repeat of B2t units, and tested them by ELISA on serum samples of CE patients and patients with related parasites. The best diagnostic values were obtained with the two tandem repeat 2B2t antigen. The influence of several clinical variables on the performance of the tests was also evaluated. Finally, the diagnostic performance of the 2B2t-ELISA was compared with that of an indirect haemagglutination commercial test. The 2B2t recombinant antigen performed better than the HF and B2t antigens, and the IHA commercial kit. Therefore, this new 2B2t-ELISA is a promising candidate test for the serodiagnosis of CE in clinical settings.


[Different evolution rule of SRY gene between hominoid and Old World monkey].
The complete SRY gene of P. nemaeus and M. arctiodes were amplified by using PCR and sequenced. Combined with the same regions of other primate animals, sequence analysis confirmed the conservation of HMGbox. In molecular phylogenetic tree constructed, we made the Ka/Ks value pairwise comparison of HMGbox flanking sequences both intra- and inter- Hominoid and the Old World monkey. To our interest, the pairwise comparison of species in Hominoid suggested a higher Ka/Ks ratio than that in the Old World monkey or between the Old World monkey and marmoset, which showed existent of different pattern of evolution. In addition, in Hominoid Ka/Ks ratio for HMGbox sequences was obviously on the increase. The rate of nonsynonymous substitutions per nonsynonymous site is faster than that of synonymous substitutions per synonymous site in Hominoid (driven by positive directional selection). Our analysis suggested that it might be resulted from functional change of this gene and the system error or the close relationship among Hominoid species. It is also supposed that the positive selection was functioned at the beginning of the divergence of Hominoid species.


Speed and heart-rate profiles in skating and classical cross-country skiing competitions.
To compare the speed and heart-rate profiles during international skating and classical competitions in male and female world-class cross-country skiers. Four male and 5 female skiers performed individual time trials of 15 km (men) and 10 km (women) in the skating and classical techniques on 2 consecutive days. Races were performed on the same 5-km course. The course was mapped with GPS and a barometer to provide a valid course and elevation profile. Time, speed, and heart rate were determined for uphill, flat, and downhill terrains throughout the entire competition by wearing a GPS and a heart-rate monitor. Times in uphill, flat, and downhill terrain were ~55%, 15-20%, and 25-30%, respectively, of the total race time for both techniques and genders. The average speed differences between skating and classical skiing were 9% and 11% for men and women, respectively, and these values were 12% and 15% for uphill, 8% and 13% for flat (all P < .05), and 2% and 1% for downhill terrain. The average speeds for men were 9% and 11% faster than for women in skating and classical, respectively, with corresponding numbers of 11% and 14% for uphill, 6% and 11% for flat, and 4% and 5% for downhill terrain (all P < .05). Heart-rate profiles were relatively independent of technique and gender. The greatest performance differences between the skating and classical techniques and between the 2 genders were found on uphill terrain. Therefore, these speed differences could not be explained by variations in exercise intensity.


Protection of mice against lethal doses of 1 beta-D-arabinofuranosylcytosine by pluripotent stem cell inhibitors.
The aim of this work was to study whether an inhibitor of pluripotent stem cell (CFU-S) recruitment, which we have shown previously to be able to increase the number of CFU-S after a fractionated treatment with 1-beta-D-arabinofuranosylcytosine, could increase the survival of mice given injections of lethal doses of the same drug. Two protocols of 1-beta-D-arabinofuranosylcytosine treatment were used in two different mouse strains, which both killed the mice within a week. An inhibitor of CFU-S was prepared by dialysis from fetal calf marrow, and a first step of purification was made by chromatography on Sephadex G-10. When given injections 2 hr before the drug, the number of surviving mice was increased significantly with the dialysate; fractions separated by chromatography appeared to be more effective to increase the animal survival. These preliminary results indicate that a factor of low molecular weight (below M.W. 3500) extracted from fetal calf marrow is able to protect animals during 1-beta-D-arabinofuranosylcytosine treatment. The inhibitor seems to be specific for CFU-S, without any inhibiting effect on tumor cell kinetics in vitro. If the absence of species specificity found for higher to lower species is confirmed for the lower to the higher species, then this inhibitor could be an effective tool during cancer chemotherapy.


AMPK exerts dual regulatory effects on the PI3K pathway.
AMP-activated protein kinase (AMPK) is a fuel-sensing enzyme that is activated when cells experience energy deficiency and conversely suppressed in surfeit of energy supply. AMPK activation improves insulin sensitivity via multiple mechanisms, among which AMPK suppresses mTOR/S6K-mediated negative feedback regulation of insulin signaling. In the present study we further investigated the mechanism of AMPK-regulated insulin signaling. Our results showed that 5-aminoimidazole-4-carboxamide-1 ribonucleoside (AICAR) greatly enhanced the ability of insulin to stimulate the insulin receptor substrate-1 (IRS1)-associated PI3K activity in differentiated 3T3-F442a adipocytes, leading to increased Akt phosphorylation at S473, whereas insulin-stimulated activation of mTOR was diminished. In 3T3-F442a preadipocytes, these effects were attenuated by expression of a dominant negative mutant of AMPK alpha1 subunit. The enhancing effect of ACIAR on Akt phosphorylation was also observed when the cells were treated with EGF, suggesting that it is regulated at a step beyond IR/IRS1. Indeed, when the cells were chronically treated with AICAR in the absence of insulin, Akt phosphorylation was progressively increased. This event was associated with an increase in levels of phosphatidylinositol -3,4,5-trisphosphate (PIP3) and blocked by Wortmannin. We then expressed the dominant negative mutant of PTEN (C124S) and found that the inhibition of endogenous PTEN per se did not affect phosphorylation of Akt at basal levels or upon treatment with AICAR or insulin. Thus, this result suggests that AMPK activation of Akt is not mediated by regulating phosphatase and tensin homologue (PTEN). Our present study demonstrates that AMPK exerts dual effects on the PI3K pathway, stimulating PI3K/Akt and inhibiting mTOR/S6K.


Amniotic Suspension Allograft and Fetal Bovine Acellular Dermal Matrix to Treat Complex, Acute, Full-thickness Wounds: A Retrospective Analysis of Safety and Treatment Efficacy.
Comorbidities; inadequate vascularity; exposure of bones, tendons, or other avascular structures; and loss or removal of significant tissue volume all complicate the clinical treatment of patients with large, acute wounds. A number of amniotic tissue products are currently available for wound healing and other applications; one of these is a human amniotic suspension allograft (ASA) consisting of particulated human amniotic membrane and cells from the amniotic fluid from the same human donor. In this study, the use of ASA with a fetal-derived bovine acellular dermal matrix to promote the healing of large, acute, full-thickness wounds is evaluated. The authors herein hypothesize treatment with ASA may allow for the healing of these wounds. This study consisted of a chart review of 33 patients, with an average age of 42.2 years, and large acute wounds that were treated with a fetal-derived bovine acellular dermal matrix adsorbed with ASA. To the best of the authors' knowledge, this is the first study to date to investigate the usefulness of ASA in wound healing for large, complex, acute wounds. In this study, 30 of 33 patients were confirmed as fully healed, and 3 patients were lost to follow-up. The average wound size was 537.4 cm2, and the average time to split-thickness skin grafting (STSG) was 30.5 days, with an average time of 6.8 days until at least 95% graft take was achieved. Of the patient population studied, 45.5% had 1 or more significant comorbidities, 30.3% had wounds larger than 500 cm2, and 39.4% had exposed bone or tendon. In a small but challenging population including a high number of patients with comorbidities and exposed bone or tendon, it was found that ASA delivery, along with a dermal matrix, was successful in treating large, complex, acute wounds.


Enhanced therapeutic effect on androgen-independent prostate cancer by depsipeptide (FK228), a histone deacetylase inhibitor, in combination with docetaxel.
To study whether a histone deacetylase inhibitor, FK228, can enhance therapeutic efficacies of other chemotherapeutic agents against androgen-independent prostate cancer (AIPCa) and to explore any possible early biomarker for predicting drug response. The therapeutic effects of FK228-based combinations on three AIPCa cell lines (PC-3, DU145, and C4-2) were evaluated by synergisms of cytotoxicity. Cell cycle analysis and Western blot assay were used to study the underlying mechanisms of drug action and search for any potential surrogate biomarker. In addition, we investigated the in vivo antitumor effects of the FK228/docetaxel combination using the PC-3 xenograft model. FK228/docetaxel surpassed other FK228-based combinations by achieving more synergism of cytotoxicity. FK228 enhanced the therapeutic effect of docetaxel against AIPCa by exhibiting markedly enhanced and prolonged inhibitory effects in vitro and better tumor regression in vivo by inducing apoptosis. It appears that p21(WAF1) induction consistently correlates with single or combination treatment. The results of our study have shown that FK228 is able to enhance the therapeutic effect of docetaxel against AIPCa. FK228 appears to be a promising second-line option in combination with docetaxel. The p21(WAF1) protein level can be used as a surrogate biomarker to predict and monitor the therapeutic response.


Quercetin reduces susceptibility to influenza infection following stressful exercise.
Exercise stress is associated with increased risk for upper respiratory tract infection. We have shown that exercise stress can increase susceptibility to infection. Quercetin, a flavonoid present in a wide variety of fruits and vegetables, has been reported to inhibit infectivity and replication of a broad spectrum of viruses and may offset the increase in susceptibility to infection associated with stressful exercise. This study examined the effects of quercetin feedings on susceptibility to the influenza virus A/Puerto Rico/8/34 (H1N1) following stressful exercise. Mice were randomly assigned to one of four treatment groups: exercise-placebo, exercise-quercetin, control-placebo, or control-quercetin. Exercise consisted of a run to fatigue (approximately 140 min) on a treadmill for 3 consecutive days. Quercetin (12.5 mg/kg) was administered via gavage for 7 days before viral challenge. At 30 min after the last bout of exercise or rest, mice (n=23-30) were intranasally inoculated with a standardized dose of influenza virus (0.04 hemagglutinating units). Mice were monitored daily for morbidity (time to sickness), symptom severity, and mortality (time to death) for 21 days. Exercise stress was associated with an increased susceptibility to infection [morbidity, mortality, and symptom severity on days 5-7 (P<0.05)]; quercetin offset the increase in susceptibility to infection [morbidity, mortality, and symptom severity on days 5-7 (P<0.05)] that was associated with stressful exercise. These data suggest that short-term quercetin feedings may prove to be an effective strategy to lessen the impact of stressful exercise on susceptibility to respiratory infection.


Binding of surface IgG by immunocompetent thymus cells.
We have previously shown that myeloma-derived or serum-derived immunoglobulin could facilitate in vitro alloantigen recognition by immunocompetent thymus cells ('subset-I thymocytes'). The present report demonstrates the isolation of surface immunoglobulins from radiolabeled splenic B lymphocytes. Solubilized radiolabeled preparations of surface immunoglobulin are shown to bind in vitro to lymphoid cells, especially to immunocompetent thymus cells. Immunocompetent thymus cells (subset-I thymocytes preferentially bind more IgG than IgM. Papain digests of IgG are also bound by subset-I thymocytes; preferential binding of the Fc piece is observed. Although the binding of IgG or its pieces per se does not activate lymphoid cells in vitro a role for the binding of IgG during alloantigen recognition in vitro between immunocompetent thymus cells is demonstrated.


Safety of selective cyclooxygenase-2 inhibitors in inflammatory bowel disease.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are relatively contraindicated in patients with inflammatory bowel disease (IBD) for fear of disease aggravation. Cyclooxygenase-2 inhibitors have fewer GI side effects than traditional NSAIDs in unselected patients. We report the safety of these agents in patients with IBD. Patients with Crohn's disease, ulcerative colitis, or pouchitis who used celecoxib or rofecoxib were identified from computerized prescription records. A retrospective chart review was conducted. Concomitant medications, past NSAID use, indication for cyclooxygenase-2 inhibitor, dose, and duration were obtained. IBD disease activity before cyclooxygenase-2 inhibitor use was graded using a modified disease activity index. Change in disease activity was graded as improved, no change, or worsened. Patients were contacted to provide data not found in the charts. The proportion of patients receiving cyclooxygenase-2 inhibitors who experienced exacerbation of IBD was determined. Eleven patients were treated with celecoxib (median dose = 200 mg/day), and 16 patients were treated with rofecoxib (median dose = 25 mg/day). Median duration of therapy was 9 months (range = 1 wk-22 months). The drug was beneficial in 14 patients, of partial benefit in eight, and of no benefit in five. Two patients (7.4%) (95% CI = 2-23%) had aggravations of IBD. Three patients (11%) had other adverse events (renal insufficiency, rash, and asymptomatic colonic ulceration). All adverse events were reversible. Our preliminary results suggest that cyclooxygenase-2 inhibitors may be safe and beneficial in most patients with IBD. A placebo-controlled trial to confirm these preliminary observations is needed.


SmTR2/4, a Schistosoma mansoni homologue of TR2/TR4 orphan nuclear receptor.
cDNA clones encoding a Schistosoma mansoni homologue of the TR2/TR4 group of nuclear receptors, SmTR2/4, were identified by screening an adult female worm cDNA library. SmTR2/4 is a 1,943 amino acid protein, the largest member of the TR2/TR4 group of nuclear receptors and also the largest nuclear receptor reported to date. SmTR2/4 retains a typical domain organisation of nuclear receptors exhibiting 69-77% sequence identity in the DNA binding domain and 16-22% sequence identity in the ligand binding domain compared with its orthologues. SmTR2/4 contains a large A/B domain and hinge region. SmTR2/4 also contains a 100 amino acid F domain, which is absent from its orthologues. SmTR2/4 mRNA is expressed in every stage of the S. mansoni life cycle, exhibiting an elevated expression level in cercariae. Western blot analysis identified two forms of SmTR2/4 protein in adult worms. Our in vitro DNA binding assay showed that SmTR2/4 binds to the DR-3 consensus hormone response element, suggesting a functional conservation among the TR2/TR4 group members in terms of DNA binding specificity. A yeast-based transactivation assay demonstrated that the A/B domain, F domain and N-terminal part of the hinge region in SmTR2/4, when tethered to a GAL4 DNA binding domain, exhibited an autonomous transcription activation function.


Solution structure of a designed amphipathic antimicrobial synthetic peptide, PGAa.
A designed peptide, PGAa showed an excellent antifungal activity as well as an efficient bactericidal activity toward gram-positive, especially in the pathogenic yeast Candida albicans 28838. The solution structures of PGAa have been determined both in 40% TFE/water solution and DPC micelle by CD and NMR spectroscopy. Based on NOEs, vicinal coupling constants, backbone amide exchange rates, and chemical shift indices, PGAa formed a long amphipathic alpha-helical conformation in both TFE and DPC micelle environments, spanning the residues Ile(2)-Ala(19) in TFE and Lys(5)-Ala(19) in DPC micelle, respectively. Solution structures suggested that the hydrophobic residues would interact with the fatty acyl chains of the lipid bilayer, while the positively charged side-chains exposed to aqueous environments. Therefore, we conclude that the alpha-helical structure as well as the highly amphiphatic nature of PGAa peptide may play a critical role in its antimicrobial activity as well as selectivities in different species.


Toluene, halothane, 1,1,1-trichloroethane and oxazepam produce ethanol-like discriminative stimulus effects in mice.
Drug discrimination procedures were used to compare the discriminative stimulus properties of ETOH and volatile inhalants. Mice were trained to discriminate between i.p. injections of ETOH (1 g/kg) and saline in a two-lever operant task. Stimulus generalization was examined after 20-min inhalation exposures to toluene (300-5400 ppm), halothane (500-8000 ppm) and 1,1,1,-trichloroethane (TCE; 125-14000 ppm). Oxazepam (10-30 mg/kg i.p.) also was tested. Concentration- or dose-related increases in ETOH-lever responding were observed with toluene, halothane, TCE and oxazepam. Shared discriminative stimulus properties among toluene, halothane, TCE and ETOH indicate that some volatile inhalants share pharmacological properties with ETOH. Previous research has shown that these inhalants also produce pentobarbital-like discriminative stimulus effects in mice. Thus, at least some inhaled anesthetics and abused solvents have behavioral effects in common with abused central nervous system depressant drugs such as ETOH and barbiturates.


[Conformational mechanism for transforming energy during active ion transport in a biological membrane].
A statistical model of active transport of ions in the biomembrane is presented. The coefficient of energy conversion for linear and non-linear processes in this system is obtained. It is shown that in the linear case this coefficient is equal to that obtained in linear thermodynamics. On the basis of the maximum of this coefficient transport parameters for ions are found. The results of the statistical model are in a good agreement with the experiments on the biological membranes.


Analysis of sea-island cotton and upland cotton in response to Verticillium dahliae infection by RNA sequencing.
Cotton Verticillium wilt is a serious soil-borne vascular disease that causes great economic loss each year. However, due to the lack of resistant varieties of upland cotton, the molecular mechanisms of resistance to this disease, especially to the pathogen Verticillium dahliae, remain unclear. We used the RNA-seq method to research the molecular mechanisms of cotton defence responses to different races of Verticillium dahliae by comparing infected sea-island cotton and upland cotton. A total of 77,212 unigenes were obtained, and the unigenes were subjected to BLAST searching and annotated using the GO and KO databases. Six sets of digital gene expression data were mapped to the reference transcriptome. The gene expression profiles of cotton infected with Verticillium dahliae were compared to those of uninfected cotton; 44 differentially expressed genes were identified. Regarding genes involved in the phenylalanine metabolism pathway, the hydroxycinnamoyl transferase gene (HCT) was upregulated in upland cotton whereas PAL, 4CL, CAD, CCoAOMT, and COMT were upregulated in sea-island cotton. Almost no differentially expressed genes in this pathway were identified in sea-island cotton and upland cotton when they were infected with V. dahliae V991 and V. dahliae D07038, respectively. Our comprehensive gene expression data at the transcription level will help elucidate the molecular mechanisms of the cotton defence response to V. dahliae. By identifying the genes involved in the defence response of each type of cotton to V. dahliae, our data not only provide novel molecular information for researchers, but also help accelerate research on genes involved in defences in cotton.


Oxidative damage, protein synthesis, and protein degradation in Alzheimer's disease.
A large number of studies has firmly established that increases in oxidative damage occurs in Alzheimer's disease (AD). Such studies have demonstrated that increased in oxidative damage selectively occurs within the brain regions involved in regulating cognitive performance. Studies from our laboratory and others have provided experimental evidence that increased levels of oxidative damage occur in subjects with Mild Cognitive Impairment (MCI), which is believed to be one of the earliest stages of AD, and is a condition which is devoid of dementia or the extensive neurofibrillary pathology and neuritic plaque deposition observed in AD. Together, these data support a role for the accrual of oxidative damage potentially serving as an early event that then initiates the development of cognitive disturbances and pathological features observed in AD. Recent studies from our laboratory have demonstrated that a decline in protein synthesis capabilities occurs in the same brain regions which exhibit increased levels of oxidative damage in MCI and AD subjects. The focus of this review is to describe the large number of studies which suggest protein synthesis may be one of the earliest cellular processes disrupted by oxidative damage in AD. Taken together, these findings have important implications for understanding the molecular and cellular basis of AD, understanding the basis for oxidative stress in AD, and may have important implications for studies involving proteomics and proteolysis in AD.


Treatment of patients with tardive dystonia with olanzapine.
Tardive dystonia represents a complication of long-term use of neuroleptics and its treatment is often unsatisfactory. Atypical neuroleptics appear to improve tardive dystonia, and cases of tardive dystonia successfully managed with clozapine have been reported. The aim of this open-label video-blinded study was to evaluate the antidystonic efficacy of olanzapine, a new atypical neuroleptic with a low risk of agranulocytosis, in a group of four patients (one man and three women) with tardive cervical dystonia. They developed severe dystonia after several years of neuroleptic treatment. Extensive laboratory evaluations, as well as neurophysiologic and neuroradiologic investigations, were negative. Olanzapine was started at a dose of 5 mg/d and increased up to 7.5 mg/d. All patients were evaluated at baseline and after 2, 4, 8, and 12 weeks of treatment, using the Toronto Western Spasmodic Torticollis Rating Scale, and videotaped. At the end of the trial, the videotapes were reviewed and scored by a blind observer. A self-rating visual analog scale completed the disability evaluation.A moderate to marked improvement in dystonia was observed in all patients, and significant differences were observed in Toronto Western Spasmodic Torticollis Rating Scale scores and videotape ratings after 8 and 12 weeks of treatment compared with the basal values (p < 0.05). The average percentage of improvement in Toronto Western Spasmodic Torticollis Rating Scale score and visual analog scale was 26.4% and 42.6%, respectively. No serious side effects were reported at the maximum dosage reached (7.5 mg/d). This study warrants a larger controlled study to conclusively demonstrate the efficacy of olanzapine in tardive dystonia.


Histone deacetylase inhibitor attenuates the effects of 27-hydroxycholesterol on the rat brain.
Hypercholesterolemia is a risk factor for Alzheimer's disease (AD). Plasma cholesterol does not pass the blood-brain barrier whereas its metabolite 27-hydroxycholesterol (27-OHC) can enter the brain. High 27-OHC in the brain has been suggested to mediate hypercholesterolemia-induced impairments of learning and memory through promoting amyloid-β accumulation and facilitating synaptic disruption. In AD brains, the activity of histone deacetylase (HDAC) is elevated. Treating AD animals with HDAC inhibitors decreases amyloid-β levels and synaptic damages, which leads to memory improvement. Whether HDAC activity is involved in the actions of 27-OHC is still uncertain. In this study, 4 weekly injections of 27-OHC/vehicle were given to rats followed by 3 daily injections of HDAC inhibitor trichostatin (TSA)/vehicle. The results of Morris water maze test reveal that all rats have intact spatial learning ability during the 5-d training phase. However, the behavioral performance during the probe trial was impaired by 27-OHC treatment, which was improved by adding TSA treatments. Furthermore, 27-OHC treatments reduced the hippocampal levels of acetylated histone H3, acetylated α tubulin, insulin-degrading enzyme and postsynaptic protein PSD-95, indicating that 27-OHC treatments may induce enhanced HDAC activity, decreased amyloid-β clearance and synaptic disruption. All reduced levels returned to the basal levels by adding TSA treatments. These findings support our hypothesis that HDAC activity is enhanced following long-term exposure to excess 27-OHC.


Iterative correction process for optical thin film synthesis with the Fourier transform method.
Several errors inherent to the Fourier transform method for optical thin film synthesis, including the inaccuracy of the spectral functions Q (sigma) used in the Fourier transforms, are compensated numerically by using successive approximations. We show that the complex phase of Q (sigma) is a key parameter which can be exploited to reduce significantly the thickness of the synthesized films and to control the shape of the refractive index profiles without affecting the spectral performance. This method is compared to other well established thin film design techniques.


Technetium-99m dithiocarbamates as potential brain agents: evaluation of aliphatic and amine-containing analogues.
Technetium-99m forms a lipophilic complex with diethyldithiocarbamate (DDC) but this complex is not retained in the brain as is thallium-201 DDC. We therefore prepared a series of aliphatic and amine-containing analogues of DDC and evaluated the properties of their 99mTc complexes in vitro and in vivo. The ethyl acetate/buffer partition coefficients of the amine-containing derivatives showed a response to pH. Four of the five complexes were less stable than DDC in vitro and showed greater retention in rabbit brain. These agents warrant further investigation for cerebral perfusion imaging with SPECT.


Over-the-scope clip as first-line therapy for ulcers with high-risk bleeding stigmata is efficient compared to standard endoscopic therapy.
Background and study aims Ulcers with high-risk stigmata have significant rebleeding rates despite standard endoscopic therapy. Data on over-the-scope clip (OTSC) for recurrent bleeding is promising but data on first line therapy is lacking. We report comparative outcomes of OTSC as first-line therapy versus standard endoscopic therapy in ulcers with high-risk stigmata. Patients and methods Consecutive adults who underwent endoscopic therapy for ulcers with high-risk stigmata between July 2019 to September 2020 were included. Patients were grouped into OTSC or standard therapy based on first-line therapy used on index endoscopy. Outcomes measured included: 1) intra-procedural hemostasis based on endoscopic documentation of adequate hemostasis; 2) 7-day rebleeding (> 2 g/dL drop in hemoglobin, hematochezia or hemorrhagic shock); 3) cost of endoscopic interventions; and 4) procedure duration measured as endoscope insertion to removal time. Cost of tools used during the index endoscopy was included. Results Sixty-eight patients were included, 47 were in standard therapy and 21 in the OTSC group. Hemostasis was achieved in 95.2 % in the OTSC group compared to 83.0 % in the standard therapy group ( P = 0.256, number needed to treat [NNT]: 9). Procedure time was shorter in the OTSC group (23 vs. 16 minutes, P = 0.002). Cost of endoscopic interventions were comparable, P = 0.203. Early rebleeding was less often in OTSC group, two (9.5 %) compared to 10 (21.3 %) in standard therapy group, NNT 9. Conclusions Use of OTSCs as first-line treatment for ulcers bleed probably improves hemostasis and decreases early rebleeding. Use of OTSC as first-line therapy shortened procedure duration without increasing the cost of endoscopic interventions.


Influence of Acetylcholinesterase Inhibitors Used in Alzheimer's Disease Treatment on the Activity of Antioxidant Enzymes and the Concentration of Glutathione in THP-1 Macrophages under Fluoride-Induced Oxidative Stress.
It has been reported that donepezil and rivastigmine, the acetylcholinesterase (AchE) inhibitors commonly used in the treatment of Alzheimer's disease (AD), do not only inhibit AChE but also have antioxidant properties. As oxidative stress is involved in AD pathogenesis, in our study we attempted to examine the influence of donepezil and rivastigmine on the activity of antioxidant enzymes and glutathione concentration in macrophages-an important source of reactive oxygen species and crucial for oxidative stress progression. The macrophages were exposed to sodium fluoride induced oxidative stress. The antioxidant enzymes activity and concentration of glutathione were measured spectrophotometrically. The generation of reactive oxygen species was visualized by confocal microscopy. The results of our study showed that donepezil and rivastigmine had a stimulating effect on catalase activity. However, when exposed to fluoride-induced oxidative stress, the drugs reduced the activity of some antioxidant enzymes (Cat, SOD, GR). These observations suggest that the fluoride-induced oxidative stress may suppress the antioxidant action of AChE inhibitors. Our results may have significance in the clinical practice of treatment of AD and other dementia diseases.


Photoaffinity labeling of benzodiazepine receptors causes altered agonist-antagonist interactions.
Previous studies have shown that [3H]flunitrazepam forms irreversible cross-links with brain tissue when exposed to ultraviolet irradiation. Comparison of the amount of [3H]flunitrazepam irreversibly incorporated and the number of benzodiazepine binding sites blocked after photolabeling has indicated that several binding sites are inactivated for each molecule of [3H]flunitrazepam incorporated. To learn the cause of this discrepancy, binding to the benzodiazepine binding sites has been examined using several radiolabeled benzodiazepine antagonists. Binding of a beta-carboline ester, CGS-8216, and Ro 15-1788 was not altered by photolabeling; however, displacement studies revealed that photolabeling converted a homogeneous set of benzodiazepine binding sites into two subsets: one of high affinity (unaltered sites) and one of low affinity. The low affinity sites could be detected by displacement studies of antagonist binding by benzodiazepines, and conversion to a low affinity form accounts for the discrepancy observed after photolabeling using [3H]flunitrazepam as ligand.


Predictors of long-term immunological outcome in rebounding patients on protease inhibitor-based HAART after initial successful virologic suppression: implications for timing to switch.
To assess predictive factors of long-term immune restoration in patients who started protease inhibitor (PI)-based HAART and experienced virological rebound after initial complete success. A retrospective longitudinal analysis of all HIV-infected patients who started their first PI-based HAART and reached viral load below 500 copies/mL was carried out in a large academic center in Italy. Patients were classified either as complete virologic responder (CR) or rebounders (REB) when confirmed plasma viremia was detected thereafter. Immunological outcome was the area under the curve (AUC) of the absolute CD4+ cell count change since the 8th month after treatment initiation (CD4+ T-cell AUC). Association between baseline characteristics, virological outcome, and CD4+ T-cell AUC was assessed by univariate and multivariate analysis. There were 374 patients who were included in the study. Mean follow-up was 30.2 months. There were 226/374 patients (60.4%) who remained CR while 148/374 (39.6%) presented at least one rebound (REB). Among REB patients, complete viral suppression was regained in 15/42 (35.7%) and 50/106 (47.1%) patients who underwent therapy changes or not, respectively. When multiple linear regression was carried out, previous nucleoside reverse transcriptase inhibitor (NRTI) experience and baseline CD4+ cell count below 350 cells/muL did not impair long-term immune restoration. The occurrence of rebound, its duration (> 18 months), and its magnitude (peak of viral load > 10,000 copies/mL) were independent negative prognostic factors. The occurrence of viral rebound is independently associated with significantly impaired long-term immunological restoration. The magnitude of viral rebound (< 10,000 copies/mL) and its duration (< 18 months) may be useful to identify those rebounding patients who may still profit from maintaining the current failing therapy if a more aggressive approach may be expected to be deleterious for tolerability reasons or lack of options.


Models of the relationship of stress, depression, and other psychosocial factors to smoking behavior: a comparison of a cohort of students in grades 6 and 8.
Six specific hypotheses regarding putative mechanisms by which stressful life events might lead to initiation of smoking among adolescents were proposed and tested on a Grade 6 cohort of students in Scarborough, Ontario, Canada. In addition, the data were used to determine the set of risk factors for initiation of smoking most pertinent to the experience of the cohort. The same relationships were examined for the 1,543 students when they were in Grade 8 and compared to the earlier Grade 6 results. The hypotheses include the effects of personal resources (coping, self-esteem, social support, and mastery), social conformity, rebelliousness, attitudes, smoking environment factors, and gender differences. The hypotheses were not unequivocally supported, except for the hypotheses about attitudes and smoking environment as well as gender effects. Males and females differ with regard to the variables and interrelationships in both years and in the final models developed. In Grade 6, there are more smoking environment items for males than for females. By Grade 8, male smoking is influenced by mastery, social conformity, and rebelliousness, while for females environmental smoking and rebelliousness are important. Male and female students differ in how stress, depression, and smoking are related in the presence of psychosocial factors.


Emergence of human rotavirus genotype G9 in metropolitan Detroit between 2007 and 2009.
Between January 2007 and April 2009, rotavirus (RV)-positive stool samples from 238 children with acute gastroenteritis, seen at Children's Hospital of Michigan in Detroit, USA, were collected and RV genotyping was performed. G and P genotypes were determined by RT-PCR and nucleotide sequencing was conducted on selected G9 and P[6] strains. Correlation between the severity of gastroenteritis episode and the infecting G genotype was done using a 14-point scoring system. The predominant G genotype was G9 (39.5 %), followed by G1 (35.3 %) and G4 (15.5 %), while P[8] was the most prevalent P genotype (66.5 %), followed by P[4] (21.9 %) and P[6] (11.2 %). The gene combinations G1P[8] and G9P[8] were the most prevalent (21.4 % and 20.6 %, respectively), followed by G4P[8] (13 %) and G9P[6] (8.8 %). Immunization data showed that only 17/238 (7.1 %) children received ≥one dose of RV vaccine (the pentavalent vaccine RotaTeq or the monovalent vaccine Rotarix) and that 10/17 were infected with G4P[8] strains. Severity of RV gastroenteritis episodes was not related to the infecting G genotype. Our results suggest a high proportion of genotype G9 strains in combination with P[8], P[6] and P[4] specificity circulating in the metropolitan Detroit area. While the protective efficacy of the RV vaccines has been demonstrated against G9P[8] strains, the level of cross-protection offered by the vaccines against G9 strains with P[6] and P[4] genotypes in the Detroit paediatric population remains to be determined.


Non-invasive tryptophan fluorescence measurements as a novel method of grading cataract.
Development of non-invasive treatments for cataract calls for a sensitive diagnostic assay. We conducted a study to test whether the ratio of folded tryptophan to non-tryptophan fluorescence emission (F-factor) may be used for grading cataracts in human lenses. The F-factor was measured on aspirated lens material from eyes undergoing femtosecond laser assisted cataract surgery (FLACS) and was compared to a preoperative optical grading of cataract using Scheimpflug imaging. The preoperative optical grading allocated the cataracts to 1 of 4 categories according to the density of the cataract. All cataracts were age-related. Lens material from 16 eyes of 14 patients was included in the study. Cataracts were preoperatively graded in categories 1, 2 and 3. No lenses were category 4. For nuclear cataracts mean values of F-factor were 52.9 (SD 12.2), 61.7 (SD 5.3) and 75.7 (SD 8.9) for categories 1, 2 and 3 respectively. Linear regression on F-factor as a function of preoperative grading category showed increasing values of F-factor with increasing preoperative grading category, R2 = 0.515. Our experiment showed that preoperative optical grading of cataracts by Scheimpflug imaging may correlate to measures of tryptophan and non-tryptophan fluorescence in human lenses. Based on our results we find that measuring the ratio between tryptophan- and non-tryptophan fluorescence may be a future tool for grading cataracts, but further research is needed.


Bacterial Diversity Associated With the Rhizosphere and Endosphere of Two Halophytes: Glaux maritima and Salicornia europaea.
Root-associated microbial communities are very important in the adaptation of halophytes to coastal environments. However, little has been reported on microbial community structures related to halophytes, or on comparisons of their compositions among halophytic plant species. Here, we studied the diversity and community structure of both rhizosphere and root endosphere bacteria in two halophytic plants: Glaux maritima and Salicornia europaea. We sampled the rhizosphere, the root endosphere, and bulk control soil samples, and performed bacterial 16S rRNA sequencing using the Illumina MiSeq platform to characterize the bacterial community diversities in the rhizosphere and root endosphere of both halophytes. Among the G. maritima samples, the richness and diversity of bacteria in the rhizosphere were higher than those in the root endosphere but were lower than those of the bulk soil. In contrast for S. europaea, the bulk soil, the rhizosphere, and the root endosphere all had similar bacterial richness and diversity. The number of unique operational taxonomic units within the root endosphere, the rhizosphere, and the bulk soil were 181, 366, and 924 in G. maritima and 126, 416, and 596 in S. europaea, respectively, implying habitat-specific patterns for each halophyte. In total, 35 phyla and 566 genera were identified. The dominant phyla across all samples were Proteobacteria and Bacteroidetes. Actinobacteria was extremely abundant in the root endosphere from G. maritima. Beneficial bacterial genera were enriched in the root endosphere and rhizosphere in both halophytes. Rhizobium, Actinoplanes, and Marinomonas were highly abundant in G. maritima, whereas Sulfurimonas and Coleofasciculus were highly abundant in S. europaea. A principal coordinate analysis demonstrated significant differences in the microbiota composition associated with the plant species and type of sample. These results strongly indicate that there are clear differences in bacterial community structure and diversity between G. maritima and S. europaea. This is the first report to characterize the root microbiome of G. maritima, and to compare the diversity and community structure of rhizosphere and root endosphere bacteria between G. maritima and S. europaea.


The effects of carbohydrate supplementation on immune responses to a soccer-specific exercise protocol.
The aim of this study was to determine the effect of carbohydrate (CHO) versus placebo (PLA) beverage consumption on the immune and plasma cortisol responses to a soccer-specific exercise protocol in 8 university team soccer players. In a randomized, counterbalanced design, the players received carbohydrate or placebo beverages before, during and after two 90 min soccer-specific exercise bouts (3 days apart) designed to mimic the activities performed and the distance covered in a typical soccer match. Blood and saliva samples were collected before, during and after the exercise protocol. Plasma lactate concentration increased to approximately 4 mmol x l(-1) at 45 and 90 min of exercise in both treatments (P<0.01). Plasma glucose concentration was significantly lower after 90 min of exercise with ingestion of the placebo than the carbohydrate (PLA: 4.57+/-0.12 mmol x l(-1); CHO: 5.49+/-0.11 mmol x l(-1); P<0.01). The pattern of change in plasma cortisol, circulating lymphocyte count and saliva immunoglobulin A secretion did not differ between the carbohydrate and placebo trials. Blood neutrophil counts were 14% higher 1 h after the placebo trial than the carbohydrate trial (PLA: 4.8+/-0.5x10(9) cells x l(-1); CHO: 4.2+/-0.5x10(9) cells x l(-1); P = 0.06), but the treatment had no effect on the degranulation response of blood neutrophils stimulated by bacterial lipopolysaccharide. We conclude that, although previous studies have shown that carbohydrate feeding is effective in attenuating immune responses to prolonged continuous strenuous exercise, the same cannot be said for a soccer-specific intermittent exercise protocol. When overall exercise intensity is moderate, and changes in plasma glucose, cortisol and immune variables are relatively small, it would appear that carbohydrate ingestion has only a minimal influence on the immune response to exercise.


The role of adenosine following pulmonary vein isolation in patients undergoing catheter ablation for atrial fibrillation: a systematic review.
Pulmonary vein reconnection is a major limitation of pulmonary vein isolation (PVI) for symptomatic atrial fibrillation (AF). Adenosine may unmask dormant PV conduction and facilitate consolidation of PV isolation. We performed a systematic review of the literature to determine the impact of routine adenosine administration on clinical outcomes in patients undergoing PVI. References and electronic databases reporting AF ablation and adenosine following PVI were searched through to 31 July 2012. Six studies included 544 patients to assess the impact of catheter ablation to target adenosine-induced PV reconnection on AF ablation outcome and 3 studies included 612 patients to assess the impact of adenosine testing on AF ablation outcome. Relative risks were calculated and combined in a meta-analysis using random effects modeling. Routine adenosine testing for PV reconnection with additional targeted ablation resulted in a significant increase in freedom from AF post-PVI (RR 1.25; 95% CI 1.12-1.40; P < 0.001). However, within the group of patients undergoing adenosine testing, those with reconnection identified a population with a trend to reduction in freedom from AF despite the use of further targeted ablation in the reconnection group (RR 0.91 with 95% CI 0.81-1.03; P = 0.15). Routine adenosine testing is associated with an improvement in freedom from AF post-PVI. Paradoxically acute adenosine-induced PV reconnection may portend a greater likelihood of AF recurrence despite additional ablation. Randomized controlled trials are required to determine the role of adenosine testing post-PVI.


Patient-reported outcomes in young adults with osteonecrosis secondary to developmental dysplasia of the hip - a longitudinal and cross-sectional evaluation.
Osteonecrosis of the femoral head is a common complication in the treatment of developmental dysplasia of the hip (DDH). While functional outcomes of affected patients are good in childhood, it is not clear how they change during the transition to young adulthood. This study determined the relationship between osteonecrosis and hip function, physical function and health status in adolescents and young adults. We performed a cross-sectional study of 169 patients with a mean age of 19.7 ± 3.8 years with and without osteonecrosis following an open or closed reduction (1995-2005). We also performed a separate longitudinal evaluation of an historical cohort of 54 patients with osteonecrosis, embedded in this sample. All completed patient-reported outcome measures in 2015/2016 to quantify hip function (maximum score 100); physical function (maximum score 100); and general health status (maximum score 1). We graded all radiographs for subtype of osteonecrosis (Bucholz-Ogden); acetabular dysplasia (centre-edge angle); subluxation (Shenton's line); and osteoarthritis (Kellgren-Lawrence). Analyses were adjusted for the number of previous surgical procedures on the hip and for the severity of residual hip dysplasia. In 149 patients (186 hips) with and without osteonecrosis, the mean differences (95% confidence interval) in hip function, physical function and quality of life were - 4.7 (- 10.26, 0.81), - 1.03 (- 9.29, 7.23) and 0.10 (- 1.15, 1.18), respectively. Adjusted analyses stratified across types of osteonecrosis showed that only patients with Bucholz-Odgen grade III had reduced hip function (p < 0.01) and physical function (p < 0.05) but no difference in health-related quality of life when compared to no osteonecrosis. Osteonecrosis secondary to DDH is a relatively benign disorder in adolescents and young adulthood. Affected patients demonstrated minimal physical disability, a normal quality of life but reduced hip function.


Bioethics in the public forum.
Public policy is, and ought to be, concerned with the welfare of human beings across a broad spectrum of circumstances and conditions. A Catholic moral theologian analyzes his involvement as a public policy consultant on in vitro fertilization. Evaluative judgements made by the Ethics Advisory Board are not edicts; they leave matters open for reconsideration and revision, while providing a basis for policy decisions for the present.


Role of extracorporeal membrane oxygenation in children with sepsis: a systematic review and meta-analysis.
The benefits of extracorporeal membrane oxygenation (ECMO) in children with sepsis remain controversial. Current guidelines on management of septic shock in children recommend consideration of ECMO as salvage therapy. We sought to review peer-reviewed publications on effectiveness of ECMO in children with sepsis. Studies reporting on mortality in children with sepsis supported with ECMO, published in PubMed, Scopus and Embase from 1972 till February 2020, were included in the review. This study was done in adherence to Preferred Reporting Items for Systematic Review and Meta-Analysis statement after registering the review protocol with PROSPERO. Study eligibility was independently assessed by two authors and disagreements resolved by a third author. Publications were reviewed for quality using Grading of Recommendations Assessment, Development, and Evaluation (GRADE) system. Random-effects meta-analyses (DerSimonian and Laird) were conducted, and 95% confidence intervals were computed using the Clopper-Pearson method. Outliers were identified by the Baujat plot and leave-one-out analysis if there was considerable heterogeneity. The primary outcome measure was survival to discharge. Secondary outcome measures included hospital length of stay, subgroup analysis of neonatal and paediatric groups, types and duration of ECMO and complications . Of the 2054 articles screened, we identified 23 original articles for systematic review and meta-analysis. Cumulative estimate of survival (13 studies, 2559 patients) in the cohort was 59% (95%CI: 51-67%). Patients had a median length of hospital stay of 28.8 days, median intensive care unit stay of 13.5 days, and median ECMO duration of 129 h. Children needing venoarterial ECMO (9 studies, 208 patients) showed overall pooled survival of 65% (95%CI: 50-80%). Neonates (< 4 weeks of age) with sepsis needing ECMO (7 studies, 85 neonates) had pooled survival of 73% (95%CI: 56- 87%). Gram positive organisms were the most common pathogens (47%) in septic children supported with ECMO. Survival rates of children with sepsis needing ECMO was 59%. Neonates had higher survival rates (73%); gram positive organisms accounted for most common infections in children needing ECMO. Despite limitations, pooled survival data from this review indicates consideration of ECMO in refractory septic shock for all pediatric age groups.


Essential Indicators Identifying Chronic Inorganic Mercury Intoxication: Pooled Analysis across Multiple Cross-Sectional Studies.
The continuous exposure to inorganic mercury vapour in artisanal small-scale gold mining (ASGM) areas leads to chronic health problems. It is therefore essential to have a quick, but reliable risk assessing tool to diagnose chronic inorganic mercury intoxication. This study re-evaluates the state-of-the-art toolkit to diagnose chronic inorganic mercury intoxication by analysing data from multiple pooled cross-sectional studies. The primary research question aims to reduce the currently used set of indicators without affecting essentially the capability to diagnose chronic inorganic mercury intoxication. In addition, a sensitivity analysis is performed on established biomonitoring exposure limits for mercury in blood, hair, urine and urine adjusted by creatinine, where the biomonitoring exposure limits are compared to thresholds most associated with chronic inorganic mercury intoxication in artisanal small-scale gold mining. Health data from miners and community members in Indonesia, Tanzania and Zimbabwe were obtained as part of the Global Mercury Project and pooled into one dataset together with their biomarkers mercury in urine, blood and hair. The individual prognostic impact of the indicators on the diagnosis of mercury intoxication is quantified using logistic regression models. The selection is performed by a stepwise forward/backward selection. Different models are compared based on the Bayesian information criterion (BIC) and Cohen`s kappa is used to evaluate the level of agreement between the diagnosis of mercury intoxication based on the currently used set of indicators and the result based on our reduced set of indicators. The sensitivity analysis of biomarker exposure limits of mercury is based on a sequence of chi square tests. The variable selection in logistic regression reduced the number of medical indicators from thirteen to ten in addition to the biomarkers. The estimated level of agreement using ten of thirteen medical indicators and all four biomarkers to diagnose chronic inorganic mercury intoxication yields a Cohen`s Kappa of 0.87. While in an additional stepwise selection the biomarker blood was not selected, the level of agreement based on ten medical indicators and only the three biomarkers urine, urine/creatinine and hair reduced Cohen`s Kappa to 0.46. The optimal cut-point for the biomarkers blood, hair, urine and urine/creatinine were estimated at 11. 6 μg/l, 3.84 μg/g, 24.4 μg/l and 4.26 μg/g, respectively. The results show that a reduction down to only ten indicators still allows a reliable diagnosis of chronic inorganic mercury intoxication. This reduction of indicators will simplify health assessments in artisanal small-scale gold mining areas.


Examining the neural targets of the AMPA receptor potentiator LY404187 in the rat brain using pharmacological magnetic resonance imaging.
Drugs that enhance alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropanoic acid (AMPA) receptor-mediated glutamatergic transmission, such as the AMPA receptor potentiator LY404187, may form treatment strategies for disorders of cognition, learning and memory. Pharmacological magnetic resonance imaging (phMRI) uses blood oxygenation level dependent (BOLD) contrast as a marker of neuronal activity and allows dynamic non-invasive in vivo imaging of the effects of CNS-active compounds. This study used phMRI to examine the effects of LY404187 in the rat brain. Groups of Sprague Dawley rats (n=7) were anaesthetised and placed in a 4.7 Tesla superconducting magnet before receiving an acute dose of LY404187 (0.5 mg/kg s.c.), either alone or after pretreatment with the selective AMPA/kainate antagonist LY293558 (15 mg/kg s.c.), or LY293558 alone (15 mg/kg s.c.). Brain images were acquired for each subject every minute for 180 min. These volumes were extensively pre-processed before being analysed for changes in BOLD contrast. LY404187 produced significant increases in BOLD contrast in brain regions including the hippocampus, lateral and medial habenulae and superior and inferior colliculi. These changes were blocked by LY293558. When administered alone, LY293558 caused widespread decreases in BOLD contrast. The known actions of LY404187 suggest the observed BOLD signal increases reflect increases in excitatory neurotransmission. The decreases in signal following LY293558 alone are harder to interpret and are discussed in terms of the negative BOLD response. This study provides the first evidence that the effects of AMPA receptor-mediating compounds can be observed using phMRI.


Pure-tone auditory threshold in school children.
To determine pure-tone auditory thresholds, 197 screened children at a typical primary school in a German town (~70,000 inhabitants) were examined. All children underwent a tympanometry and an audiometry at 17 frequencies from 125 to 16 kHz. Regarding age effects, two groups (6-8 and 9-12 years) were analyzed. The cross-sectional research was supplemented by a follow-up study with 35 children of the first graders 3 years later. School children have the poorest hearing sensitivity at low frequencies (below 1 kHz) and the best sensitivity at the extended high frequencies above 8 kHz. Hearing thresholds are rising significantly with age. Through all frequencies, averaged improvements were 3.8 dB (right ear) and 3.7 dB (left ear) at the cross-sectional study and 3.7 dB (right ear) and 5.1 dB (left ear) at the longitudinal study. The overall deviation (left and right) from the standard thresholds for adults were 7.4 and 3.6 dB for the younger and older age groups, respectively. The ear canal volume (ECV) measured by tympanometric tests was at mean with 1.06 cm(3) for the 6- to 8-year age group significantly lower (p < 0.001) in comparison with 1.18 cm(3) for the 9- to 12-year age group. Also, girls had significant (p < 0.001) smaller ECV (mean 1.07 cm(3)) than boys (1.17 cm(3)). Auditory performance improves with rising age in school children.


[Importance of vascular factors for cognitive rehabilitation in Parkinson's disease patients].
The examination of the Parkinson's disease patients, patients with subcortical encephalopathy and their combination, using neuropsychological testings and definitions of the gait and balance disturbance degree, the dependence of balance impairment and cognitive functions from concomitant cerebrovascular patology has been revealed.


HER2-targeted hybrid peptide that blocks HER2 tyrosine kinase disintegrates cancer cell membrane and inhibits tumor growth in vivo.
HER2 is a transmembrane oncoprotein encoded by the HER2/neu gene and is overexpressed in approximately 20% to 30% of breast cancers. We have recently designed a novel class of drug, the hybrid peptide, which is chemically synthesized and is composed of a target-binding peptide and a lytic peptide containing cationic-rich amino acid components that disintegrate the cell membrane, leading to cancer cell death via membrane lysis. In this study, we designed a HER2-binding peptide linked to this novel lytic peptide, which we termed the HER2-lytic hybrid peptide and assessed the cytotoxic activity of this hybrid peptide in vitro and in vivo. The HER2-lytic hybrid peptide showed high cytotoxic activity against all ovarian and breast cancer cell lines, even trastuzumab- and/or lapatinib-resistant cells, but not against normal cells. Competition assays using anti-HER2 antibody and knockdown of this receptor by siRNA confirmed the specificity of the HER2-lytic hybrid peptide. In addition, it was shown that the HER2-lytic hybrid peptide can disintegrate the cancer cell membrane of HER2-overexpressing SK-BR-3 cancer cells in only 5 minutes, but not normal cells, and block HER2 signaling. Intravenous administration of the HER2-lytic peptide in the athymic mouse implanted with BT-474 and MDA-MB-453 cells significantly inhibited tumor progression. The HER2-lytic hybrid peptide was effective even in breast cancer cell lines that are resistant to trastuzumab and/or lapatinib in vitro and in vivo. Therefore, this hybrid peptide may provide a potent treatment option for patients with cancer.


Comparative proteomics uncovers the signature of natural selection acting on the ejaculate proteomes of two cricket species isolated by postmating, prezygotic phenotypes.
Two of the most well-supported patterns to have emerged over the past two decades of research in evolutionary biology are the occurrence of divergent natural selection acting on many male and female reproductive tract proteins and the importance of postmating, prezygotic phenotypes in reproductively isolating closely related species. Although these patterns appear to be common across a wide variety of taxa, the link between them remains poorly documented. Here, we utilize comparative proteomic techniques to determine whether or not there is evidence for natural selection acting on the ejaculate proteomes of two cricket species (Allonemobius fasciatus and A. socius) which are reproductively isolated primarily by postmating, prezygotic phenotypes. In addressing this question, we compare the degree of within-species polymorphism and between-species divergence between the ejaculate and thorax proteomes of these two species. We found that the ejaculate proteomes are both less polymorphic and more divergent than the thorax proteomes. Additionally, we assessed patterns of nucleotide variation for two species-specific ejaculate proteins and found evidence for both reduced levels of variation within species and positive selection driving divergence between species. In contrast, non-species-specific proteins exhibited higher levels of within-species nucleotide variation and no signatures of positive selection. Nucleotide and putative functional data for the two species-specific proteins, along with data for a third protein (ejaculate serine protease), suggest that all three of these genes are candidate speciation genes in need of further study. Overall, these patterns of proteome and nucleotide divergence provide support for the hypothesis that there is a causative link between selection-driven divergence of male ejaculate proteins and the evolution of postmating, prezygotic barriers to gene flow within Allonemobius.


A randomized double-blind placebo-controlled clinical trial on efficacy and safety of association of simethicone and Bacillus coagulans (Colinox®) in patients with irritable bowel syndrome.
Irritable bowel syndrome (IBS) is a chronic gastrointestinal (GI) disorder that affects 15-20% of the Western population. There are currently few therapeutic options available for the treatment of IBS. The aim of this study is to evaluate the efficacy and the safety of a medical device containing a combination of Simethicone and Bacillus coagulans in the treatment of IBS. This is a monocentric double-blind, placebo-controlled parallel group clinical trial. Adult subjects suffering from IBS as defined by Rome III criteria were enrolled. Bloating, discomfort, abdominal pain were assessed as primary end point. Subjects received the active treatment or placebo 3 time a day after each meal for 4 weeks of study period. Subjects were submitted to visit at Day 0 (T1), at Days 14 (T2) and 29 (T3). Fifty-two patients were included into the study. Intragroup analysis showed a significant reduction of the bloating, discomfort and pain in Colinox® group (CG) compared to placebo group (PG). Between group analysis confirmed, at T1-T3, significant differences between CG and PG in bloating and discomfort. Simethicone is an inert antifoaming able to reduce bloating, abdominal discomfort. Literature offers increasing evidence linking alterations in the gastrointestinal microbiota and IBS and it is well known that probiotics are important to restore the native gut microbiota. The Colinox medical device is specifically targeted against most intrusive symptom of IBS (bloating) and it is also able to counteract the most accredited ethiopathogenetic factor in IBS (alterations of intestinal microbiota). This is the first randomized double-blind placebo-controlled clinical trial demonstrating the efficacy and safety of a combination of simethicone and Bacillus coagulans in treatment of IBS.


On the design and analysis of multicenter trials in acute renal failure.
Improving outcomes for patients with acute renal failure (ARF) is one of the most urgent tasks for 21st century nephrologists and other critical care physicians. The incidence of hospital-acquired ARF is rising (to 5% or more), partly because of changes in the demographic and clinical characteristics of the hospitalized patient population. When ARF is severe enough to require dialysis, in-hospital mortality rates are distressingly high, in excess of 50% in most published studies. In the past several years, a number of attempts have been made to influence the course of ARF, either relatively early (eg, atrial natriuretic peptide [ANP], insulin growth factor-1 [IGF-1]) or coincident with the dialysis procedure in severe cases (eg, variations in dialysis membrane and modality). Several lessons can be learned from these efforts. This article will briefly address issues in the design and analysis of clinical trials, focusing on elements of special interest to practitioners of renal medicine.


Heart failure management at home: a non-randomised prospective case-controlled trial (HeMan at Home).
Heart failure (HF) is a growing clinical and economic burden for patients and health systems. The COVID-19 pandemic has led to avoidance and delay in care, resulting in increased morbidity and mortality among many patients with HF. The increasing burden of HF during the COVID-19 pandemic led us to evaluate the quality and safety of the Hospital at Home (HAH) for patients presenting to their community providers or emergency department (ED) with symptoms of acute on chronic HF (CHF) requiring admission. A non-randomised prospective case-controlled of patients enrolled in the HAH versus admission to the hospital (usual care, UC). Primary outcomes included length of stay (LOS), adverse events, discharge disposition and patient satisfaction. Secondary outcomes included 30-day readmission rates, 30-day ED usage and ED dwell time. Sixty patients met inclusion/exclusion criteria and were included in the study. Of the 60 patients, 40 were in the HAH and 20 were in the UC group. Primary outcomes demonstrated that HAH patients had slightly longer LOS (6.3 days vs 4.7 days); however, fewer adverse events (12.5% vs 35%) compared with the UC group. Those enrolled in the HAH programme were less likely to be discharged with postacute services (skilled nursing facility or home health). HAH was associated with increased patient satisfaction compared with Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) score in North Carolina. Secondary outcomes of 30-day readmission and ED usage were similar between HAH and UC. The HAH pilot programme was shown to be a safe and effective alternative to hospitalisation for the appropriately selected patient presenting with acute on CHF.


Comparative study of Fe(II)/sulfite, Fe(II)/PDS and Fe(II)/PMS for p-arsanilic acid treatment: Efficient organic arsenic degradation and contrasting total arsenic removal.
As a widely used feed additives, p-arsanilic acid (p-AsA) frequently detected in the environment poses serious threats to aquatic ecology and water security due to its potential in releasing more toxic inorganic arsenic. In this work, the efficiency of Fe(II)/sulfite, Fe(II)/PDS and Fe(II)/PMS systems in p-AsA degradation and simultaneous arsenic removal was comparatively investigated for the first time. Efficient p-AsA abatement was achieved in theses Fe-based systems, while notable discrepancy in total arsenic removal was observed under identical acidic condition. By using chemical probing method, quenching experiments, isotopically labeled water experiments, p-AsA degradation was ascribed to the combined contribution of high-valent Fe(IV) and SO4•-in these Fe(II)-based system. In particular, the relative contribution of Fe(IV) and SO4•- in the Fe(II)/sulfite system was highly dependent on the molar ratio of [Fe(II)] and [sulfite]. Negligible arsenic removal was observed in the Fe(II)/sulfite and Fe(II)/PDS systems, while ∼80% arsenic was removed in the Fe(II)/PMS system under identical acidic condition. This interesting phenomenon was due to that ferric precipitation only occurred in the Fe(II)/PMS system. As(V) was further removed via adsorption onto the iron precipitate or the formation of ferric arsenate-sulfate compounds, which was confirmed by particle diameter measurements, fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Through tuning solution pH, complete removal of total arsenic could achieve in all three systems. Among these three Fe-based technologies, the hybrid oxidation-coagulation Fe(II)/PMS system demonstrated potential superiority for arsenic immobilization by not requiring pH adjustment for coagulation and facilitating the in-situ generation of ferric arsenate-sulfate compounds with comparably low solubility levels like scorodite. These findings would deepen the understanding of these three Fe-based Fenton-like technologies for decontamination in water treatment.


Suspected stonefish envenomation in Queensland over eight years: a retrospective series.
Stonefish envenomation results in localized severe pain and swelling and systemic features, including vomiting, arrhythmia, pulmonary oedema, and possibly death. There are limited data regarding the effectiveness of the available antivenom. The aim of this series is to characterize presentations of patients with suspected stonefish envenomation and investigate treatment, including antivenom. This is a retrospective observational series of suspected stonefish envenomation as reported to the Queensland Poisons Information Centre or Princess Alexandra Hospital Clinical Toxicology Unit from July 2015 to January 2023. Patients were identified through the databases held by both the Centre and Unit, and data on clinical features and investigations were collected from the patient's electronic medical record. There were 87 suspected stonefish envenomations from July 2015 to January 2023. The median age was 26 (range: 5-69) years, and 69 (79 per cent) patients were male. Pain was reported in 85 (98 per cent) with a median peak pain score of 10 (range 4-12; three rated their pain greater than 10/10). A clear wound was documented in 64 (74 per cent), with local swelling in 63 (72 per cent). A foreign body was retained in eight (9 per cent) presentations. Systemic symptoms were rare, with vomiting in four (5 per cent) and dizziness in two (2 per cent) presentations. There were no instances of hypotension, arrhythmia, or pulmonary oedema. Hot water was administered in 72 (83 per cent) presentations. Oral analgesia was given in 55 (63 per cent). Parenteral analgesia was given in 53 (61 per cent), most commonly opioids. Local anaesthetic block was performed in 19 presentations (22 per cent), with effectiveness documented in 16/19 (84 per cent). Five patients received antivenom for intractable pain, and all received subsequent parenteral analgesia or local anaesthetic block. Stonefish envenomation is characterized by severe pain. Systemic symptoms were rare and not severe in this series. Local anaesthetic block appeared to be the most effective intervention for severe pain when performed. Antivenom appeared to be ineffective in managing pain.


State of a sustainable drainage system at end-of-life: assessment of potential water pollution by leached metals from recycled pervious pavement materials when used as secondary aggregate.
Sustainable drainage systems (SuDS) have emerged as an effective and attractive approach for stormwater management, prevention of water pollution and flood control due to its sustainable, environmentally friendly and cost-effective approaches. One of the SuDS devices widely used to infiltrate, store and treat surface runoff which allows it to recharge groundwater is the pervious paving systems (PPS). Previous studies have demonstrated relatively high pollution removal efficiencies typically ranging from 98.7% for total hydrocarbons to 89% of COD. Although a small number of the studies have assessed the performance characteristics of the PPS system in long-established installations in terms of retention of pollutants, hydrological features, biodegradation of pollutants etc., none has assessed the risk of potential groundwater and soil pollution by pollutants such as metals retained in the PPS materials either as a disposed waste material (in the case of used geotextiles) or during re-use as secondary aggregates. Thus, this study evaluated potential risks associated with the decommissioning and beneficial use of wastes produced during the disassembly of a PPS. The authors believe that this was the first PPS to be addressed in this way. The method involved the determination of leachable concentrations of 14 metals in the PPS samples made up of extracts from the model profile which included the geotextile fibre (G), dust alone (D), aggregates and dust (AD), aggregates alone (AA) and pavement blocks (P) which were analysed and compared with two different groups of regulatory threshold limits. The results showed that the measured concentrations of all the metals were below the appropriate threshold values for irrigation purposes as specified by FAO and USEPA. Furthermore, results all indicated that the dismantled materials were all below EU LFD WAC limits for inert waste, indicating relative ease of disposal and suitability for use as recycled aggregate. This, admittedly limited data, indicates that recycling of aggregates from demolition wastes arising from end of life PPS would not be limited by the potential leaching of heavy metals, including re-use within another PPS. This would minimise dependence on virgin aggregates and hence reduce rate of exploitation of natural resources and improve sustainability score card of SuDS.


Transmission of diverse oral bacteria to murine placenta: evidence for the oral microbiome as a potential source of intrauterine infection.
Microbial infection of the intrauterine environment is a major cause of preterm birth. The current paradigm indicates that intrauterine infections predominantly originate from the vaginal tract, with the organisms ascending into the sterile uterus. With the improvements in technology, an increasing number of bacterial species have been identified in intrauterine infections that do not belong to the vaginal microflora. We have demonstrated previously that intrauterine infections can originate from the oral cavity following hematogenous transmission. In this study, we begin to systemically examine what proportion of the oral microbiome can translocate to the placenta. Pooled saliva and pooled subgingival plaque samples were injected into pregnant mice through tail veins to mimic bacteremia, which occurs frequently during periodontal infections. The microbial species colonizing the murine placenta were detected using 16S rRNA gene-based PCR and clone analysis. A diverse group of bacterial species were identified, many of which have been associated with adverse pregnancy outcomes in humans although their sources of infection were not determined. Interestingly, the majority of these species were oral commensal organisms. This may be due to a dose effect but may also indicate a unique role of commensal species in intrauterine infection. In addition, a number of species were selectively "enriched" during the translocation, with a higher prevalence in the placenta than in the pooled saliva or subgingival plaque samples. These observations indicate that the placental translocation was species specific. This study provides the first insight into the diversity of oral bacteria associated with intrauterine infection.


Monte Carlo calculation of dose rate distributions around 192Ir wires.
Monte Carlo calculations of absolute dose rate in liquid water are presented in the form of away-along tables for 1 and 5 cm 192Ir wires of 0.3 mm diameter. Simulated absolute dose rate values can be used as benchmark data to verify the calculation results of treatment planning systems or directly as input data for treatment planning. Best fit value of attenuation coefficient suitable for use in Sievert-integrals-type calculations has been derived based on Monte Carlo calculation results. For the treatment planning systems that are based on TG43 formalism we have also calculated the required dosimetry parameters.


TMEM70 functions in the assembly of complexes I and V.
Protein complexes from the oxidative phosphorylation (OXPHOS) system are assembled with the help of proteins called assembly factors. We here delineate the function of the inner mitochondrial membrane protein TMEM70, in which mutations have been linked to OXPHOS deficiencies, using a combination of BioID, complexome profiling and coevolution analyses. TMEM70 interacts with complex I and V and for both complexes the loss of TMEM70 results in the accumulation of an assembly intermediate followed by a reduction of the next assembly intermediate in the pathway. This indicates that TMEM70 has a role in the stability of membrane-bound subassemblies or in the membrane recruitment of subunits into the forming complex. Independent evidence for a role of TMEM70 in OXPHOS assembly comes from evolutionary analyses. The TMEM70/TMEM186/TMEM223 protein family, of which we show that TMEM186 and TMEM223 are mitochondrial in human as well, only occurs in species with OXPHOS complexes. Our results validate the use of combining complexome profiling with BioID and evolutionary analyses in elucidating congenital defects in protein complex assembly.


Can We Improve Our Physical Health by Altering Our Social Networks?
Persons with more types of social relationships live longer and have less cognitive decline with aging, greater resistance to infectious disease, and better prognoses when facing chronic life-threatening illnesses. We have known about the importance of social integration (engaging in diverse types of relationships) for health and longevity for 30 years. Yet, we still do not know why having a more diverse social network would have a positive influence on our health, and we have yet to design effective interventions that influence key components of the network and in turn physical health. Better understanding of the role of social integration in health will require research on how integrated social networks influence health relevant behaviors, regulate emotions and biological responses, and contribute to our expectations and worldviews.


Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice.
Mice are a widely used pre-clinical model system in large part due to their potential for genetic manipulation. The ability to manipulate gene expression in specific cells under temporal control is a powerful experimental tool. The liver is central to metabolic homeostasis and a site of many diseases, making the targeting of hepatocytes attractive. Adeno-associated virus 8 (AAV8) vectors are valuable instruments for the manipulation of hepatocellular gene expression. However, their off-target effects in mice have not been thoroughly explored. Here, we sought to identify the short-term off-target effects of AAV8 administration in mice. To do this, we injected C57BL/6J wild-type mice with either recombinant AAV8 vectors expressing Cre recombinase or control AAV8 vectors and characterised the changes in general health and in liver physiology, histology and transcriptomics compared to uninjected controls. We observed an acute and transient trend for reduction in homeostatic liver proliferation together with induction of the DNA damage marker γH2AX following AAV8 administration. The latter was enhanced upon Cre recombinase expression by the vector. Furthermore, we observed transcriptional changes in genes involved in circadian rhythm and response to infection. Notably, there were no additional transcriptomic changes upon expression of Cre recombinase by the AAV8 vector. Overall, there was no evidence of liver injury, and only mild T-cell infiltration was observed 14 days following AAV8 infection. These data advance the technique of hepatocellular genome editing through Cre-Lox recombination using Cre expressing AAV vectors, demonstrating their minimal effects on murine physiology and highlight the more subtle off target effects of these systems.


Assay using succinyldithiocholine as substrate: the method of choice for the measurement of cholinesterase catalytic activity in serum to diagnose succinyldicholine sensitivity.
No comparative information is available concerning the ability of various cholinesterase (ChE) methods to identify succinyldicholine-sensitive patients, purely on the basis of the enzyme activity recorded in serum. Here, we evaluated six different methods for the measurement of ChE activity; 131 subjects were subdivided according to ChE phenotype and, therefore, to succinyldicholine sensitivity. ChE phenotype was determined by measuring dibucaine and fluoride numbers. DNA analysis was also performed to confirm correlation between the phenotype classification used in the study and the ChE genotype. The tested methods were significantly different in their ability to discriminate between the subjects with and without succinyldicholine-sensitive phenotypes. The succinyldithiocholine/5,5'-dithio-bis(2-nitrobenzoate) (DTNB) method showed the highest accuracy (area under the receiver operating characteristic (ROC) curve 0.97) followed by the propionylthiocholine/DTNB method (area under the ROC curve 0.94). On the other hand, the two methods using butyrylthiocholine as substrate and that employing benzoylcholine showed limited clinical utility in discriminating subjects at risk of prolonged apnea (area under the ROC curve < or = 0.9). Using the succinyldithiocholine method, a value < or = 23 U/l was approximately five times as likely to occur in a sensitive individual as in a normal one.