A systematic review of early phase studies for children and young people with relapsed and refractory rhabdomyosarcoma: The REFoRMS-SR project.
Rhabdomyosarcoma is the commonest soft tissue sarcoma in children. Around one-third of children with rhabdomyosarcoma experience relapse or have refractory disease, which is associated with a poor prognosis. This systematic review of early phase studies in pediatric relapsed/refractory rhabdomyosarcoma was conducted to inform future research and provide accurate information to families and clinicians making difficult treatment choices. Nine databases and five trial registries were searched in June 2021. Early phase studies of interventions for disease control in patients under 18 years old with relapsed/refractory rhabdomyosarcoma were eligible. No language/geographic restrictions were applied. Studies conducted after 2000 were included. Survival outcomes, response rates, quality of life and adverse event data were extracted. Screening, data extraction and quality assessment (Downs and Black Checklist) were conducted by two researchers. Owing to heterogeneity in the included studies, narrative synthesis was conducted. Of 16,965 records screened, 129 published studies including over 1100 relapsed/refractory rhabdomyosarcoma patients were eligible. Most studies evaluated systemic therapies. Where reported, 70% of studies reported a median progression-free survival ≤6 months. Objective response rate was 21.6%. Adverse events were mostly hematological. One-hundred and seven trial registry records of 99 studies were also eligible, 63 of which report they are currently recruiting. Study quality was limited by poor and inconsistent reporting. Outcomes for children with relapsed/refractory rhabdomyosarcoma who enroll on early phase studies are poor. Improving reporting quality and consistency would facilitate the synthesis of early phase studies in relapsed/refractory rhabdomyosarcoma (PROSPERO registration: CRD42021266254).


Empirical Calibration of a Cylindrical Ion Trap for Mass-Selected Gas-Phase Fluorescence Spectroscopy.
The ion motion in a quadrupole ion trap of hyperbolic geometry is well described by the Mathieu equations. A simpler cylindrical ion trap has also gained significance and has been used by us for fluorescence-spectroscopy experiments. This design allows for the easy replacement of the end-cap with a mesh, enhancing the photon collection. It is crucial to obtain a firm understanding of the ion motion in cylindrical ion traps and their capability as mass spectrometers. We present here an empirical method of calibrating a cylindrical ion trap based on fluorescence detection. This can be done nearly background-free in a pulsed experiment. The ions are located at the center of the trap, where the field is primarily quadrupolar, and here an effective Mathieu description is found through an effective geometry parameter. In spectroscopy experiments, high buffer-gas pressures are needed to efficiently cool the ions, which complicates the ions' motion and hence their stability. Still, simulations show that the stability diagram closely aligns with the Mathieu diagram, albeit shifted due to collisions. We map the stability diagram for six molecular ions by fluorescence collection from four cations and two anions spanning m/z from 212 to 647. The stability diagram is parametrized through the Mathieu functions with an m/z-dependent effective geometry parameter and a q-dependent shrinkage of the diagram. Based on the calibration, we estimate the mass resolution to be +7/-3 Da for ions with masses in the hundreds of Da.


[Automation of composite processes for biochemical studies].
Approximate division of biochemical investigations into three components: experimental research, results treatment and modeling, as well as external information filling is made. Computerization of biochemical researches for these components are analyzed and authors developments for experimental researches computerization on IBM-compatible computer basis are presented, as well as the local computer net of biochemistry institute.


Ultrastructural relationship between the mu opioid receptor and its interacting protein, GPR177, in striatal neurons.
GPR177, the mammalian ortholog of Drosophila Wntless/Evi/Sprinter, was recently identified as a novel mu-opioid receptor (MOR) interacting protein. GPR177 is a trans-membrane protein pivotal to mediating the secretion of Wnt signaling proteins. Wnt proteins, in turn, are essential in regulating neuronal development, a phenomenon inhibited upon chronic exposure to MOR agonists such as morphine and heroin. We previously showed that GPR177 and MOR are co-localized in the mouse dorsolateral striatum; however, the nature of this interaction was not fully elucidated. Therefore, in the present study, we examined cellular substrates for interactions between GPR177 and MOR using a combined immunogold-silver and peroxidase detection approach in coronal sections in the dorsolateral segment of the striatum. Semi-quantitative analysis of the ultrastructural distribution of GPR177 and MOR in striatal somata and in dendritic processes showed that, of the somata and dendritic processes exhibiting GPR177, 32% contained MOR immunolabeling while for profiles exhibiting MOR, 37% also contained GPR177 immunoreactivity. GPR177-labeled particles were localized predominantly along both the plasma membrane and within the cytoplasm of MOR-labeled dendrites. Somata and dendritic processes that contained both GPR177 and MOR more often received symmetric (inhibitory-type) synapses from unlabeled axon terminals. To further define the phenotype of GPR177 and MOR-containing cellular profiles, triple immunofluorescence detection showed that GPR177 and MOR are localized in neurons containing the opioid peptide, enkephalin, within the dorsolateral striatum. The results provide an anatomical substrate for interactions between MOR and its interacting protein, GPR177, in striatal opioid-containing neurons that may underlie the morphological alterations produced in neurons by chronic opiate use.


Increased cell surface hydrophobicity of a Serratia marcescens NS 38 mutant lacking wetting activity.
The cell surface hydrophobicity of Serratia marcescens appears to be an important factor in its adhesion to and colonization of various interfaces. The cell surface components responsible for mediating the hydrophobicity of S. marcescens have not been completely elucidated, but may include prodigiosin and other factors. In the present report we have investigated the potential role of serratamolide, an amphipathic aminolipid present on the surfaces of certain S. marcescens strains, in modulating cell surface hydrophobicity. The hydrophobic properties of a serratamolide-producing strain (NS 38) were compared with those of a serratamolide-deficient mutant (NS 38-9) by monitoring the kinetics of adhesion to hexadecane. Serratamolide production was monitored by thin-layer chromatography and the wetting activity of washed-cell suspensions on polystyrene. Wild-type NS 38 cells were far less hydrophobic than the serratamolide-deficient mutant cells were; the removal coefficients were 48 min-1 for the mutant, as compared with only 18 min-1 for the wild type. The data suggest that the presence of serratamolide on S. marcescens cells results in a reduction in hydrophobicity, presumably by blocking hydrophobic sites on the cell surface.


Challenges to optimal medicines use in people living with dementia and their caregivers: A literature review.
Dementia is fast becoming a global concern due to a demographic shift towards an older population. Many studies have shown that caring for a family member or friend has a profound and negative impact on the physical, emotional and psychosocial aspects of the caregivers' life. One significant activity that a family caregiver undertakes is assistance with the management of medicines. This review was undertaken to ascertain what the issues are that affect optimal medicines use from the perspectives of people living with dementia and their caregivers, both in the community and care home settings. A literature search was conducted using electronic databases, employing a combination of search terms. A total of 16 studies met the inclusion criteria. Six broad themes were identified, together with some recommendations to improve medicines use in people with dementia. Challenges to medicines use centred on medicines management and administration, the impact on the caregiver and care recipient, their partnership and interface with formal care. Future research should focus on developing targeted interventions that can overcome these challenges to achieve optimal medicines use.


Effect of endoleaks on changes in aortoiliac volume after endovascular repair for abdominal aortic aneurysm.
To evaluate changes in aortoiliac volume after endovascular repair (EVAR) for abdominal aortic aneurysm (AAA) in patients with and without endoleaks. We retrospectively analyzed 137 patients who underwent EVAR for AAA. We manually measured the aortoiliac volume on pre-procedural baseline CT angiograms (CTAs) and post-procedural follow-up CTAs. All post-procedural CTAs were evaluated for the presence of endoleaks. Follow-up examinations were grouped into five time points relative to the date of the EVAR procedure and mean aortoiliac volume changes from the baseline were calculated. In 51 patients (37.2%), endoleaks were detected during follow-up. In patients without any endoleaks, mean aortoiliac volume decreased by 21.1% from the pre-interventional baseline examination to the last follow-up examination. In patients with any endoleak during follow-up aortoiliac volume increased by 12.2% and in patients with only transient, post-procedural endoleaks (n = 18), aortoiliac volume decreased by 13.4% over the same time period. After EVAR for AAA, aortoiliac volume on CT angiography decreases by approximately 20% over time in the absence of endoleaks and increases in the presence of endoleaks. Transient post-procedural endoleaks, however, do not influence long-term volume regression.


What data do California HMOs use to select hospitals for contracting?
To explore whether health maintenance organization (HMO) executives in a mature market are familiar with hospital report cards, whether they find the report cards useful (and if not, why not), and how they weight such data relative to other factors. Cross-sectional survey of HMO executives in 1999. We contacted all 47 licensed HMOs and the sponsors of all 90 employee medical benefit plans in California with at least 1000 participants. Thirty of the 47 (63.8%) eligible HMOs provided usable responses: 19 in writing, 11 by telephone. HMO executives reported basing their contracting decisions primarily on hospital accreditation, location, and price. Although hospital quality is considered important, HMO executives rely primarily on accreditation, government disciplinary actions, reputation, and member satisfaction as measures of quality. Respondents voiced multiple concerns about the validity and usefulness of currently available process and outcome data. Accredited plans are more likely than unaccredited plans to perform independent analyses of hospital performance. Although HMO executives are interested in information on hospital quality, and are confident that such information will improve care, they are concerned about the limitations of available data and uncomfortable weighting these data heavily in selecting network hospitals. Prior empirical evidence suggests that HMOs may rely on surrogate quality measures and informal evaluation mechanisms to steer their members toward better-performing hospitals. Policy makers and producers of hospital report cards will need to address these problems by providing more timely data with longitudinal follow-up and external validation.


HIV-1-discordant couples in sub-Saharan Africa: explanations and implications for high rates of discordancy.
In sub-Saharan Africa, approximately 1 in 2 HIV-1-infected persons living in a couple have a serodiscordant partner. Recent data suggest a large proportion of new HIV-1 infections in mature epidemics occur within discordant couples, making discordancy a major contributor to the spread of HIV/AIDS in Africa. What accounts for high rates of HIV-1 discordance and why some individuals remain uninfected despite repeated sexual exposure to HIV-1 is unknown. Studying HIV-1-discordant couples may contribute to understanding correlates of HIV-1 immunity and acute infection. Additionally, HIV-1-discordant couples are an important population for prevention efforts. Consequently, HIV-1-discordant couples are increasingly viewed as a valuable source of participants for HIV vaccine and prevention trials. This review summarizes and critiques existing data on HIV-1-discordant couples in developing countries, including an analysis of transmission rates within discordant couples, description of biological and behavioral characteristics important in planning HIV-1 vaccine and prevention trials, and challenges faced when carrying out such studies.


Alcohol-Related Liver Disease: Is There a Safe Alcohol Consumption Limit for Liver Disease?
This review is to evaluate how much alcohol is safe in the context of alcohol-related liver disease (ALD). In patients without an established diagnosis of ALD consuming alcohol at quantities below 12 to 20 g daily with alcohol-free days is associated with a very low risk of developing disease. This risk is mediated by the presence of cofactors such as sex, medical comorbidity, obesity, and genetic factors. A threshold effect below which liver disease will not occur is not seen, instead a dose-response relationship where risk ranges from low to high. Once ALD is present, natural history studies confirm that continued alcohol consumption is clearly associated with an increased risk of ill health and premature death. In conclusion, low-level alcohol consumption in the absence of liver disease is associated with a very small risk of developing ALD, but once ALD is present patients should be supported to achieve complete abstinence from alcohol.


Antioxidant and antiangiogenic activities of the essential oils of Myristica fragrans and Morinda citrifolia.
Toinvestigate the anti-angiogenic activity and antioxidant properties of Myristica fragrans (M. fragrans) (nutmeg) and Morinda citrifolia (M. citrifolia)(mengkudu) oils. The nutmeg and megkudu essential oils were obtained by steam distillation. The antioxidant activities of both essential oils were determined by beta-carotene/linoleic acid bleaching assay and reducing power while the anti-angiogenic activity was investigated using rat aortic ring assay using various concentrations. The results showed that nutmeg oil has higher antioxidant activity than mengkudu oil. The nutmeg oil effectively inhibited the oxidation of linoleic acid with (88.68±0.1)% while the inhibition percentage of oxidation of linoleic acid of the mengkudu oil is (69.44±0.4)%. The nutmeg oil and mengkudu oil showed reducing power with an EC(50) value of 181.4 μg/mL and 3 043.0 μg/mL, respectively. The antiangiogenic activity of nutmeg oil showed significant antiangiogenic activity with IC(50) of 77.64 μg/mL comparing to mengkudu oil which exhibits IC(50) of 109.30 μg/mL. Bioactive compound(s) will be isolated from the nutmeg essential oil to be developed as antiangiogenic drugs.


[Importance of determination of the content of lactate dehydrogenase and its isoenzymes in patients with myocardial infarct complicated by rupture of the heart].
In 18 patients with myocardial infarction. who had died due to the rupture of the heart muscle the activity of lactate-dehydrogenase (LDG) and of its isoenzymes in the blood serum and cardiac tissue was studied. Comparisons were made with the results of investigations in donors, in persons dead as a result of accidents, in patients with uncomplicated transmural myocardial infarction, with aneurysm of the heart, severe cardiogenic shock and with other lethal complications of myocardial infarction. The highest levels of the blood LDG activity were registered in cases involving rupture of the heart. The determination of the LDG and its isoenzymes activity mat be used in assessing the degree of the resorption-necrotic syndrome intensity and of factors indicating the risk of a developing aneurysm and rupture of the heart in patients with myocardial infarction.


Magnoliae flos inhibits mast cell-dependent immediate-type allergic reactions.
The mast cell plays a pivotal role in initiating allergic response by secreting intracytoplasmic granular mediators such as histamine. Magnoliae flos has been used for the treatment of allergic disease in Korea. However, its effect in experimental models remains unknown. The present report describes an inhibitory effect of Magnoliae flos on mast cell-mediated immediate-type allergic reactions. Topical application of compound 48/80 can induce an ear swelling response in normal (WBB6F1-+/+) mice but not in the congenic mast cell-deficient WBB6F1-W/Wv mice. Magnoliae flos inhibited concentration-dependently mast cell-dependent ear swelling response induced by compound 48/80 by topical application. Magnoliae flos inhibited concentration-dependently passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) IgE in rats by topical application. Magnoliae flos also inhibited concentration-dependently the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, Magnoliae flos had a significant inhibitory effect on compound 48/80-induced systemic anaphylactic reaction. These results indicate that Magnoliae flos inhibits immediate-type allergic reactions by inhibition of mast cell degranulation in vivo and in vitro.


Transferability of features for neural networks links to adversarial attacks and defences.
The reason for the existence of adversarial samples is still barely understood. Here, we explore the transferability of learned features to Out-of-Distribution (OoD) classes. We do this by assessing neural networks' capability to encode the existing features, revealing an intriguing connection with adversarial attacks and defences. The principal idea is that, "if an algorithm learns rich features, such features should represent Out-of-Distribution classes as a combination of previously learned In-Distribution (ID) classes". This is because OoD classes usually share several regular features with ID classes, given that the features learned are general enough. We further introduce two metrics to assess the transferred features representing OoD classes. One is based on inter-cluster validation techniques, while the other captures the influence of a class over learned features. Experiments suggest that several adversarial defences decrease the attack accuracy of some attacks and improve the transferability-of-features as measured by our metrics. Experiments also reveal a relationship between the proposed metrics and adversarial attacks (a high Pearson correlation coefficient and low p-value). Further, statistical tests suggest that several adversarial defences, in general, significantly improve transferability. Our tests suggests that models having a higher transferability-of-features have generally higher robustness against adversarial attacks. Thus, the experiments suggest that the objectives of adversarial machine learning might be much closer to domain transfer learning, as previously thought.


A microRNA-mediated decrease in eukaryotic initiation factor 2α promotes cell survival during PS-341 treatment.
MicroRNAs (miRs) play pivotal roles in carcinogenesis and endoplasmic reticulum (ER) that performs the folding, modification and trafficking of proteins targeted to the secretory pathway. Cancer cells often endure ER stress during tumor progression but use the adaptive ER stress response to gain survival advantage. Here we report: (i) A group of miRs, including miR-30b-5p and miR-30c-5p, are upregulated by proteasome inhibitor PS-341 treatment, in HepG2 and MDA-MB-453 cells. (ii) Two representative PS-341-induced miRs: miR-30b-5p and miR-30c-5p are found to promote cell proliferation and anti-apoptosis in both tumor cells. (iii) eIF2α is confirmed as the congenerous target of miR-30b-5p and miR-30c-5p, essential to the anti-apoptotic function of these miRs. (iv) Upregulation of miR-30b-5p or miR-30c-5p, which occurs latter than the increase of phosphorylated eIF2α (p-eIF2α) in the cell under ER stress, suppresses the p-eIF2α/ATF4/CHOP pro-apoptotic pathway. (v) Inhibition of the miR-30b-5p or miR-30c-5p sensitizes the cancer cells to the cytotoxicity of proteasome inhibition. In conclusion, we unravels a new miRs-based mechanism that helps maintain intracellular proteostasis and promote cell survival during ER stress through upregulation of miR-30b-5p and miR-30c-5p which target eIF2α and thereby inhibit the p-eIF2α/ATF4/CHOP pro-apoptotic pathway, identifying miR-30b-5p and miR-30c-5p as potentially new targets for anti-cancer therapies.


A nonmusical paradigm for identifying absolute pitch possessors.
The ability to identify and reproduce sounds of specific frequencies is remarkable and uncommon. The etiology and defining characteristics of this skill, absolute pitch (AP), have been very controversial. One theory suggests that AP requires a specific type of early musical training and that the ability to encode and remember tones depends on these learned musical associations. An alternate theory argues that AP may be strongly dependent on hereditary factors and relatively independent of musical experience. To date, it has been difficult to test these hypotheses because all previous paradigms for identifying AP have required subjects to employ knowledge of musical nomenclature. As such, these tests are insensitive to the possibility of discovering AP in either nonmusicians or musicians of non-Western training. Based on previous literature in pitch memory, a paradigm is presented that is intended to distinguish between AP possessors and nonpossessors independent of the subjects' musical experience. The efficacy of this method is then tested with 20 classically defined AP possessors and 22 nonpossessors. Data from these groups strongly support the validity of the paradigm. The use of a nonmusical paradigm to identify AP may facilitate research into many aspects of this phenomenon.


Evaluation of the metabolism of high energy phosphates in patients with Chagas' disease.
Abnormalities in myocardial metabolism have been observed in patients with heart failure of different etiologies. Magnetic resonance spectroscopy (MRS) with phosphorus-31 is a noninvasive technique that allows detection of myocardial metabolic changes. To determine the resting metabolism of high-energy phosphates in patients with Chagas' disease (CD) by MRS with phosphorus-31. We studied 39 patients with CD, 23 with preserved ventricular function (PF Group) and 16 with ventricular dysfunction (VD Group), assessed by Doppler echocardiography. MRS of the anterosseptal region was performed in 39 patients and 8 normal subjects (C Group) through a Phillips 1.5 Tesla device, obtaining the phosphocreatine/beta-adenosine triphosphate myocardial ratio (PCr/β-ATP). The levels of cardiac PCr/β-ATP were reduced in VD Group in relation to PF Group, and the latter presented reduced levels compared to C Group (VD Group: 0.89 ± 0.31 vs PF Group: 1.47 ± 0.34 vs C Group: 1.88 ± 0.08, p < 0.001). A correlation was found between left ventricular ejection fraction and PCr/β-ATP in 39 patients (r = 0.64, p < 0.001). Patients under functional class I (n = 22) presented PCr/β-ATP of 1.45 ± 0.35, and those in functional classes II and III (n = 17), PCr/β-ATP of 0.94 ± 0.36 (p < 0.001). The 31-phosphorus MRS was able to detect non-invasively changes in the rest energy metabolism of patients with Chagas' disease, with and without systolic dysfunction. These changes were related to the severity of heart impairment.


[Performance of nursing students in the intramuscular administration of drugs].
The goal of the present study was to compare the performance of two groups of nursing undergraduate students, submitted to two different teaching methods, in their ability to perform the intramuscular injection on clients needing the procedure. Both, control (CG) and experimental group (EG) (n = 35 each) were submitted to traditional method and instructional method respectively, according to the methodological procedure described by Nogueira (1995). The performance of the students was evaluated by a nurse who had no knowledge of the group each student belong. The nurse used the "check-list" in order to evaluate the performance of student. The results indicate that the time spent by the students of each group was approximately the same i.e. 9.4 min for CG and 9.3 for EG. The students of the control group showed ordinary performance (42.9%), good performance (48.9%) and very good performance (8.5%) whereas those of the EG showed ordinary performance (5.7%), good performance (60%) and very good performance (34.3%). The present results shows that the students are responsible by their own learning, motivating us to use new teaching methods on nursing undergraduate course.


[Contribution of oxidative stress to pulmonary hypertension induced by chronic hypoxia].
Chronic hypoxia causes pulmonary hypertension and right ventricular hypertrophy associated with media wall thickening of pulmonary arteries in rats. Platelet-activating factor plays an important role in the pulmonary vascular remodeling induced by chronic hypoxia, and reactive oxygen species are involved in tissue injury induced by platelet activating factor. We therefore hypothesized that reactive oxygen species contribute to the pulmonary hypertension induced by chronic hypoxia, and examined the effect of N-acetyl-L-cysteine (NAC), a free radical scavenger and the precursor of glutathione sulfhydryl (GSH), on hypoxia-induced pulmonary hypertension. We used a chemiluminescence-HPLC assay to measure the levels of phosphatidylcholine hydroperoxide (PCOOH) in the rat lungs exposed to hypoxia. Three weeks of normobaric hypoxia (FiO2 = 0.1) with NAC significantly reduced pulmonary hypertension, right ventricular hypertrophy, and media wall thickening of the pulmonary arteries. NAC had no effect on the hematocrit of normoxic or chronically normobaric hypoxic rats. Lung PCOOH levels were significantly higher in the hypoxic rats than in the control rats, and those in the NAC-treated rats were significantly lower than those in the hypoxic rats that were not given NAC. Lung PCOOH levels were significantly higher in the hypoxic rats than in the control rats, and those in the NAC-treated rats were significantly lower than those in the hypoxic rats that were not given NAC. These results indicate that hypoxia induces oxidative stress in the lung tissue, and that oxidative stress may have a role in the development of pulmonary hypertension induced by chronic hypoxia in rats.


Measurements of mitochondrial volumes are affected by the amount of mitochondria used in the determinations.
Mitochondrial water spaces were determined by centrifugal filtration, by using 3H2O and [14C]-sucrose, -mannitol, -inulin and -dextran. The volume (in microliter/mg of mitochondrial protein) of each of the spaces was inversely proportional to the amount of mitochondria (mg of protein) centrifuged. The dextran space (representing extramitochondrial water carried down with the mitochondria) decreased the most, and accounted for most of the changes observed in the other spaces. However, the calculated matrix and intermembrane spaces also decreased when increasing amounts of mitochondria were centrifuged. For each space, the same value was obtained when centrifugal filtration was done at 8000 and at 15,600 g, and when the mitochondria were incubated with the markers for 15 s to 5 min, indicating that sucrose, mannitol and inulin do not penetrate the matrix, nor does dextran penetrate the intermembrane space, under the incubation and centrifugation conditions generally used to measure mitochondrial spaces.


Investigational prostaglandin D2 receptor antagonists for airway inflammation.
By activating DP1 and DP2 receptors on immune and non-immune cells, prostaglandin D2 (PGD2), a major metabolic product of cyclo-oxygenase pathway released after IgE-mediated mast cell activation, has pro-inflammatory effects, which are relevant to the pathophysiology of allergic airway disease. At least 15 selective, orally active, DP2 receptor antagonists and one DP1 receptor antagonist (asapiprant) are under development for asthma and/or allergic rhinitis. In this review, the authors cover the pharmacology of PGD2 and PGD2 receptor antagonists and look at the preclinical, phase I and phase II studies with selective DP1 and DP2 receptor antagonists. Future research should aim to develop once daily compounds and increase the drug clinical potency which, apart from OC000459 and ADC-3680, seems to be relatively low. Further research and development of DP2 receptor antagonists is warranted, particularly in patients with severe uncontrolled asthma, whose management is a top priority. Pediatric studies, which are not available, are required for assessing the efficacy and safety of this novel drug class in children with asthma and allergic rhinitis. Studies on the efficacy of DP2 receptor antagonists in various asthma phenotypes including: smokers, obese subjects, early vs late asthma onset, fixed vs reversible airflow limitation, are required for establishing their pharmacotherapeutic role.


Novel therapeutic targets in primary biliary cirrhosis.
Primary biliary cirrhosis (PBC) is a chronic immune-mediated liver disease characterized by progressive cholestasis, biliary fibrosis and eventually cirrhosis. It results in characteristic symptoms with marked effects on life quality. The advent of large patient cohorts has challenged the view of PBC as a benign condition treated effectively by the single licensed therapy-ursodeoxycholic acid (UDCA). UDCA nonresponse or under-response has a major bearing on outcome, substantially increasing the likelihood that liver transplantation will be required or that patients will die of the disease. In patients with high-risk, treatment-unresponsive or highly symptomatic disease the need for new treatment approaches is clear. Evolution in our understanding of disease mechanisms is rapidly leading to the advent of new and re-purposed therapeutic agents targeting key processes. Notable opportunities are offered by targeting what could be considered as the 'upstream' immune response, 'midstream' biliary injury and 'downstream' fibrotic processes. Combination therapy targeting several pathways or the development of novel agents addressing multiple components of the disease pathway might be required. Ultimately, PBC therapeutics will require a stratified approach to be adopted in practice. This Review provides a current perspective on potential approaches to PBC treatment, and highlights the challenges faced in evaluating and implementing those treatments.


Investigation into the catalytic role for the tryptophan residues within domain III of Pseudomonas aeruginosa exotoxin A.
The role of the tryptophan residues in the substrate-binding and catalytic mechanism of an enzymatically active C-terminal fragment of Pseudomonas aeruginosa exotoxin A was studied by individually or jointly replacing these residues with phenylalanine. Substitution of W-466 decreased the ADP-ribosyltransferase and NAD(+)-glycohydrolase activities by 20- and 3-fold, respectively. In contrast, substitution of W-417 or W-558 with phenylalanine both resulted in a 3-fold decrease in ADP-ribosyltransferase activity with, however, only a decrease by 40% and 70% in NAD(+)-glycohydrolase activity, respectively. Simultaneous replacement of W-466 and W-558 resulted in a 200-fold decrease in ADP-ribosyltransferase and an 6-fold decrease in NAD(+)-glycohydrolase activities, suggesting that W-466 may play a minor role in the transfer of ADP-ribose to the eEF-2 protein. Chemical modification of the tryptophan residues in the wild-type toxin fragment by N-bromosuccinimide revealed the presence of a single residue important for enzymatic activity, W-466, with a minor contribution from W-558. Additionally, tryptophan residues, W-305 and W-417, were refractory to oxidation by N-bromosuccinimide, which likely indicated the buried nature of these residues within the protein structure. Titration of the wild-type toxin fragment with NAD+ resulted in the quenching of the intrinsic tryptophan fluorescence to 58% of the initial value. Titration of the various single and a double tryptophan replacement mutant protein(s) indicated that W-558 and W-466 are responsible for the substrate-induced fluorescence quenching, with the former being responsible for the largest fraction of the observed quenching in the wild-type toxin. Consequently, a molecular mechanism is proposed for the substrate-induced fluorescence quenching of both W-466 and W-558. Furthermore, molecular modeling of the recent crystal structures for both exotoxin A (domain III fragment) and diphtheria toxin, combined with a variety of previous results, has led to the proposal for a catalytic mechanism for the ADP-ribosyltransferase reaction. This mechanism features a SN1 attack (instead of the previously purported SN2 mechanism) by the diphthamide residue (nucleophile) of eukaryotic elongation factor 2 on the C-1 of the nicotinamide ribose of NAD+, which results in an inversion of configuration likely due to steric constraints within the NAD(+)-toxin-elongation factor 2 complex.


Nutrition and nail disease.
The nail is a specialized keratinous skin appendage that grows approximately 2 to 3 mm per month, with complete replacement achieved in 6 to 9 months. Although this structure can be easily overlooked, nail disorders comprise approximately 10% of all dermatologic conditions. This contribution first provides an overview on the basic anatomy of the nail that will delineate between the nail unit (eg, hyponychium, nail bed, proximal nail fold, and matrix) and anatomic components not part of the nail unit (eg, lateral nail folds, nail plate, and eponychium). The function of each nail structure will also be presented. The chemical profile of the normal nail plate is reviewed with a discussion of its keratin content (hair type keratin vs epithelial type keratin), sulfur content, and mineral composition, including magnesium, calcium, iron, zinc, sodium, and copper. The remainder will focus on nail manifestations seen in states of malnutrition. Virtually every nutritional deficiency can affect the growth of the nail in some manner. Finally, the discussion will include anecdotal use of nutritional and dietary supplements in the setting of brittle nail syndrome as well as a brief overview of biotin and its promising utility in the treatment of nail disorders.


Demonstration of pollinator-mediated competition between two native Impatiens species, Impatiens noli-tangere and I. textori (Balsaminaceae).
Plant-plant interspecific competition via pollinators occurs when the flowering seasons of two or more plant species overlap and the pollinator fauna is shared. Negative sexual interactions between species (reproductive interference) through improper heterospecific pollen transfer have recently been reported between native and invasive species demonstrating pollination-driven competition. We focused on two native Impatiens species (I. noli-tangere and I. textori) found in Japan and examined whether pollinator-mediated plant competition occurs between them. We demonstrate that I. noli-tangere and I. textori share the same pollination niche (i.e., flowering season, pollinator fauna, and position of pollen on the pollinator's body). In addition, heterospecific pollen grains were deposited on most stigmas of both I. noli-tangere and I. textori flowers that were situated within 2 m of flowers of the other species resulting in depressed fruit set. Further, by hand-pollination experiments, we show that when as few as 10% of the pollen grains are heterospecific, fruit set is decreased to less than half in both species. These results show that intensive pollinator-mediated competition occurs between I. noli-tangere and I. textori. This study suggests that intensive pollinator-mediated competition occurs in the wild even when interacting species are both native and not invasive.


Precautionary practices of healthcare workers who disinfect medical and dental devices using high-level disinfectants.
BACKGROUND High-level disinfectants (HLDs) are used throughout the healthcare industry to chemically disinfect reusable, semicritical medical and dental devices to control and prevent healthcare-associated infections among patient populations. Workers who use HLDs are at risk of exposure to these chemicals, some of which are respiratory and skin irritants and sensitizers. OBJECTIVE To evaluate exposure controls used and to better understand impediments to healthcare workers using personal protective equipment while handling HLDs. DESIGN Web-based survey. PARTICIPANTS A targeted sample of members of professional practice organizations representing nurses, technologists/technicians, dental professionals, respiratory therapists, and others who reported handling HLDs in the previous 7 calendar days. Participating organizations invited either all or a random sample of members via email, which included a hyperlink to the survey. METHODS Descriptive analyses were conducted including simple frequencies and prevalences. RESULTS A total of 4,657 respondents completed the survey. The HLDs used most often were glutaraldehyde (59%), peracetic acid (16%), and ortho-phthalaldehyde (15%). Examples of work practices or events that could increase exposure risk included failure to wear water-resistant gowns (44%); absence of standard procedures for minimizing exposure (19%); lack of safe handling training (17%); failure to wear protective gloves (9%); and a spill/leak of HLD during handling (5%). Among all respondents, 12% reported skin contact with HLDs, and 33% of these respondents reported that they did not always wear gloves. CONCLUSION Findings indicated that precautionary practices were not always used, underscoring the importance of improved employer and worker training and education regarding HLD hazards.


Targeted delivery of probiotics to enhance gastrointestinal stability and intestinal colonisation.
The aim of this work was to assess the viability of some commercial probiotics after exposure to gastric acid and the possibility of modifying these formulations for delivery into the distal parts of the intestines. Gastrointestinal tolerance testing was conducted for three commercial probiotics and an in-house freeze-dried Lactobacillus acidophilus strain. The contents of the commercial products and the in-house freeze-dried strain were then loaded into capsules for site-specific delivery into the colon using the Phloral® coating technology; the viability upon release was then ascertained. An assessment of the potential of these products to adhere to intestinal cells was also conducted. The results showed that all the commercial products contained the minimum number of probiotic strains as indicated on their respective packages. When gastric acid tolerance tests were performed on these products, all the commercial probiotics and the prepared freeze-dried strain demonstrated over 106 CFU reductions within 5min. When these were encapsulated for site-specific delivery into the distal parts of the gut, viabilities of approximately 90% were obtained after these capsules had been initially deposited in gastric acid for 2h. An evaluation of the ability of the probiotic formulations to adhere to intestinal cells demonstrated adhesion in the range 64-76% for the products evaluated. The need to target the delivery of probiotics into the intestines has been demonstrated here as this offers a greater potential for colonisation of the intestines once the harshness of the stomach has been overcome.


Pre-notification increases uptake of colorectal cancer screening in all demographic groups: a randomized controlled trial.
To assess whether pre-notification is effective in increasing uptake of colorectal cancer screening for all demographic groups. Scottish national colorectal cancer screening programme. Males and females aged 50-74 years received a faecal occult blood test by post to complete at home. They were randomized to receive in addition: the pre-notification letter, the pre-notification letter + information booklet, or the usual invitation. Overall, 59,953 subjects were included in the trial between 13/04/09 and 29/05/09 and followed to 27/11/09. Pre-notification letters were posted two weeks ahead of the screening test kit. Uptake was defined as the return of a screening test and chi-squared tests compared uptake between the trial arms. Logistic regression assessed the impact of the letter and letter + booklet on uptake independently of gender, age, deprivation and screening round. Uptake was higher with both the letter (59.0%) and the letter + booklet (58.5%) compared with the usual invitation (53.9%, p < 0.0001). This increased uptake was seen for males, females, all age groups and all deprivation categories including least deprived females (letter 69.9%, usual invitation 66.6%) and most deprived males (42.6% vs. 36.1%), the groups with the highest and lowest levels of uptake respectively in the pilot screening rounds conducted prior to the roll out of the programme. Uptake with the pre-notification letter compared with the usual invitation was higher both unadjusted and adjusted for demographic factors (odds ratio 1.24, 95% CI 1.193-1.294). Pre-notification is an effective method of increasing uptake in colorectal cancer screening for both genders and all age and deprivation groups.


[Hygienic evaluation of the provision of the population of the Russian Federation with drinking water and measures for its improvements].
In accordance with the sanitary-and-epidemiological requirements, drinking water should be epidemiologically, radiationally, and chemically safe and have good organoleptic properties. In 2007 versus 2006, the situation associated with the state of both underground and surface sources of centralized drinking water supply and the quality of water in the areas of hydraulic works substantially unchanged and continues to remain poor. In 2007, the proportion of water pipes from the underground sources unconformable to the sanitary regulations and standards was 20%, including those due to the absence of sanitary control areas (13.1%), the required complex of waste disposal plants (6.1%), or sterilizing units (2.6%). After water preparation, the quality of water continues to remain poor in terms of sanitary and chemical parameters. In 2007 in the Russian Federation, 17.5 and 5.8% of water samples from the water-supply network were unconformable to the hygienic requirements in sanitary-and-chemical and microbiological parameters, respectively. As for rural water supply, in 2007 in the Russian Federation, rural areas exploited 59,362 water pipes, i.e. 82.5% of the water pipers in Russia as a whole. The proportion of water pipes in the rural areas, which were unconformable to the sanitary regulations and standards, was 21.2% in 2007, including those due to the absence of sanitary control areas (13.7%), a required complex of waste disposal plants (6.8%), or sterilizing units (3.3%).


High estrogen in men after injectable testosterone therapy: the low T experience.
Testosterone replacement improves quality of life and is aromatized in men in adipose tissues to estrogen. Hyperestrogenism is believed to be harmful to male sexuality. This is a description of our experience of screening 34,016 men in the Low T Centers, of which approximately 50% were converted to treatment. Men were treated with injectable testosterone, and we have available data from 2009 to 2014. The data were extracted from our electronic health record (AdvancedMD) of 35 Low T Centers across the United States. In all, 7,215 (20.2%) out of the 34,016 patients had high estradiol levels defined as ≥42.6 pg/ml. Estradiol was measured using electro-chemiluminescence immunoassay. Of the patients who had high estradiol levels, the age distribution was as follows: 132/989 (13.3%) were older than 65 years, 3,753/16,955 (22.1%) were between 45 and 65 years; 2,968/15,857 (18.7%) were between 25 and 44 years, 7/215 (3.3%) were younger than 25 years. The difference between extreme age groups (<25 and ≥65) was statistically significant using a chi-square test (p = .013). The correlation coefficient of serum estradiol to age was .53, SD = 8.21. It was observed that practitioners used aromatase inhibitor and selective estrogen receptor modulator to treat symptoms of hyperestrogenism, irrespective of blood estradiol levels. Gynecomastia was rarely documented as a reason for the prescription. Our finding was that high estradiol levels were not associated with higher rates of low libido but established higher rates of documented low libido with those with normal or lower estradiol levels. The difference was statistically significant (p < .05).


Hereditary sideroblastic anaemia and ataxia: an X linked recessive disorder.
We report two families in which a non-progressive spinocerebellar syndrome and a sideroblastic anaemia are segregating together in an X linked recessive fashion. Four males in two generations of one family and a fifth male from an unrelated family had both conditions. Both the sideroblastic anaemia and the spinocerebellar syndrome differ from those which have previously been reported to be inherited in an X linked recessive manner. The association of these two clinically distinct disorders in two unrelated families suggests that they are either two closely linked loci which have undergone simultaneous mutation or pleiotropic effects of an altered allele at a single locus. All the heterozygous women had normal neurological examinations and normal haematocrits and red cell indices. Some had ring sideroblasts on bone marrow examination, a dimorphic peripheral blood smear, and raised serum free erythrocyte protoporphyrin, suggesting that a proportion of heterozygotes can be detected by appropriate haematological studies.


The Challenge of Reorganizing Rehabilitation Services at the Time of COVID-19 Pandemic: A New Digital and Artificial Intelligence Platform to Support Team Work in Planning and Delivering Safe and High Quality Care.
Introduction: The COVID-19 pandemic has posed great challenges in inpatient rehabilitation services, not only to implement the preventative measures to avoid the spreading of the virus in a highly interactive, multidisciplinary setting but also to create a rehabilitation pathway for post-COVID-19 patients. The aim of this retrospective study was to describe the role of a digital and artificial intelligence platform (DAIP) in facilitating the implementation of changes in a rehabilitation service during the COVID-19 pandemic. Materials and Methods: We gathered qualitative and quantitative descriptors of the DAIP, including measures to assess its efficiency in scheduling therapy sessions, and staff satisfaction using two simple numeric rating scales and the System Usability Scale. We describe how the volume of activity and the quality of care of our rehabilitation service have changed when the DAIP was implemented by comparing the pre-COVID-19 and the pandemic periods for patients' [sex, age, co-morbidities, diagnosis, and Functional Independence Measure (FIM) gain] and service's (bed occupancy, patients' length of stay, and staff capacity) characteristics. Results: Bed occupancy and the impact of rehabilitation on patients' outcome remained stable between the two periods. The DAIP provided a qualitative support for goal setting from remote; 95% of the planned sessions were delivered; the time for scheduling and registering sessions dropped by 50%. Staff satisfaction was about 70% for the easiness and 60% for the usefulness, and the mean "usability" score was close to the cut off for sufficient usability (mean score 65 where 68 is the cut off). Conclusion: By applying the DAIP to rehabilitation treatment, it was shown that the management of rehabilitation can be efficiently performed even in the COVID-19 pandemic. Staff satisfaction reflected a good acceptance of the changes considering the turbulent changes and the stress burden occurring at the time of the pandemic.


Association of Transcobalamin II (TCN2) and Transcobalamin II-Receptor (TCblR) Genetic Variations With Cobalamin Deficiency Parameters in Elderly Women.
Cobalamin (vitamin B12) deficiency is a subtle progressive clinical disorder, affecting nearly 1 in 5 individuals > 60 years old. This deficiency is produced by age-related decreases in nutrient absorption, medications that interfere with vitamin B12 absorption, and other comorbidities. Clinical heterogeneity confounds symptom detection for elderly adults, as deficiency sequelae range from mild fatigue and weakness to debilitating megaloblastic anemia and permanent neuropathic injury. A better understanding of genetic factors that contribute to cobalamin deficiency in the elderly would allow for targeted nursing care and preventive interventions. We tested for associations of common variants in genes involved in cobalamin transport and homeostasis with metabolic indicators of cobalamin deficiency (homocysteine and methylmalonic acid) as well as hematologic, neurologic, and functional performance features of cobalamin deficiency in 789 participants of the Women's Health and Aging Studies. Although not significant when corrected for multiple testing, eight single nucleotide polymorphisms (SNPs) in two genes, transcobalamin II (TCN2) and the transcobalamin II-receptor (TCblR), were found to influence several clinical traits of cobalamin deficiency. The three most significant findings were the identified associations involving missense coding SNPs, namely, TCblR G220R (rs2336573) with serum cobalamin, TCN2 S348F (rs9621049) with homocysteine, and TCN2 P259R (rs1801198) with red blood cell mean corpuscular volume. These SNPs may modify the phenotype in older adults who are more likely to develop symptoms of vitamin B12 malabsorption.


A novel P2X4 receptor-selective antagonist produces anti-allodynic effect in a mouse model of herpetic pain.
Accumulating evidence indicates that purinergic P2X4 receptors (P2X4R: cation channels activated by extracellular ATP) expressed in spinal microglia are crucial for pathological chronic pain caused by nerve damage, suggesting a potential target for drug discovery. We identified NP-1815-PX (5-[3-(5-thioxo-4H-[1,2,4]oxadiazol-3-yl)phenyl]-1H-naphtho[1, 2-b][1,4]diazepine-2,4(3H,5H)-dione) as a novel antagonist selective for P2X4R with high potency and selectivity compared with other P2XR subtypes. In in vivo assay for acute and chronic pain, intrathecal administration of NP-1815-PX produced an anti-allodynic effect in mice with traumatic nerve damage without affecting acute nociceptive pain and motor function (although its oral administration did not produce the effect). Furthermore, in a mouse model of herpetic pain, P2X4R upregulation in the spinal cord exclusively occurred in microglia, and intrathecal NP-1815-PX suppressed induction of mechanical allodynia. This model also showed K(+)/Cl(-) cotransporter 2 (KCC2) downregulation, which is implicated in dorsal horn neuron hyperexcitability; this downregulation was restored by intrathecal treatment with NP-1815-PX or by interfering with brain-derived neurotrophic factor (BDNF) signaling, a P2X4R-activated microglial factor implicated in KCC2 downregulation. Taken together, the newly developed P2X4R antagonist NP-1815-PX produces anti-allodynic effects in chronic pain models without altering acute pain sensitivity, suggesting that microglial P2X4R could be an attractive target for treating chronic pain.


[Surgical treatment of perforated and bleeding gastroduodenal ulcers].
Twelve-year experience of surgical treatment of gastroduodenal ulcers complicated by perforation and bleeding is analyzed. Modern antisecretion and antihelicobacter drugs have changed conception of pathogenesis and treatment of ulcers. Choice of surgical method is the key moment of surgical policy. Perforation and bleeding in ulcer disease are life-threatening complications especially in elderly patients with concomitant diseases. In these patients technical simplicity and tolerance to surgery are very important.


[Prognostic evaluation of the flying longevity of pilots with idiopathic mitral valve prolapse].
In the period of 1995-2001, 576 17- to 49-yr. old pilots of fighters and fighter-bombers with the idiopathic mitral valve prolapse (IMVP) were investigated at the Seventh Central military aviation clinical hospital. In this group, a very feebly marked mitral valve prolapse was found in 98.7% of cases. In 96.7% class-1 IMVP was accompanied by a mitral reguragitation, most often the first-degree one. Based on results of comprehensive examination, medically qualified to fly were 560 out of 576 pilots from the main group (97.0%) and 144 out of 150 pilots from the group of control (97.2%). In the next 5 years the flying career was continued by 76.3% and 78.7% of the pilots, respectively. A clinical-diagnostic and expert algorithm for medical certification of pilots of highly maneuverable aircraft with IMVP was developed which will reduce time of in-hospital medical certification and facilitate more accurately determination of the criteria for immediate and long-range estimation of the flying longevity.


Developing Global Leaders for Research, Regulation, and Stewardship of Crop Protection Chemistry in the 21st Century.
To provide sufficient food and fiber to the increasing global population, the technologies associated with crop protection are growing ever more sophisticated but, at the same time, societal expectations for the safe use of crop protection chemistry tools are also increasing. The goal of this perspective is to highlight the key issues that face future leaders in crop protection, based on presentations made during a symposium titled "Developing Global Leaders for Research, Regulation and Stewardship of Crop Protection Chemistry in the 21st Century", held in conjunction with the IUPAC 13th International Congress of Pesticide Chemistry in San Francisco, CA, USA, during August 2014. The presentations highlighted the fact that leaders in crop protection must have a good basic scientific training and understand new and evolving technologies, are aware of the needs of both developed and developing countries, and have good communication skills. Concern is expressed over the apparent lack of resources to meet these needs, and ideas are put forward to remedy these deficiencies.


Hip Arthroscopy for Femoroacetabular Impingement in a Military Population.
Femoroacetabular impingement (FAI) can lead to hip pain and early joint degeneration. There have been few reports to date on the outcomes of hip arthroscopy for the treatment of FAI in the military population. Purpose/Hypothesis: The purpose of this study was to compare patient demographics with postoperative outcomes after hip arthroscopy for symptomatic FAI and to identify preoperative risk factors for poor outcomes. The hypothesis was that certain preoperative patient characteristics will be predictive of poorer outcomes and that lower outcomes scores will be associated with a higher likelihood of medical separation from the military. Case series; Level of evidence, 4. Retrospective chart review of active-duty and dependent patients older than 18 years who underwent hip arthroscopy for symptomatic FAI from 2009 to 2014 at a single institution. A total of 469 (309 males and 160 females) surgeries were performed on 456 active-duty personnel and 13 dependent civilians, with a mean 2.5-year follow-up. Overall, 39% (n = 179) were able to return to duty (RTD), 18% (n = 82) were medically cleared to return to normal daily activities but did not remain on active duty, and 43% (n = 195) required referral to the Disability Evaluation System (DES). Increasing rank and male sex were positive predictors and Axis 1 psychiatric diagnosis, revision surgery, concomitant psoas tenotomy, multiple medical comorbidities, and complaints of generalized pelvic pain were negative predictors for returning to duty. US Marine Infantry and Special Forces showed improved RTD rates (50%-86%) compared with administrative, more sedentary, occupations (22%). On average, Single Alpha Numeric Evaluation (SANE) and visual analog scale (VAS) scores improved after surgery, with SANE scores improving 37 ± 28 points and VAS scores improving 2.6 ± 2.5 points. The mean postoperative SANE and VAS scores differed significantly between the RTD group and those not returning to duty; 87 and 1.2 points compared with 69 and 3.6 points, respectively ( P < .0001). Hip arthroscopy for the treatment of symptomatic FAI effectively improves pain symptoms and self-reported overall function but shows a much lower than expected return to full, unrestricted active duty in the general active-duty military population. Underlying psychiatric diagnoses, female sex, and more sedentary occupations are associated with lower RTD rates. Furthermore, lower postoperative SANE and VAS scores are associated with lower RTD rates. Only the more active and elite components of the military study population showed RTD rates consistent with previously reported outcomes of return to competitive sports after hip arthroscopy for FAI.


The Influence of Pitch Size on Running Performance and Physiological Responses During Hurling-Specific Small-Sided Games.
The current study examined how the impact of pitch dimensions influences physiological and running performance during 4-minute small-sided games (SSGs). Twenty-four (n = 24) hurling players were monitored with global positioning system and heart rate monitors during the in-season training period. Total distance (in meters), high-speed running distance (in meters) (≥17 km·h), very high-speed running distance (≥22 km·h) (in meters), total accelerations (n), acceleration distance (in meters), and peak and mean velocity (in kilometers per hour) were calculated. Additionally, SSGs rate of perceived exertion (RPESSG; AU), % maximum heart rate, and individualized training impulse (iTRIMP; AU) were collected. The current results show that the manipulation of SSGs pitch size has an impact on the running performance and physiological responses. The data showed that SSGs played on large pitches (SSG80 × 20 m) had greater running demands than medium (SSG60 × 20 m) or small (SSG40 × 20 m) pitches, with significantly more distance covered in all movement categories. Total distance covered at high speed was 354 ± 111 m on a large pitch, 254 ± 72 m on a medium pitch, and 198 ± 62 m on a small pitch. Large pitch dimensions resulted in greater physiological and perceptual demands on players (higher %HRmax, iTRIMP [AU], and RPESSG [AU]) compared with medium and small pitches. The current data help applied practitioners to understand further how modifying different aspects of SSGs can alter the running and physiological responses of players. Moreover, applied practitioners now have consistent information to design and optimize their training time in mixing the physical, technical, and tactical elements within specific SSGs pitch dimensions.


Epidemiology and health-economic burden of urinary-catheter-associated infection in English NHS hospitals: a probabilistic modelling study.
Catheter-associated urinary tract infection (CAUTI) and bloodstream infection (CABSI) are leading causes of healthcare-associated infection in England's National Health Service (NHS), but health-economic evidence to inform investment in prevention is lacking. To quantify the health-economic burden and value of prevention of urinary-catheter-associated infection among adult inpatients admitted to NHS trusts in 2016/17. A decision-analytic model was developed to estimate the annual prevalence of CAUTI and CABSI, and their associated excess health burdens [quality-adjusted life-years (QALYs)] and economic costs (£ 2017). Patient-level datasets and literature were synthesized to estimate population structure, model parameters and associated uncertainty. Health and economic benefits of catheter prevention were estimated. Scenario and probabilistic sensitivity analyses were conducted. The model estimated 52,085 [95% uncertainty interval (UI) 42,967-61,360] CAUTIs and 7529 (UI 6857-8622) CABSIs, of which 38,084 (UI 30,236-46,541) and 2524 (UI 2319-2956) were hospital-onset infections, respectively. Catheter-associated infections incurred 45,717 (UI 18,115-74,662) excess bed-days, 1467 (UI 1337-1707) deaths and 10,471 (UI 4783-13,499) lost QALYs. Total direct hospital costs were estimated at £54.4M (UI £37.3-77.8M), with an additional £209.4M (UI £95.7-270.0M) in economic value of QALYs lost assuming a willingness-to-pay threshold of £20,000/QALY. Respectively, CABSI accounted for 47% (UI 32-67%) and 97% (UI 93-98%) of direct costs and QALYs lost. Every catheter prevented could save £30 (UI £20-44) in direct hospital costs and £112 (UI £52-146) in QALY value. Hospital catheter prevention is poised to reap substantial health-economic gains, but community-oriented interventions are needed to target the large burden imposed by community-onset infection.


The genetic basis of kidney cancer: why is tuberous sclerosis complex often overlooked?
Fifty years ago, the Eker rat was identified as the first animal model of hereditary renal adenoma and carcinoma, with histopathology resembling human renal carcinoma. Ten years ago, a mutation in the TSC2 gene was identified in the Eker rat at Fox Chase Cancer Center by Yeung and Knudson, and in Tokyo by Kobayashi and Hino. The literature contains dozens of reports of renal cell carcinoma (RCC) in tuberous sclerosis complex (TSC) patients, including tumors in children as young as five and one report in an infant. Despite these facts, the association between TSC and RCC is under-recognized, and sometimes completely omitted from discussions of inherited renal carcinoma. Here, we will review the clinical association of RCC in TSC, consider the factors that have led to its under-emphasis within the RCC field, address the cellular and biochemical mechanisms that may contribute to RCC in cells with TSC1 or TSC2 mutations, and finally discuss the ways in which the TSC signaling pathways may be linked to sporadic RCC in the general population.


Potential effect of skull thickening on the associations between cognition and brain atrophy in ageing.
intracranial volume (ICV) is commonly used as a marker of premorbid brain size in neuroimaging studies as it is thought to remain fixed throughout adulthood. However, inner skull table thickening would encroach on ICV and could mask actual brain atrophy. we investigated the effect that thickening might have on the associations between brain atrophy and cognition. the sample comprised 57 non-demented older adults who underwent structural brain MRI at mean age 72.7 ± 0.7 years and were assessed on cognitive ability at mean age 11 and 73 years. Principal component analysis was used to derive factors of general cognitive ability (g), information processing speed and memory from the recorded cognitive ability data. The total brain tissue volume and ICV with (estimated original ICV) and without (current ICV) adjusting for the effects of inner table skull thickening were measured. General linear modelling was used to test for associations. all cognitive ability variables were significantly (P < 0.01) associated with percentage total brain volume in ICV measured without adjusting for skull thickening (g: η(2) = 0.177, speed: η(2) = 0.264 and memory: η(2) = 0.132). After accounting for skull thickening, only speed was significantly associated with percentage total brain volume in ICV (η(2) = 0.085, P = 0.034), not g or memory. not accounting for skull thickening when computing ICV can distort the association between brain atrophy and cognitive ability in old age. Larger samples are required to determine the true effect.


Microbacterium hydrothermale sp. nov., an actinobacterium isolated from hydrothermal sediment.
A Gram-stain-positive, aerobic, non-motile, rod-shaped bacterium, strain 0704C9-2(T), was isolated from hydrothermal sediment of the Indian Ocean. The organism grew with 0-5% (w/v) NaCl and at 10-37 °C, with optimal growth occurring with 1% NaCl and at 28-30 °C. Comparative 16S rRNA gene sequence analysis revealed that strain 0704C9-2(T) belonged to the genus Microbacterium. It exhibited highest 16S rRNA gene sequence similarity with Microbacterium testaceum DSM 20166(T) (98.4%). Levels of similarity with the type strains of all other recognized species of the genus Microbacterium were less than 98.0%. DNA-DNA hybridization experiments with strain 0704C9-2(T) and its closest relative, M. testaceum DSM 20166(T), revealed a low reassociation value of 42.9%. The DNA G+C content of strain 0704C9-2(T) was 73.3 mol%. The cell-wall peptidoglycan contained ornithine and the acyl type was glycolyl. The major whole-cell sugars were mannose, galactose, rhamnose and glucose. The major cellular fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C16:0 and iso-C15:0. The predominant menaquinones were MK-11, MK-10 and MK-12. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids and an unknown phospholipid. On the basis of phenotypic, phylogenetic and genotypic data, strain 0704C9-2(T) represents a novel species within the genus Microbacterium, for which the name Microbacterium hydrothermale sp. nov. is proposed. The type strain is 0704C9-2(T) ( = LMG 27542(T) = CGMCC 1.12512(T)).


Plasma LTBP2 as a potential biomarker in differential diagnosis of connective tissue disease-associated interstitial lung disease and idiopathic pulmonary fibrosis: a pilot study.
Few biomarkers distinguish connective tissue disease-associated interstitial lung disease (CTD-ILD) from idiopathic pulmonary fibrosis (IPF). Latent transforming growth factor-β binding protein-2 (LTBP2), a secreted extracellular matrix protein, is involved in pulmonary fibrosis. However, the role of LTBP2 in differentially diagnosing CTD-ILD and IPF is unclear. In this study, enzyme-linked immunosorbent assays quantified plasma LTBP2 concentrations in 200 individuals (35 healthy controls, 42 CTD patients without ILD, 89 CTD-ILD patients, and 34 IPF patients). CTD-ILD and IPF were further classified based on chest imaging pattern and pulmonary function test results. Plasma LTBP2 levels were significantly elevated in the IPF group compared with the CTD-ILD group. ROC analysis further suggested the possible value of LTBP2 in differentially diagnosing CTD-ILD and IPF. Additionally, CTD-ILD patients with progressive lung fibrosis had higher plasma LTBP2 concentrations than those who did not. Similarly, patients with IPF developing acute exacerbation showed higher plasma LTBP2 levels than those with stable IPF. This is the first study showing that LTBP2 was closely associated with the usual interstitial pneumonia (UIP) pattern in rheumatoid arthritis-associated ILD (RA-ILD). Moreover, the optimal cutoff values of LTBP2 for distinguishing IPF from CTD-UIP/RA-UIP were 33.75 and 38.33 ng/mL with an AUC of 0.682 and 0.681, respectively. Our findings suggest that plasma LTBP2 levels may differentially diagnose CTD-ILD and IPF, and assess their fibrotic activity. Additionally, clinical LTBP2 evaluation may be a great aid to identifying the presence of the UIP pattern in RA-ILD and to discriminating IPF from CTD-UIP, particularly RA-UIP.


Interaction of the aryl hydrocarbon receptor ligand 6-methyl-1,3,8-trichlorodibenzofuran with estrogen receptor alpha.
The polycyclic aromatic hydrocarbon 6-methyl-1,3,8-trichlorodibenzofuran (MCDF) is related to the industrial byproduct dioxin and is a weak agonist and partial antagonist at the aryl hydrocarbon receptor (AhR). Tamoxifen is used for the treatment and prevention of breast cancer and interferes with the interaction of estrogen with estrogen receptor alpha (ER). The combination of MCDF and tamoxifen lowered the effective dose of both drugs required to inhibit 7,12-dimethylbenz(a)anthracene-induced mammary tumor growth in rats and protected against the estrogenic effects of tamoxifen on the uterus in rats (A. McDougal et al., Cancer Res 2001;61:3902-7), pointing to the potential use of MCDF in breast cancer treatment. Potential AhR-ER cross-talk is evidenced by the antiestrogenic activity of MCDF and the degradative effect of MCDF on ER protein levels. Our studies confirmed that MCDF degraded the ER. MCDF displayed antiestrogenic activity at higher concentrations in MCF-7 human breast cancer cells, but MCDF alone (10(-6) M) stimulated the growth of MCF-7 cells. MCDF also activated an estrogen response element (ERE)-luciferase reporter and increased mRNA levels of the estrogen-responsive gene transforming growth factor (TGF)-alpha. The estrogenic effects of MCDF are ER dependent because they were blocked by the pure antiestrogen ICI 182,780. MCDF induced ER-coactivator interaction in glutathione S-transferase pull-down assays and the formation of an ER.ERE complex in gel mobility shift assays, further indicating that the estrogenic actions of MCDF are mediated by the ER. In addition, knockdown of the AhR with small interfering RNA did not affect MCDF-induced ERE-luciferase activity. Overall, these data support the conclusion that MCDF is a partial agonist at the ER. This study provides the first evidence for the direct interaction of the ER with MCDF and challenges the view that MCDF is simply an AhR-specific ligand.


Suppression of oxidative stress by β-hydroxybutyrate, an endogenous histone deacetylase inhibitor.
Concentrations of acetyl-coenzyme A and nicotinamide adenine dinucleotide (NAD(+)) affect histone acetylation and thereby couple cellular metabolic status and transcriptional regulation. We report that the ketone body d-β-hydroxybutyrate (βOHB) is an endogenous and specific inhibitor of class I histone deacetylases (HDACs). Administration of exogenous βOHB, or fasting or calorie restriction, two conditions associated with increased βOHB abundance, all increased global histone acetylation in mouse tissues. Inhibition of HDAC by βOHB was correlated with global changes in transcription, including that of the genes encoding oxidative stress resistance factors FOXO3A and MT2. Treatment of cells with βOHB increased histone acetylation at the Foxo3a and Mt2 promoters, and both genes were activated by selective depletion of HDAC1 and HDAC2. Consistent with increased FOXO3A and MT2 activity, treatment of mice with βOHB conferred substantial protection against oxidative stress.


[Prenatal determination of fetal RHD in maternal plasma: two-years experience of routine clinical use].
To evaluate the predictive value of RHD fetal genotype in maternal plasma of Rh D negative mothers after 10 weeks of gestation in a clinical use. Prospective, comparative study between fetal RHD genotyping in maternal plasma, with amplification of exons 4,5,10 of the RHD gene, by real-time multiplex PCR, and Rh D serology at birth, in 218 pregnancy and their 223 babies, between November 2002 and 2004. Combining the amplification of three exons, the concordance rate of fetal Rh D genotyping in maternal plasma and baby phenotyping at delivery was 100%. Four women whose the babies were Rh D negative were positive for RHD exon 10 during pregnancy. This positivity was, in three cases, correlated with the presence of RHDpsi pseudogene and in last case, with a haplotype Cdes (r's). RHD genotyping was performed for five twin pregnancies. Multiplex PCR using maternal plasma provides perfect prenatal prediction of fetal RHD gene. These results confirm that this non invasive procedure is the preferred method for assessing Rh D fetal status in Rh negative mothers. Using this method for two years in routine practice has led us to modify our management scheme for sensitized Rh D-negative pregnant women.


Functional conservation between rodents and chicken of regulatory sequences driving skeletal muscle gene expression in transgenic chickens.
Regulatory elements that control expression of specific genes during development have been shown in many cases to contain functionally-conserved modules that can be transferred between species and direct gene expression in a comparable developmental pattern. An example of such a module has been identified at the rat myosin light chain (MLC) 1/3 locus, which has been well characterised in transgenic mouse studies. This locus contains two promoters encoding two alternatively spliced isoforms of alkali myosin light chain. These promoters are differentially regulated during development through the activity of two enhancer elements. The MLC3 promoter alone has been shown to confer expression of a reporter gene in skeletal and cardiac muscle in transgenic mice and the addition of the downstream MLC enhancer increased expression levels in skeletal muscle. We asked whether this regulatory module, sufficient for striated muscle gene expression in the mouse, would drive expression in similar domains in the chicken. We have observed that a conserved downstream MLC enhancer is present in the chicken MLC locus. We found that the rat MLC1/3 regulatory elements were transcriptionally active in chick skeletal muscle primary cultures. We observed that a single copy lentiviral insert containing this regulatory cassette was able to drive expression of a lacZ reporter gene in the fast-fibres of skeletal muscle in chicken in three independent transgenic chicken lines in a pattern similar to the endogenous MLC locus. Reporter gene expression in cardiac muscle tissues was not observed for any of these lines. From these results we conclude that skeletal expression from this regulatory module is conserved in a genomic context between rodents and chickens. This transgenic module will be useful in future investigations of muscle development in avian species.


Identification of a novel synonymous mutation in the human β -Ureidopropionase Gene UPB1 affecting pre-mRNA splicing.
β-Ureidopropionase is the third enzyme of the pyrimidine degradation pathway and it catalyzes the conversion of N-carbamyl-β-alanine and N-carbamyl-β-aminoisobutyric acid to β-alanine and β-aminoisobutyric acid, respectively, and ammonia and CO2. To date, only 16 genetically confirmed patients with a complete ß-ureidopropionase deficiency have been reported. Here, we report the clinical, biochemical, and molecular analysis of a newly identified patient with β-ureidopropionase deficiency. Mutation analysis of the UPB1 gene showed that the patient was compound heterozygous for a novel synonymous mutation c.93C >T (p.Gly31Gly) in exon 1 and a previously described missense mutation c.977G >A (p.Arg326Gln) in exon 9. The in silico predicted effect of the synonymous mutation p.Gly31Gly on pre-mRNA splicing was investigated using a minigene approach. Wild-type and the mutated minigene constructs, containing the entire exon 1, intron 1, and exon 2 of UPB1, yielded different splicing products after expression in HEK293 cells. The c.93C >T (p.Gly31Gly) mutation resulted in altered pre-mRNA splicing of the UPB1 minigene construct and a deletion of the last 13 nucleotides of exon 1. This deletion (r.92_104delGCAAGGAACTCAG) results in a frame shift and the generation of a premature stop codon (p.Lys32SerfsX31). Using a minigene approach, we have thus identified the first synonymous mutation in the UPB1 gene, creating a cryptic splice-donor site affecting pre-mRNA splicing.


Reliability and validity of the questionnaire to determine the biosocial rhythms of daily living in the disabled elderly.
The questionnaire to determine the biosocial rhythms of daily living in the disabled elderly was newly developed. This questionnaire was aimed to evaluate a state of synchronization of biological rhythms in the disabled elderly. Eighteen items of the questionnaire relating to the synchronization of biological rhythms were finally selected by the test-retest method that was conducted for 68 disabled elderly living in a community with a duration of one year. The factor analysis showed that the questionnaire consisted of five factors: outdoor activities, ultradian rhythms, subjective evaluation of health status, social support, and sleep habits. The cumulative contribution rate of five factors was 53.2%. Reliability of the questionnaire was confirmed by a calculation of the Equal-length Spearman-Brown coefficients ranging from 0.60 to 0.80. Regarding the construct validity of the questionnaire, results of factor analysis showed five factors that were consistent with the synchronizers known in chronobiology. The total score of the questionnaire was significantly correlated to Barthel Index score and the competence score, suggesting that it partly reflects the activities of daily living of the disabled elderly. We conclude that a new questionnaire to determine the biosocial rhythm of daily living in the disabled elderly is useful to evaluate the biosocial synchronization of the disabled elderly because of its high reliability and validity.


Glucose production and substrate cycle activity in a fasting adapted animal, the northern elephant seal.
During prolonged fasting physiological mechanisms defend lean tissue from catabolism. In the fasting state, glucose is derived solely from gluconeogenesis, requiring some catabolism of amino acids for gluconeogenic substrates. This creates a conflict in animals undergoing fasts concurrently with metabolically challenging activities. This study investigated glucose metabolism in fasting and developing neonatal elephant seals. Glucose production and glucose cycle activity were measured early (2 weeks) and late (6 weeks) in the postweaning fasting period. Additionally the role of regulatory hormones on glucose production and glucose cycle activity were investigated. Glucose cycle activity was highly variable throughout the study period, did not change over the fasting period, and was not correlated with insulin or glucagon level. Endogenous glucose production (EGP) was 2.80+/-0.65 mg kg(-1) min(-1) early and 2.21+/-0.12 during late fasting. Insulin to glucagon molar ratio decreased while cortisol levels increased over the fast (t=5.27, 2.84; P=0.003, 0.04; respectively). There was no relationship between EGP and hormone levels. The glucose production values measured in this study were high and exceeded the estimated gluconeogenic substrate available. These data suggest extensive glucose recycling via Cori cycle activity occurring in northern elephant seals, and we propose a possible justification for this recycling.


The Association between ABO and Rh Blood Groups and Risk of Endometriosis in Iranian Women.
Endometriosis is a common gynaecological disease that affects quality of life for women. Several studies have revealed that both environmental and genetic factors contribute to the development of endometriosis. The aim of this study was to investigate the distribution of ABO and Rh blood groups in Iranian women with endometriosis who presented to two referral infertility centers in Tehran, Iran. In this case-control study, women who referred to Royan Institute and Arash Women's Hospital for diagnostic laparoscopy between 2013 and 2014 were assessed. Based on the laparoscopy findings, we categorized the women into two groups: endometriosis and control (women without endometriosis and normal pelvis). Chi-square and logistic regression tests were used for data analysis. In this study, we assessed 433 women, of which 213 patients were assigned to the endometriosis group while the remaining 220 subjects comprised the control group. The most frequent ABO blood group was O (40.6%). The least frequent blood group was AB (4.8%). In terms of Rh blood group, Rh+ (90.1%) was more frequent than Rh- (9.9%). There was no significant correlation between ABO (P=0.091) and Rh (P=0.55) blood groups and risk of endometriosis. Also, there was no significant difference between the two groups with regards to the stage of endometriosis and distribution of ABO and Rh blood groups (P>0.05). Although the O blood group was less dominant in Iranian women with endometriosis, we observed no significant correlation between the risk of endometriosis and the ABO and Rh blood groups. Endometriosis severity was not correlated to any of these blood groups.


Reliability of Symptoms and Dipstick for Postoperative Catheter-Associated Urinary Tract Infections.
The aim of the study was to determine the diagnostic reliability of patient symptoms and urine dipstick results, including blood, leukocyte esterase, and nitrite, in diagnosing postoperative catheter-associated urinary tract infections (CAUTIs) after gynecologic surgery. A prospective cohort study of patients undergoing gynecologic surgeries who required short-term (>24 hours) postoperative catheterization was conducted. Patients completed a questionnaire regarding symptoms. Urine dipstick analysis was completed on catheterized urine samples and urine was sent for culture for all patients. Positive likelihood ratios (LRs) were used to examine diagnostic reliability of patient symptoms and urine dipstick results in diagnosing postoperative CAUTIs. Fifty-seven patients with postoperative short-term indwelling catheterization were recruited, 25 (44%) of whom had CAUTIs diagnosed by urine culture at recruitment and 32 (56%) of whom did not have CAUTIs. Urine dipstick parameters were found to successfully diagnose CAUTIs, with positive LRs of 1.44 (95% confidence interval [CI], 1.04-1.99), 6.77 (95% CI, 2.23-20.52), and 9.47 (95% CI, 1.23-72.69) for blood, leukocyte esterase, and nitrite, respectively. The combination of leukocyte esterase and nitrite yielded a positive LR of 9.48 (95% CI, 2.62-34.25). Individual symptoms, alone or in combination, did not successfully diagnose positive urine culture (positive LRs <1.8). Urine dipstick parameters are reliable diagnostic tests for diagnosing postoperative CAUTIs after gynecologic surgery, particularly when in combination. Patient symptoms have little diagnostic value for positive urine cultures in catheterized patients after gynecologic surgery.


Effect of glucose on glycerol metabolism by Clostridium butyricum DSM 5431.
The levels of 1,3-propanediol dehydrogenase and of the glycerol dehydrogenase in Clostridium butyricum grown on glucose-glycerol mixtures were similar to those found in extracts of cells grown on glycerol alone, which can explain the simultaneous glucose-glycerol consumption. On glycerol, 43% of glycerol was oxidized to organic acids to obtain energy for growth and 57% to produce 1,3-propanediol. With glucose-glycerol mixtures, glucose catabolism was used by the cells to produce energy through the acetate-butyrate production and NADH, whereas glycerol was used chiefly in the utilization of the reducing power since 92-93% of the glycerol flow was converted through the 1,3-propanediol pathway. The apparent K(m)s for the glycerol dehydrogenase was 16-fold higher for the glycerol than that for the glyceraldehyde in the case of the glyceraldehyde-3-phosphate dehydrogenase and fourfold higher for the NAD+, providing an explanation for the shift of the glycerol flow toward 1,3-propanediol when cells were grown on glucose-glycerol mixtures.


G-quadruplex ligands targeting telomeres do not inhibit HIV promoter activity and cooperate with latency reversing agents in killing latently infected cells.
Altered telomere maintenance mechanism (TMM) is linked to increased DNA damage at telomeres and telomere uncapping. We previously showed that HIV-1 latent cells have altered TMM and are susceptible to ligands that target G-quadruplexes (G4) at telomeres. Susceptibility of latent cells to telomere targeting could potentially be used to support approaches to eradicate HIV reservoirs. However, G4 ligands also target G-quadruplexes in promoters blocking gene transcription. Since HIV promoter sequence can form G-quadruplexes, we investigated whether G4 ligands interfere with HIV-1 promoter activity and virus reactivation from latency, and whether telomere targeting could be combined with latency reversing agents (LRAs) to promote elimination of HIV reservoirs. Our results indicate that Sp1 binding region in HIV-1 promoter can adopt G4 structures in duplex DNA, and that in vitro binding of Sp1 to G-quadruplex is blocked by G4 ligand, suggesting that agents targeting telomeres interfere with virus reactivation. However, our studies show that G4 agents do not affect HIV-1 promoter activity in cell culture, and do not interfere with latency reversal. Importantly, primary memory CD4 + T cells infected with latent HIV-1 are more susceptible to combined treatment with LRAs and G4 ligands, indicating that drugs targeting TMM may enhance killing of HIV reservoirs. Using a cell-based DNA repair assay, we also found that HIV-1 infected cells have reduced efficiency of DNA mismatch repair (MMR), and base excision repair (BER), suggesting that altered TMM in latently infected cells could be associated with accumulation of DNA damage at telomeres and changes in telomeric caps.


Spectral character of sunlight modulates photosynthesis of previtamin D3 and its photoisomers in human skin.
The photosynthesis of previtamin D3 from 7-dehydrocholesterol in human skin was determined after exposure to narrow-band radiation or simulated solar radiation. The optimum wavelengths for the production of previtamin D3 were determined to be between 295 and 300 nanometers. When human skin was exposed to 295-nanometer radiation, up to 65 percent of the original 7-dehydrocholesterol content was converted to previtamin D3. In comparison, when adjacent skin was exposed to simulated solar radiation, the maximum formation of previtamin D3 was about 20 percent. Major differences in the formation of lumisterol3, and tachysterol3 from previtamin D3 were also observed. It is concluded that the spectral character of natural sunlight has a profound effect on the photochemistry of 7-dehydrocholesterol in human skin.


Multiple monovalent ion-dependent pathways for the folding of the L-21 Tetrahymena thermophila ribozyme.
Synchrotron hydroxyl radical (*OH) footprinting is a technique that monitors the local changes in solvent accessibility of the RNA backbone on milliseconds to minutes time-scales. The Mg(2+)-dependent folding of the L-21 Sca 1 Tetrahymena thermophila ribozyme has been followed using this technique at an elevated concentration of monovalent ion (200 mM NaCl) and as a function of the initial annealing conditions and substrate. Previous studies conducted at low concentrations of monovalent ion displayed sequential folding of the P4-P6 domain, the peripheral helices and the catalytic core, with each protection displaying monophasic kinetics. For ribozyme annealed in buffer containing 200 mM NaCl and folded by the addition of 10 mM MgCl(2), multiple kinetic phases are observed for *OH protections throughout the ribozyme. The independently folding P4-P6 domain is the first to fold with its protections displaying 50-90% burst phase amplitudes. That the folding of P4-P6 within the ribozyme does not display the 100% burst phase of isolated P4-P6 at 200 mM NaCl shows that interactions with the remainder of the ribozyme impede this domain's folding. In addition, *OH protections constituting each side of a tertiary contact are not coincident in some cases, consistent with the formation of transient non-native interactions. While the peripheral contacts and triple helical scaffold exhibit substantial burst phases, the slowest protection to appear is J8/7 in the catalytic core, which displays a minimal burst amplitude and whose formation is coincident with the recovery of catalytic activity. The number of kinetic phases as well as their amplitudes and rates are different when the ribozyme is annealed in low-salt buffer and folded by the concomitant addition of monovalent and divalent cations. Annealed substrate changes the partitioning of the ribozyme among the multiple folding populations. These results provide a map of the early steps in the ribozyme's folding landscape and the degree to which the preferred pathways are dependent upon the initial reaction conditions.


Effects of rare sugar D-allulose on acid production and probiotic activities of dairy lactic acid bacteria.
It has recently been reported that the rare sugar d-allulose has beneficial effects, including the suppression of postprandial blood glucose elevation in humans, and can be substituted for sucrose as a low-calorie food ingredient. To examine the applications of d-allulose in the dairy industry, we investigated the effects of d-allulose on the acid production of 8 strains of yogurt starter (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and 4 strains of lactococci, including potential probiotic candidates derived from dairy products. Acid production by 2 L. delbrueckii ssp. bulgaricus yogurt starter strains in milk was suppressed by d-allulose, but this phenomenon was also observed in some strains with another sugar (xylose), a sugar alcohol (sorbitol), or both. In contrast, among the dairy probiotic candidates, Lactococcus lactis H61, which has beneficial effects for human skin when drunk as part of fermented milk, was the only strain that showed suppression of acid production in the presence of d-allulose. Strain H61 did not metabolize d-allulose. We did not observe suppression of acid production by strain H61 with the addition of xylose or sorbitol, and xylose and sorbitol were not metabolized by strain H61. The acid production of strain H61 after culture in a constituted medium (tryptone-yeast extract-glucose broth) was also suppressed with the addition of d-allulose, but growth efficiency and sugar fermentation style were not altered. Probiotic activities-such as the angiotensin-converting enzyme inhibitory activity of H61-fermented milk and the superoxide dismutase activity of H61 cells grown in tryptone-yeast extract-glucose broth-were not affected by d-allulose. d-Allulose may suppress acid production in certain lactic acid bacteria without altering their probiotic activity. It may be useful for developing new probiotic dairy products from probiotic strains such as Lactococcus lactis H61.


Involvement of Ca2+ sensitization in ropivacaine-induced contraction of rat aortic smooth muscle.
The mechanisms of amino-amide local anesthetic agent-induced vasoconstriction remain unclear. The current study was designed to examine the roles of the protein kinase C (PKC), Rho kinase, and p44/42 mitogen-activated protein kinase (p44/42 MAPK) signaling pathways in calcium (Ca2+)-sensitization mechanisms in ropivacaine-induced vascular contraction. Endothelium-denuded rat aortic rings, segments, and strips were prepared. The cumulative dose-response relations of contraction and intracellular Ca2+ concentration to ropivacaine were tested, using isometric force transducers and a fluorometer, respectively. The dose-dependent ropivacaine-induced phosphorylation of PKC and p44/42 MAPK and the membrane translocation of Rho kinase were also detected using Western blotting. Ropivacaine induced a dose-dependent biphasic contractile response and an increase in intracellular Ca2+ concentration of rat aortic rings, increasing at concentrations of 3 x 10 m to 3 x 10 m and decreasing from 10 m to 3 x 10 m, with a greater tension/intracellular Ca2+ concentration ratio than that induced with potassium chloride. The contraction was attenuated in a dose-dependent manner, by the PKC inhibitors bisindolylmaleimide I and calphostin C, the Rho-kinase inhibitor Y 27632, and the p44/42 MAPK inhibitor PD 098059. Ropivacaine also induced an increase in phosphorylation of PKC and p44/42 MAPK, and membrane translocation of Rho kinase in accordance with the contractile responses, which were also significantly inhibited by bisindolylmaleimide I and calphostin C, Y 27632, and PD 098059, correspondingly. These findings demonstrated that PKC-, Rho kinase-, and p44/42 MAPK-mediated Ca2+-sensitization mechanisms are involved in the ropivacaine-induced biphasic contraction of rat aortic smooth muscle.


Activation of Dendritic Cells in Tonsils Is Associated with CD8 T Cell Responses following Vaccination with Live Attenuated Classical Swine Fever Virus.
Classical swine fever (CSF) is a highly contagious disease caused by the classical swine fever virus (CSFV). The live attenuated C-strain vaccine is highly efficacious, initiating protection within several days of delivery. The vaccine strain is detected in the tonsil early after inoculation, yet little is known of the role that tonsillar immune cells might play in initiating protection. Comparing the C-strain vaccine with the pathogenic CSFV Alfort-187 strain, changes in the myeloid cell compartment of the tonsil were observed. CSFV infection led to the emergence of an additional CD163+CD14+ cell population, which showed the highest levels of Alfort-187 and C-strain infection. There was also an increase in both the frequency and activation status (as shown by increased MHC-II expression) of the tonsillar conventional dendritic cells 1 (cDC1) in pigs inoculated with the C-strain. Notably, the activation of cDC1 cells coincided in time with the induction of a local CSFV-specific IFN-γ+ CD8 T cell response in C-strain vaccinated pigs, but not in pigs that received Alfort-187. Moreover, the frequency of CSFV-specific IFN-γ+ CD8 T cells was inversely correlated to the viral load in the tonsils of individual animals. Accordingly, we hypothesise that the activation of cDC1 is key in initiating local CSFV-specific CD8 T cell responses which curtail early virus replication and dissemination.


Synergistic effect of ginger and nifedipine on human platelet aggregation: a study in hypertensive patients and normal volunteers.
In this study, we evaluated the synergistic effect of ginger and nifedipine on anti-platelet aggregation in normal human volunteers and hypertensive patients. The results showed that the percentage of platelet aggregation induced by collagen, ADP and epinephrine in hypertensive patients was larger than that in normal volunteers. Either aspirin or ginger could potentiate the anti-platelet aggregation effect of nifedipine in normal volunteer and hypertensive patients. These results suggested that ginger and nifedipine possessed synergistic effect on anti-platelet aggregation. A combination of 1 g ginger with 10 mg nifedipine per day could be valuable for cardiovascular and cerebrovascular complication due to platelet aggregation.


Geochemical Fractionation and Assessment of Probabilistic Ecological Risk of Potential Toxic Elements in Sediments Using Monte Carlo Simulations.
The need for further research into potentially toxic elements in Serbian rivers led to an investigation of distributions, sources, and ecological risks in a sample base of sediments from 15 rivers. The analyses were carried out through both experimental and theoretical methods. Geochemical fractionation of Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, As, V, and Zn in sediments was studied using a sequential extraction procedure. Both a Håkanson risk index (RI) and a Monte Carlo simulation (MCS) were used in order to estimate ecological risk, applying the probability distribution of RI values instead of single-point calculations. In order to both further the development of the used method and include additional processes, software for the simulations was developed instead of using proprietary solutions. Metal fractionation showed high percentage recoveries of Cd, Cr, Co, Cu, Fe, Ni, and V in residual fractions. The high content of Pb, Mn, and Zn in mobile fractions might cause serious environmental concerns. In some localities, Cu and Cd could be problematic elements, since their mobility was high. An environmental assessment based on the described criteria provided risk levels varying from low to median (mainly contributed by Cd and Cu).


Effect of restorative, endodontic, and fatigue treatments on the cuspal deflection of maxillary premolars subjected to different cyclic occlusal forces: an in vitro study.
This study aimed to determine the effect of adhesive direct composite restorations, endodontic treatments, and fatigue treatments on the cuspal deflection of maxillary premolars subjected to different cyclic occlusal forces. Thirty intact maxillary second premolars were selected. Ten teeth were left untreated (group IN), 10 teeth were subjected to endodontic and restorative treatment (group FL), and the remaining 10 teeth were subjected to endodontic, restorative, and fatigue treatments (group FT). All teeth were subjected to 5 occlusal compressive loading forces (98, 147, 196, 245, and 294 N) with a universal testing device. A total of 15 experimental groups were obtained with 3 tooth conditions (IN, FL, FT) and 5 different occlusal loading values. Deflection amounts (μm) were measured with laser sensors and recorded, and obtained data were statistically analyzed with one-way analysis of variance at a significance level of .05. Mean cuspal deflection values (μm) and SDs of experimental groups ranged as follows: IN-98 (24.4 ± 19.8), IN-147 (34.8 ± 28.9), IN-196 (43.8 ± 34.7), IN-245 (54.5 ± 46.4), IN-294 (60.3 ± 50.6), FL-98 (56 ± 49.1), FL-147 (62.6 ± 49.6), FL-196 (72.4 ± 52.1), FL-245 (81.3 ± 56), FL-294 (92.2 ± 60.9), FT-98 (77.2 ± 80.9), FT-147 (83.4 ± 81.3), FT-196 (92.6 ± 83.7), FT-245 (102.7 ± 85.4), and FT-294 (124.2 ± 89.5). Mean values of three main experimental groups were as follows: IN (43.5 μm), FL (72.9 μm) and FT (96.0 μm). Significant differences were found between the three main groups and relevant subgroups (P < .001). Highest cuspal deflection values (CDV) were obtained in FT groups. Lowest CDV were obtained in IN groups. FL groups showed higher deflection values than IN groups. CDV increased progressively as the teeth were restored and subjected to fatigue treatment.