How health service delivery guides the allocation of major trauma patients in the intensive care units of the inclusive (hub and spoke) trauma system of the Emilia Romagna Region (Italy). A cross-sectional study.
To evaluate cross-sectional patient distribution and standardised 30-day mortality in the intensive care units (ICU) of an inclusive hub and spoke trauma system. ICUs of the Integrated System for Trauma Patient Care (SIAT) of Emilia-Romagna, an Italian region with a population of approximately 4.5 million. 5300 patients with an Injury Severity Score (ISS) >15 were admitted to the regional ICUs and recorded in the Regional Severe Trauma Registry between 2007 and 2012. Patients were classified by the Abbreviated Injury Score as follows: (1) traumatic brain injury (2) multiple injuriesand (3) extracranial lesions. The SIATs were divided into those with at least one neurosurgical level II trauma centre (TC) and those with a neurosurgical unit in the level I TC only. A higher proportion of patients (out of all SIAT patients) were admitted to the level I TC at the head of the SIAT with no additional neurosurgical facilities (1083/1472, 73.6%) compared with the level I TCs heading SIATs with neurosurgical level II TCs (1905/3815; 49.9%). A similar percentage of patients were admitted to level I TCs (1905/3815; 49.9%) and neurosurgical level II TCs (1702/3815, 44.6%) in the SIATs with neurosurgical level II TCs. Observed versus expected mortality (OE) was not statistically different among the three types of centre with a neurosurgical unit; however, the best mean OE values were observed in the level I TC in the SIAT with no neurosurgical unit. The Hub and Spoke concept was fully applied in the SIAT in which neurosurgical facilities were available in the level I TC only. The performance of this system suggests that competition among level I and level II TCs in the same Trauma System reduces performance in both. The density of neurosurgical centres must be considered by public health system governors before implementing trauma systems.


Details of Toll-like receptor:adapter interaction revealed by germ-line mutagenesis.
The immunovariant N-ethyl-N-nitrosourea-induced mutations Pococurante (Poc) and Lackadaisical were found to alter MyD88, creating striking receptor-selective effects. Poc, in particular, prevented sensing of all MyD88-dependent Toll-like receptor (TLR) ligands except diacyl lipopeptides. Furthermore, Poc-site and classical BB loop mutations caused equivalent phenotypes when engrafted into any TLR/IL-1 receptor/resistance (TIR) domain. These observations, complemented by data from docking studies and site-directed mutagenesis, revealed that BB loops and Poc sites interact homotypically across the receptor:adapter signaling interface, whereas the C-terminal alpha(E)-helices support adapter:adapter and receptor:receptor oligomerization. We have thus defined the TIR domain surface that mediates association between TLRs and MyD88 and the surface required for MyD88 or TLR oligomerization. Moreover, MyD88 engages individual TLRs differently, suggesting the feasibility of selective pharmacologic TIR domain receptor blockade.


Factors associated with increased risk for dementia in individuals age 80 years or older with congestive heart failure.
An increasing body of evidence shows that individuals diagnosed with congestive heart failure (CHF) are at a higher risk for dementia. However, the prevalence rate of dementia among persons with CHF in very old individuals has not been previously reported, and little is known about the comorbidities that place old persons with CHF at a higher risk for dementia. The aim of this study was to compare the prevalence of dementia in individuals 80 years or older who have CHF with that in individuals without CHF and to identify factors related to dementia in individuals diagnosed with CHF. A total of 702 participants from a Swedish population-based longitudinal study (Octogenerian Twin) were included. The group consisted of same-sex twin pairs, age 80 years or older, and 138 participants had CHF. Dementia was diagnosed according to criteria in the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised. Generalized estimating equations including gender, age and educational level, waist circumference, diabetes, hypertension, smoking, depression, and blood values were used in a case-control analysis. Individuals with CHF had a significantly higher prevalence of vascular dementia, 16% vs 6% (P < 0.001), and of all types of dementia, 40% vs 30% (P < 0.01), than those not diagnosed with CHF. The generalized estimating equation models showed that depression, hypertension, and/or increased levels of homocysteine were all associated with a higher risk for dementia in individuals with CHF. Diabetes was specifically associated with an increased risk for vascular dementia. The prevalence of dementia was higher among individuals with CHF than in those without CHF. Diabetes, depression, and hypertension in patients with CHF require special attention from healthcare professionals because these conditions are associated with an elevated risk for dementia. Higher levels of homocysteine were also found to be a marker of dementia in patients with CHF. Further research is needed to identify the factors related to dementia in individuals 80 years or older diagnosed with CHF.


Study of the short- and long-term outcomes of 65 horses with peritonitis.
The records of 65 horses with peritonitis examined at two UK referral centres over a period of 12 years were reviewed. Peritonitis was defined in terms of the horse's peritoneal fluid containing more than 5 x 10(9) nucleated cells/l. Horses that had developed peritonitis after abdominal surgery or a rupture of the gastrointestinal tract were excluded. Of the 65 horses, 56 (86 per cent) survived to be discharged. Follow-up information was obtained from practice records and telephone calls to the owners for 38 of the horses. Of these, 32 (84 per cent) had survived for at least 12 months and were considered to be long-term survivors; the others six were euthanased within 12 months. Thirteen (34 per cent) of the horses discharged had experienced complications that could have been sequelae to peritonitis and eight of the 13 were euthanased. The cause of the peritonitis was identified in 15 cases; survival rates were lowest in horses with peritonitis secondary to urinary tract involvement or intra-abdominal masses. Of the other 50 cases, 47 (94 per cent) survived to discharge, but two were euthanased owing to recurrent colic.


Ultraweak intrinsic Fabry-Perot cavity array for distributed sensing.
This Letter reports on an ultraweak intrinsic Fabry-Perot interferometer (IFPI) array fabricated by a femtosecond (fs) laser for distributed sensing applications. Ultralow reflectors (<-60 dB) were obtained. IFPIs with different physical lengths showed identical temperature sensitivity (-1.5 GHz/°C). A distributed temperature sensing test was conducted. No crosstalk between IFPI elements in the array was observed, implying the device's utility as a distributed sensing system. The possibility of using smaller bandwidths for sensor interrogation was experimentally proven. A small-scale temperature distribution test was conducted on a continuously cascaded ultraweak IFPI array, demonstrating its high spatial resolution. The temperature detection limit of this system was measured to be less than 0.0667°C.


Effects of Foliar Insecticides on Leaf-Level Spectral Reflectance of Soybean.
Pest-induced changes in plant reflectance are crucial for the development of pest management programs using remote sensing. However, it is unknown if plant reflectance data is also affected by foliar insecticides applied for pest management. Our study assessed the effects of foliar insecticides on leaf reflectance of soybean. A 2-yr field trial and a greenhouse trial were conducted using randomized complete block and completely randomized designs, respectively. Treatments consisted of an untreated check, a new systemic insecticide (sulfoxaflor), and two representatives of the most common insecticide classes used for soybean pest management in the north-central United States (i.e., λ-cyhalothrin and chlorpyrifos). Insecticides were applied at labeled rates recommended for controlling soybean aphid; the primary insect pest in the north-central United States. Leaf-level reflectance was measured using ground-based spectroradiometers. Sulfoxaflor affected leaf reflectance at some red and blue wavelengths but had no effect at near-infrared or green wavelengths. Chlorpyrifos affected leaf reflectance at some green, red, and near-infrared wavelengths but had no effect at blue wavelengths. λ-cyhalothrin had the least effect on spectral reflectance among the insecticides, with changes to only a few near-infrared wavelengths. Our results showing immediate and delayed effects of foliar insecticides on soybean reflectance indicate that application of some insecticides may confound the use of remote sensing for detection of not only insects but also plant diseases, nutritional and water deficiencies, and other crop stressors.


A dose-response study with formoterol Turbuhaler as maintenance therapy in asthmatic patients.
The aim of this randomized, double-blind, parallel group study was to determine the lowest effective dose of 6, 12 and 24 microg formoterol fumarate dihydrate Turbuhaler b.i.d. compared with placebo. The 4 week treatment was preceded by a 1 week run-in period. Morning peak expiratory flow (PEF) before intake of the study drug was the primary variable. Patients recorded PEF, prior to and 15 min after intake of the study drug (immediate response), asthma symptoms, and use of rescue medication morning and evening. Of 221 patients (71 females and 150 males), 194 were included in the efficacy per protocol (PP) analysis; mean age 47 yrs, mean forced expiratory volume in one second (FEV1) 2.01 L (58% of predicted), mean FEV1 reversibility 27% at entry. Ninety percent used inhaled steroids. Compared with placebo, 6 microg formoterol b.i.d was found to be the lowest effective dose in the morning (p=0.008) and evening p=0.0041) PEF. The mean increases in PEF were 22 and 23 L x min-1 respectively, compared with placebo. After 6 microg formoterol, the mean immediate increase in morning PEF was 42 L x min-1 compared to an increase of only 9 L x min-1 after placebo (<0.0001). All doses produced a statistically significant decrease in asthma symptoms, day and night, and the need for rescue medication at night. All doses were well-tolerated. In conclusion, the lowest effective dose in this study was formoterol Turbuhaler 6 microg b.i.d.


Pertussis toxin inhibits chemotactic peptide-stimulated generation of inositol phosphates and lysosomal enzyme secretion in human leukemic (HL-60) cells.
The binding of the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine to its cell surface receptor rapidly elicits the hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase C to form the putative second messengers inositol 1,4,5-trisphosphate and sn-1,2-diacylglycerol. To investigate the possible role of a guanine nucleotide binding protein in transduction of this membrane signal, we examined the effects of pertussis toxin on chemotactic peptide-stimulated inositol phospholipid metabolism in differentiated HL-60 cells labeled with [3H]inositol. Pertussis toxin inhibited the chemotactic tripeptide-stimulated production of inositol mono-, bis-, and trisphosphates and secretion of N-acetyl-beta-D-glucosaminidase in a time- and concentration-dependent manner. Treatment with pertussis toxin did not alter the total incorporation or the distribution of [3H]inositol in inositol phospholipid. Chemotactic peptide receptor number was unchanged, although a slight decrease in binding affinity was observed. These findings suggest a role for a guanine nucleotide binding protein in coupling the chemotactic peptide receptor to phospholipase C.


Cost Effectiveness of Environmental Lead Risk Mitigation in Low- and Middle-Income Countries.
Environmental remediation efforts in low- and middle-income countries have yet to be evaluated for their cost effectiveness. To address this gap we calculate a cost per Disability Adjusted Life Year (DALY) averted following the environmental remediation of the former lead smelter and adjoining residential areas in Paraiso de Dios, Haina, the Dominican Republic, executed from 2009 to 2010. The remediation had the effect of lowering surface soil lead concentrations to below 100 mg/kg and measured geometric mean blood lead levels (BLLs) from 20.6 μg/dL to 5.34 ug/dL. Because BLLs for the entire impacted population were not available, we use environmental data to calculate the resulting disease burden. We find that before the intervention 176 people were exposed to elevated environmental lead levels at Paraiso de Dios resulting in mean BLLs of 24.97 (95% CI: 24.45-25.5) in children (0-7 years old) and 13.98 μg/dL (95% CI: 13.03-15) in adults. We calculate that without the intervention these exposures would have resulted in 133 to 1,096 DALYs and that all of these were averted at a cost of USD 392 to 3,238, depending on assumptions made. We use a societal perspective, meaning that we include all costs regardless of by whom they were incurred and estimate costs in 2009 USD. Lead remediation in low- and middle-income countries is cost effective according to World Health Organization thresholds. Further research is required to compare the approach detailed here with other public health interventions.


Health-Related Quality of Life Among Jordanian Adolescents.
Adolescence is an unpredictable stage of life with varied and rapid changes. In Jordan, health-related quality of life (HRQoL) has been examined among diabetic and obese children and adolescents. The purpose of this study was to assess the HRQoL of Jordanian healthy adolescents. Three hundred fifty-four male and female adolescents whose ages ranged from 12 to 19 participated in the study. A descriptive comparative design was employed to investigate adolescents' HRQoL. The results revealed statistically significant differences in physical well-being, psychosocial well-being, and autonomy in favor of male adolescents. In addition, statistically significant differences were observed in favor of nonsmoker adolescents in psychosocial well-being, self-perception, parent relations and home life, financial resources, social relations and peers and school environment. In conclusion, the creation of a school health nurse role in Jordanian schools is crucial for helping adolescents improve their health.


Antiamnesic and neuroprotective effects of donepezil against learning impairments induced in mice by exposure to carbon monoxide gas.
Donepezil is a potent acetylcholinesterase inhibitor that also interacts with the sigma1 receptor, an intracellular neuromodulatory protein. In the present study, we analyzed the antiamnesic and neuroprotective activities of donepezil in a mouse hypoxia model induced by repetitive CO exposure, comparing donepezil's pharmacological profile with other cholinesterase inhibitors tacrine, rivastigmine, and galanthamine, and the reference sigma1 agonist igmesine. CO exposure induced, after 7 days, hippocampal neurodegeneration, analyzed by Cresyl violet staining, and behavioral alterations, measured using spontaneous alternation and passive avoidance responses. When injected 20 min before the behavioral tests, i.e., 7 to 8 days after CO, all drugs showed antiamnesic properties. Preadministration of the sigma1 receptor antagonist N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(dimethylamino)ethylamine (BD1047) blocked only the igmesine and donepezil effects. The neuroprotective activity of the drugs was tested by injection 20 min before the first CO exposure (preinsult protection) or by injection 1 h after the last CO exposure (postinsult protection). All drugs alleviated the hypoxia-induced neurodegeneration and behavioral impairments when injected before CO exposure. Preadministration of BD1047 blocked both the igmesine and donepezil effects. However, when injected after CO exposure, only igmesine and donepezil induced effective neuroprotection, and the morphological and behavioral effects were BD1047-sensitive. These results showed that donepezil is a potent antiamnesic and neuroprotective compound against the neurodegeneration induced by excitotoxic insult, and its pharmacological actions as both an acetylcholinesterase inhibitor and sigma1 receptor agonist contribute to its marked efficacy. In particular, the drug is a more potent postinsult protecting agent compared with more selective cholinesterase inhibitors.


Mps1 phosphorylates Borealin to control Aurora B activity and chromosome alignment.
Maintenance of chromosomal stability relies on coordination between various processes that are critical for proper chromosome segregation in mitosis. Here we show that monopolar spindle 1 (Mps1) kinase, which is essential for the mitotic checkpoint, also controls correction of improper chromosome attachments. We report that Borealin/DasraB, a member of the complex that regulates the Aurora B kinase, is directly phosphorylated by Mps1 on residues that are crucial for Aurora B activity and chromosome alignment. As a result, cells lacking Mps1 kinase activity fail to efficiently align chromosomes due to impaired Aurora B function at centromeres, leaving improper attachments uncorrected. Strikingly, Borealin/DasraB bearing phosphomimetic mutations restores Aurora B activity and alignment in Mps1-depleted cells. Mps1 thus coordinates attachment error correction and checkpoint signaling, two crucial responses to unproductive chromosome attachments.


Geostatistical Borehole Image-Based Mapping of Karst-Carbonate Aquifer Pores.
Quantification of the character and spatial distribution of porosity in carbonate aquifers is important as input into computer models used in the calculation of intrinsic permeability and for next-generation, high-resolution groundwater flow simulations. Digital, optical, borehole-wall image data from three closely spaced boreholes in the karst-carbonate Biscayne aquifer in southeastern Florida are used in geostatistical experiments to assess the capabilities of various methods to create realistic two-dimensional models of vuggy megaporosity and matrix-porosity distribution in the limestone that composes the aquifer. When the borehole image data alone were used as the model training image, multiple-point geostatistics failed to detect the known spatial autocorrelation of vuggy megaporosity and matrix porosity among the three boreholes, which were only 10 m apart. Variogram analysis and subsequent Gaussian simulation produced results that showed a realistic conceptualization of horizontal continuity of strata dominated by vuggy megaporosity and matrix porosity among the three boreholes.


Evidence for induction of interferon-alpha and interferon-beta in retinal glial cells of Müller.
Studies were performed to determine if retinal glial cells of Müller transcribe the genes for interferon-alpha (IFNalpha) or IFNbeta upon exposure to virus. Responses to herpes simplex virus type 1 (HSV-1) were tested with cultured murine Müller cells and, in vivo, with retinas obtained after bilateral injection of either HSV-1 or buffer into the anterior chamber of the eyes of BALB/c mice. Induction of IFN transcription and relative temporal changes in transcript levels occurred over time after either in vitro or in vivo exposure to HSV-1. Transcription of both IFN genes was induced in cultured glia within 1 hr after exposure to virus. IFN transcripts were detected in retinas by 24 hr postinfection and these were maximal at 3 days. By in situ hybridization (ISH), IFNalpha2 mRNA localized to focal areas in the intact retinas of virus-injected eyes and was consistent with our previous report of a transient, focal appearance of viral antigens in those retinas. Uninfected cells and ocular tissues were negative for IFN transcripts. Combined ISH and immunohistochemistry on retinal impression smears confirmed that glial fibrillary acidic protein-positive Müller cells are an intraretinal source of IFNalpha and IFNbeta transcripts after ocular exposure to HSV-1. Our results support a role for Muller cells as participants in intraretinal antiviral or immunomodulatory responses via type 1 IFN production and may have implications for future therapeutic interventions.


Alcohol use and intimate partner violence as predictors of separation among U.S. couples: a longitudinal model.
The purpose of this study was to investigate the relationship between partner violence (male-to-female and female-to-male) and heavy drinking (consuming five or more drinks per occasion) and the effect of this relationship on marital separation. Married and cohabiting couples were selected through multistage random probability sampling from U.S. households. In 1995, 1,635 couples were interviewed; 1,392 of these couples were re-interviewed in 2000. The final analysis included couples who were still married or living with the same partner (intact n = 1,136) from the baseline, and couples who had broken up (nonintact n = 191) sometime during the 5-year follow-up period. The findings indicate that couples who reported female-perpetrated violence, female alcohol-related problems and male heavy-drinking episodes (one to three times a month) were 2.5-3 times more at risk for separation. On the other hand, female heavy drinking (one to three times a month) is less likely to predict separation. Cohabiting couples are also more at risk for separation than married couples. Couples with longer relationships are less at risk for separation. Marital counseling, therapy or intervention programs must address issues pertaining to partner violence and alcohol use among couples while concurrently providing information about healthy relationships and helping couples develop skills to attain them. The importance of preventing partner violence is increasingly recognized in the field of public health.


Proteolytic activation of recombinant pro-memapsin 2 (pro-beta-secretase) studied with new fluorogenic substrates.
Memapsin 2 (beta-secretase), a membrane-anchored aspartic protease, is involved in the cleavage of beta-amyloid precursor protein to form beta-amyloid peptide. The primary structure of memapsin 2 suggests that it is synthesized in vivo as pro-memapsin 2 and converted to memapsin 2 by an activating protease [Lin et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 1456-1460]. To simulate this activation mechanism and to produce stable mature memapsin 2 for kinetic/specificity studies, we have investigated the activation of recombinant pro-memapsin 2 by several proteases with trypsin-like specificity. Clostripain, kallikrein, and trypsin increased the activity of pro-memapsin 2. Clostripain activation was accompanied by the cleavage of the pro region to form mainly two activation products, Leu(30p)- and Gly(45p)-memapsin 2. Another activation product, Leu(28p)-memapsin 2, was also purified. Kinetics of the activated memapsin 2 were compared with pro-memapsin 2 using two new fluorogenic substrates, Arg-Glu(5-[(2-aminoethyl)amino]naphthalene-1-sulfonic acid (EDANS))-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys(4-(4-dimethylaminophe nyl azo)benzoic acid (DABCYL))-Arg and (7-methoxycoumarin-4-yl)acetyl (MCA))-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys(2,4-dinitrophenyl (DNP)). These results establish that the activity of pro-memapsin 2 stems from a part-time and reversible uncovering of its active site by its pro region. Proteolytic removal of part of the pro-peptide at Leu(28p) or Gly(45p), which diminishes the affinity of the shortened pro-peptide to the active site, results in activated memapsin 2. These results also suggest that Glu(33p)-memapsin 2 observed in the cells expressing this enzyme [Vassar et al. (1999) Science 286, 735-741; Yan et al. (1999) Nature 402, 533-537] is an active intermediate of in vivo activation, or that the peptide Glu(33p)-Arg(44p) may serve a regulatory role.


Adaptive contact networks change effective disease infectiousness and dynamics.
Human societies are organized in complex webs that are constantly reshaped by a social dynamic which is influenced by the information individuals have about others. Similarly, epidemic spreading may be affected by local information that makes individuals aware of the health status of their social contacts, allowing them to avoid contact with those infected and to remain in touch with the healthy. Here we study disease dynamics in finite populations in which infection occurs along the links of a dynamical contact network whose reshaping may be biased based on each individual's health status. We adopt some of the most widely used epidemiological models, investigating the impact of the reshaping of the contact network on the disease dynamics. We derive analytical results in the limit where network reshaping occurs much faster than disease spreading and demonstrate numerically that this limit extends to a much wider range of time scales than one might anticipate. Specifically, we show that from a population-level description, disease propagation in a quickly adapting network can be formulated equivalently as disease spreading on a well-mixed population but with a rescaled infectiousness. We find that for all models studied here--SI, SIS and SIR--the effective infectiousness of a disease depends on the population size, the number of infected in the population, and the capacity of healthy individuals to sever contacts with the infected. Importantly, we indicate how the use of available information hinders disease progression, either by reducing the average time required to eradicate a disease (in case recovery is possible), or by increasing the average time needed for a disease to spread to the entire population (in case recovery or immunity is impossible).


Efficient Cas9 multiplex editing using unspaced sgRNA arrays engineering in a Potato virus X vector.
Systems based on the clustered, regularly interspaced, short palindromic repeat (CRISPR) and CRISPR-associated proteins (Cas) have revolutionized genome editing in many organisms, including plants. Most CRISPR-Cas strategies in plants rely on genetic transformation using Agrobacterium tumefaciens to supply the gene editing reagents, such as Cas nucleases or the synthetic guide RNA (sgRNA). While Cas nucleases are constant elements in editing approaches, sgRNAs are target-specific and a screening process is usually required to identify those most effective. Plant virus-derived vectors are an alternative for the fast and efficient delivery of sgRNAs into adult plants, due to the virus capacity for genome amplification and systemic movement, a strategy known as virus-induced genome editing. We engineered Potato virus X (PVX) to build a vector that easily expresses multiple sgRNAs in adult solanaceous plants. Using the PVX-based vector, Nicotiana benthamiana genes were efficiently targeted, producing nearly 80% indels in a transformed line that constitutively expresses Streptococcus pyogenes Cas9. Interestingly, results showed that the PVX vector allows expression of arrays of unspaced sgRNAs, achieving highly efficient multiplex editing in a few days in adult plant tissues. Moreover, virus-free edited progeny can be obtained from plants regenerated from infected tissues or infected plant seeds, which exhibit a high rate of heritable biallelic mutations. In conclusion, this new PVX vector allows easy, fast and efficient expression of sgRNA arrays for multiplex CRISPR-Cas genome editing and will be a useful tool for functional gene analysis and precision breeding across diverse plant species, particularly in Solanaceae crops.


Linear free energy correlations for fission product release from the Fukushima-Daiichi nuclear accident.
This paper extends the preliminary linear free energy correlations for radionuclide release performed by Schwantes et al., following the Fukushima-Daiichi Nuclear Power Plant accident. Through evaluations of the molar fractionations of radionuclides deposited in the soil relative to modeled radionuclide inventories, we confirm the initial source of the radionuclides to the environment to be from active reactors rather than the spent fuel pool. Linear correlations of the form In χ = −α ((ΔGrxn°(TC))/(RTC)) + β were obtained between the deposited concentrations, and the reduction potentials of the fission product oxide species using multiple reduction schemes to calculate ΔG°rxn (TC). These models allowed an estimate of the upper bound for the reactor temperatures of TC between 2015 and 2060 K, providing insight into the limiting factors to vaporization and release of fission products during the reactor accident. Estimates of the release of medium-lived fission products 90Sr, 121mSn, 147Pm, 144Ce, 152Eu, 154Eu, 155Eu, and 151Sm through atmospheric venting during the first month following the accident were obtained, indicating that large quantities of 90Sr and radioactive lanthanides were likely to remain in the damaged reactor cores.


Detection and expression of DNA homologous to the tox gene in nontoxinogenic isolates of Corynebacterium diphtheriae.
Three probes have been described which can be used to detect the presence of DNA sequences homologous to the tox gene of Corynebacterium diphtheriae. Probes "A" and "B" detected sequences coding for A and B fragments of diphtheria toxin, respectively. The third "A-B" probe contained both the A and B coding sequences. The B probe was completely unambiguous in detecting only toxin-related sequences, and the A probe was only slightly less so. The efficacy of the probes was tested on a series of toxinogenic and nontoxinogenic isolates of C. diphtheriae. All isolates which were toxinogenic as characterized by the gel immunodiffusion technique gave positive reactions with the probes. Of particular interest was the finding that 14 of 43 nontoxinogenic isolates also carried DNA homologous to both the A and B probes. All 14 isolates were nontoxinogenic by the rabbit intracutaneous test as well as by the gel immunodiffusion test; however, 12 of them produced ADP-ribosylating activity, whereas two were negative. The isolates producing ADP-ribosylating activity belonged to a cohort of cultures, of which 11 were isolated in South Dakota and 1 was isolated in Montana. Genomic DNAs of all 12 appeared to be identical when restriction enzyme digest patterns were compared, and the same fragment carried the tox gene in all of them. The tox-bearing nontoxinogenic isolates from Alaska and Florida each had unique restriction patterns and did not produce ADP-ribosylating activity. A number of genomic fragments of all the tox-bearing nontoxinogenic isolates hybridized with beta converting phage DNA. The significance of these observations to the natural history of diphtheria was discussed.


When reduction strategies are put in place and mental health consumers are still secluded: an analysis of clinical and sociodemographic characteristics.
Internationally, seclusion practices continue to be the subject of intense clinical health service and academic scrutiny. Despite extensive efforts to reduce and eliminate this controversial practice, seclusion remains a clinical intervention widely used in contemporary mental health service settings. Early identification of people who are at risk for seclusion and the timely application of alternative evidence-based interventions are critical for reducing incidents of seclusion in real-world practice settings. This retrospective study aimed to determine the relationship between sociodemographic and clinical characteristics, and the use of seclusion for those mental health consumers for whom evidence-based seclusion-reduction initiatives had little impact. A 12-month centred moving average was fitted to seclusion data from a psychiatric inpatient unit over 2 years to determine stabilization in seclusion reduction. The number of consumers admitted was calculated from the point of stabilization for 1 year (n = 469). In this cohort, univariate analysis sought to compare the characteristics of those who were secluded and those who were not. A multivariate logistic regression model was undertaken to associate future seclusion based on significant independent variables. Of those people admitted, 88 (19%) were secluded. The majority of seclusions occurred in the first 5 days (70/88, 79%). Multivariate logistic regression indicated that three variables maintained their independent associative risk of seclusion: (i) age less than 35 years; (ii) assessment of risk of violence to others; and (iii) a history of seclusion. The implications of these findings for nursing practice are discussed.


[Clinico-laboratory and morphological parallels in hepatic echinococcosis].
The trial included 50 patients with hydatid echinococcosis of the liver and 5 control patients operated urgently for extrahepatic pathology. The patients underwent clinical and biochemical tests, ultrasound investigation of the liver. Structural and functional changes in the liver were analysed with reference to the stage of the echinococcal cyst. Morphological picture of the liver was studied by light microscopy, its functional picture--by activity of AsAT and AlAT in hepatic tissue. Morphological and biochemical tests were made on hepatic biopsies obtained from the pericystic and far from the cyst zones. The most severe histological changes affecting all structures of the liver were registered in the parasite's life cycles II and III. AsAT and AlAT changes in the liver were not related to the biological cycle of the parasite; echinococcal patients' liver contained them in lower concentrations than that of control subjects. Moreover, blood serum levels of AsAT and AlAT are higher than in liver tissue. Thus, growth of the echinococcal cyst in the liver is accompanied with marked morphofunctional disorders. This necessitates early detection and surgical treatment of hepatic echinococcosis. To prevent postoperative complications, adequate preoperative preparation and postoperative intensive treatment must be conducted.


What Factors Are Associated With Implant Breakage and Revision After Intramedullary Nailing for Femoral Metastases?
Actual and impending pathologic fractures of the femur are commonly treated with intramedullary nails because they provide immediate stabilization with a minimally invasive procedure and enable direct weightbearing. However, complications and revision surgery are prevalent, and despite common use, there is limited evidence identifying those factors that are associated with complications. Among patients treated with intramedullary nailing for femoral metastases, we asked the following questions: (1) What is the cumulative incidence of local complications? (2) What is the cumulative incidence of implant breakage and what factors are associated with implant breakage? (3) What is the cumulative incidence of revision surgery and what factors are associated with revision surgery? Between January 2000 and December 2015, 245 patients in five centers were treated with intramedullary nails for actual and impending pathologic fractures of the femur caused by bone metastases. During that period, the general indications for intramedullary nailing of femoral metastases were impending fractures of the trochanter region and shaft and actual fractures of the trochanter region if sufficient bone stock remained; nails were used for lesions of the femoral shaft if they were large or if multiple lesions were present. Of those treated with intramedullary nails, 51% (117) were actual fractures and 49% (111) were impending fractures. A total of 60% (128) of this group were women; the mean age was 65 years (range, 29-93 years). After radiologic followup (at 4-8 weeks) with the orthopaedic surgeon, because of the palliative nature of these treatments, subsequent in-person followup was performed by the primary care provider on an as-needed basis (that is, as desired by the patient, without any scheduled visits with the orthopaedic surgeon) throughout each patient's remaining lifetime. However, there was close collaboration between the primary care providers and the orthopaedic team such that orthopaedic complications would be reported. A total of 67% (142 of 212) of the patients died before 1 year, and followup ranged from 0.1 to 175 months (mean, 14.4 months). Competing risk models were used to estimate the cumulative incidence of local complications (including persisting pain, tumor progression, and implant breakage), implant breakage separately, and revision surgery (defined as any reoperation involving the implant other than débridement with implant retention for infection). A cause-specific multivariate Cox regression model was used to estimate the association of factors (fracture type/preoperative radiotherapy and fracture type/use of cement) with implant breakage and revision, respectively. Local complications occurred in 12% (28 of 228) of the patients and 6-month cumulative incidence was 8% (95% confidence interval [CI], 4.7-11.9). Implant breakage occurred in 8% (18 of 228) of the patients and 6-month cumulative incidence was 4% (95% CI, 1.4-6.5). Independent factors associated with increased risk of implant breakage were an actual (as opposed to impending) fracture (cause-specific hazard ratio [HR_cs], 3.61; 95% CI, 1.23-10.53, p = 0.019) and previous radiotherapy (HR_cs, 2.97; 95% CI, 1.13-7.82, p = 0.027). Revisions occurred in 5% (12 of 228) of the patients and 6-month cumulative incidence was 2.2% (95% CI, 0.3-4.1). The presence of an actual fracture was independently associated with a higher risk of revision (HR_cs, 4.17; 95% CI, 0.08-0.82, p = 0.022), and use of cement was independently associated with a lower risk of revision (HR_cs, 0.25; 95% CI, 1.20-14.53, p = 0.025). The cumulative incidence of local complications, implant breakage, and revisions is low, mostly as a result of the short survival of patients. Based on these results, surgeons should consider use of cement in patients with intramedullary nails with actual fractures and closer followup of patients after actual fractures and preoperative radiotherapy. Future, prospective studies should further analyze the effects of adjuvant therapies and surgery-related factors on the risk of implant breakage and revisions. Level III, therapeutic study.


The effect of fruits and vegetables on urinary stone risk factors.
The overall effect of fruit and vegetable intake on urinary stone risk profile is not yet known. We studied the effect of a two-week period of fruit and vegetable elimination on urinary stone risk profile in 12 normal adults, and of supplementing the diet with a fair quantity of low-oxalate fruits and vegetables in 26 idiopathic calcium stone formers characterized by hypocitraturia and a very low fruit and vegetable intake in their usual diet. In the normal subjects, the elimination of fruits and vegetables from the diet decreased the urinary excretion of potassium (-62%), magnesium (-26%), citrate (-44%) and oxalate (-31%), and increased that of calcium (+49%) and ammonium (+12%) (P < 0.05 for all). The relative saturation for calcium oxalate and calcium phosphate increased from 6.33 to 8.24 (P = 0.028), and from 0.68 to 1.58 (P = 0.050), respectively. In the hypocitraturic stone formers, the introduction of these foods in the diet increased urinary volume (+64%), pH (from 5.84 to 6.19), excretion of potassium (+68%), magnesium (+23%), and citrate (+68%), while it decreased the excretion of ammonium (-18%) (P < 0.05 for all). The relative saturation for calcium oxalate and uric acid fell from 10.17 to 4.96 (P < 0.001), and from 2.78 to 1.12 (P = 0.003), respectively. The total elimination of fruits and vegetables in normal subjects brings about adverse changes in the urinary stone risk profile that are only partially counterbalanced by a reduction in oxalate. In contrast, the addition of these foods to the diet of hypocitraturic stone formers not used to eating them not only significantly increases citrate excretion without affecting oxalate excretion, but also decreases calcium oxalate and uric acid relative saturation.


Biosynthesis of vitamins and cofactors in bacterium-harbouring trypanosomatids depends on the symbiotic association as revealed by genomic analyses.
Some non-pathogenic trypanosomatids maintain a mutualistic relationship with a betaproteobacterium of the Alcaligenaceae family. Intensive nutritional exchanges have been reported between the two partners, indicating that these protozoa are excellent biological models to study metabolic co-evolution. We previously sequenced and herein investigate the entire genomes of five trypanosomatids which harbor a symbiotic bacterium (SHTs for Symbiont-Haboring Trypanosomatids) and the respective bacteria (TPEs for Trypanosomatid Proteobacterial Endosymbiont), as well as two trypanosomatids without symbionts (RTs for Regular Trypanosomatids), for the presence of genes of the classical pathways for vitamin biosynthesis. Our data show that genes for the biosynthetic pathways of thiamine, biotin, and nicotinic acid are absent from all trypanosomatid genomes. This is in agreement with the absolute growth requirement for these vitamins in all protozoa of the family. Also absent from the genomes of RTs are the genes for the synthesis of pantothenic acid, folic acid, riboflavin, and vitamin B6. This is also in agreement with the available data showing that RTs are auxotrophic for these essential vitamins. On the other hand, SHTs are autotrophic for such vitamins. Indeed, all the genes of the corresponding biosynthetic pathways were identified, most of them in the symbiont genomes, while a few genes, mostly of eukaryotic origin, were found in the host genomes. The only exceptions to the latter are: the gene coding for the enzyme ketopantoate reductase (EC:22.214.171.124) which is related instead to the Firmicutes bacteria; and two other genes, one involved in the salvage pathway of pantothenic acid and the other in the synthesis of ubiquinone, that are related to Gammaproteobacteria. Their presence in trypanosomatids may result from lateral gene transfer. Taken together, our results reinforce the idea that the low nutritional requirement of SHTs is associated with the presence of the symbiotic bacterium, which contains most genes for vitamin production.


Diagnostic evaluation of neonatal ascites.
We present the case of a premature infant noted to have massive abdominal distension at birth. Obstetric ultrasound at delivery was significant for abdominal distension, oligohydramnios, and restricted chest wall motion. He was diagnosed to have posterior urethral valves with intraperitoneal rupture of the bladder. We present this case with a brief review of the literature.


Health and evolution.
Darwinian medicine may shed new light on the notion of health and many current health problems. In this paper, health, as an ability to realize one's own welfare, is compared with health as an ability--either being developed or actually present--to perform a reproductive function of one's species. It is argued that knowledge about the conditions for health in the latter sense may enhance our efforts to promote health in the former sense.


Gorlin-Goltz syndrome with familial manifestation.
The detection of odontogenic keratocysts (OKC) in the oral cavity is one of the main criteria for the clinical manifestation of Gorlin-Goltz syndrome (Nevoid Basal Cell Carcinoma Syndrome - NBCCS). From a clinical point of view, we distinguish between "syndromic" and "sporadic" OKC. Syndromic cysts, often multifocal, may be an accidental finding on X-ray examination. They can manifest gradually depending on the development of permanent dentition. Sporadic cysts are rather solitary lesions with clinical manifestation in adulthood. Mutations in the PTCH1 gene are thought to be the cause of the clinical manifestation of NBCCS. These abnormalities can be transmitted from one generation to another and lead to a familial occurrence of the disease. In 35-50% of cases, these are a newly arising mutations. It is necessary to take into account the typical manifestations which in the next generation begin at a younger age and the disease usually has a more serious course. We found a familial manifestation of NBCCS in two pairs of patients (mother and daughter and two siblings). Odontogenic keratocysts and cutaneous basal cell carcinomas were diagnosed and genetic testing revealed mutations in the PTCH 1 gene in all four individuals. With regard to the possibility of familial occurrence of NBCCS, it is necessary to pay increased attention to family history and, if necessary, to ensure clinical and genetic examination of parents and other family members. Patients of childbearing potential with evidence of NBCCS should be informed of the increased likelihood of the disease in the offspring.


Shunt infusion studies: impact on patient outcome, including health economics.
The diagnosis of shunt malfunction is often not straightforward. We have explored, in symptomatic shunted patients with hydrocephalus or pseudotumour cerebri syndrome (PTCS), the accuracy of CSF infusion tests in differentiating a functioning shunt from one with possible problems, and the health economic consequences. Participants: hydrocephalus/PTCS patients with infusion tests performed from January 2013 until December 2015. We followed patients up after 6 and 12 months from the test to determine whether they had improved, had persisting symptoms or had required urgent revision. We calculated the total cost savings of revision versus infusion tests and standard protocol of revision and ICP monitoring versus infusion tests. Three hundred sixty-five shunt infusion tests had been performed where a shunt prechamber/reservoir was present. For hydrocephalus patients, more than half of the tests (~ 55%, 155 out of 280) showed no shunt malfunction versus 125 with possible malfunction (ages 4 months to 90 years old). For PTCS patients aged 10 to 77 years old, 47 had possible problems and 38 no indication for shunt malfunction. Overall, > 290 unnecessary revisions were avoided over 3 years' time. Two hundred fifty-eight (> 85%) of those non-surgically managed, remained well, did not deteriorate and did not require surgery. No infections were associated with infusion studies. For Cambridge, the overall savings from avoiding revisions was £945,415 annually. Our results provide evidence of the importance of shunt testing in vivo to confirm shunt malfunction. Avoiding unnecessary shunt revisions carries a strong health benefit for patients that also translates to a significant financial benefit for the National Health Service and potentially for other healthcare systems worldwide.


Evolutionary dynamics and preferential expression of homeologous 18S-5.8S-26S nuclear ribosomal genes in natural and artificial glycine allopolyploids.
Polyploidy is an important evolutionary process in plants, but much remains to be learned about the evolution of gene expression in polyploids. Evolution and expression of the 18S-5.8S-26S ribosomal gene family was investigated at homeologous loci in the Glycine subgenus Glycine perennial soybean polyploid complex, which consists of several diploid genomes that have formed allopolyploids in various combinations, often recurrently. A semiquantitative PCR method targeting the internal transcribed spacer (ITS) of the 18S-5.8S-26S nuclear ribosomal DNA (nrDNA) was used to survey the ratio between homeologous repeats in polyploid genomes and to test for preferential expression of homeologous nrDNA loci. Most natural polyploids possess one predominant nrDNA homeolog in their genome. Analysis of F2 segregation in an artificial cross suggested that in some plants, most or all repeats at one homeologous locus have been lost, whereas in other plants two loci remain, but both have been homogenized by concerted evolution. In most natural allopolyploids harboring a relatively balanced ratio of homeologs, one homeolog was expressed preferentially, but in the majority of plants, low levels of transcription could be detected from the other homeolog. Individuals within some tetraploid taxa varied as to which homeolog was expressed preferentially. In some plants, the degree of preferential expression also varied among tissues. Preferential expression was absent in synthetic polyploids and in some artificial diploid hybrids, suggesting that nucleolar dominance is not necessarily a direct result of hybridization or polyploidization. The establishment of preferential expression in Glycine allopolyploids appears to be either stochastic within lineages or genotype specific.


Immune Modulation Properties of Zoledronic Acid on TcRγδ T-Lymphocytes After TcRαβ/CD19-Depleted Haploidentical Stem Cell Transplantation: An analysis on 46 Pediatric Patients Affected by Acute Leukemia.
TcRαβ/CD19-cell depleted HLA-haploidentical hematopoietic stem cell transplantation (haplo-HSCT) represents a promising new platform for children affected by acute leukemia in need of an allograft and lacking a matched donor, disease recurrence being the main cause of treatment failure. The use of zoledronic acid to enhance TcRγδ+ lymphocyte function after TcRαβ/CD19-cell depleted haplo-HSCT was tested in an open-label, feasibility, proof-of-principle study. Forty-six children affected by high-risk acute leukemia underwent haplo-HSCT after removal of TcRαβ+ and CD19+ B lymphocytes. No post-transplant pharmacological graft-versus-host disease (GvHD) prophylaxis was given. Zoledronic acid was administered monthly at a dose of 0.05 mg/kg/dose (maximum dose 4 mg), starting from day +20 after transplantation. A total of 139 infusions were administered, with a mean of 3 infusions per patient. No severe adverse event was observed. Common side effects were represented by asymptomatic hypocalcemia and acute phase reactions (including fever, chills, malaise, and/or arthralgia) within 24-48 h from zoledronic acid infusion. The cumulative incidence of acute and chronic GvHD was 17.3% (all grade I-II) and 4.8% (all limited), respectively. Patients given 3 or more infusions of zoledronic acid had a lower incidence of both acute GvHD (8.8 vs. 41.6%, p = 0.015) and chronic GvHD (0 vs. 22.2%, p = 0.006). Transplant-related mortality (TRM) and relapse incidence at 3 years were 4.3 and 30.4%, respectively. Patients receiving repeated infusions of zoledronic acid had a lower TRM as compared to those receiving 1 or 2 administration of the drug (0 vs. 16.7%, p = 0.01). Five-year overall survival (OS) and disease-free survival (DFS) for the whole cohort were 67.2 and 65.2%, respectively, with a trend toward a better OS for patients receiving 3 or more infusions (73.1 vs. 50.0%, p = 0.05). The probability of GvHD/relapse-free survival was significantly worse in patients receiving 1-2 infusions of zoledonic acid than in those given ≥3 infusions (33.3 vs. 70.6%, respectively, p = 0.006). Multivariable analysis showed an independent positive effect on outcome given by repeated infusions of zoledronic acid (HR 0.27, p = 0.03). These data indicate that the use of zoledronic acid after TcRαβ/CD19-cell depleted haploHSCT is safe and may result in a lower incidence of acute GvHD, chronic GvHD, and TRM.


[Quality of life of sarcopenic patients: contribution of the SarcoPhAge study].
The consequences of sarcopenia on quality are difficult to evaluate and consequently are quite poorly studied. The few studies assessing the quality of life in sarcopenic subjects currently use generic quality of life questionnaires such as SF-36 and EQ-5D. The SarcoPhAge study, conducted on a cohort developed in Liège including 534 subjects of 65 years or older, suggested that sarcopenic subjects present a significant worse quality of life in the domains of physical function compared to non-sarcopenic subjects. Generic tools do not cover exhaustively all the areas of potential dysfunction concerned in this geriatric syndrome. Yet, there is no specific and validated quality of life questionnaire for sarcopenia. It would be useful to have at our disposal a sarcopenia specific quality of life questionnaire to assess not only the prospective quality of life of sarcopenic subjects but also to assess the efficacy and the relevance of new therapeutic and preventive strategies developed in the field of sarcopenia.


Extracellular gelsolin binds lipoteichoic acid and modulates cellular response to proinflammatory bacterial wall components.
The various functions of gelsolin in extracellular compartments are not yet clearly defined but include actin scavenging and antiinflammatory effects. Gelsolin was recently reported to bind endotoxin (LPS) from various Gram-negative bacteria with high affinity. In this study we investigate whether gelsolin also interacts with bacterial wall molecules of Gram-positive bacteria such as lipoteichoic acid (LTA) and whether gelsolin's interaction with bacterial lipids from Gram-negative or Gram-positive bacteria affects their cellular inflammatory responses. A peptide based on the PPI binding site of gelsolin (160-169) binds purified LTA at the same molecular ratio that it binds phosphatidylinositol 4,5-bisphosphate. The OD of recombinant human plasma gelsolin was found to decrease following the addition of purified LTA, and the binding of gelsolin to LTA inhibits F-actin depolymerization by gelsolin. Simultaneously, the ability of LTA to activate translocation of NF-kappaB, E-selectin expression, and adhesion of neutrophils to LTA-treated human aortic endothelial cells was compromised by gelsolin. Gelsolin was able to partially inhibit LPS- or LTA-induced release of IL-8 from human neutrophils but was unable to prevent Gram-positive Bacillus subtilis or Gram-negative Pseudomonas aeruginosa growth and had no effect on the antibacterial activity of the cathelicidin-derived antibacterial peptide LL37. These data suggest that extracellular gelsolin is involved in the host immune recognition of LTA or LPS following release of these molecules from the bacterial outer membrane during cell division or attack by drugs and immune components.


A comparison of Lagrangian model estimates to light detection and ranging (LIDAR) measurements of dust plumes from field tilling.
A Lagrangian particle model has been adapted to examine human exposures to particulate matter < or = 10 microm (PM10) in agricultural settings. This paper reports the performance of the model in comparison to extensive measurements by elastic LIDAR (light detection and ranging). For the first time, the LIDAR measurements allowed spatially distributed and time dynamic measurements to be used to test the predictions of a field-scale model. The model outputs, which are three-dimensional concentration distribution maps from an agricultural disking operation, were compared with the LIDAR-scanned images. The peak cross-correlation coefficient and the offset distance of the measured and simulated plumes were used to quantify both the intensity and location accuracy. The appropriate time averaging and changes in accuracy with height of the plume were examined. Inputs of friction velocity, Monin-Obukhov length, and wind direction (1 sec) were measured with a three-axis sonic anemometer at a single point in the field (at 1.5-m height). The Lagrangian model of Wang et al. predicted the near-field concentrations of dust plumes emitted from a field disking operation with an overall accuracy of approximately 0.67 at 3-m height. Its average offset distance when compared with LIDAR measurements was approximately 38 m, which was 6% of the average plume moving distance during the simulation periods. The model is driven by weather measurements, and its near-field accuracy is highest when input time averages approach the turbulent flow time scale (3-70 sec). The model accuracy decreases with height because of smoothing and errors in the input wind field, which is modeled rather than measured at heights greater than the measurement anemometer. The wind steadiness parameter (S) can be used to quantify the combined effects of wind speed and direction on model accuracy.


Applications of PNA-Based Artificial Restriction DNA Cutters.
More than ten years ago, artificial restriction DNA cutters were developed by combining two pseudo-complementary peptide nucleic acid (pcPNA) strands with either Ce(IV)/EDTA or S1 nuclease. They have remarkably high site-specificity and can cut only one predetermined site in the human genome. In this article, recent progress of these man-made tools have been reviewed. By cutting the human genome site-selectively, desired fragments can be clipped from either the termini of chromosomes (telomeres) or from the middle of genome. These fragments should provide important information on the biological functions of complicated genome system. DNA/RNA hybrid duplexes, which are formed in living cells, are also site-selectively hydrolyzed by these cutters. In order to further facilitate the applications of the artificial DNA cutters, various chemical modifications have been attempted. One of the most important successes is preparation of PNA derivatives which can form double-duplex invasion complex even under high salt conditions. This is important for in vivo applications, since the inside of living cells is abundant of metal ions. Furthermore, site-selective DNA cutters which require only one PNA strand, in place of a pair of pcPNA strands, are developed. This progress has opened a way to new fields of PNA-based biochemistry and biotechnology.


A benchmark for chromatin binding measurements in live cells.
Live-cell measurement of protein binding to chromatin allows probing cellular biochemistry in physiological conditions, which are difficult to mimic in vitro. However, different studies have yielded widely discrepant predictions, and so it remains uncertain how to make the measurements accurately. To establish a benchmark we measured binding of the transcription factor p53 to chromatin by three approaches: fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS) and single-molecule tracking (SMT). Using new procedures to analyze the SMT data and to guide the FRAP and FCS analysis, we show how all three approaches yield similar estimates for both the fraction of p53 molecules bound to chromatin (only about 20%) and the residence time of these bound molecules (∼1.8 s). We also apply these procedures to mutants in p53 chromatin binding. Our results support the model that p53 locates specific sites by first binding at sequence-independent sites.


Chromosome mapping of retrotransposable elements Rex1 and Rex3 in three fish species in the subfamily Hypoptopomatinae (Teleostei, Siluriformes, Loricariidae).
Transposons and retrotransposons represent a great portion of the eukaryotic genome. These repeated elements can be defined as genetic entities capable of self-inserting into different positions within the genome modifying the functions of the genes with which they are associated. The purpose of the present work was to isolate, identify, and map the sequences of Rex1 and Rex3 transposable elements present in the genomes of fishes of the family Loricariidae. The retrotransposable elements were identified through amplification with Rex1 and Rex3 primers, and the nucleotide composition of the retrotransposons was determined by automatic sequencing. In all the species analyzed it was possible to isolate Rex1 partial sequences from 520 to 563 bp in size and Rex3 partial sequences from 407 to 454 bp in size. Comparisons with the available literature showed that Loricariidae retrotransposons Rex1 and Rex3 have a high sequence similarity to putative homologous genes in different fish orders. Fluorescence in situ hybridization experiments conducted in 3 species, Hisonotus leucofrenatus, Pseudotocinclus tietensis, and Parotocinclus maculicauda, using the retrotransposable elements isolated for each species as probes, revealed that Rex1 and Rex3 share a similar dispersion pattern and are both found on all chromosomes of the 3 species organized in small clusters and dispersed on all the chromosomes, including euchromatic and heterochromatic regions, although a larger concentration was observed in the heterochromatic regions of H. leucofrenatus. The data found in the present study contradict much of the literature that substantiates that the retrotranspon elements are preferentially accumulated in the heterochromatin regions in fishes.


Context specific protein function prediction.
Although whole-genome sequencing of many organisms has been completed, numerous newly discovered genes are still functionally unknown. Using high-throughput data such as protein-protein interaction (PPI) information to assign putative protein function to the unknown genes has been proposed, since in many cases it is not feasible to annotate the newly discovered genes by sequence-based approaches alone. In addition to PPI data, information such as protein localization within a cell may be employed to improve protein function prediction in two ways: 1) By using such localization information as a direct indicator of protein function (e.g. nucleolus localized proteins might be involved in ribosome biogenesis), and 2) by refining noisy PPI data by localization information. In the latter case, localization information may be used to distinguish different types of PPIs: Namely, interactions between co-localized proteins (more reliable), and interactions between differently localized proteins (potentially less reliable). In this paper, we propose a probabilistic method to predict protein function from PPI data and localization information. A Bayesian network is used to model dependencies between protein function, PPI data and localization information. We showed in our cross-validation experiment that in some cases, our method (conditioning PPI data by localization information) significantly improves prediction precision, as compared to a simple Naive Bayes method that assumes PPI data and localization information are conditionally independent given protein function. Finally, we predicted 57 unknown genes as "ribosome biogenesis" proteins.


Catecholamine storage vesicle protein expression in genetic hypertension.
Chromogranin A expression is heritable in humans, and both plasma chromogranin A concentration and its releasable adrenal and sympathetic neuronal pools are augmented in established essential (hereditary) hypertension. To evaluate chromogranin A further as a simpler or "intermediate phenotype" in the complex trait of hypertension, we studied chromogranin A expression in the spontaneously hypertensive rat (SHR), a rodent model of essential hypertension. Both plasma (p < 0.0001) and adrenal medullary (p = 0.003 to p < 0.0001) chromogranin A were elevated in the SHR, even at the earliest stages (3-4 weeks of age). In the adult adrenal gland, both chromogranin A (p=0.005) and norepinephrine (p=0.011) were increased in the SHR, while dopamine beta-hydroxylase activity was diminished (p < 0.0001). Chromogranin A mRNA expression was also elevated in the SHR adrenal medulla (p = 0.017). Differences in chromogranin A processing were not noted between SHR and Wistar Kyoto control (WKY) rats. In an SHR x WKY genetic intercross, control of the adrenal chromogranin A phenotype by a single major locus was suggested by comparison of phenotypic variance of the F2 vs F1 generations, and by bimodal frequency histogram (3:1 ratio), confirmed by maximum likelihood analysis (chi2 = 74.6, p < 0.000001) in the F2 generation. However, microsatellite alleles at a surrogate locus (Ighe) 12.7 cM from chromogranin A (Chga), on rat chromosome 6, failed to co-segregate with blood pressure in an F2 generation (F = 0.06, p = 0.94). In another rodent model of hereditary hypertension, the genetically hypertensive mouse (BPH/2), adrenal chromogranin A (p=0.018) and norepinephrine (p = 0.004) were actually diminished. We conclude that over-expression of chromogranin A is a variable feature of mammalian genetic hypertension. In one rodent model (the SHR), over-expression of chromogranin A is largely controlled by a single genetic locus, but the chromogranin A locus itself is not directly linked to determination of the blood pressure elevation of the SHR.


Pyridine/Pyrimidine Substituted Imidazol-5-one Analogs as HIV-1 RT Inhibitors: Design, Synthesis, Docking and Molecular Dynamic Simulation Studies.
This paper reports the synthesis, Non-nucleoside reverse transcriptase inhibitory (NNRTIs) activity and computational studies of 2-((4-chloro-2-subtitutedphenoxy) methyl)-4-(furan-2-ylmethylene)-1-substituted Pyridine/-pyrimidine-1H-imidazol-5(4H)-ones. The imidazol-5-one analogs were synthesized by conventional method and characterized by FT-IR, NMR and mass spectral data. All compounds were evaluated for in-vitro NNRTI activity by using reverse transcriptase (RT) assay kit (Roche). The in-silico docking studies were conducted on RT enzyme to investigate binding site interactions of synthesized compounds. The MMGBSA method was also used to calculate the binding free energy between the inhibitors and RT enzyme. The MD simulation was further performed for the apo form of the RT enzyme and docked complex of compound A6-RT enzyme to better understand the stability of the protein-ligand complex. The bioactivity analysis revealed that most of the synthesized compounds showed significant inhibitory activity against RT enzyme and the IC50 value was found to be in the range of 1.76-3.88 μM. The computational studies suggest that the docked compounds form the H-bonding with amino acid residue Lys101 and hydrophobic interactions with amino acid residues Tyr188, Tyr181, Trp229, and Tyr318, which act as the primary driving forces for protein-ligand interaction. The reported imidazol-5-one analogs can act as lead for further development of prospective RT inhibitors.


Metabolic engineering for improved fermentation of pentoses by yeasts.
The fermentation of xylose is essential for the bioconversion of lignocellulose to fuels and chemicals, but wild-type strains of Saccharomyces cerevisiae do not metabolize xylose, so researchers have engineered xylose metabolism in this yeast. Glucose transporters mediate xylose uptake, but no transporter specific for xylose has yet been identified. Over-expressing genes for aldose (xylose) reductase, xylitol dehydrogenase and moderate levels of xylulokinase enable xylose assimilation and fermentation, but a balanced supply of NAD(P) and NAD(P)H must be maintained to avoid xylitol production. Reducing production of NADPH by blocking the oxidative pentose phosphate cycle can reduce xylitol formation, but this occurs at the expense of xylose assimilation. Respiration is critical for growth on xylose by both native xylose-fermenting yeasts and recombinant S, cerevisiae. Anaerobic growth by recombinant mutants has been reported. Reducing the respiration capacity of xylose-metabolizing yeasts increases ethanol production. Recently, two routes for arabinose metabolism have been engineered in S. cerevisiae and adapted strains of Pichia stipitis have been shown to ferment hydrolysates with ethanol yields of 0.45 g g(-1) sugar consumed, so commercialization seems feasible for some applications.


Main and interaction effects of childhood trauma and the MAOA uVNTR polymorphism on psychopathy.
Psychopathy is characterized by callous affect, interpersonal manipulation, a deviant lifestyle, and antisocial behavior. Previous research has linked psychopathic traits to childhood trauma, but also to the upstream variable number tandem repeat (uVNTR) polymorphism of the monoamine oxidase A (MAOA) gene. An interaction between childhood trauma and MAOA genotype has been associated with antisocial behavior, but so far little is known about interaction effects of childhood trauma and the MAOA uVNTR on psychopathy. In order to bridge this gap, we used data of 1531 male and 1265 female twins and their siblings from a Finnish community sample to estimate structural equation models. The psychopathy and childhood trauma constructs were conceptualized as bifactor models with one general and two orthogonal group factors. Data comprised self-reports on childhood trauma and psychopathic traits as well as MAOA uVNTR genotype. In both genders, childhood trauma was associated with the general factor that represents the overarching psychopathy construct, and with the group factor that captures social deviance, but not with the group factor capturing psychopathic core personality traits. Women with a low activity variant of the MAOA uVNTR reported slightly higher levels of psychopathy than those with a high activity allele, but only with respect to the general psychopathy factor. There was no evidence for an interaction effect between MAOA uVNTR genotype and childhood trauma on psychopathy in either gender. Our results suggest that psychopathy in general and social deviance in particular are associated with childhood trauma in men and women, and that psychopathic traits are subject to variation in the MAOA uVNTR genotype in women.


Disparities in current cigarette smoking among US adults, 2002-2016.
To assess disparities in current (past 30 days) cigarette smoking among US adults aged ≥ 18 years during 2002-2016. Nine indicators associated with social disadvantage were analysed from the 2002 to 2016 National Survey on Drug Use and Health: education, annual family income, sex, race/ethnicity, urbanicity, serious psychological distress, health insurance, public assistance, and employment status. Using descriptive and multivariable analyses, we measured trends in smoking overall and within the assessed variables. We also evaluated effect of interactions on disparities and estimated the excess number of smokers attributable to disparities. During 2002-2016, current cigarette smoking prevalence declined overall (27.5%-20.7%; p trend < 0.01), and among all subgroups except Medicare insurees and American Indians/Alaska Natives (AI/ANs). Overall inequalities in cigarette smoking grew even wider or remained unchanged for several indicators during the study period. In 2016, comparing groups with the least versus the most social advantage, the single largest disparity in current smoking prevalence was seen by race/ethnicity (prevalence ratio = 5.1, AI/ANs vs Asians). Education differences alone explained 38.0% of the observed racial/ethnic disparity in smoking prevalence. Interactions were also present; compared with the population-averaged prevalence among all AI/AN individuals (34.0%), prevalence was much higher among AI/ANs with <high school diploma (53.0%), unemployed (58.0%), or with serious psychological distress (66.9%). The burden of smoking attributable to race/ethnic disparities in smoking prevalence was an estimated 27.6 million smokers. Overall smoking inequality increased or remained unchanged because of slower declines in smoking prevalence among disadvantaged groups. Targeted interventions among high-risk groups can narrow disparities.


Considerable delay in diagnosis and acute management of subarachnoid haemorrhage.
Rebleeding from subarachnoid haemorrhage (SAH) usually occurs within the first six hours after the initial bleeding. Rebleeding can be prevented effectively with tranexamic acid (TXA). Although a broad consensus has evolved that SAH should be treated as an emergency, it is likely that delays do exist in the diagnosis and treatment of SAH patients. The aim of this study was to prospectively assess the interval between symptom onset, emergency room (ER) admission, initial diagnosis and treatment, and final closure of the aneurysm. We prospectively studied the time course from the initial bleeding to ER admission, computed tomography (CT), TXA treatment, referral to the neurosurgical department, and to the final closure of the aneurysm. A total of 133 patients with SAH due to ruptured intracranial aneurysms were admitted to two neurosurgical units in Copenhagen, Denmark, during a one-year period. The median time to admission was 60 min. The median delay from admission to CT scan was 55 min. Long pre-hospital delay (p = 0.03) and high Glasgow Coma Scale score on arrival (p = 0.0006) were associated with a longer time to CT scan. The median time from CT scan to initiation of TXA treatment was 50 min. The median time from initial insult to final closure of the aneurysm was 30 hours. The present study demonstrates that considerable diagnostic delays exist in connection with CT and TXA treatment after patients' arrival to the ER.


Barefoot running: an evaluation of current hypothesis, future research and clinical applications.
Barefoot running has become a popular research topic, driven by the increasing prescription of barefoot running as a means of reducing injury risk. Proponents of barefoot running cite evolutionary theories that long-distance running ability was crucial for human survival, and proof of the benefits of natural running. Subsequently, runners have been advised to run barefoot as a treatment mode for injuries, strength and conditioning. The body of literature examining the mechanical, structural, clinical and performance implications of barefoot running is still in its infancy. Recent research has found significant differences associated with barefoot running relative to shod running, and these differences have been associated with factors that are thought to contribute to injury and performance. Crucially, long-term prospective studies have yet to be conducted and the link between barefoot running and injury or performance remains tenuous and speculative. The injury prevention potential of barefoot running is further complicated by the complexity of injury aetiology, with no single factor having been identified as causative for the most common running injuries. The aim of the present review was to critically evaluate the theory and evidence for barefoot running, drawing on both collected evidence as well as literature that have been used to argue in favour of barefoot running. We describe the factors driving the prescription of barefoot running, examine which of these factors may have merit, what the collected evidence suggests about the suitability of barefoot running for its purported uses and describe the necessary future research to confirm or refute the barefoot running hypotheses.


Loss of P53 Function Activates JAK2-STAT3 Signaling to Promote Pancreatic Tumor Growth, Stroma Modification, and Gemcitabine Resistance in Mice and Is Associated With Patient Survival.
One treatment strategy for pancreatic ductal adenocarcinoma is to modify, rather than deplete, the tumor stroma. Constitutive activation of the signal transducer and activator of transcription 3 (STAT3) is associated with progression of pancreatic and other solid tumors. We investigated whether loss of P53 function contributes to persistent activation of STAT3 and modification of the pancreatic tumor stroma in patients and mice. Stat3, Il6st (encodes gp130), or Trp53 were disrupted, or a mutant form of P53 (P53R172H) or transgenic sgp130 were expressed, in mice that developed pancreatic tumors resulting from expression of activated KRAS (KrasG12D, KC mice). Pancreata were collected and analyzed by immunohistochemistry, in situ hybridization, quantitative reverse-transcription polymerase chain reaction (qPCR), or immunoblot assays; fluorescence-activated cell sorting was performed to identify immune cells. We obtained frozen pancreatic tumor specimens from patients and measured levels of phosphorylated STAT3 and P53 by immunohistochemistry; protein levels were associated with survival using Kaplan-Meier analyses. We measured levels of STAT3, P53, ligands for gp130, interleukin 6, cytokines, sonic hedgehog signaling, STAT3 phosphorylation (activation), and accumulation of reactive oxygen species in primary pancreatic cells from mice. Mice with pancreatic tumors were given gemcitabine and a Janus kinase 2 (JAK2) inhibitor; tumor growth was monitored by 3-dimensional ultrasound. STAT3 was phosphorylated constitutively in pancreatic tumor cells from KC mice with loss or mutation of P53. Tumor cells of these mice accumulated reactive oxygen species and had lower activity of the phosphatase SHP2 and prolonged phosphorylation of JAK2 compared with tumors from KC mice with functional P53. These processes did not require the gp130 receptor. Genetic disruption of Stat3 in mice, or pharmacologic inhibitors of JAK2 or STAT3 activation, reduced fibrosis and the numbers of pancreatic stellate cells in the tumor stroma and altered the types of immune cells that infiltrated tumors. Mice given a combination of gemcitabine and a JAK2 inhibitor formed smaller tumors and survived longer than mice given control agents; the tumor stroma had fewer activated pancreatic stellate cells, lower levels of periostin, and alterations in collagen production and organization. Phosphorylation of STAT3 correlated with P53 mutation and features of infiltrating immune cells in human pancreatic tumors. Patients whose tumors had lower levels of phosphorylated STAT3 and functional P53 had significantly longer survival times than patients with high levels of phosphorylated STAT3 and P53 mutation. In pancreatic tumors of mice, loss of P53 function activates JAK2-STAT3 signaling, which promotes modification of the tumor stroma and tumor growth and resistance to gemcitabine. In human pancreatic tumors, STAT3 phosphorylation correlated with P53 mutation and patient survival time. Inhibitors of this pathway slow tumor growth and stroma formation, alter immune cell infiltration, and prolong survival of mice. Transcript profiling: ArrayExpress accession number: E-MTAB-3278.


Antigen recognition during progression from acute to chronic infection with a cagA-positive strain of Helicobacter pylori.
We have previously published two reports on acute infection with Helicobacter pylori, one of an adult male and one of a family of four. In the present study, we have isolated H.pylori from each of twin boys in the family and compared these by use of random amplified polymorphic DNA PCR. In addition, we have monitored the antibody response over time of the family and the adult male by Western blotting (immunoblotting) with two different strains of H. pylori as the antigen and by use of a commercial kit. The acutely infected twin boys were infected by an identical strain of H. pylori. The twin boys responded to antigens of 19, 26.5, and 29 kDa 30 days after the initial diagnosis, with recognition of 43-to 49-, 66-, 69-, and 87-kDa antigens by day 63. One twin responded to the CagA antigen on day 63, whereas the other responded on day 857. Antibody to the CagA antigen was not detected by use of the infecting strain, UNSW-RU1. Investigation of UNSW-RU1 revealed the presence of cagA. In two acutely infected adults (one, the father of the boys), the initial response to a 45-kDA antigen was later followed by responses to 19-, 29-, 49-, 60-, 77-, and 84-kDa antigens. Sera from the twins' younger sister, born 17 months after the twins acute episode, indicated that she also had become infected. This report supports intrafamilial transmission of H. pylori. Initial antibody responses in the children were to small-molecular-size antigens; in the adults, the initial responses were to larger-molecular-size antigens. The pattern of the serological response differs according to the antigen used. This has implications in regard to international data comparisons.


[Influence of long-term lactation on 7,12-dimethylbenz(a)anthracene (DMBA) induced rat mammary tumor (author's transl)].
Female rats of the Sprague-Dawley strain were mated 60 days after a single intravenous injection with 5 mg of 7,12-dimethylbenz(a)anthracene. In order to maintain the lactation for more than 3 weeks, 5 pups (5 to 10 days old) were attached to each mother rat throughout the experiment. Experimental rats were divided into 6 groups: 1. non-lactation (4 rats); 2. 3-week lactation (8 rats); 3. 6-week lactation (10 rats); 4. 8-week lactation (7 rats); 5. 12-week lactation (9 rats); and 6. 17-week lactation (4 rats). Lactating condition was judged from the body weight curves of the 5 pups. The appearance, number and diameter of tumors were recorded every day. The average of the tumor induction time was 140+/-74 days. The tumors were divided into 3 types: 1. progressive; 2. static; and 3. regressive. During pregnancy few tumors of the regressive type were noted. However, tumors of the progressive type increased significantly with the weeks of pregnancy. In the first week postpartum, although the total number of tumors was approximately the same as in the last week of pregnancy, the tumors of the progressive type decreased significantly and the tumors of the regressive type increased. In this period most tumors showed a tendency to regress. In the non-lactating group, the tumors of the progressive type began to increase from the fifth to seventh week postpartum without relation to the lactating period (3 to 17 weeks). Vaginal smears were examined every day in all mother rats to determine the recurrence of the estrous cycle. The first estrus appeared 7 days on an average after delivery in the non-lactating group; 27 days on an average in the 3-week lactation group; and 35 to 38 days on an average in the 6- to 17-week lactation groups. Under the condition of high prolactin level, the behavior of the tumors of the progressive type seemed to be related to estrogen. This was also supported by histological findings in the ovaries.


Agranulocytosis monitoring with Clozapine patients: to follow guidelines or to attempt therapeutic controversies?
Clozapine is an atypical antipsychotic with superior efficacy in the treatment of refractory schizophrenia. But it can cause agranulocytosis, which occurs in one to two percent of patients. This paper was prepared to discuss the condoned and controversial issues of therapy with this drug, but only within a haematological context. The feasibility of attempting therapeutically controversial blood monitoring regimes, as opposed to following standardised Western guidelines, given the differences in terms of accessibility, convenience and financial considerations between the public and private sector medical care will also be discussed. The proposal of adopting a structured pro forma, with a risk-benefit assessment, in the event of unavoidable veering from the guidelines may allay medicolegal implications, especially in countries where blood monitoring is not mandatory. It is hoped that this article will stimulate further research in our region, bearing in mind the increasing awareness and focus on genetic polymorphism, and the possibility of drawing up our own monitoring guidelines in the near future.


Recombinant human TNF alpha stimulates production of granulocyte colony-stimulating factor.
Tumor necrosis factor alpha (TNF alpha) stimulates production of granulocyte colony-stimulating factor (G-CSF) protein and mRNA in fibroblast cells. In contrast, TNF beta is at least 1,000-fold less potent than TNF alpha in increasing levels of G-CSF and GM-CSF activity and mRNA. The fibroblasts produce G-CSF and GM-CSF mRNA in a coordinate fashion after exposure to TNF. The ability of TNF alpha to stimulate production of CSF in fibroblasts may be pivotal to regulation of hematopoiesis.


Reclaimed and Up-Cycled Cathodes for Lithium-Ion Batteries.
As electric vehicles become more widely used, there is a higher demand for lithium-ion batteries (LIBs) and hence a greater incentive to find better ways to recycle these at their end-of-life (EOL). This work focuses on the process of reclamation and re-use of cathode material from LIBs. Black mass containing mixed LiMn2O4 and Ni0.8Co0.15Al0.05O2 from a Nissan Leaf pouch cell are recovered via two different recycling routes, shredding or disassembly. The waste material stream purity is compared for both processes, less aluminium and copper impurities are present in the disassembled waste stream. The reclaimed black mass is further treated to reclaim the transition metals in a salt solution, Ni, Mn, Co ratios are adjusted in order to synthesize an upcycled cathode, LiNi0.6Mn0.2Co0.2O2 via a co-precipitation method. The two reclamation processes (disassembly and shredding) are evaluated based on the purity of the reclaimed material, the performance of the remanufactured cell, and the energy required for the complete process. The electrochemical performance of recycled material is comparable to that of as-manufactured cathode material, indicating no detrimental effect of purified recycled transition metal content. This research represents an important step toward scalable approaches to the recycling of EOL cathode material in LIBs.


Reliability of a method for the localization of displaced maxillary canines using a single panoramic radiograph.
The present study was initiated to determine the validity and reproducibility of a method previously reported for localization of displaced maxillary canines by panoramic radiographs. Eleven dental specialists (five orthodontists, five surgeons, and one radiologist) and five (final year) dental students were asked to interpret initial panoramic radiographs of 20 patients with 26 unerupted canines. The method contends that, provided that the radiographic image of the unerupted canines superimposes on the root of the lateral incisor at a height coronal to the apical third, a ratio between the widest mesiodistal dimension of the affected canine and the homolateral central incisor [Canine-Incisor Index (CII)] of more than 1.15 indicates palatal displacement. Among the observers, the measurement error was small, insufficient to cause overlap between the ranges of values for the CII of buccal and palatal canines. Inter-observer reproducibility was high. Without exception, all observers correctly diagnosed the location of each of the impacted canines. The CII cut-off point of 1.15, which was arbitrarily chosen in the previous work, was shown to be valid to differentiate buccal from palatal displacement. The present method is rapid, simple, accurate, and easily taught to dental students or dental specialists, with the simplest of initial instruction. While confirmations with other radiographic techniques is strongly advised before a definitive treatment is undertaken, this method has merit in providing more information than has been available from the panoramic radiograph hitherto, to satisfy the needs of an initial orthodontic consultation.


Inhibition of cyclooxygenase activity reduces rotavirus infection at a postbinding step.
Elevated levels of prostaglandins (PGs), products of cyclooxygenases (COXs), are found in the plasma and stool of rotavirus-infected children. We sought to determine the role of COXs, PGs, and the signal transduction pathways involved in rotavirus infection to elucidate possible new targets for antiviral therapy. Human intestinal Caco-2 cells were infected with human rotavirus Wa or simian rotavirus SA-11. COX-2 mRNA expression and secreted PGE2 levels were determined at different time points postinfection, and the effect of COX inhibitors on rotavirus infection was studied by an immunofluorescence assay (IFA). To reveal the signal transduction pathways involved, the effect of MEK, protein kinase A (PKA), p38 mitogen-activated protein kinase (MAPK), and NF-kappaB inhibitors on rotavirus infection was analyzed. In infected Caco-2 cells, increased COX-2 mRNA expression and secreted PGE2 levels were detected. Indomethacin (inhibiting both COX-1 and COX-2) and specific COX-1 and COX-2 inhibitors reduced rotavirus infection by 85 and 50%, respectively, as measured by an IFA. Indomethacin reduced virus infection at a postbinding step early in the infection cycle, inhibiting virus protein synthesis. Indomethacin did not seem to affect viral RNA synthesis. Inhibitors of MEK, PKA, p38 MAPK, and NF-kappaB decreased rotavirus infection by at least 40%. PGE2 counteracted the effect of the COX and PKA inhibitors but not of the MEK, p38 MAPK, and NF-kappaB inhibitors. Conclusively, COXs and PGE2 are important mediators of rotavirus infection at a postbinding step. The ERK1/2 pathway mediated by PKA is involved in COX induction by rotavirus infection. MAPK and NF-kappaB pathways are involved in rotavirus infection but in a PGE2-independent manner. This report offers new perspectives in the search for therapeutic agents in treatment of severe rotavirus-mediated diarrhea in children.


Characterization of the ultrasonic attenuation coefficient and its frequency dependence in a polymer gel dosimeter.
Research on polymer-gel dosimetry has been driven by the need for three-dimensional dosimetry, and because alternative dosimeters are unsatisfactory or too slow for that task. Magnetic resonance tomography is currently the most well-developed technique for determining radiation-induced changes in polymer structure, but quick low-cost alternatives remain of significant interest. In previous work, ultrasound attenuation and speed of sound were found to change as a function of absorbed radiation dose in polymer-gel dosimeters, although the investigations were restricted to one ultrasound frequency. Here, the ultrasound attenuation coefficient mu in one polymer gel (MAGIC) was investigated as a function of radiation dose D and as a function of ultrasonic frequency f in a frequency range relevant for imaging dose distributions. The nonlinearity of the frequency dependence was characterized, fitting a power-law model mu = af(b); the fitting parameters were examined for potential use as additional dose readout parameters. In the observed relationship between the attenuation coefficient and dose, the slopes in a quasi-linear dose range from 0 to 30 Gy were found to vary with the gel batch but lie between 0.0222 and 0.0348 dB cm(-1) Gy(-1) at 2.3 MHz, between 0.0447 and 0.0608 dB cm(-1) Gy(-1) at 4.1 MHz and between 0.0663 and 0.0880 dB cm(-1) Gy(-1) at 6.0 MHz. The mean standard deviation of the slope for all samples and frequencies was 15.8%. The slope was greater at higher frequencies, but so were the intra-batch fluctuations and intra-sample standard deviations. Further investigations are required to overcome the observed variability, which was largely associated with the sample preparation technique, before it can be determined whether any frequency is superior to others in terms of accuracy and precision in dose determination. Nevertheless, lower frequencies will allow measurements through larger samples. The fit parameter a of the frequency dependence, describing the attenuation coefficient at 1 MHz, was found to be dose dependent, which is consistent with our expectations, as polymerization is known to be associated with increased absorption of ultrasound. No significant dose dependence was found for the fit parameter b, which describes the nonlinearity with frequency. This is consistent with the increased absorption being due to the introduction of new relaxation processes with characteristic frequencies similar to those of existing processes. The data presented here will help with optimizing the design of future 3D dose-imaging systems using ultrasound methods.


The effects of vitamin D supplementation on muscle function among postmenopausal women: a systematic review and meta-analysis of randomized controlled trials.
The loss of muscle mass and its strength is one of the most critical changes in aging which is associated with an increased risk of falls, osteoporotic fractures and mobility disability. Vitamin D, with its extra-skeletal benefits, might improve muscle function in elderly. The current systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted to summarize available relevant data and determine the effect of vitamin D supplementation on muscle function among postmenopausal women. We reached databases including; Cochrane library, Embase, PubMed, and Web of Science database until the end of May 2018 to identify relevant published RCTs. Heterogeneity among included studies was assessed using Q-test and I2 statistics. Random-effect model was applied to pool data and weighted mean difference (WMD) was calculated representing summary effect size. Outcomes of interest included the effects of vitamin D supplementation on hand grip strength (HGS), back muscle strength (BMS), and Timed Up and Go (TUG). Twelve RCTs out of 1739 potential reports were included in our meta-analysis. The pooled findings showed that vitamin D supplementation had no significant effect on HGS (WMD -0.03 kilogram (Kg); 95 % CI, -0.26, 0.20; P=0.78), BMS (WMD 7.21 newton (N); 95 % CI, -5.98, 20.40; P=0.28), and TUG (WMD 0.01 second (S); 95 % CI, -0.17, 0.18; P=0.93) in postmenopausal women. Overall, the current meta-analysis showed that taking vitamin D supplementation by postmenopausal women did not affect markers of muscle function. Further studies are required to confirm the effect of vitamin D supplementation on markers of muscle function.


Usefulness of N-terminal pro-brain natriuretic peptide to predict postoperative cardiac complications and long-term mortality after emergency lower limb orthopedic surgery.
After emergency orthopedic-geriatric surgery, cardiac complications are an important cause of morbidity and mortality. The utility of N-terminal pro-brain natriuretic peptide (NT-pro-BNP) for the prediction of cardiac complications and mortality was evaluated. NT-pro-BNP was tested pre- and postoperatively in 89 patients >60 years of age. They were followed for 2 years for cardiac complications (defined as acute myocardial infarction, congestive cardiac failure, atrial fibrillation or major arrhythmia) or death. Receiver operating characteristic curves were constructed to determine the optimal discriminatory level for cardiac events and death using NT-pro-BNP. Twenty-three patients (25.8%) sustained an in-hospital postoperative cardiac complication. Total all-cause mortality was 3 of 89 (3.4%) in hospital, 21 of 89 (23.6%) at 1 year, and 27 of 89 (30.3%) at 2 years. Median preoperative and postoperative NT-pro-BNP levels were higher in patients who had an in-hospital cardiac event compared to those without (387 vs 1,969 pg/ml, p <0.001; and 676 vs 7,052 pg/ml, p <0.001 respectively). The optimal discriminatory level for preoperative NT-pro-BNP was 842 pg/ml and that for postoperative NT-pro-BNP was 1,401 pg/ml for the prediction of in-hospital cardiac events and 1- and 2-year mortality. Preoperative NT-pro-BNP >/=842 pg/ml (odds ratio 11.6, 95% confidence interval 2.1 to 65.0, p = 0.005) was an independent predictor of in-hospital cardiac complications using multivariate analysis and pre- and postoperative NT-pro-BNP levels were independent predictors of 2-year cardiovascular events. Patients who had preoperative NT-pro-BNP >/=842 pg/ml or postoperative NT-pro-BNP >/=1,401 pg/ml had significantly worse survival using log-rank testing (p <0.001) and these variables independently predicted 2-year mortality. In conclusion, increase pre- and postoperative NT-pro-BNP levels are independent predictors of in-hospital cardiac events and 1- and 2-year mortality in older patients undergoing emergency orthopedic surgery.


Immunoperoxidase detection of baculovirus antigens in insect cells.
The sequence of events in the infection of TN-368-10 and TN-368-13 cells by Autographa californica nuclear polyhedrosis virus (AcMNPV) was investigated by using the indirect immunoperoxidase technique. Antisera raised against enveloped nucleocapsids detected homologous antigens at 6 to 8 h post infection which was about 2 h before the appearance of both intracellular and extracellular infectious virus. Similar tests using polyhedrin antiserum showed that polyhedrin is first synthesized at 12 h post infection, 2 to 4 h after the appearance of infectious nonoccluded virus. The immunoperoxidase technique was also applied to four other invertebrate cell lines after inoculation with AcMNPV. The most significant result was that 90% of AcMNPV-inoculated Bombyx mori 5 cells produced enveloped nucleocapsid antigens and infectious virus but only 1% or less of the cells produced polyhedrin. This disparity emphasizes the need for assays for NPV infection that are independent of polyhedron production.


Outcome of landmark-guided percutaneously inserted tunneled central venous catheters in infants and children under 3 years with cancer.
There is a paucity of information on procedural and long-term outcomes of tunneled central venous catheters (TCVC) in infants and children younger than 3 years undergoing anticancer therapy. This study aims to evaluate the success, safety, and complications leading to surgical revision or premature removal of TCVC in this particular patient group. The clinical course of pediatric patients with percutaneous inserted TCVC, including Groshong (GC) and Hickman/Broviac (HB) catheters, has been analyzed retrospectively. The data analysis includes patient and device characteristics, adverse events during insertion, and dwell period complications. A consecutive series of 238 children undergoing implantation of 273 TCVC, including 148 (54.2%) GC and 125 (45.8%) HB catheters, with a total of 38,209 catheter days at risk (cdr) were reviewed. The patient cohort consisted of 65 (23.8%) infants, 77 (28.2%) children aged 1-2 years, and 131 (48.0%) aged 2-3 years. The overall rate of adverse events during catheter insertion was 12.8% (n = 35) with no differences between age groups or devices. The overall rate of long-term complication was 28.2% (n = 77, catheter risk [CR] per 1,000 cdr = 1.75), with the highest prevalence in infants (P = 0.01). The most common complication was late dislocation (n = 24, 8.8%, CR = 0.47), followed by early dislocation (n = 20, 7.3%) and infection (n = 18, 7.4%, CR = 0.42). Percutaneous landmark-guided insertion of TCVC in neonates and small children with cancer is safe. Patterns of long-term complications are different from those for older children and should be prevented through appropriate management.


Phase II trial of ifosfamide and mesna in previously treated patients with non-Hodgkin's lymphoma: Cancer and Leukemia Group B study 8552.
Ifosfamide (1.25 g/m2 intravenously/day x 5) with mesna (20 per cent of the ifosfamide dose x six doses on each day of ifosfamide therapy) was administered to 46 previously treated patients with non-Hodgkin's lymphoma of which 31 were eligible and evaluable. A 29 per cent response rate (9/31) was observed (two CR and seven PR) with a median duration of response of 2.5 months. Myelosuppression was dose-limiting. Hemorrhagic cystitis was observed in three patients (10 per cent). Nausea and vomiting was generally mild or moderate. One patient developed transient neurotoxic symptoms with confusion and disorientation. An additional patient developed an anaphylactic-type reaction with shortness of breath and respiratory stridor during the fourth course of therapy. Ifosfamide, as a single agent, produces remissions of limited duration in non-Hodgkin's lymphoma in patients in second or third relapse.