Colchicine suppresses the release of fibroblast growth factors from alveolar macrophages in vitro. The basis of a possible therapeutic approach ot the fibrotic disorders.
Fibrosis is the accumulation of fibroblasts and the connective tissue products secreted by these cells, usually subsequent to tissue injury. While fibrosis can be useful in preserving the general structural integrity of a tissue, it often alters cell-cell and cell-connective tissue interactions, which leads to loss of tissue function. On the basis of the concept that mononuclear phagocytes can direct the development of fibrosis through the release of specific mediators that stimulate fibroblast proliferation, we propose a therapeutic strategy to prevent fibrosis by preventing the release of these specific mediators. The present study demonstrated that colchicine, a widely used and well-tolerated drug, can block alveolar macrophage release of 2 mediators associated with the development of fibrosis in interstitial lung diseases, fibronectin, and the alveolar-macrophage-derived growth factor (AMDGF). Colchicine blocked the spontaneous release of fibronectin by alveolar macrophages obtained from patients with fibrotic lung disease by 23 +/- 4% after 24 h and by greater than 90% after 72 h. AMDGF release was blocked by 68 +/- 10% after 4 h (p less than 0.01, all comparisons). The effect of colchicine was not due to nonspecific toxicity since [14C]proline tracer studies demonstrated that macrophages treated with colchicine were capable of de novo protein synthesis and the secretion of several protein products, despite the fact that fibronectin and AMDGF release were suppressed. The effect of colchicine on the spontaneous release of both fibronectin and AMDGF could be observed at concentrations less than 10 ng/ml, levels that can be achieved in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)


A comparison of methods for estimating the kinetic parameters of two simple types of transport process.
Sets of experimental data, with known characteristics and error structures, have been simulated for the Michaelis-Menten equation plus a second term, either for linear transport or for competitive inhibition. The Michaelis-Menten equation plus linear term was fitted by several methods and the accuracy and the precision of the parameter estimates from the several methods were compared. The model-fitting methods were: three for least-squares non-linear regression, computer versions of two graphical methods and of two non-parametric methods. The most precise and accurate method was that of D.W. Marquardt (J. Soc. Ind. Appl. Math. 11 (1963) 431--441). The Michaelis-Menten equation with competitive inhibition was also fitted by several methods, viz., two for least-squared non-linear regression, non-parametric method and four variants of the Preston-Schaeffer-Curran plot (Preston, R.L. et al. (1974) J. Gen. Physiol. 64, 443--467). The most precise and accurate of these was the non-linear regression method of W.W. Cleland (Adv. Enzymol. 29 (1967) 1--32). For both these models, the various graphical methods and non-parametric methods gave poor results and are not recommended.


Exposed hydroxyapatite orbital implants. Report of six cases.
Six patients with complications of primary or secondary hydroxyapatite implants were studied. Complications included socket infection and/or conjunctival dehiscence. Complications were detected during regular follow-up examinations, and various treatment approaches were used. The hydroxyapatite implant exposure occurred 4 to 6 weeks (mean, 4.5 weeks) after implantation. Three of the six implants were wrapped in preserved donor sclera before implantation. One of the implants showed wide exposure and chronic infection and was removed. In two cases, scleral patch grafts with a conjunctival pedicle graft were performed, resulting in successful coverage of the implant without further conjunctival dehiscence. In one of the patients, a Tenon's conjunctival flap was advanced to cover the defect, and was unsuccessful with the spicules of the hydroxyapatite eroding through the vascular flap after 1 month. Three of the patients demonstrate a persistent conjunctival epithelial defect. These three patients with chronically exposed hydroxyapatite have remained stable with follow-up intervals ranging from 8 to 12 months. Early exposure of hydroxyapatite orbital implants is a potential problem despite meticulous technique. Implant coverage is difficult, although chronic exposure seems to be tolerated often in the hydroxyapatite orbital implant without migration or extrusion.


Comparison of analgesic effect of direct breastfeeding, oral 25% dextrose solution and placebo during 1st DPT vaccination in healthy term infants: a randomized, placebo controlled trial.
To compare analgesic effect of direct breast feeding, 25% dextrose solution and placebo as we give 1st intramuscular whole cell DPT injection to 6week - 3month old infants. Randomized, placebo controlled trial. Immunization clinic of Department of Pediatrics, LLRM Medical College. Infants coming for their 1st DPT vaccination were randomized in to three groups of 40 each. The primary outcome variable was the duration of cry after vaccination. Secondary outcome variables were Modified Facial Coding Score (MFCS) and latency of onset of cry. 120 babies were equally enrolled in breast feed group, 25% dextrose fed group and distilled water fed group. Median (interquartile range) of duration of cry was significantly lower in breast fed (33.5 (17-54) seconds) and 25% dextrose fed babies (47.5 (31-67.5) seconds) as compared to babies given distilled water (80.5 (33.5-119.5) seconds) (P<0.001). MFCS at 1 min and 3 min was significantly lower in direct breast fed and dextrose fed babies. Direct breastfeeding and 25% dextrose act as analgesic in young infants undergoing DPT vaccination in young infants less than 3 month of age.


Pushing Photochemistry into Water: Acceleration of the Di-π-Methane Rearrangement and the Paternó-Büchi Reaction "On-Water".
Two representative organic photoreactions, namely a bimolecular photocycloaddition and a monomolecular photorearrangement, are presented that are accelerated when the reaction is performed "on-water", that is, at the water-substrate interface with no solvation of the reaction components. According to the established models of ground-state reactions "on-water", the enhanced efficiency of the photoreactions is explained by hydrophobic effects (Paternó-Büchi reaction) or specific hydrogen bonding (di-π-methane rearrangement) at the water-substrate interface that decrease the energy of the respective transition state. These results point to the potential of this approach to conduct photoreactions more efficiently in an ecologically favorable medium.


Imaging organelle membranes in live cells at the nanoscale with lipid-based fluorescent probes.
Understanding how organelles interact, exchange materials, assemble, disassemble, and evolve as a function of space, time, and environment is an exciting area at the very forefront of chemical and cell biology. Here, we bring attention to recent progress in the design and application of lipid-based tools to visualize and interrogate organelles in live cells, especially at super resolution. We highlight strategies that rely on modification of natural lipids or lipid-like small molecules ex cellula, where organelle specificity is provided by the structure of the chemically modified lipid, or in cellula using cellular machinery, where an enzyme labels the lipid in situ. We also describe recent improvements to the chemistry upon which lipid probes rely, many of which have already begun to broaden the scope of biological questions that can be addressed by imaging organelle membranes at the nanoscale.


Co2+ anchored on surface-functionalized PET non-woven fabric and used as high efficiency monoatom-like catalyst for activating Oxone in water.
Fenton-like processes have emerged as most promising techniques for generating reactive oxygen-containing radicals to deal with increasing levels of environmental pollution. Developing novel catalysts with simple manufacturing requirements, excellent activity levels, and stability remains a long-term goal in terms of practical application. So herein, a new polyethylene terephthalate (PET) non-woven fabric based composite catalyst has been fabricated, using radiation-induced graft polymerization of a functionalized group to chelate Co2+ ions as heterogeneous catalysts in peroxymonosulfate (Oxone) activation. Several impact factors, including catalyst dosage, Oxone concentration, reaction temperature, pH value, Co2+ precipitation ratio (of Co@PET at different pH values), and highly concentrated NaCl have been investigated here. Notably, Co@PET has shown the lowest activation energy of any reported catalyst, for degrading RhB by activating Oxone. Interestingly, as experimental RhB and Oxone solutions were passed through single Co@PET sheets, the RhB was decomposed into a colorless solution in the penetration process. Based on radical trapping and quenching experiments, a channel was determined to dominate RhB degradation, and furthermore, Co@PET could be re-used for RhB degradation by activating Oxone. These results showed that Co@PET effectively provided improved Fenton-like catalytic performance and stability, and was suitable for practical applications.


Improved anticoagulation of titanium by sequential immobilization of oligo(ethylene glycol) and 2-methacryloyloxyethyl phosphorylcholine.
Titanium and its alloys have been widely used for blood-contacting biomedical devices; however, their blood compatibility needs to be improved. In this study, titanium surface was modified by sequential immobilization of oligo(ethylene glycol) (OEG) and 2-methacryloyloxyethyl phosphorylcholine (MPC) to improve its anticoagulation. Water contact angle results showed an excellent hydrophilic surface after the immobilization. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) confirmed that OEG and MPC were successfully immobilized on titanium surface. Static platelet adhesion and APTT (activated partial thromboplastin time) experiments suggested that the anticoagulation of titanium was significantly enhanced by the immobilization of OEG and further by subsequent MPC grafting. The approach in the present study opens up a window of promising an effective and efficient method to improve the anticoagulation of blood-contact biomedical devices such as coronary stents.


Encephalomyelitis with rigidity complicating human immunodeficiency virus infection.
A 34-year-old man with human immunodeficiency virus type 1 (HIV-1) presented with axial rigidity, painful spasms, and delayed hemiparesis and dementia. Cerebrospinal fluid analysis showed no antiglutamic acid dehydrogenase antibodies but viral genome from Epstein-Barr virus was detected by polymerase chain reaction. Clinical features and possible viral aetiology of progressive encephalomyelitis with rigidity are briefly discussed.


Accuracy of centrally recorded OPCS codes for vascular surgery in the United Kingdom.
Centrally recorded OPCS codes are based upon district returns. The aim of this study is to determine the accuracy of this system with regard to vascular surgery. Prospectively recorded audit data for vascular and endovascular procedures were compared with those obtained from the Department of Health and Welsh Office. Five U.K. hospitals were involved in the study. Data were obtained for the twelve months, 1 April 1994-30 March 1995 (these being the most up to date figures available). The total number of arterial reconstructions based on audit data was 1082. Those recorded by the OPCS codes were 743. This represents a discrepancy of -31.3% (range for the five hospitals: -13.1% to -63.8%). When examining specific codes similar discrepancies were seen. For example, in one hospital 38 AAA repairs were carried out but only two were centrally recorded. However, examination of ICD9 codes (relating to hospital admissions) for that hospital showed that 38 patients with AAA were admitted. A similar wide variation was seen when examining iliac and superficial femoral artery endovascular procedures. Despite the discrepancies of audit and OPCS data, the codes for reconstructions did reflect relative workload of the different hospitals. This study shows that there is a marked underestimate of vascular workload when comparing central recorded data with that obtained from local audit. Marked variation is seen in the accuracy of data submitted from different hospitals.


Mesenchymal stromal cell spheroids in sulfated alginate enhance muscle regeneration.
The therapeutic efficacy of mesenchymal stromal cells (MSCs) for tissue regeneration is critically linked to the potency of the complex mixture of growth factors, cytokines, exosomes, and other biological cues that they secrete. The duration of cell-based approaches is limited by rapid loss of cells upon implantation, motivating the need to prolong cell viability and extend the therapeutic influence of the secretome. We and others demonstrated that the secretome is upregulated when MSCs are formed into spheroids. Although the efficacy of the MSC secretome has been characterized in the literature, no studies have reported the therapeutic benefit of in situ sequestration of the secretome within a wound site using engineered biomaterials. We previously demonstrated the capacity of sulfated alginate hydrogels to sequester components of the MSC secretome for prolonged presentation in vitro, yet the efficacy of this platform has not been evaluated in vivo. In this study, we used sulfated alginate hydrogels loaded with MSC spheroids to aid in the regeneration of a rat muscle crush injury. We hypothesized that the use of sulfated alginate to bind therapeutically relevant growth factors from the MSC spheroid secretome would enhance muscle regeneration by recruiting host cells into the tissue site. The combination of sulfated alginate and MSC spheroids resulted in decreased collagen deposition, improved myogenic marker expression, and increased neuromuscular junctions 2 weeks after injury. These data indicate that MSC spheroids delivered in sulfated alginate represent a promising approach for decreased fibrosis and increased functional regeneration of muscle. STATEMENT OF SIGNIFICANCE: The therapeutic efficacy of mesenchymal stromal cells (MSCs) for tissue regeneration is attributed to the complex diversity of the secretome. Cell-based approaches are limited by rapid cell death, motivating the need to extend the availability of the secretome. We previously demonstrated that sulfated alginate hydrogels sequester components of the MSC secretome for prolonged presentation in vitro, yet no studies have reported the in situ sequestration of the secretome. Herein, we transplanted MSC spheroids in sulfated alginate hydrogels to promote muscle regeneration. MSC spheroids in sulfated alginate decreased collagen deposition, improved myogenic marker expression, and increased neuromuscular junctions. These data indicate that MSC spheroids delivered in sulfated alginate represent a promising approach for decreasing fibrosis and increasing functional muscle regeneration.


Common mtDNA variations at C5178a and A249d/T6392C/G10310A decrease the risk of severe COVID-19 in a Han Chinese population from Central China.
Mitochondria have been shown to play vital roles during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and coronavirus disease 2019 (COVID-19) development. Currently, it is unclear whether mitochondrial DNA (mtDNA) variants, which define mtDNA haplogroups and determine oxidative phosphorylation performance and reactive oxygen species production, are associated with COVID-19 risk. A population-based case-control study was conducted to compare the distribution of mtDNA variations defining mtDNA haplogroups between healthy controls (n = 615) and COVID-19 patients (n = 536). COVID-19 patients were diagnosed based on molecular diagnostics of the viral genome by qPCR and chest X-ray or computed tomography scanning. The exclusion criteria for the healthy controls were any history of disease in the month preceding the study assessment. MtDNA variants defining mtDNA haplogroups were identified by PCR-RFLPs and HVS-I sequencing and determined based on mtDNA phylogenetic analysis using Mitomap Phylogeny. Student's t-test was used for continuous variables, and Pearson's chi-squared test or Fisher's exact test was used for categorical variables. To assess the independent effect of each mtDNA variant defining mtDNA haplogroups, multivariate logistic regression analyses were performed to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) with adjustments for possible confounding factors of age, sex, smoking and diseases (including cardiopulmonary diseases, diabetes, obesity and hypertension) as determined through clinical and radiographic examinations. Multivariate logistic regression analyses revealed that the most common investigated mtDNA variations (> 10% in the control population) at C5178a (in NADH dehydrogenase subunit 2 gene, ND2) and A249d (in the displacement loop region, D-loop)/T6392C (in cytochrome c oxidase I gene, CO1)/G10310A (in ND3) were associated with a reduced risk of severe COVID-19 (OR = 0.590, 95% CI 0.428-0.814, P = 0.001; and OR = 0.654, 95% CI 0.457-0.936, P = 0.020, respectively), while A4833G (ND2), A4715G (ND2), T3394C (ND1) and G5417A (ND2)/C16257a (D-loop)/C16261T (D-loop) were related to an increased risk of severe COVID-19 (OR = 2.336, 95% CI 1.179-4.608, P = 0.015; OR = 2.033, 95% CI 1.242-3.322, P = 0.005; OR = 3.040, 95% CI 1.522-6.061, P = 0.002; and OR = 2.890, 95% CI 1.199-6.993, P = 0.018, respectively). This is the first study to explore the association of mtDNA variants with individual's risk of developing severe COVID-19. Based on the case-control study, we concluded that the common mtDNA variants at C5178a and A249d/T6392C/G10310A might contribute to an individual's resistance to developing severe COVID-19, whereas A4833G, A4715G, T3394C and G5417A/C16257a/C16261T might increase an individual's risk of developing severe COVID-19.


Systematic review and meta-analysis of strategies for the diagnosis of suspected pulmonary embolism.
To assess the likelihood ratios of diagnostic strategies for pulmonary embolism and to determine their clinical application according to pretest probability. Medline, Embase, and Pascal Biomed and manual search for articles published from January 1990 to September 2003. Studies that evaluated diagnostic tests for confirmation or exclusion of pulmonary embolism. DATA EXTRACTED: Positive likelihood ratios for strategies that confirmed a diagnosis of pulmonary embolism and negative likelihood ratios for diagnostic strategies that excluded a diagnosis of pulmonary embolism. 48 of 1012 articles were included. Positive likelihood ratios for diagnostic tests were: high probability ventilation perfusion lung scan 18.3 (95% confidence interval 10.3 to 32.5), spiral computed tomography 24.1 (12.4 to 46.7), and ultrasonography of leg veins 16.2 (5.6 to 46.7). In patients with a moderate or high pretest probability, these findings are associated with a greater than 85% post-test probability of pulmonary embolism. Negative likelihood ratios were: normal or near normal appearance on lung scan 0.05 (0.03 to 0.10), a negative result on spiral computed tomography along with a negative result on ultrasonography 0.04 (0.03 to 0.06), and a d-dimer concentration < 500 mug/l measured by quantitative enzyme linked immunosorbent assay 0.08 (0.04 to 0.18). In patients with a low or moderate pretest probability, these findings were associated with a post-test probability of pulmonary embolism below 5%. Spiral computed tomography alone, a low probability ventilation perfusion lung scan, magnetic resonance angiography, a quantitative latex d-dimer test, and haemagglutination d-dimers had higher negative likelihood ratios and can therefore only exclude pulmonary embolism in patients with a low pretest probability. The accuracy of tests for suspected pulmonary embolism varies greatly, but it is possible to estimate the range of pretest probabilities over which each test or strategy can confirm or rule out pulmonary embolism.


Coupling of electrochemical-temperature-mechanical processes in marine clay during electro-osmotic consolidation.
Electro-osmotic consolidation has been applied in several geotechnical engineering applications that contain a series of complex processes, including electrochemical processes, temperature changes, and mechanical evolution. To explore the combination of electrochemical-temperature-mechanical processes in marine clay, electro-osmotic consolidation experiments were conducted using a self-made electro-osmotic consolidation system under various durations and voltages. The following findings was obtained: (1) the change in the pH value increased during electro-osmotic consolidation and as the voltage rise; (2) the temperature increased with a rise in voltage in the initial stage of the experiments, which was induced by Joule heating; (3) the temperature rise promoted the electro-osmotic consolidation process, which included a rise in the coefficient of consolidation and a reduction in water content; (4) horizontal shrinkage occurred when the horizontal stress increment was greater than the critical stress condition. In addition, the volume difference reached a constant value, and was proportional to the voltage rise. After the discussion, a coupling analysis was conducted, which can help to better understand the mechanism of electro-osmotic consolidation and can provide reference for engineering applications.


Feasibility of deploying a U.S. simulation-based gynecological brachytherapy educational workshop to an international setting.
A decline in brachytherapy (BT) use for cervical cancer has negatively affected cure rates in the United States and abroad. To improve trainee exposure to BT, a simulation-based educational curriculum incorporating a pelvic mannequin was developed and implemented at several U.S. residency programs. We sought to describe an initial experience with deployment of this curriculum to an international setting. The setting was in Israel, a middle eastern country with cervical cancer incidence of 5-8 cases per 100,000 women. Israel was selected for this pilot because of its desire to increase exposure to trainees, lack of mandatory BT case requirements, and few residencies nationally. In determining the feasibility of deployment to this setting, a partnership was formed between a U.S. and Israeli brachytherapist to understand cultural context and institutional and logistical needs. Feasibility was defined as successful completion of the workshop. Trainee comfort and knowledge with BT was assessed with preworkshop and postworkshop surveys, with changes compared. The curriculum was incorporated into a 1-day course on gynecologic malignancies, with adaptation to local setting and routine. Among 15 attendees, eight were residents, from four programs. All completed the workshop. All domains assessed by the surveys improved and all respondents found the program to be helpful. International deployment of the simulation-based educational BT curriculum was feasible and well-received. Further collaboration is needed to deploy and adapt the curriculum to countries of high cervical cancer incidence that could benefit from increased education.


Vitamin D and skin cancer: a problem in gene regulation.
The skin is the major source of Vitamin D(3) (cholecalciferol), and ultraviolet light (UV) is critical for its formation. Keratinocytes, the major cell in the epidermis, can further convert Vitamin D(3) to its hormonal form, 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] (calcitriol). 1,25(OH)(2)D(3) in turn stimulates the differentiation of keratinocytes, raising the hope that 1,25(OH)(2)D(3) may prevent the development of malignancies in these cells. Skin cancers (squamous cell carcinoma (SCC), basal cell carcinoma (BCC), and melanomas) are the most common cancers afflicting humans. UV exposure is linked to the incidence of these cancers-UV is thus good and bad for epidermal health. Our focus is on the mechanisms by which 1,25(OH)(2)D(3) regulates the differentiation of keratinocytes, and how this regulation breaks down in transformed cells. Skin cancers produce 1,25(OH)(2)D(3), contain ample amounts of the Vitamin D receptor (VDR), and respond to 1,25(OH)(2)D(3) with respect to induction of the 24-hydroxylase, but fail to differentiate in response to 1,25(OH)(2)D(3). Why not? The explanation may lie in the overexpression of the DRIP complex, which by interfering with the normal transition from DRIP to SRC as coactivators of the VDR during differentiation, block the induction of genes required for 1,25(OH)(2)D(3)-induced differentiation.


Measurement of methotrexate in human cerebrospinal fluid using a chemiluminescence immunoassay intended for serum and plasma matrices.
The concentration of MTX in blood is often measured quickly and easily by immunoassays. Thus, immunoassays may facilitate the easy determination of the concentration of MTX in the cerebrospinal fluid (CSF). In this study, we measured methotrexate (MTX) concentrations in the CSF using a high-performance liquid chromatography (HPLC) method intended for analyzing CSF matrices and a chemiluminescence immunoassay (CLIA) method intended for assessing serum and plasma matrices and verified the differences in the results of the two methods. HPLC analysis for MTX in the CSF was performed using a Prominence UFLC system with a C18 column. The HPLC method was validated in accordance with the 2018 FDA guideline. The CLIA method was performed using an ARCHITECT i1000SR system intended for serum and plasma matrices. A total of 47 CSF samples (14 clinical and 33 spiked specimens) were analyzed using the two methods. The HPLC method passed the validation criteria. The concentration of MTX in the same sample, determined using the HPLC and CLIA methods, differed proportionally; the percent difference in the concentrations averaged -23.0% (95% confidence interval: -36.9% to -9.1%) as revealed by the Bland-Altman plot. The relationship between the measured values, evaluated using the Passing-Bablok regression, was as follows: HPLC = 1.205 × CLIA - 0.024. The equation deduced in this study can be used to correct the concentration of MTX measured using the CLIA method.


Comparison of cumulative low back loads of caregivers when transferring patients using overhead and floor mechanical lifting devices.
Mechanical lifting devices are recommended as an important intervention for reducing lifting injuries among nursing personnel; previous research suggests that spinal loads are not minimized for all device types. The purpose of this study was to describe the spinal loading pattern while performing a bed to chair transfer comparing Overhead and Floor powered lifting devices. A Latin Square design was employed to evaluate five lifting devices while performing a heavy patient transfer. The primary outcomes were spinal compression, and anterior shear (across median and cumulative loading conditions), and ratings of perceived exertion. An inverse dynamic approach was used to calculate the net joint forces and moments about the L5/S1 spinal level. The transfer was partitioned into seven distinct phases for biomechanical analysis. The proportion of time spent and the mean loads sustained in each phase of the transfer were described. Significant differences in loads were observed between the differing lifting devices, particularly during the transport phases for the Overhead devices. Nurse subjects consistently ranked Overhead lifting devices as most preferred. A large proportion of the time to complete the transfer and cumulative loads occurred during phases that involved only the sling and not the mechanical component of the device. Overhead lifting devices were shown to have lower spinal loads during the transport phases. The results of this study have implications for the use and selection of mechanical lifting devices as part of a strategy to reduce back injuries.


Prediction of postoperative pulmonary complications in oesophagogastric cancer surgery.
In a series of 102 patients with cancer of the oesophagus or gastric cardia in whom surgery had been performed, postoperative respiratory infection, respiratory failure and mortality secondary to respiratory complication occurred in 55.8, 34.3 and 21.6 per cent respectively. The incidence of complications correlated well with the pre-operative peak expiratory flow rate when it was less than 65 per cent of the predicted normal value. However, the predictive ability of peak expiratory flow rate was much less than the patient's age, pre-operative serum albumin level, pre-operative Pa,O2 and number of surgical wounds when all risk factors were considered. Significantly more complications occurred when the patient was greater than 65 years old, Pa,O2 was less than 10 kPa, serum albumin was less than 39 g/l and when the operation required three incisions. These findings may serve as guidelines for identification of high-risk patients in the future.


Subcutaneous delivery of daratumumab in Japanese patients with relapsed/refractory multiple myeloma.
Subcutaneous daratumumab (DARA SC; daratumumab co-formulated with recombinant human hyaluronidase PH20) is administered in ~ 5 min and demonstrates safety and efficacy comparable to intravenous daratumumab, with low infusion-related reaction (IRR) rates in global populations. This open-label, multicenter, phase 1 study is the first evaluation of DARA SC in Japanese patients. Eligible patients had relapsed/refractory multiple myeloma (RRMM; ≥ 2 prior lines of therapy including a proteasome inhibitor and immunomodulatory drug). Patients (N = 6) received DARA SC 1,800 mg until progression (weekly for Cycles 1-2; every 2 weeks for Cycles 3-7; monthly for Cycles 7 + [28-day cycles]). The primary objective was to evaluate safety. Secondary objectives included efficacy and pharmacokinetics. Median time of administration was 3-4 min for all injections. No dose-limiting toxicity occurred, and no treatment-emergent adverse events were serious or led to discontinuation. No IRRs were observed; 4 (67%) patients had injection-site reactions (all grade 1). Overall response rate was 67%. Pharmacokinetics of DARA SC in Japanese patients were similar to findings from the global phase 1b PAVO study (NCT02519452). DARA SC at a flat dose of 1,800 mg was well tolerated in Japanese RRMM patients with comparable efficacy and pharmacokinetics to intravenous daratumumab. ClinicalTrials.gov identifier NCT03242889.


Classes of depression, anxiety, and functioning in acute coronary syndrome patients.
To describe change in subgroups characterized by patterns of depression, anxiety, and functional impairment; examine treatment effects on subgroup membership; examine effects of sex and age on subgroup membership. Latent class models were used to meet the first 2 objectives using 79 patients with depression/anxiety. Generalized estimating equations were used to meet the third objective. Three subgroups characterized by different combinations of psychiatric disorders and functioning were identified. Patients who received treatment were more likely to transition to a less impaired subgroup. Unique information about holistic treatment effects can be gained when multiple outcomes are considered simultaneously.


Trauma and multiple sclerosis. An hypothesis.
An obligatory event in the pathogenesis of the multiple sclerosis plaque appears to be an increase in the permeability of the blood-brain barrier. Neuropathological observations of the brain of persons suffering from concussion after relatively minor head injury, as well as of animals subjected to experimental brain injury, have shown that alterations of the blood-brain barrier constitute a common result of such trauma. It is postulated that the alterations of the blood-brain barrier secondary to trauma of the brain or spinal cord of patients with already established multiple sclerosis may result in an exacerbation or recurrence of a previously symptomatic plaque, in the appearance of symptoms from a silent lesion, or in the formation of a new plaque in such an area of selected vulnerability. In other persons injury to the nervous system may cause the development of multiple sclerosis plaques in the previously damaged areas when the disease has its onset after the trauma. There is no evidence to support the idea that trauma ever causes multiple sclerosis.


High efficacy of two artemisinin-based combinations (artemether-lumefantrine and artesunate plus amodiaquine) for acute uncomplicated malaria in Ibadan, Nigeria.
To test the hypothesis that artesunate plus amodiaquine (ASAQ) is as effective as artemether-lumefantrine (AL) in the treatment of acute uncomplicated malaria in Nigerian children. In an open label, randomized controlled clinical trial, children aged 6 months to 10 years were randomized to receive artesunate (4 mg/kg daily) plus amodiaquine (10 mg/kg daily) or AL (5-14 kg, one tablet; 15-24 kg, two tablets and 25-34 kg, three tablets twice daily). Both drug regimens were given for 3 days and follow-up was for 28 days. A total of 132 children (66 in each group) were randomized to receive either ASAQ or AL. Day 28 cure rates in the per protocol (PP) population were 93% for ASAQ and 95% for AL (OR = 0.71, 95% CI = 0.12-3.99, rho = 0.66). Using Kaplan-Meier product-limit estimates of failure, the median survival time for ASAQ was 21 days and for AL 28 days (P = 0.294). PCR corrected day 28 cure rate for PP populations were 98.4% for ASAQ and 100% for AL. Both drugs were well-tolerated. ASAQ is as effective as AL and both combinations were efficacious and safe.


Long-lasting enhancement in the intrinsic excitability of deep dorsal horn neurons.
Intrinsic excitability (IE) can be defined as an output of action potentials from a given input signal. Changes to the IE of a neuron are an important aspect of the cellular plasticity that underlies learning and memory process. In this study, long-term plastic change in IE of deep dorsal horn neurons (DHNs) was investigated. Associative spike pairing stimulation (PS) induced a long-lasting increase in IE. Buffering intracellular calcium with BAPTA (10mM) prevented the induction of a long-lasting increase in IE. PS failed to induce a long-lasting increase in IE in the presence of either D-APV (50 microM) or cadmium chloride (100 microM). Apamin (100 nM) partially blocked the induction of a long-lasting increase in IE. This intrinsic plasticity requires a rise in postsynaptic Ca(2+) and NMDA receptor activation during the induction period, and this process might be mediated by the down-regulation of small-conductance calcium-dependent potassium (SK) channels. In deep DHNs, PS induced excitatory postsynaptic potential (EPSP)-spike (E-S) potentiation, which increases the firing probability and the number of spikes, by consistent dorsal rootlet stimulation. Under bath application of bicuculline (10 microM) and strychnine (1 microM), PS induced E-S potentiation and long-lasting increases in IE. These results suggest that an increase in IE might underlie E-S potentiation, while a reduction in inhibitory transmission does not contribute to E-S potentiation and long-lasting increases in IE. We conclude that PS enhances the IE of deep DHNs, which may play an important role in spinal processing of nociceptive information.


Isolation and characterization of growth hormone from Atlantic cod (Gadus morhua).
Growth hormone was purified from cod pituitary extract by a simple two-step procedure involving gel filtration and reversed-phase high-performance liquid chromatography (rpHPLC). At each stage of purification, fractions were monitored by rpHPLC, SDS-polyacrylamide gel electrophoresis, and immunoblotting using anti-chum salmon growth hormone (GH) antiserum. The yield of purified hormone was 1.3 mg/g pituitary. Cod GH was found to exist in two monomeric forms (Mr = 20K and 22K) and dimeric forms (Mr = 40K and 42K). The two monomeric forms have a pI of 5.8, an identical amino acid composition, histidine as the N-terminal residue, and an identical lysyl endopeptidase peptide map. Staining with concanavalin A was observed on the 20K component only, but analysis for total reducing sugar did not confirm these results. Cod GH was found to be a potent stimulator of growth in juvenile rainbow trout which received intraperitoneal injections of the hormone. The partial amino acid sequence has been determined.


Extracellular Vesicles and Their Roles in Cancer Progression.
Extracellular vesicles (EVs) produced by cancer cells function as a unique form of intercellular communication that can promote cell growth and survival, help shape the tumor microenvironment, and increase invasive and metastatic activity. There are two major classes of EVs, microvesicles (MVs) and exosomes, and they differ in how they are formed. MVs are generated by the outward budding and fission of the plasma membrane. On the other hand, exosomes are derived as multivesicular bodies (MVBs) fuse with the plasma membrane and release their contents. What makes EVs especially interesting is how they mediate their effects. Both MVs and exosomes have been shown to contain a wide-variety of bioactive cargo, including cell surface, cytosolic, and nuclear proteins, as well as RNA transcripts, micro-RNAs (miRNAs), and even fragments of DNA. EVs, and their associated cargo, can be transferred to other cancer cells, as well as to normal cell types, causing the recipient cells to undergo phenotypic changes that promote different aspects of cancer progression. These findings, combined with those demonstrating that the amounts and contents of EVs produced by cancer cells can vary depending on their cell of origin, stage of development, or response to therapies, have raised the exciting possibility that EVs can be used for diagnostic purposes. Moreover, the pharmaceutical community is aggressively pursuing the use of EVs as a potential drug delivery platform. Here, in this chapter, we will highlight what is currently known about how EVs are generated, how they impact cancer progression, and the different ways they are being exploited for clinical applications.


Fosfomycin modifies the replication kinetics of bovine alphaherpesvirus-1 and reduces the timing of its protein expression on bovine (MDBK) and human (SH-SY5Y) cell lines.
Bovine alphaherpesvirus 1 (BoAHV-1) predisposes cattle to respiratory secondary bacterial infections, which can be treated with the broad-spectrum antibiotic fosfomycin. This drug also suppresses NF-kB activity and pro-inflammatory responses. Therefore, cattle may be exposed to an interaction between the virus and the antibiotic which may have effects on it. The aim of this study was to determine the effect of calcium fosfomycin (580 µg/mL) on BoAHV-1 (moi = 0.1) replication. Two cell lines (MDBK and SH-SY5Y) were used in this study. Our results show that fosfomycin has novel properties. By MTT assay we have shown that it is non-cytotoxic for any of the cell lines. Extracellular and intracellular viral titers demonstrated that fosfomycin has a cell-type and time-dependent effect on BoAHV-1 replication. By direct immunofluorescence it was shown that it reduces the timing of BoAHV-1 protein expression, and by qPCR, we found that its effect on NF-kB mRNA expression depends on the cell type.


A qualitative analysis of the barriers and enablers faced by Australian rural general practitioners in the non-pharmacological management of congestive heart failure in community dwelling patients.
Congestive heart failure (CHF) is a significant health problem in Australia, and disproportionately affects rural Australians. Management of CHF in Australia is heavily centred around the general practitioner (GP). Australian and international literature indicates there is a gap between current and best practice in CHF management. There is little known about the non-pharmacological aspects of management, or CHF management in a rural Australian context. This study aimed to identify what Australian GPs practicing in the Northern Rivers Region of New South Wales, Australia, perceived were the barriers and enablers in the non-pharmacological management of CHF amongst community dwelling patients, to inform healthcare access, resourcing and delivery in Australian rural environments. Qualitative study involving a realist thematic analysis of data collected from semi-structured face-to-face interviews. Fifteen GPs and GP trainees participated. Four interlinked key themes underpinning GPs' experiences with non-pharmacological management of CHF were interpreted from the interview data: (1) resources, (2) complexity of heart failure, (3) relationships, and (4) patient demographics, priorities and views affect how patients engage with non-pharmacological management of CHF. Rural Australian GPs face considerable barriers to non-pharmacological management of CHF. The data suggests that increased rural Australian health services and community transportation, multidisciplinary management, and stronger professional networks have the potential to be invaluable enablers of CHF management. Further research exploring non-pharmacological management of CHF in other rural contexts may provide additional insights to better inform rural healthcare access and resourcing.


[Argon laser iridectomy and gonioplasty in the treatment of angle-closure glaucoma].
Treatment of the narrow angle in angle-closure glaucoma depends on iridectomy and mechanical opening of the angle if an argon laser is used for photocoagulation. The effect of the heat is to shrink the iris tissue (gonioplasty) and to free the angle. Maximum power and safety of the laser beam are achieved by utilizing convex contact lenses on the cornea.


Low intensity vibration mitigates tumor progression and protects bone quantity and quality in a murine model of myeloma.
Myeloma facilitates destruction of bone and marrow. Since physical activity encourages musculoskeletal preservation we evaluated whether low-intensity vibration (LIV), a means to deliver mechanical signals, could protect bone and marrow during myeloma progression. Immunocompromised-mice (n=25) were injected with human-myeloma cells, while 8 (AC) were saline-injected. Myeloma-injected mice (LIV; n=13) were subjected to daily-mechanical loading (15min/d; 0.3g @ 90Hz) while 12 (MM) were sham-handled. At 8w, femurs had 86% less trabecular bone volume fraction (BV/TV) in MM than in AC, yet only a 21% decrease in LIV was observed in comparison to AC, reflecting a 76% increase versus MM. Cortical BV was 21% and 15% lower in MM and LIV, respectively, than in AC; LIV showing 30% improvement over MM. Similar outcomes were observed in the axial skeleton, showing a 35% loss in MM with a 27% improved retention of bone in the L5 of LIV-treated mice as compared to MM. Transcortical-perforations in the femur from myeloma-induced osteolysis were 9× higher in MM versus AC, reduced by 57% in LIV. Serum-TRACP5b, 61% greater in MM versus AC, rose by 33% in LIV compared to AC, a 45% reduction in activity when compared to MM. Histomorphometric analyses of femoral trabecular bone demonstrated a 70% elevation in eroded surfaces of MM versus AC, while measures in LIV were 58% below those in MM. 72% of marrow in the femur of MM mice contained tumor, contrasted by a 31% lower burden in LIV. MM mice (42%) presented advanced-stage necrosis of tibial marrow while present in just 8% of LIV. Myeloma infiltration inversely correlated to measures of bone quality, while LIV slowed the systemic, myeloma-associated decline in bone quality and inhibited tumor progression through the hindlimbs.


The exosomal secretomes of mesenchymal stem cells extracted via 3D-printed lithium-doped calcium silicate scaffolds promote osteochondral regeneration.
The development of surface modification techniques has brought about a major paradigm shift in the clinical applications of bone tissue regeneration. Biofabrication strategies enable the creation of scaffolds with specific microstructural environments and biological components. Lithium (Li) has been reported to exhibit anti-inflammatory, osteogenic, and chondrogenic properties by promoting several intracellular signaling pathways. Currently, research focuses on fabricating scaffolds with simultaneous dual bioactivities to enhance osteochondral regeneration. In this study, we modified the surface of calcium silicate (CS) scaffolds with Li using a simple immersion technique and evaluated their capabilities for bone regeneration. The results showed that Li ions could be easily coated onto the surfaces of CS scaffolds without affecting the microstructural properties of CS itself. Furthermore, the modifications did not affect the printing capabilities of the CS, and porous scaffolds could be fabricated via extrusion. Moreover, the presence of Li improved the surface roughness and hydrophilicity, thus leading to enhanced secretion of osteochondral-related regeneration factors, such as alkaline phosphatase (ALP), bone sialoprotein (BSP), and collagen II (Col II) proteins. Subsequent in vivo studies, including histological and micro-CT analyses, confirmed that the Li-modified CS scaffolds promoted osteochondral regeneration. The transcriptome analysis suggested that the enhanced osteochondrogenic capabilities of our scaffolds were influenced by paracrine exosomes. We hope this study will inspire further research on osteochondral regeneration.


Undertreatment of nausea and other symptoms in hospitalized cancer patients.
This survey aimed to provide a symptom profile, in particular for nausea and its treatment, of advanced cancer patients prior to implementation of a clinical practice guideline for nausea. An audit of 82 advanced cancer patients admitted to a major Australian teaching hospital was undertaken. While nausea was present in 26 (32%) of respondents, antiemetics were prescribed in 52 (68%), a higher proportion than in the past, and being taken by 32 (39%). Patients reporting moderate-to-severe nausea often missed out on treatment. These findings underscore the need for more attention to education of hospital staff in the assessment and treatment of this frequently bothersome symptom, and raise issues for study design of symptom control research.


GLUT 4 and insulin receptor binding and kinase activity in trained human muscle.
1. Physical training enhances sensitivity and responsiveness of insulin-mediated glucose uptake in human muscle. This study examines if this effect of physical training is due to increased insulin receptor function or increased total concentration of insulin-recruitable glucose transporter protein (GLUT 4). 2. Seven healthy young subjects carried out single leg bicycle training for 10 weeks at 70% of one leg maximal oxygen uptake (VO2,max). Subsequently biopsies were taken from the vastus lateralis muscle of both legs. 3. Single leg VO2,max increased for the trained leg (46 +/- 3 to 52 +/- 2 ml min-1 kg-1 (means +/- S.E.M., P < 0.05), and cytochrome c oxidase activity was higher in this compared to the untrained leg (2.0 +/- 0.1 vs. 1.4 +/- 0.1 nmol s-1 (mg muscle)-1, P < 0.05). Insulin binding as well as basal- and insulin-stimulated receptor kinase activity did not differ between trained and untrained muscle. The concentration of GLUT 4 protein was higher in the former (14.9 +/- 1.9 vs. 11.6 +/- 1.0 arbitrary units (micrograms protein)-1 in crude membranes, P < 0.05). The training-induced increase in GLUT 4 (26 +/- 11%) matched a previously reported increase in maximum insulin-stimulated leg glucose uptake (25 +/- 7%) in the same subjects, and individual values of the two variables correlated (correlation coefficient (r) = 0.84, P < 0.05). 4. In conclusion, in human muscle training induces a local contraction-dependent increase in GLUT 4 protein, which enhances the effect of insulin on glucose uptake. On the other hand, insulin receptor function in muscle is unlikely to be affected by training.


Genome wide classification and characterisation of CpG sites in cancer and normal cells.
This study identifies common methylation patterns across different cancer types in an effort to identify common molecular events in diverse types of cancer cells and provides evidence for the sequence surrounding a CpG to influence its susceptibility to aberrant methylation. CpG sites throughout the genome were divided into four classes: sites that either become hypo or hyper-methylated in a variety cancers using all the freely available microarray data (HypoCancer and HyperCancer classes) and those found in a constant hypo (Never methylated class) or hyper-methylated (Always methylated class) state in both normal and cancer cells. Our data shows that most CpG sites included in the HumanMethylation450K microarray remain unmethylated in normal and cancerous cells; however, certain sites in all the cancers investigated become specifically modified. More detailed analysis of the sites revealed that majority of those in the never methylated class were in CpG islands whereas those in the HyperCancer class were mostly associated with miRNA coding regions. The sites in the Hypermethylated class are associated with genes involved in initiating or maintaining the cancerous state, being enriched for processes involved in apoptosis, and with transcription factors predicted to bind to these genes linked to apoptosis and tumourgenesis (notably including E2F). Further we show that more LINE elements are associated with the HypoCancer class and more Alu repeats are associated with the HyperCancer class. Motifs that classify the classes were identified to distinguish them based on the surrounding DNA sequence alone, and for the identification of DNA sequences that could render sites more prone to aberrant methylation in cancer cells. This provides evidence that the sequence surrounding a CpG site has an influence on whether a site is hypo or hyper methylated.


Dietary fish oil inhibits cholesterol monohydrate crystal nucleation and gallstone formation in the prairie dog.
Epidemiologic studies suggest that populations consuming a diet rich in fish oil have lower rates of both atherosclerotic heart disease and gallstones. The mechanisms underlying this inhibitory effect on cholesterol gallstone formation remain unclear. We therefore studied the effect of dietary fish oil on bile composition and cholesterol precipitation in an animal model of gallstone disease. Adult male prairie dogs were fed a standard control diet (n = 12) or a lithogenic 1.2% cholesterol diet (n = 16). One half of the animals in each group had their diet supplemented with concentrated fish oil. After 14 days animals receiving the cholesterol diet all developed biliary cholesterol monohydrate crystals and gallstones. When fish oil was added to this high cholesterol diet, solid cholesterol crystal precipitation and gallstone formation were completely inhibited. This inhibition of gallstone formation was accompanied by a significant decrease in biliary calcium and total protein concentration. Microscopic cholesterol liquid crystals were evident in the bile of all of the animals fed the cholesterol plus fish oil diet. Dietary fish oil also significantly prolonged cholesterol monohydrate crystal observation time in animals receiving the lithogenic diet. These data suggest that dietary fish oil exerts a potent antilithogenic effect on cholesterol gallstone disease and may induce a stable liquid crystalline phase retarding nucleation.


Effective and safe immunizations with live-attenuated vaccines for children after living donor liver transplantation.
Immunizations using live-attenuated vaccines are not recommended for post-liver transplant children due to its theoretical risks. However, they will encounter vaccine-preventable viral diseases upon returning to real-life situations. We performed a total of 70 immunizations with four individual live-attenuated vaccines to 18 pediatric post-living donor liver transplant (LDLT) recipients who fulfilled a clinical criteria including humoral and cell-mediated immunity. The seroconversion rates at the first dose for measles (strain AIK-C), rubella (strain TO-336), varicella (strain Oka), and mumps (strains Hoshino) were 100% (15/15), 100% (15/15), 82% (9/11), and 82% (9/11), respectively. During observed period (-5 years 11 months), a few cases with waning immunity (antibodies were once produced but the levels fell over time) were seen except after rubella immunization. Clinical diseases after seroconversion or definite serious adverse effects due to immunization were not observed. Immunizations using selected live-attenuated vaccines were safe and effective for post-LDLT children who were not severely immunosuppressed.


A Latent Class Analysis of HIV Risk Factors Among Men and Women with Opioid Use Disorder in Pre-trial Detention.
Adults entering pre-trial detention who inject drugs are at high risk for acquiring HIV/AIDS. In the current study, we examined pre-incarceration HIV risk behaviors among 114 people with opioid use disorder who inject drugs. Participants were recruited from the Baltimore City Detention Center as part of a randomized controlled trial of pre-release methadone treatment. Using latent class analysis, we found three separate latent classes, which we identified as the sex exchange class (14.2%), drug equipment sharing class (36.8%) and lower risk class (49.0%). Women in the sex exchange class (n = 16) reported having multiple male partners and selling sex for money or drugs; however, this group also reported more consistent condom use and less frequent injection drug and equipment sharing than participants in the drug equipment sharing class. Our findings highlight distinct profiles of jail detainees with OUD based on their risks for HIV, and could inform more targeted interventions for each group.Clinical Trials Registration: Clinicaltrials.gov NCT02334215.


Processing of yeast exoglucanase (beta-glucosidase) in a KEX2-dependent manner.
We have detected proteolytic processing of a form of exoglucanase representative of the endoplasmic reticulum (form A). This processing did not take place when form A was obtained from protoplasts lysed in the presence of either EDTA or leupeptin, two wel-characterized inhibitors of KEX2 endoprotease from Saccharomyces cerevisiae. Sequencing of the amino terminus of an A-like form of enzyme secreted by a kex2 mutant indicated the presence of 4 amino acids, with a pair of basic residues (Lys-Arg) at their carboxyl side, preceding the amino terminus of the wild-type external exoglucanase.


Silk fibroin and silk-based biomaterial derivatives for ideal wound dressings.
Silk fibroin (SF) is derived from Bombyx mori silkworm cocoons and has been used in textiles and as a suture material for decades. More recently, SF has been used for various new biomedical applications, including as a wound dressing, owing to its excellent biological and mechanical properties. Specifically, the mechanical stiffness, versatility, biocompatibility, biodegradability, water vapour permeability and slight bactericidal properties make SF an excellent candidate biomaterial for wound dressing applications. The effectiveness of SF as a wound dressing has been tested and well-documented in vitro as well as in-vivo, as described here. Dressings based on SF are currently used for treating a wide variety of chronic and acute (e.g. burn) wounds. SF and its derivatives prepared as biomaterials are available as sponges, hydrogels, nanofibrous matrices, scaffolds, micro/nanoparticles, and films. The present review discusses the potential role of SF in wound dressing and its modulation for wound dressing applications. The comparison of SF based dressings with other natural polymers understands the readers, the scope and limitation of the subject in-depth.


Long-Term Impact of Daily E-cigarette Exposure on the Lungs of Asthmatic Mice.
Although the greater popularity of electronic cigarettes (EC) among asthmatics is alarming, there is limited knowledge of the long-term consequences of EC exposure in asthmatics. Mild asthmatic C57/BL6J adult male and female mice were established by intranasal insufflation with three combined allergens. The asthmatic and age and sex-matched' naïve mice were exposed to air, nicotine-free (propylene glycol [PG]/vegetable glycerin [VG]-only), or PG/VG+Nicotine, 4 hours daily for 3 months. The effects of EC exposure were accessed by measuring cytokines in bronchoalveolar lavage, periodic acid-schiff (PAS) staining, mitochondrial DNA copy numbers (mtCN), and the transcriptome in the lung. Significance was false discovery rate <0.2 for transcriptome and 0.05 for the others. In asthmatic mice, PG/VG+Nicotine increased PAS-positive cells and IL-13 compared to mice exposed to air and PG/VG-only. In naïve mice exposed to PG/VG+Nicotine and PG/VG-only, higher INF-γ was observed compared to mice exposed only to air. PG/VG-only and PG/VG+Nicotine had significantly higher mtCN compared to air exposure in asthmatic mice, while the opposite pattern was observed in non-asthmatic naïve mice. Different gene expression patterns were profoundly found for asthmatic mice exposed to PG/VG+Nicotine compared to PG/VG-only, including genes involved in mitochondrial dysfunction, oxidative phosphorylation, and p21-activated kinase (PAK) signaling. This study provides experimental evidence of the potential impact of nicotine enhancement on the long-term effects of EC in asthmatics compared to non-asthmatics. The findings from this study indicate the potential impact of EC in asthmatics by addressing multiple biological markers. The long-term health outcomes of EC in the susceptible group can be instrumental in supporting policymaking and educational campaigns and informing the public, healthcare providers, and EC users about the underlying risks of EC use.


Ageing and hepatic steatosis exacerbate ischemia/reperfusion-accelerated outgrowth of colorectal micrometastases.
Ischemia/reperfusion (I/R) injury is frequently encountered during hepatic surgery. We recently showed that I/R accelerates the outgrowth of pre-established colorectal micrometastases. The aim of this study was to assess the influence of ischemia time, gender, age, and liver steatosis on the accelerated outgrowth of colorectal metastases following I/R. Five days after tumor cell inoculation, mice were subjected to 20, 30 or 45 min of left lobar I/R. To assess the influence of age, gender, and liver steatosis on I/R-accelerated tumor growth, we compared old with young mice, male with female mice, and mice with healthy livers with mice with steatotic livers. Endpoints were extent of tissue necrosis and tumor growth. With increasing ischemia times, tissue necrosis and I/R-accelerated tumor growth increased, with a significant stimulatory effect at 30 and 45 min of ischemia. I/R-stimulated outgrowth of micrometastases was further increased by 33% in aged mice and by 42% in steatotic livers and was associated with increased tissue necrosis. In female mice tissue necrosis had decreased by 47% and tumor growth was reduced in both control and clamped liver lobes. The stimulatory effect of I/R on metastasis outgrowth was similar in male and female mice. I/R-accelerated outgrowth of colorectal micrometastases largely depends on the duration of the ischemic period, with a safe upper limit of 20 min in mice. The stimulatory effects of I/R on tumor growth are exacerbated in aged mice and in steatotic livers.


[Passive cocaine inhalation by a 25-month-old child].
Cocaine and its principal metabolite were isolated from the urine, confirmed by urine chromatographic analysis, in a 25-month-old girl who had unsteady gait and a third fever seizure. She was premature, had intrauterine growth restriction (IUGR), and a psychomotor delay. The hair examination confirmed prolonged cocaine exposure. The concentration was significant and showed that she had been intoxicated for the 10 previous months. Passive cocaine inhalation may have caused and contributed to these symptoms.


Transition-Metal-, Additive-, and Solvent-Free [3 + 3] Annulation of RCF2-Imidoyl Sulfoxonium Ylides with Cyclopropenones to Give Multifunctionalized CF3-Pyridones.
An efficient and practical strategy was developed to synthesize 1,3,4-triaryl-6-trifluoromethylpyridones from CF3-imidoyl sulfoxonium ylides and cyclopropenones in good to excellent yields. This stepwise [3 + 3] annulation reaction was carried out under transition-metal-, additive-, and solvent-free conditions, generating 1 equiv of dimethyl sulfoxide as byproduct and tolerating a series of functional groups.


General Relativity and the AKSZ Construction.
In this note the AKSZ construction is applied to the BFV description of the reduced phase space of the Einstein-Hilbert and of the Palatini-Cartan theories in every space-time dimension greater than two. In the former case one obtains a BV theory for the first-order formulation of Einstein-Hilbert theory, in the latter a BV theory for Palatini-Cartan theory with a partial implementation of the torsion-free condition already on the space of fields. All theories described here are BV versions of the same classical system on cylinders. The AKSZ implementations we present have the advantage of yielding a compatible BV-BFV description, which is the required starting point for a quantization in presence of a boundary.


Activation of percarbonate by water treatment sludge-derived biochar for the remediation of PAH-contaminated sediments.
Sludge from a groundwater treatment plant was used to prepare biochar by pyrolysis. The Fe-Mn rich biochar was used to activate percarbonate for the remediation of polycyclic aromatic hydrocarbons (PAHs) contaminated aquatic sediments. Results showed that the sludge-derived biochar (SBC) produced at a pyrolysis temperature of 700 °C was the most effective in activating percarbonate, which exhibited significant oxidative removal of PAHs. PAHs degradation took place via a Fenton-like oxidation manners, contributed from the Fe3+/Fe2+ and Mn3+/Mn2+ redox pairs, and achieved the highest degradation efficiency of 87% at pH0 6.0. Reactions between oxygenated functional groups of biochar and H2O2 generated of O2•- and HO• radicals in abundance under neutral and alkaline pH was responsible for the catalytic degradation of PAHs. Our results provided new insights into the environmental applications of SBC for the green sustainable remediation of organics-contaminated sediments and aided in reduction of associated environmental and health risk.


Free-standing hybrid film for separation of dye pollutant with self-cleaning ability under visible light.
The development of low cost and environmental-friendly materials has long been an ambition for effective removal of dye pollutants in complex water environments. In this study, a free-standing separation film of bacterial cellulose reinforced/functionalized by graphitic phase carbon nitride is developed by a facile suction filtration strategy, of which the former is precoated by polypyrrole, and the latter is pre-doped by oxygen to endow the as-obtained film an enhanced photocatalytic performance and self-cleaning ability. The as-obtained film exhibits a high tensile stress of 51.8 ± 1.1 MPa, and a high resistance to cold, heat, acid and alkali. For typical dyes of methylene blue and rhodamine B, a high dye rejection rate of 99.9% at 138 L/m2•h•bar feed flux is obtained by the as-obtained film. Even at a salt concentration higher than 5%, it still maintained high dye removal rates and achieves effective separation of dye and salt. Simultaneously, a high dye photocatalytic degradation of the composite films rates up to 98% in only 90 min, and a high self-cleaning ability demonstrated by recovery of flux after light treatment in cyclic tests. The density functional theory calculation validates the beneficial effects of improved light response range and separated photogenerated electron/holes for the effective degradation of dyes by oxygen-doped carbon nitride coupled with one-dimensional polypyrrole chains. Overall, this study proposes a new direction for the separation of dye pollutants with a high visible-light self-cleaning capacity by structural tailoring of bacterial cellulose with carbon nitride.


Incidence of delayed ipsilateral intraparenchymal hemorrhage after stent-assisted coiling of intracranial aneurysms in a high-volume single center.
Delayed ipsilateral intraparenchymal hemorrhage (IPH) has been reported following technically successful treatment of intracranial aneurysms using flow-diverting stents in up to 8.5% of patients. We report a similar, though less frequent phenomenon in the setting of stent-assisted coil embolization. Institutional review board approval was obtained. A retrospective analysis of a prospective neurointerventional procedure registry was performed to review all IPHs that occurred in aneurysm patients within 90 days of endovascular treatment performed between November 2002 and November 2014 at one institution. Age, sex, hypertension, dual antiplatelet therapy, and technical details of the procedure were recorded. A total of 1697 patients underwent endovascular treatment of an intracranial aneurysm without a flow diverter at our institution during the study period. Among these, 138 patients underwent stent-assisted coiling (8.1%). Of these, three patients (2.2%) suffered a delayed IPH within the vascular territory distal to the treated lesion (one woman, median age 60 years). Recently described in the setting of flow diversion, delayed ipsilateral IPH is not limited to flow-diverting stents. Though less frequent, a potential for this complication may exist following any intracranial stenting procedure, possibly related to hemorrhagic conversion of microembolic phenomena in the setting of dual antiplatelet or anticoagulation therapy.


Cystic dysplasia of the testis: sonographic diagnosis and differentiation from testicular microlithiasis.
Cystic dysplasia of the testis is an uncommon benign cause of an intratesticular mass in children. It is often associated with renal abnormalities, which reflects the close developmental association between the kidneys and testes. The diagnosis is usually suspected based on sonographic findings. We present a case of cystic dysplasia of the testis occurring in a boy with a multicystic dysplastic kidney, in whom the sonographic findings were initially misinterpreted as testicular microlithiasis.


Manufacturing of monolithic superelastic rods with variable properties for spinal correction: feasibility study.
A new concept of monolithic spinal rod with variable flexural stiffness is proposed to reduce the risk of adjacent segment degeneration and fracture associated with rigid spinal fixation techniques while providing adequate stability to the spine. The concept is based on the use of Ti-Ni shape memory alloy rods subjected to different processing schedules implying local annealing, cold work, or a combination of both. A feasibility study of the concurrent technological routes is performed by comparing their potential to locally control material microstructure and properties.


The future of obesity: new drugs versus lifestyle interventions.
Obesity causes serious medical complications and impairs quality of life. However, its management remains challenging. To assist health professionals who counsel patients who are overweight or obese by discussing the possible add-on value of new drugs over lifestyle interventions. A critical analysis is made of the available evidence of the long-term efficacy of diet and exercise and/or anti-obesity agents such as orlistat, sibutramine and rimonabant. Lifestyle interventions remain the cornerstone of the treatment of obesity, but adherence is poor and long-term success is modest. Pharmacological agents may be useful adjuncts for improving weight loss and maintenance, and health outcomes, and should be continued in good responders. Drug therapy and lifestyle intervention are not opponent strategies, but should probably be combined to tackle obesity.


Anomia in cerebral diseases.
Naming performance was studied in demented patients, age-matched controls, left-hemisphere stroke patients with aphasia and right-hemisphere stroke patients. The experimental naming test compared four levels of perceptual difficulty and two language variables: word frequency and word length. Naming accuracy and error types were compared among subject groups. Perceptual difficulty influenced naming in the demented and right-hemisphere stroke patients, but not in aphasics and controls. Visual errors, likewise, characterized the former two groups. Semantically-related errors and circumlocutions characterized the naming of aphasic and demented patients, while phonemic errors were common only in aphasics. The results suggested differing patterns of anomia in different patient groups.


Studies on aspartase. II. Role of sulfhydryl groups in aspartase from Escherichia coli.
Aspartase (L-aspartate ammonia-lyase, EC 220.127.116.11) of Escherichia coli W contains 38 half-cystine residues per tetrameric enzyme molecule. Two sulfhydryl groups were modified with N-ethylmaleimide or 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) per subunit, while 8.3 sulfhydryl groups were titrated with p-mercuribenzoic acid. In the presence of 4 M guanidine - HCl, 8.6 sulfhydryl groups reacted with DTNB per subunit. Aspartase was inactivated by various sulfhydryl reagents following pseudo-first-order kinetics. Upon modification of one sulfhydryl group per subunit with N-Ethylmaleimide, 85% of the original activity was lost; a complete inactivation was attained concomitant with the modification of two sulfhydryl groups. These results indicate that one or two sulfhydryl groups are essential for enzyme activity. L-Aspartate and DL-erythro-beta-hydroxyaspartate markedly protected the enzyme against N-ethylmaleimide-inactivation. Only the compounds having an amino group at the alpha-position exhibited protection, indicating that the amino group of the substrate contributes to the protection of sulfhydryl groups of the enzyme. Examination of enzymatic properties after N-ethylmaleimide modification revealed that 5-fold increase in the Km value for L-aspartate and a shift of the optimum pH for the activity towards acidic pH were brought about by the modification, while neither dissociation into subunits nor aggregation occurred. These results indicate that the influence of the sulfhydryl group modification is restricted to the active site or its vicinity of the enzyme.


Relation between medication-related jaw necrosis and blood glucose levels A retrospective epidemiological study
Maslach and Jackson. Results: In the field of examined dimensions of burnout syndrome, the most characteristic was emotional exhaustion, which has occurred with a frequency of nearly 30% and with typically high intensity among professionals working in neonatal intensive care units (N = 278). In addition, one-fifth were characterized by a high frequency and intensity of depersonalization. At the same time, nearly a third of them experienced a decrease in personal effectiveness. Workplace and time spent in healthcare did not show a significant connection with the incidence of burnout syndrome, however, burnout syndrome occurred with a higher frequency and intensity among those working in level III. neonatal intensive care unit. Conclusion: This is the first comprehensive burnout survey among premature and new born intensive care units in


Heparin formation and mechanism of its action as growth promoter.
Heparin was prepared and purified by two methods. Rf value, elemental and chemical analyses of our isolate coincided with the clinically used heparin. Heparin stimulated mitotic activities in Escherichia coli B and Aspergillus ochraceus. This was evident from the increase in DNA, RNA, and protein synthesis. The mechanism of growth stimulation by heparin probably takes place as follows: Heparin associates with nuclear protein, releasing DNA from DNP and thus stimulates DNA synthesis. Heparin was taken up by the cells of A. flavus, A. niger, and A. ochraceus. The uptake of heparin increased with time and was affected by several factors, such as pH, different salts, and heparin concentration.


The novel HLA-A*02:1068Q allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.
HLA-A*02:1068Q differs from HLA-A*02:03:01:01 by one nucleotide in exon 1.


Self-sustaining limbic status epilepticus induced by 'continuous' hippocampal stimulation: electrographic and behavioral characteristics.
A model of status epilepticus centered in the limbic system and elicited by 'continuous' focal electrical stimulation of the hippocampus is presented. Under appropriate conditions, the status epilepticus persisted for many hours after discontinuing the electrical stimulus. The critical determinant for the establishment of this self-sustaining limbic status epilepticus (SSLSE) was the length of stimulation, rather than the side (left vs. right) of stimulation or kindling before stimulation. Observations, obtained from stimulus-free intervals spaced regularly during the stimulus protocol and from the period after stimulation had been completed, revealed a distinct and stereotyped electrographic progression of SSLSE though several stages. Brief monitoring periods throughout the stimulus protocol yielded electrographic criteria that predicted which animals would experience experience SSLSE. The presence of synchronous, stimulus-independent seizure activity bilaterally in the hippocampi during stimulation was necessary to establish SSLSE. Intense motor seizure activity, like that seen with kindled motor seizures, occurred intermittently during SSLSE. However, 'limbic' behavioral seizures identical to those seen after low doses of kainic acid or during the early stages of kindling were nearly continuous. These studies indicate that there is a predictable course to limbic status epilepticus and point to the hippocampus as a key element involved in initiating and maintaining this syndrome.


Aging-Related Oxidative Stress: Positive Effect of Memory Training.
The cognitive impairment characterizing the phenotype of older adults has been related to the efficiency of the antioxidant system. This study aimed at investigating the effect of memory training (MT) on memory, global cognitive functioning, and the oxidant and antioxidant capacity of plasma. We recruited 52 healthy subjects aged over 60. Twenty-nine subjects were submitted to 6-months of MT (Experimental Group, EG), and 23 were used as a Control Group (CG). Global cognitive functioning was assessed by the Mini-Mental State Examination (MMSE) and Short- and Long-Term Memory (STM and LTM, respectively) by the Rey Auditory Verbal Learning Test (RAVLT) at baseline (T0) and after 6-months (T1). Meanwhile, Reactive Oxygen Metabolites derivative compounds (d-ROMs), Biological Antioxidant Potential (BAP), and their ratio were evaluated on plasma. Results showed that the MMSE and RAVLT scores improved in EG at T1. At the same time, the d-ROMs levels significantly decreased, while the BAP and BAP/d-ROMs ratio showed an opposite trend. In both groups, the MMSE and LTM scores were negatively associated with d-ROMs levels, and positively correlated with BAP levels and the BAP/d-ROMs ratio. When we considered the Δvalue (Δvariable = variable post-MT minus variable pre-MT) in EG, the ΔMMSE and ΔLTM scores were negatively associated to Δd-ROMs, and positively to ΔBAP and ΔBAP/dROM. In conclusion, our results suggest that MT improves memory and global cognitive functioning. These processes were significantly associated to increase in resistance against oxidative stress at the plasma level in healthy older adults.


Clinical application of spinal robot in cervical spine surgery: safety and accuracy of posterior pedicle screw placement in comparison with conventional freehand methods.
The novel robot-assisted (RA) technique has been utilized increasingly to improve the accuracy of cervical pedicle screw placement. Although the clinical application of the RA technique has been investigated in several case series and comparative studies, the superiority and safety of RA over conventional freehand (FH) methods remain controversial. Meanwhile, the intra-pedicular accuracy of the two methods has not been compared for patients with cervical traumatic conditions. This study aimed to compare the rate and risk factors of intra-pedicular accuracy of RA versus the conventional FH approach for posterior pedicle screw placement in cervical traumatic diseases. A total of 52 patients with cervical traumatic diseases who received cervical screw placement using RA (26 patients) and FH (26 patients) techniques were retrospectively included. The primary outcome was the intra-pedicular accuracy of cervical pedicle screw placement according to the Gertzbin-Robbins scale. Secondary outcome parameters included surgical time, intraoperative blood loss, postoperative drainage, postoperative hospital stay, and complications. Moreover, the risk factors that possibly affected intra-pedicular accuracy were assessed using univariate analyses. Out of 52 screws inserted using the RA method, 43 screws (82.7%) were classified as grade A, with the remaining 7 (13.5%) and 2 (3.8%) screws classified as grades B and C. In the FH cohort, 60.8% of the 79 screws were graded A, with the remaining screws graded B (21, 26.6%), C (8, 10.1%), and D (2, 2.5%). The RA technique showed a significantly higher rate of optimal intra-pedicular accuracy than the FH method (P = 0.008), but there was no significant difference between the two groups in terms of clinically acceptable accuracy (P = 0.161). Besides, the RA technique showed remarkably longer surgery time, less postoperative drainage, shorter postoperative hospital stay, and equivalent intraoperative blood loss and complications than the FH technique. Furthermore, the univariate analyses showed that severe obliquity of the lateral atlantoaxial joint in the coronal plane (P = 0.003) and shorter width of the lateral mass at the inferior margin of the posterior arch (P = 0.014) were risk factors related to the inaccuracy of C1 screw placement. The diagnosis of HRVA (P < 0.001), severe obliquity of the lateral atlantoaxial joint in the coronal plane (P < 0.001), short pedicle width (P < 0.001), and short pedicle height (P < 0.001) were risk factors related to the inaccuracy of C2 screw placement. RA cervical pedicle screw placement was associated with a higher rate of optimal intra-pedicular accuracy to the FH technique for patients with cervical traumatic conditions. The severe obliquity of the lateral atlantoaxial joint in the coronal plane independently contributed to high rates of the inaccuracy of C1 and C2 screw placements. RA pedicle screw placement is safe and useful for cervical traumatic surgery.


Characterization of a monoclonal antibody identifying a CD45RA antigen on feline leukocytes.
The antibody produced by a murine hybridoma obtained from the fusion of SP2/0 plasmacytoma cells with splenocytes of a mouse immunized with feline bone marrow was found to react with 60% of bone marrow cells and 80% of peripheral blood leukocytes (PBL); reactivity in the latter tissue was restricted almost entirely to mononuclear cells. Two-color FACScan analyses of this antibody with mAbs specific for feline lymphocytes revealed positive and negative populations of CD4 and CD8 cells. The reactivity for CD4 and CD8 cells was animal age dependent, binding to a higher percentage of the cells in young (2-9 months) versus older animals (> 4 years). In a mitogen driven assay for IgG production by PBL the addition of this antibody to the cultures enhanced the suppressor activity of CD8 cells, a function attributed to activation of a CD4 suppressor-inducer population; removal of CD8 cells negated any induction of suppression. Mild papain digestion of bone marrow and PBL completely removed the antigen detected by this antibody while not affecting reactivity of a pan-T antibody. Western blot analysis showed binding of the antibody to polypeptides of approximately 200 kDa on feline bone marrow and PBL. The data suggest that this mAb is identifying the feline homologue of the leukocyte common antigen of cells with a functional specificity characteristic of a CD45RA isoform.


Morphological and molecular-biological features of glioblastoma progression in tolerant and susceptible to hypoxia Wistar rats.
Hypoxia is a major pathogenetic factor in many cancers. Individual resistance to suboptimal oxygen availability is subject to broad variation and its possible role in tumorigenesis remains underexplored. This study aimed at specific characterization of glioblastoma progression in male tolerant and susceptible to hypoxia Wistar rats. Hypoxia resistance was assessed by gasping time measurement in an 11,500 m altitude-equivalent hypobaric decompression chamber. Based on the outcome, the animals were assigned to three groups termed 'tolerant to hypoxia' (n = 13), 'normal', and 'susceptible to hypoxia' (n = 24). The 'normal' group was excluded from subsequent experiments. One month later, the animals underwent inoculation with rat glioblastoma 101.8 followed by monitoring of survival, body weight dynamics and neurological symptoms. The animals were sacrificed on post-inoculation days 11 (subgroup 1) and 15 (subgroup 2). Relative vessels number, necrosis areas and Ki-67 index were assessed microscopically; tumor volumes were determined by 3D reconstruction from histological images; serum levels of HIF-1α, IL-1β, and TNFα were determined by ELISA. None of the tolerant to hypoxia animals died of the disease during observation period, cf. 85% survival on day 11 and 55% survival on day 15 in the susceptible group. On day 11, proliferative activity of the tumors in the tolerant animals was higher compared with the susceptible group. On day 15, proliferative activity, necrosis area and volume of the tumors in the tolerant to hypoxia animals were higher compared with the susceptible group. ELISA revealed no dynamics in TNFα levels, elevated levels of IL-1β in the susceptible animals on day 15 in comparison with day 11 and tolerant ones. Moreover, there were elevated levels of HIF-1α in the tolerant animals on day 15 in comparison with day 11. Thus, the proliferative activity of glioblastoma cells and the content of HIF-1α were higher in tolerant to hypoxia rats, but the mortality associated with the tumor process and IL-1β level in them were lower than in susceptible animals. Specific features of glioblastoma 101.8 progression in tolerant and susceptible to hypoxia rats, including survival, tumor growth rates and IL-1β level, can become the basis of new personalized approaches for cancer diseases treatment in accordance to individual hypoxia resistance.


Effect of surgery-induced acute muscle wasting on postoperative outcomes and quality of life.
Gastrectomy results in a significant loss of body composition in the long term, but the acute skeletal muscle wasting after gastrectomy has been rarely investigated. Moreover, the association between postoperative muscle wasting and quality of life (QOL) has never been reported. In the present study, we aimed to investigate the risk factors for acute muscle wasting after gastric cancer surgery and its effect on QOL and short-term postoperative outcomes. We conducted a prospective study of patients who underwent curative gastrectomy for gastric cancer between June 2015 and December 2015. Skeletal muscle mass was measured by computed tomography within 1 month before and 1 week after surgery. QOL was assessed 1, 3, and 6 months postoperatively. Univariate and multivariate analyses were performed to identify the risk factors for clinically relevant muscle wasting (muscle wasting ≥10%). A total of 110 patients were included, in which 35 patients had muscle wasting ≥10% within 1 week after surgery. Age ≥65 years and diabetes were independent risk factors for muscle wasting ≥10%. Patients with muscle wasting ≥10% had a poorer QOL in terms of fatigue and physical functioning at 1 and 3 months postoperatively, as well as a higher incidence of postoperative complications, a higher incidence of handgrip strength reduction ≥10%, longer hospital stays, and higher costs. Age ≥65 years and diabetes were independently associated with clinically relevant muscle wasting within 1 week after gastric cancer surgery. Clinically relevant muscle wasting was associated with a poorer QOL and short-term outcomes after surgery.


Cardiovascular outcomes during extended follow-up of the AIM-HIGH trial cohort.
Epidemiologic studies have shown that low levels of high-density lipoprotein-cholesterol (HDL-C) and elevated triglycerides are independent predictors of cardiovascular (CV) events, though randomized trials of HDL-C-raising therapies to reduce clinical events have been largely disappointing. The Atherothrombosis Intervention in Metabolic Syndrome with Low HDL/High Triglycerides and Impact on Global Health Outcomes (AIM-HIGH) trial failed to show that extended release niacin (ERN) reduced CV events in patients with atherogenic dyslipidemia who were on statin-based therapy. We sought to determine whether extended follow-up of AIM-HIGH participants changed these null results. AIM-HIGH was a placebo-controlled trial of 3414 patients with established CV disease, low baseline HDL-C, and elevated triglycerides levels randomized to ERN 1500-2000 mg/d vs placebo. Participants also received simvastatin with or without ezetimibe to attain on-treatment low-density lipoprotein cholesterol levels of 40-80 mg/dL. The trial was halted after a mean 3-year follow-up because of futility. Among 3236 participants alive at the end of blinded study, 2613 (81%; ERN = 1,312, placebo = 1301) were followed a mean 1.1 additional years. Ninety-five percent of subjects remained on statin, but only 4% on ERN. At a mean total follow-up of 4.1 years, there were 343 primary CV endpoints in the ERN arm and 305 CV endpoints in placebo participants (HR 1.11, 95% CI 0.96, 1.30). Ischemic stroke was also not significantly different after extended follow-up in the two groups (2.2% vs 1.5%, P = .13). In patients with CV disease and atherogenic dyslipidemia on statin-based therapy, 3 years of ERN treatment did not lower CV event rates. An additional year of follow-up off assigned treatment did not alter these findings.


Pharmacogenetic effect of an endothelin-1 haplotype on response to bucindolol therapy in chronic heart failure.
Beta-blocker therapy has become a mainstay therapy for the over 5 million patients with chronic heart failure in the United States. Variation in clinical response to beta-blockers is a well-known phenomenon and may be because of genetic differences between patients. We hypothesized that variation in genes of the endothelin system mediate the clinical response to beta-blockers in heart failure. Single nucleotide polymorphisms (SNPs) in six endothelin system genes were genotyped in 309 heart failure patients in a randomized trial of bucindolol versus placebo therapy. We adjusted for multiple comparisons and tested for association between genotype and time to two prospective endpoints. Nine SNPs were sufficiently common to undergo statistical analysis. The SNPs had no significant effect on prospective outcomes in the placebo group, or on the primary endpoint of time to death in either arm. Two SNPs (IVS-4 G/A and Lys198Asn) in the endothelin-1 gene, however, predicted time to the combined endpoint of heart failure hospitalization or all-cause death in bucindolol-treated patients. The alleles at these SNPs were in tight linkage disequilibrium appearing on either of two complementary haplotypes. A 'dose-response' trend was observed, with participants carrying the rarer haplotype having the highest hazard ratios as compared to the relative 'protective' effect of the common haplotype. A common endothelin-1 gene haplotype may be a pharmacogenetic predictor of a favorable clinical response to beta-blocker therapy in heart failure patients. The existence of a less common 'high-risk' haplotype could identify a subpopulation of heart failure patients destined to respond poorly to beta-blocker therapies.


Quality of care and resource use among mechanically ventilated patients before and after an intervention to assist nurse-nonvocal patient communication.
Implement and test unit-wide patient-nurse assisted communication strategies (SPEACS). SPEACS improved nurse-patient communication outcomes; effects on patient care quality and resource use are unknown. Prospective, randomized stepped-wedge pragmatic trial of 1440 adults ventilated ≥2 days and awake for at least one shift in 6 ICUs at 2 teaching hospitals 2009-2011 with blinded retrospective medical record abstraction. 323/383 (84%) nurses completed training; their communication knowledge (p < .001) and satisfaction and comfort (p < .001) increased. ICU days with physical restraint use (p = .44), heavy sedation (p = .73), pain score documentation (p = .97), presence of ICU-acquired pressure ulcers (p = .78), coma-free days (p = .76), ventilator-free days (p = .83), ICU length of stay (p = .77), hospital length of stay (p = .22), and median costs (p = .07) did not change. SPEACS improved ICU nurses' knowledge, satisfaction and comfort in communicating with nonvocal MV patients but did not impact patient care quality or resource use.


Immobilized enzyme nylon tube reactors: creatininase and creatine kinase linked to the pyruvate kinase-lactate dehydrogenase indicator system in the detection of creatinine and creatine.
A four enzyme system catalyzing a sequence of reactions was immobilized onto the inside surfaces of nylon tubes and studied as a possible method of analysis for the first substrate, creatinine in the sequence of reactions. The system was first studied in solution as a guide to optimizing conditions for preparation and use of the immobilized enzyme nylon tube reactors. The scope and limitations of this approach to routine analysis is discussed.


Evaluation of pretreatments for solubilisation of components and recovery of fermentable monosaccharides from microalgae biomass grown in piggery wastewater.
Microalgae-bacteria biomass cultured in wastewater is an interesting renewable material capable of metabolising nutrients from wastes into carbohydrates, proteins, and lipids through photosynthesis. Despite the interest in the valorisation of this biomass to improve the viability of microalgae-based wastewater treatment processes, very scarce research has been devoted to the fractional recovery of its components. This work evaluates the effect of different pretreatments coupled with enzymatic hydrolysis on the solubilisation of biomass components and on the recovery of fermentable monosaccharides (glucose and xylose) from Scenedesmaceae based biomass grown in a thin layer reactor feed with piggery wastewater. Chemical pretreatments generated high concentrations of byproducts, mainly organic acids. No bacterial DNA was found in these pretreated biomasses. The acid pretreatment provided the highest carbohydrate solubilisation (98%) and monosaccharide recovery (81%). Enzymatic hydrolysis coupled with alkaline NaOH 2 M pretreatment achieved almost complete solubilisation of the biomass components, but high carbohydrate losses. Physical pretreatments remarkably increased the solubilisation of the biomass components during the enzymatic hydrolysis step, especially bead milling, which achieved solubilisation yields of 83% of carbohydrates, 43% of proteins, and 60% of lipids. The presence of viable bacteria in these pretreated biomasses could be related to the high carbohydrate losses and the generation of methanol and ethanol in addition to organic acids as byproducts.


Major stoma related morbidity in young children following stoma formation and closure: A retrospective cohort study.
Little is known about stoma related morbidity in young children. Therefore, the aim of this study is to assess major morbidity after stoma formation and stoma closure and its associated risk factors. All consecutive young children (age ≤ three years) who received a stoma between 1998 and 2018 at our tertiary referral center were retrospectively included. The incidence of major stoma related morbidity (Clavien-Dindo grade ≥III) was the primary outcome. This was separately analysed for stoma formation alone, stoma closure alone and all stoma interventions combined. Non-stoma related morbidity was excluded. Risk factors for major morbidity were identified using multivariable logistic regression analysis. In total 336 young children were included with a median follow-up of 6 (IQR:2-11) years. Of these young children, 5% (n = 17/336) received a jejunostomy, 57% (n = 192/336) an ileostomy, and 38% (n = 127/336) a colostomy. Following stoma formation, 27% (n = 92/336) of the young children experienced major stoma related morbidity, mainly consisting of high output stoma, prolapse and stoma stenosis. The major morbidity rate was 23% (n = 66/292) following stoma closure, most commonly comprising anastomotic leakage/stenosis, incisional hernia and adhesive obstructions. For combined stoma interventions, major stoma related morbidity was 39% (n = 130/336). Ileostomy was independently associated with a higher risk of developing major morbidity following stoma formation (OR:2.5; 95%-CI:1.3-4.7) as well as following closure (OR:2.7; 95%-CI:1.3-5.8). Major stoma related morbidity is a frequent and severe clinical problem in young children, both after stoma formation and closure. The risk of morbidity should be considered when deliberating a stoma.


The change of alkaline phosphatase binding conditions with trophoblast membrane at different stages of human placenta.
Alkaline phosphatase was brought into solution from microsomal fractions of placentas of varying gestational age by using gradually increasing concentrations of proteinase papain. When the activity of the soluble alkaline phosphatase (S) was compared with that of the non-soluble residue (R), the S/R ratio rose as pregnancy progressed. The electrophoretic pattern showed that in serum from pregnant women the papain-soluble alkaline phosphatase corresponded to the heat-stable one. These results indicate that the cytoplasmic membrane of the trophoblast changes with the growth of the placenta so that this enzyme is easily dissolved by papain. It is probable that alkaline phosphatase molecules easily enter the maternal blood stream in late pregnancy.


A novel anti-inflammatory role links the CARS2 locus to protection from coronary artery disease.
Genome-wide association studies (GWAS) identified a coronary artery disease (CAD) risk locus on 13.q34 tagged by rs61969072 (T/G). This variant lies in an intergenic region, proximal to ING1, CARKD and CARS2 but its causal relationship to CAD is unknown. We first demonstrated that rs61969072 and tightly linked single nucleotide polymorphisms (SNPs) associate with CARS2 but not ING1 or CARKD expression in carotid endarterectomy samples, with reduced CARS2 abundance in carriers of the CAD risk allele (G). THP-1 monocytes were differentiated and polarized to proinflammatory (M1) and anti-inflammatory (M2) macrophages. CARS2 gene expression decreased in M1 and increased in M2 macrophages, consistent with a role for CARS2 in inflammation. Gene expression profiling revealed an increase in pro-inflammatory markers in response to CARS2 siRNA knockdown in THP-1 derived macrophages, accompanied by an increased abundance of inflammatory cytokines in the cell supernatant. Functional enrichment analysis of impacted transcripts identified the anti-inflammatory IL10 signalling pathway. Western blot analysis of CARS2 silenced macrophages revealed reduced STAT3 phosphorylation in response to IL-10 and increased expression of LPS-induced genes that are repressed by IL-10, indicating a role for CARS2 in anti-inflammatory signalling. Finally, to simulate vessel wall conditions, macrophages, and smooth muscle cells (SMC) were maintained in co-culture. Significantly, CARS2 silencing in macrophages altered the SMC phenotype, decreasing expression of contractile genes and increasing expression of inflammatory genes. These data highlight a novel anti-inflammatory novel role for CARS2 in human macrophages and SMCs that may underlie the protective effect of a common GWAS-identified variant.


Central serous chorioretinopathy resolution after traumatic cyclodialysis repair.
To report a rare case of central serous chorioretinopathy resolution after traumatic cyclodialysis repair. Observation: A 66-year-old Japanese woman was referred to our hospital with a visual disturbance in her right eye (OD). She had a history of blunt ocular injury when hit by a carton box 3 years previously, but the best-corrected visual acuity (BCVA) was 1.0. At the initial visit, the BCVA was 0.3 OD and 1.2 in the left eye (OS). Optical coherence tomography (OCT) showed a serous retinal detachment (SRD) in the macula; the submacular choroid was thicker OD (316 μm) than OS (246 μm). Fluorescent fundus angiography showed a subretinal macular leak. Gonioscopy and anterior-segment OCT showed angle recession and a cyclodialysis cleft at the temporal angle and cyclodialysis in the four quadrants. One month after focal photocoagulation was applied to the leakage point, the retinal detachment had not improved and the BCVA worsened to 0.2. After subsequent surgical repair of the cyclodialysis using an ab interno approach, the ciliochoroidal detachment resolved by 1 month with a simultaneous rapid decrease in the SRD and complete resolution by 2 months. At the final visit, 5 months postoperatively, the BCVA was 1.0 OD. During this period, the choroidal thickness decreased to 264 μm OD but was unchanged at 247 μm OS. Conclusion and Importance: Traumatic cyclodialysis, presumably via choroidal venous overload, can cause CSC. Since the presence of mild cyclodialysis and/or ciliochoroidal detachment may be difficult to find, post-traumatic CSC during the chronic phase of ocular trauma might be overlooked.


Contact printing of protein microarrays.
A review is provided of contact-printing technologies for the fabrication of planar protein microarrays. The key printing performance parameters for creating protein arrays are reviewed. Solid pin and quill pin technologies are described and their strengths and weaknesses compared.