Baseline assessment of patient safety culture in primary care centres in Kuwait: a national cross-sectional study.
Assessments of the culture surrounding patient safety can inform healthcare settings on how their structures and processes impact patient outcomes. This study investigated patient safety culture in Primary Health Care Centres in Kuwait, and benchmarked the findings against regional and international results. This study also examined the association between predictors and outcomes of patient safety culture in these settings. This cross-sectional quantitative study used the Medical Office Survey on Patient Safety Culture. The study was targeted at staff of all the Primary Health Care Centres in Kuwait with at least one year of experience. Data were analysed using SPSS 23 at a significance level of ≤ .05. Univariate (means, standard deviations, frequencies, percentages) and bivariate (chi-squared tests, student t-tests, ANOVA F-tests, Kruskal-Wallis tests, Spearman's correlation) analyses provided an overview of participant socio-demographics and the association between patient safety culture composites and outcomes. We undertook a multivariate regression analysis to predict the determinants of patient safety culture. Results were benchmarked against similar local (Kuwait, 2014), regional (Yemen, 2015) and international (US, 2018) studies. The responses of 6602 employees from 94 centres were included in the study, with an overall response rate of 78.7%. The survey revealed Teamwork (87.8% positive ratings) and Organisational Learning (78.8%) as perceived areas of strength. Communication about Error (57.7%), Overall Perceptions of Patient Safety and Quality (57.4%), Communication Openness (54.4%), Owner/Managing Partner/Leadership Support for Patient Safety (53.8%) and Work Pressure and Pace (28.4%) were identified as areas requiring improvement. Benchmarking analysis revealed that Kuwait centres are performing at benchmark levels or better on four and six composites when compared to international and regional findings, respectively. Regression modelling highlighted significant predictions regarding patient safety outcomes and composites. This is the first major study addressing the culture of patient safety in public Primary Health Care Centres regionally. Improving patient safety culture is critical for these centres to improve the quality and safety of the healthcare services they provide. The findings of this study can guide country-level strategies to develop the systems that govern patient safety practices.


Comparative proteomics of related symbiotic mussel species reveals high variability of host-symbiont interactions.
Deep-sea Bathymodiolus mussels and their chemoautotrophic symbionts are well-studied representatives of mutualistic host-microbe associations. However, how host-symbiont interactions vary on the molecular level between related host and symbiont species remains unclear. Therefore, we compared the host and symbiont metaproteomes of Pacific B. thermophilus, hosting a thiotrophic symbiont, and Atlantic B. azoricus, containing two symbionts, a thiotroph and a methanotroph. We identified common strategies of metabolic support between hosts and symbionts, such as the oxidation of sulfide by the host, which provides a thiosulfate reservoir for the thiotrophic symbionts, and a cycling mechanism that could supply the host with symbiont-derived amino acids. However, expression levels of these processes differed substantially between both symbioses. Backed up by genomic comparisons, our results furthermore revealed an exceptionally large repertoire of attachment-related proteins in the B. thermophilus symbiont. These findings imply that host-microbe interactions can be quite variable, even between closely related systems.


Impact of non invasive and beat-to-beat arterial pressure monitoring on intraoperative hemodynamic management.
The Nexfin device allows for non-invasive beat-to-beat blood pressure monitoring (BP(NXF)). Perioperative hypotension and hypertension have been shown to be associated with poor clinical outcomes. The goal of the present study was to assess the ability of this device to decrease the duration of significant intraoperative hypo- or hypertension compared to standard BP monitoring by cuff (BP(CUFF)). We studied25 patients (ASA I-III) undergoing either abdominal or orthopedic surgery. BP(CUFF) was monitored every 5 min from the introduction of anesthesia, while BP(NXF) was monitored continuously on the opposite arm. When systolic BP(NXF) (SBP(NXF)) decreased or increased more than 20% relative to baseline SBP(NXF), a standard BP(CUFF) measurement was taken to compare values. In addition, the time interval between the 20% change in SBP(NXF) and the next scheduled standard SBP(CUFF) measurement was recorded for each event. The mean length of surgery was 3.0 ± 0.3 h. Patients presented with 11 ± 4 episodes of hypotension and 12 ± 4 episodes of hypertension during the surgery. If BP(CUFF) had been used, this would have resulted in 21 ± 7 min of hypotension and 20 ± 10 min of hypertension. If hemodynamic changes seen by SBP(NXF) were appropriately treated, an average of 7 ± 1 min/h of hypotension time, 7 ± 2 min/h of hypertension time and 14 ± 3 min per hour of hypo- or hypertension time may have been identified. The Nexfin BP has the potential to decrease the time of hypotension and hypertension compared to conventional intermittent BP(CUFF) monitoring. Therefore, this device has the potential to positively impact clinical outcomes.


Antimicrobial activity of amazonian medicinal plants.
The aqueous extracts of currently utilized Amazonian medicinal plants were assayed in vitro searching for antimicrobial activity against human and animal pathogenic microorganisms. Medium resuspended lyophilized aqueous extracts of different organs of Amazonian medicinal plants were assayed by in vitro screening for antimicrobial activity. ATCC and standardized microorganisms obtained from Oswaldo Cruz Foundation/Brazil were individually and homogeneously grown in agar plate, and holes previously perforated in the gel were filled with diluted plant aqueous extracts. Inhibition halos were evaluated and controlled by the use of the fluoroquinolone ciprofloxacin. The Amazonian medicinal plants, Hymenelobium petraeum showed inhibitory activity over Staphylococcus aureus, Enterococcus faecalis, Salmonella enterica serovar Typhi, Acinetobacter baumannii and Candida albicans, while Vatairea guianensis and Symphonia globulifera presented inhibitory activity exclusively for Staphylococcus aureus. Also, Ptychopetalum olacoides and Pentaclethra macroloba inhibited the growth of Klebsiella ozaenae and Acinetobacter baumannii. The aqueous botanic extracts that showed activity against microroganisms of ATTC and Osvaldo Cruz strains had at least 40% of antimicrobial activity when compared to halo inhibition produced by the commercial antibiotic ciprofloxacin utilized as a control. Of all plants extracts assayed, the Hymenelobium petraeum had the best performance, sometimes exhibiting higher activity than ciprofloxacin. It is not well-defined by the physicians the exact indication of the majority of medicinal plants in the Amazon area in Brazil. Natives utilize the plants according to their symptoms, based on the traditional knowledge transmitted orally from generation to generation, among Amerindians, Afrodescendents and ethnic mixed populations. A significant number of Amazonian medicinal plants are totally unknown related to their medicinal properties including mechanism of action and therapeutic effects, as very few information is reported in the scientific literature. A tiny amount of data is presented, as the preliminary antimicrobial properties of the medicinal plants here accessed, under the urgent necessity of new antibiotics in the market and in face of the increased resistance of infectious microorganisms to antimicrobials.


Isolated thickening of one aortic cusp: preferential thickening of the noncoronary cusp.
Over a 2-year period we identified 197 patients with the previously undescribed finding of isolated thickening of one aortic cusp. The noncoronary cusp was involved most frequently in 56% (p less than 0.01), followed by the right coronary cusp in 35%, and the left coronary cusp in 9%. Isolated thickening of one aortic cusp occurs more commonly in male patients (64%) (p less than 0.01) and at an earlier age (65 years) than mitral annular calcification (70 years) (p less than 0.001).


Adipose-derived stem cells combined with neuregulin-1 delivery systems for heart tissue engineering.
Myocardial infarction (MI) is the leading cause of death worldwide, and extensive research has therefore been performed to find a cure. Neuregulin-1 (NRG) is a growth factor involved in cardiac repair after MI. We previously described how biocompatible and biodegradable microparticles, which are able to release NRG in a sustained manner, represent a valuable approach to avoid problems related to the short half-life after systemic administration of proteins. The effectiveness of this strategy could be improved by combining NRG with several cytokines involved in cardiac regeneration. The present study investigates the potential feasibility of using NRG-releasing particle scaffold combined with adipose-derived stem cells (ADSC) as a multiple growth factor delivery-based tissue engineering strategy for implantation in the infarcted myocardium. NRG-releasing particle scaffolds with a suitable size for intramyocardial implantation were prepared by TROMS. Next, ADSC were adhered to particle scaffolds and their potential for heart administration was assessed in a MI rat model. NRG was successfully encapsulated reaching encapsulation efficiencies of 92.58 ± 3.84%. NRG maintained its biological activity after the microencapsulation process. ADSCs adhered efficiently to particle scaffolds within a few hours. The ADSC-cytokine delivery system developed proved to be compatible with intramyocardial administration in terms of injectability through a 23-gauge needle and tissue response. Interestingly, ADSC-scaffolds were present in the peri-infarted tissue 2 weeks after implantation. This proof of concept study provides important evidence required for future effectiveness studies and for the translation of this approach.


ERBB2 amplification status in 67 salivary duct carcinomas assessed by immunohistochemistry, fluorescence in situ hybridization, and targeted exome sequencing.
Salivary duct carcinoma (SDC) is an aggressive salivary gland malignancy with poor survival. Approximately 30% SDC harbor HER2 amplification and response to trastuzumab has been reported. However, a systematic approach for HER2 status assessment in this tumor type has not been established. A total of 67 tumor samples were evaluated for HER2 protein overexpression or ERBB2 gene amplification using at least 2 methods: immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and/or targeted exome next-generation sequencing (NGS). NGS assessed ERBB2 copy number fold change (FC) and total copy number (TCN). HER2 status was first determined by IHC/FISH according to the 2018 ASCO/CAP breast cancer guidelines. FISH results, the "gold standard", were compared with the NGS results. All (15/15) IHC positive, 35% (6/17) equivocal, and no (0/19) IHC negative SDC were HER2 amplified by FISH. HER2 FISH signal/cell showed a good correlation with FC (Spearman correlation: 0.708, R2: 0.501, p < 0.0001) and TCN (Spearman correlation: 0.763, R2: 0.582, p < 0.0001). Receiver operating characteristics curve estimation showed an area under curve (AUC) of 0.975 for ERBB2 FC. FC cutoff of ≥1.8 corresponded to an accuracy of 95.2% for ERBB2 amplification (Youden's index: 0.84, sensitivity: 89.47%, specificity: 100%). FC < 1.3 could be reliably classified as ERBB2 not amplified and FC ≥ 1.3 and <1.8 as equivocal. TCN estimation showed AUC of 0.981. TCN cutoff of >6.0 corresponded to an accuracy of 92% for HER2 amplification (Youden's index: 0.81, sensitivity: 81.2%, specificity: 100%). TCN < 4 could be reliably classified as ERBB2 not amplified and TCN ≥ 4.0 and ≤6.0 as equivocal. FC and TCN were binarized with respective cutoffs of ≥1.8 and ≥6.0 and the proportion of agreement with FISH were 95% and 92%, respectively. The assessment of ERBB2 copy number by NGS is accurate and reliable with FC or TCN nearly equivalent to FISH in identifying HER2 amplified SDC.


Experimental Study of the Shear Performance of Combined Concrete-ECC Beams without Web Reinforcement.
Shear damage of beams is typically brittle damage that is significantly more detrimental than flexural damage. Based on the super-high toughness and good crack control ability of engineered cementitious composites (ECC), the shear performance of concrete-ECC beams was investigated by replacing a portion of the concrete in the tensile zone of reinforced concrete beams with ECC and employing high-strength reinforcing bars to design concrete-ECC beams. The purpose of this investigation is to elucidate and clarify the shear performance of concrete-ECC beams. Experimental and FE analyses were conducted on the shear performance of 36 webless reinforced concrete-ECC composite beams with varied concrete strengths, shear-to-span ratios, ECC thicknesses, and interfacial treatments between the layers. The results indicate that the effect of the shear-to-span ratio is greater, the effect of the form of interface treatment is smaller, the effect is weakened after the ECC thickness is greater than 70 mm (i.e., the ratio of the replacement height to section height is approximately 0.35), the shear resistance is reduced when the hoop rate is greater, and the best shear resistance is obtained when the ECC 70 mm thickness and the hoop rate of 0.29% are used together. This study can serve as a technical reference for enhancing the problems of low durability and inadequate fracture control performance of RC beams in shear and as a guide for structural design research.


Insight into the failure mechanism of large-scale cylindrical lithium-sulphur cells.
Li-S batteries with a sulphur loading content of 5 mg cm-2 were produced as large-scale 18 650 cylindrical cells. We have found that a key failure mode of cylindrical Li-S battery cells is the severe capacity fading during the galvanostatic charge-discharge process due to the corrosion of the electrodes, the electrolyte decomposition, and the severe polysulphide shuttling effect.


Normal insulin receptor tyrosine kinase activity and glucose transporter (GLUT 4) levels in the skeletal muscle of hyperinsulinaemic hypertensive rats.
The spontaneous hypertensive rat is an animal model characterized by a syndrome of hypertension, insulin resistance and hyperinsulinaemia. To elucidate whether in analogy to other insulin resistant animal models an inactivity of the insulin receptor kinase or an alteration of the glucose transporter (GLUT 4) level in the skeletal muscle might contribute to the pathogenesis of insulin resistance we determined insulin receptor kinase activity and GLUT 4 level in the hindlimbs of spontaneous hypertensive rats and normotensive control rats. Normotensive normoinsulinaemic Lewis and Wistar rats were used as insulin sensitive controls, obese Zucker rats were used as an insulin resistant control with known reduced skeletal muscle insulin receptor kinase activity. Binding of 125I-insulin, crosslinking of 125I-B26-insulin, autophosphorylation in vitro with 32P-ATP and phosphorylation of the synthetic substrate Poly (Glu 4: Tyr 1) were performed after partial purification of solubilized receptors on wheat germ agglutinin columns. GLUT 4 levels were determined by Western blotting of subcellular muscle membranes. Insulin receptors from spontaneous hypertensive rats compared to those from Lewis and Wistar rats showed no difference of the binding characteristics or the in vitro auto- and substrate phosphorylation activity of the receptor, while in the Zucker rats the earlier described insulin receptor kinase defect was clearly evident. Western blots of subcellular muscle membrane fractions with antibodies against GLUT 4 revealed no difference in transporter levels. These data suggest that insulin resistance in spontaneous hypertensive rats is caused neither by an insulin receptor inactivity nor by a decreased number of glucose transporters in the skeletal muscle.


Transient light-induced conformational changes in rhodopsin.
Arguments are presented which support the possibility that the unfolding of the rhodopsin molecule during photolysis up to the stage of metarhodopsin II is followed by a spontaneous refolding of the protein, once the isomerized retinaldehyde has left its original binding site. Such a transient conformational change might imply a very similar conformation for rhodopsin and opsin, apart from the presence of the chromophore.


Impact of predation by spiders on a territorial damselfly (Odonata: Calopterygidae).
Predation by orb-weaving spiders and crab spiders on the damselfly Calopteryx haemorrhoidalis was studied at a small stream in Southern France. One species of orb-weaving spider, Larinioides folium, caught 76% of the damselflies which fell prey to spiders. Displacement experiments on spiders on sections of bank and the positioning of webs in male territories show that the density and distribution of damselflies is not influenced by orbweb density or by the position of webs. Predation rates corresponded to orb-web density, but neither for sex nor for stage was there a relationship with damselfly density. Mean daily predation rates ranged between 0.9% for females and 4.1% for adult males. Predation risk to adult damselflies by orb-weaving spiders was male biased, whereas among tenerals there was no bias. Males were captured more frequently at territories near the water. Captures show a maximum at noon when territorial disputes of adult males were most frequent. After orbwebs were placed within territories predation rate of males was strongly increased. Predation risk to adult females in the direct vicinity of the stream was less than in the bank vegetation where they perch close to orbwebs. The risk of predation by crab spiders, which catch damselflies at their perching sites, was not sex-biased.


Changes in glycosylation alter the affinity of the human transferrin receptor for its ligand.
When transferrin receptors of human erythroleukemic cells were pulse-labeled with [35S]methionine and then chased in the absence of radioactive precursor, the first detectable immunoprecipitable form of the receptor had a molecular mass of 85 kDa. This form of the receptor was converted to the mature form of 93 kDa with a half-time of about 40-60 min. Both the immature (85 kDa) and mature (93 kDa) receptors associated as dimers, the native form of the receptor. The 85-kDa, as well as the 93-kDa, receptors bound to a monoclonal antibody raised against the transferrin receptor or to transferrin-Sepharose. In order to determine whether glycosylation was necessary for ligand binding, purified receptors were isolated from cells grown in the presence of tunicamycin. When K562 cells were grown in the presence of tunicamycin, an 80-kDa nonglycosylated form of the receptor was synthesized. This nonglycosylated receptor was also capable of dimer formation; however, much less of it reached the cell surface than the fully glycosylated form, although both untreated and tunicamycin-grown cells appeared to synthesize transferrin receptors at similar rates. Although the number of receptor molecules/cell was similar in control and tunicamycin-treated cells, the nonglycosylated receptors exhibited a much lower affinity for transferrin than those of untreated cells; in contrast, when receptors were purified by immunoprecipitation and digested with bacterial alkaline phosphatase, no difference was observed between the affinity of these receptors and undigested immunoprecipitated receptors. These results suggest that glycosylation is not necessary for specific binding of transferrin to its receptor, but the affinity of this binding can be influenced greatly by the presence or absence of carbohydrate residues.


Effects of Omega-3 as an adjuvant in the treatment of periodontal disease: A systematic review and meta-analysis.
BACKGROUND AND OBJECTIVES: Promoting resolution of inflammation using new classes of lipids mediators has been proposed for the management of inflammatory disease. This systematic review and meta-analysis aimed to evaluate the benefits of the use of omega-3 fatty acids as an adjuvant in the nonsurgical treatment of periodontitis. The data search was conducted into three main databases: PubMed, Embase, and Cochrane. The search equation was built around the PICO framework in which the population was constituted by human adults suffering from chronic periodontitis that had to be treated with conventional SRP with the adjunction of omega-3 fatty acids (I) or without the adjunction of omega-3 fatty acids (C), with, as a first outcome the probing pocket depth reduction (PPD) and as a second outcome the clinical attachment loss reduction (CAL). Risk of bias within studies was evaluated for each included study using the Cochrane collaboration tool for randomized studies (RoB Tool). A meta-analysis was performed using REVMAN 5.3. After a global search, 117 studies were selected but only seven of them were eligible for the systematic review and meta-analysis. Six out of seven studies showed a significantly better PPD reduction in the omega-3 fatty acids group compared to the control group and five out of seven studies showed a significantly better CAL reduction in the omega-3 fatty acids group compared to the control group. The meta-analysis showed a statistically significant difference for PPD reduction (SMD: -0.78 [95% CI: -1.02, -0.54, p < .0001]) and CAL reduction (SMD: -0.80 [95% CI: -1.04, -0.56, p < .0001]) in favor of the test group. After scaling and root planning, PPD reduction and CAL reduction were observed in both control and test groups, but with statistically significant better values for the omega-3 fatty acids group. Patients suffering from periodontitis could benefit from the use of omega-3 fatty acids to increase the effectiveness of a nonsurgical treatment.


Amblyopia prevalence and risk factors in Australian preschool children.
To determine the prevalence of and factors associated with amblyopia in a sample of Australian preschool children. Population-based, cross-sectional study. The Sydney Paediatric Eye Disease Study examined 2461 (73.8% participation) children aged between 6 and 72 months from 2007 to 2009. Visual acuity (VA) was assessed in children aged ≥ 30 months using the Electronic Visual Acuity system, and a subset using the logarithm of the minimum angle of resolution chart. Amblyopia was categorized into unilateral and bilateral subtypes: Unilateral amblyopia was defined as a 2-line difference in reduced VA between the 2 eyes, in addition to strabismus, anisometropia, and/or visual axis obstruction; bilateral amblyopia was defined as bilateral reduced VA with either bilateral visual axis obstruction or significant bilateral ametropia. Information on ethnicity, birth parameters, and measures of socioeconomic status were collected in questionnaires completed by parents. Amblyopia. We included 1422 children aged 30 to 72 months, of whom 27 (1.9%) were found to have amblyopia or suspected amblyopia. Mean spherical equivalent for the amblyopic eyes was +3.57 diopters, with a mean VA of 20/50. Only 3 of the 27 amblyopic children had previous diagnoses or treatments for amblyopia. In regression analysis controlling for age, gender, and ethnicity, amblyopia was significantly associated with hyperopia (odds ratio [OR], 15.3; 95% confidence interval [CI], 6.5-36.4), astigmatism (OR, 5.7; 95% CI, 2.5-12.7), anisometropia (OR, 27.8; 95% CI, 11.2-69.3), and strabismus (OR, 13.1; 95% CI, 4.3-40.4). There were no significant associations of amblyopia with low birthweight (<2500 g), preterm birth (<37 weeks), maternal smoking, age, gender, ethnicity, or measures of socioeconomic status (all P>0.05). Amblyopia was found in 1.9% of this Australian preschool sample, which is comparable with prevalence rates reported by other recent studies in preschool children. Refractive errors, particularly significant hyperopia and astigmatism, in addition to anisometropia and strabismus, were the major amblyogenic factors. There was a low amblyopia detection rate in this preschool population, which suggests that different strategies are required to improve current vision screening strategies in preschoolers. The authors have no proprietary or commercial interest in any of the materials discussed in this article.


Determination of LOX-1-ligand activity in mouse plasma with a chicken monoclonal antibody for ApoB.
Oxidized LDL (OxLDL) is implicated in endothelial dysfunction as well as the formation and progression of atherosclerosis. It has become evident that the atherogenic properties induced by OxLDL are mainly mediated via lectin-like OxLDL receptor-1 (LOX-1). Over the past decade, much research has been performed to investigate lipid metabolism and atherogenesis using genetically engineered mice. To understand the significance of OxLDL, methods to measure the levels of OxLDL in these experimental animals should be established. Utilizing a chicken monoclonal antibody technique, here, we generated anti-human ApoB antibodies that are able to recognize mouse VLDL/LDL. These antibodies were selected from single chain fragment of variable region (scFv) phage library constructed from chickens immunized with human LDL. One of these antibodies, HUC20, was reconstructed into IgY form. Immunohistochemical analysis revealed that this novel antibody specifically stains atherosclerotic lesions of ApoE-deficient mice, associated with Oil red O positive and macrophage-antigen-positive regions. Furthermore, in combination with recombinant LOX-1, a sandwich enzyme immunoassay was developed to measure the levels of LOX-1 ligands in mouse plasma. The sandwich enzyme immunoassay revealed a dramatic increase in the level of LOX-1 ligands in the plasma of ApoE-deficient mice fed high-fat diet, suggesting a link between the level of LOX-1-ligands and the progression of atherosclerosis in mice. Hence, the chicken anti-ApoB monoclonal antibody HUC20 developed here, could be a useful tool to analyze the role of ApoB-containing lipoprotein in atherogenesis in mice.


miSSING LINKS: miRNAs and plant development.
The discovery of hundreds of plant micro RNAs (miRNAs) has triggered much speculation about their potential roles in plant development. The search for plant genes involved in miRNA processing has revealed common factors such as DICER, and new molecules, including HEN1. Progress is also being made toward identifying miRNA target genes and understanding the mechanisms of miRNA-mediated gene regulation in plants. This work has lead to a reexamination of many previously characterized mutations that are now known to affect components or targets of miRNA-mediated pathways.


FAM3D is essential for colon homeostasis and host defense against inflammation associated carcinogenesis.
The physiological homeostasis of gut mucosal barrier is maintained by both genetic and environmental factors and its impairment leads to pathogenesis such as inflammatory bowel disease. A cytokine like molecule, FAM3D (mouse Fam3D), is highly expressed in mouse gastrointestinal tract. Here, we demonstrate that deficiency in Fam3D is associated with impaired integrity of colonic mucosa, increased epithelial hyper-proliferation, reduced anti-microbial peptide production and increased sensitivity to chemically induced colitis associated with high incidence of cancer. Pretreatment of Fam3D-/- mice with antibiotics significantly reduces the severity of chemically induced colitis and wild type (WT) mice co-housed with Fam3D-/- mice phenocopy Fam3D-deficiency showing increased sensitivity to colitis and skewed composition of fecal microbiota. An initial equilibrium of microbiota in cohoused WT and Fam3D-/- mice is followed by an increasing divergence of the bacterial composition after separation. These results demonstrate the essential role of Fam3D in colon homeostasis, protection against inflammation associated cancer and normal microbiota composition.


[Quantitative and qualitative investigations of salivary gland function in dependence on irradiation dose and volume for reduction of xerostomia in patients with head-and-neck cancer].
Radiation treatment of head-and-neck tumors mostly leads to a damage to the salivary glands and a consequential permanent loss of saliva. The aim of this investigation was to establish a modern three-dimensional conformal radiotherapy (3D-CRT) to show a decrease in severe xerostomia in contrast to the proven conventional technique (K-RT) with photons and electrons. Between April 2002 and September 2003, 32 patients (25 male, seven female, mean age: 58 years) with malignant tumors of the head and neck were included-after surgery or in case of inoperability with curative intent-in a prospective, nonrandomized study. 10/32 patients (31%) received K-RT with photons and electrons, and 22/32 patients (69%) 3D-CRT (six to eight photon portals). The quantity of saliva was measured as stimulated saliva flow rate (ml/5 min) prior to treatment, at the end, and 1, 6, and 12 months after termination of treatment. To find out the resulting mean dose of both parotid glands for every patient in Gray (D(mean) doses), the D(mean) doses of the ipsilateral and the contralateral parotid gland, determined by dose-volume histograms (DVHs), were averaged over. For calculation of the NTCP (normal tissue complication probability), the logistic model was used. In the trend the stimulated salivary flow rates were higher in the group with 3D-CRT than in the group with K-RT during the whole observation period (at 10 weeks after the start of radiotherapy 3D-CRT vs. K-RT with 1.56 +/- 1.6 vs. 0.82 +/- 1.2 ml/5 min; p < 0.1). The patients treated with the K-RT had, on average, significantly higher averaged D(mean) values than those irradiated with 3D-CRT (p < 0.012). Patients, who were irradiated with 3D-CRT for tumors of the larynx or hypopharynx, showed, on average, significantly lower D(mean) values than patients, who were treated with 3D-DRT because of oral cavity or oropharynx carcinomas or with K-RT irrespective of the primary tumor site (p < 0,003). The resulting dose for 50% complication probability (TD(50)) of the salivary glands was 36.9 Gy (30.9-43.5 Gy; 95% confidence interval). The gradient k of the curve located in point TD(50) was 7.7 (4.8-15.8; 95% confidence interval). Basically, 3D-CRT seems to be suitable as a standard for all patients with carcinomas of the oral cavity, oro- and hypopharynx. Especially in patients with tumors located in the larynx and hypopharynx, averaged D(mean) doses of both parotids during irradiation can be reached, to conserve salivary flow rates, which are similar to baseline flow rates.


Indole-3-acetic acid improves the hepatic mitochondrial respiration defects by PGC1a up-regulation.
Recent evidences have linked indole-3-acetic acid (I3A), a gut microbiota-derived metabolite from dietary tryptophan, with the protection against non-alcoholic fatty liver disease (NAFLD). However, the values of I3A on mitochondrial homeostasis in NAFLD have yet to be analyzed. In this study, we verified that I3A alleviated dietary-induced metabolic impairments, particularly glucose dysmetabolism and liver steatosis. Importantly, we expanded the understanding of I3A further to enhance mitochondrial oxidative phosphorylation in the liver by RNA-seq. Consistently, I3A restored the deficiency of mitochondrial respiration complex (MRC) capacity in palmitic acid (PA)-induced HepG2 without initiating oxidative stress in vitro. These changes were dependent on peroxisome proliferator-activated receptor γ coactivator 1 (PGC1)-a, a key regulator of mitochondrial biogenesis. Silencing of PGC1a by siRNA and pharmacologic inhibitor SR-18292, blocked the restoration of I3A on mitochondrial oxidative phosphorylation. In addition, pre-treatment of I3A guarded against the deficiency of MRC capacity. In conclusion, our findings uncovered that I3A increased hepatic PGC1a expression, contributing to mitochondrial respiration improvement in NAFLD.


Lewis acid-catalyzed one-pot, three-component route to chiral 3,3'-bipyrroles.
3,3'-Bipyrroles 3 could be synthesized using a double Michael addition reaction involving diaroyl acetylene 1 and the appropriate 1,3-dicarbonyls 2 using ammonium acetate as a nitrogen source. The axial chirality of bipyrrole was anticipated from the X-ray crystal structure and DFT calculations and confirmed by separating the racemates on a chiral column and subsequent CD spectra of the enantiomers. The absolute configuration of the enantiomers was achieved by theoretical CD spectra calculation using the ZINDO method.


Warfarin-induced impairment of bone material quality in a patient undergoing maintenance hemodialysis: A case report.
The use of warfarin in patients undergoing hemodialysis is associated with decreased bone mineral density and an increased incidence of bone fracture. However, no studies to date have directly estimated bone quality with bone histomorphometry in patients with bone abnormalities who are taking warfarin and undergoing hemodialysis. A 47-year-old female with Noonan syndrome presented with progressive bilateral lower extremity pain on walking, and skin sclerosis. She had been undergoing maintenance hemodialysis for 25 years following 2 years of peritoneal dialysis for chronic glomerulonephritis. She had been taking warfarin as an anticoagulant agent for 13 years after she underwent an aortic valve replacement. Warfarin-induced impairment of bone material quality. Histomorphometric analysis of the bone biopsy specimens showed impairment in bone calcification processes, a high turnover of bone remodeling, low bone volume, and mild fibrosis. The bone abnormality could not be categorized into any type of representative bone disease classification such as osteitis fibrosa, osteomalacia, adynamic bone disease, uremic osteodystrophy, or hyperparathyroidism, but was consistent with warfarin-induced impairment of bone material quality. Warfarin can induce impairment of bone material quality in a patient undergoing hemodialysis.


Targeted exosome-mediated delivery of opioid receptor Mu siRNA for the treatment of morphine relapse.
Cell-derived exosomes have been demonstrated to be efficient carriers of small RNAs to neighbouring or distant cells, highlighting the preponderance of exosomes as carriers for gene therapy over other artificial delivery tools. In the present study, we employed modified exosomes expressing the neuron-specific rabies viral glycoprotein (RVG) peptide on the membrane surface to deliver opioid receptor mu (MOR) siRNA into the brain to treat morphine addiction. We found that MOR siRNA could be efficiently packaged into RVG exosomes and was associated with argonaute 2 (AGO2) in exosomes. These exosomes efficiently and specifically delivered MOR siRNA into Neuro2A cells and the mouse brain. Functionally, siRNA-loaded RVG exosomes significantly reduced MOR mRNA and protein levels. Surprisingly, MOR siRNA delivered by the RVG exosomes strongly inhibited morphine relapse via the down-regulation of MOR expression levels. In conclusion, our results demonstrate that targeted RVG exosomes can efficiently transfer siRNA to the central nervous system and mediate the treatment of morphine relapse by down-regulating MOR expression levels. Our study provides a brand new strategy to treat drug relapse and diseases of the central nervous system.


Methodological issues in the recruitment of ethnic minority subjects to research via the Internet: a discussion paper.
In this paper, issues in Internet recruitment of ethnic minorities in the US are explored through an analysis of an Internet survey study. The issues include those related to: (a) the difficulties in ensuring authenticity; (b) a lack of cooperation by gate keepers; (c) the flexibility required in the recruitment process; (d) a very low response rate; and (e) selected groups of ethnic minorities. Based on the discussions on the issues, we propose regular updates of knowledge and skills related to Internet interactions and technologies, usage of multiple recruitment sources, pluralistic recruitment approaches, and a quota sampling method.


Quality of Life of Mothers and Fathers 4 to 6 Months After Birth: The Effect of a Very Preterm Delivery.
The sparse literature on the effect of a preterm delivery on parents' quality of life (QoL) yields inconsistent results, restricting their analysis to mothers. The present study aimed to assess the effect of a very preterm delivery on parents' gender-specific perception of QoL, 4 to 6 months after birth. A total of 117 parents of very preterm infants hospitalized at birth in a level III Neonatal Intensive Care Unit, and 214 parents of never hospitalized full-term infants born in a public maternity, both located in the North of Portugal, participated in the study, 4 to 6 months after delivery (November 2013-June 2015). The Portuguese version of the World Health Organization Quality of Life - BREF Inventory was applied and scores were transformed to reflect a 0 to 100 scale. The mean [standard deviation (SD)] of overall QoL ranged between 72.1 (13.3) among mothers of full-term infants and 74.6 (12.5) among mothers of very preterm infants. The perception of QoL was not significantly different among parents of very preterm and full-term infants, according to gender. The highest scores were observed in the psychological and physical dimensions, for both mothers and fathers. The lack of differences on the perception of QoL among mothers and fathers of very preterm and full-term infants, highlights the need to deeply understand and explore the influence of accommodation mechanisms, the extended family/community and health policies on parental QoL trajectories.


Comparison of the growth-inhibitory effect of Hypericum perforatum L. extracts, differing in the concentration of phloroglucinols and flavonoids, on leukaemia cells.
In this study we compared, simultaneously, the growth-inhibitory effect of Hypericum perforatum L. extracts, containing various amounts of hyperforin (A 3.25%; B 2.21%; C 0.21% w/w) and flavonoids (A and B 5.3%; C 10% w/w), but closely same amounts of naphthodiantrones (0.3%) on two leukaemic cell lines K562 and U937 in the WST-1 assay. The GI50 (concentration of extracts which caused 50% of cell growth inhibition) for H. perforatum extracts analysed and characterized by HPLC for their biologically active constituents was 248.3-621.3 microg mL(-1) in K562 and 378.2-911.7 microg mL(-1) in U937 cells. The corresponding values of the three main groups were 1.6-3.9 microM naphthodianthrones, 1.0-40.7 microM phloroglucinols and 30.5-68.5 microM flavonoids. The results of this study supported the hypothesis that, apart from hyperforin and flavonoids, other components of the extract could be involved in its growth-inhibitory effect that it exerts without light activation.


Mapping and insertional mutagenesis of a vaccinia virus gene encoding a 13,800-Da secreted protein.
The objective of this study was to identify and characterize the gene encoding a protein of approximately 12 kDa that is secreted from cells infected with the vaccinia virus. The absence of this protein from the medium of cells infected with a spontaneous deletion mutant (6/2) suggested that the open reading frame (ORF) was located within a 12,800-base pair segment near the left end of the genome (G. Kotwal and B. Moss, Nature (London) 335, 176-178, 1988). Antibody to the 12-kDa protein immunoprecipitated an appropriate size in vitro translation product of mRNA that hybridized to a DNA segment containing an ORF (N1L) that could encode a 13.8-kDa polypeptide. The similarity in the sizes of the in vitro translation product and the secreted protein was consistent with the absence of processing. Transcriptional analysis revealed major and minor early RNA start sites preceding the N1L ORF as well as a late RNA start site with an atypical TAAAAT sequence. The N1L gene was interrupted by replacing a segment of the ORF with the Escherichia coli beta-galactosidase gene. When two-dimensional polyacrylamide gel electrophoretic patterns of [35S]methionine-labeled proteins secreted from cells infected with parental and recombinant viruses were compared, a spot missing from the latter corresponded in molecular weigh and isoelectric point with that predicted from the N1L ORF. The latter analysis revealed the presence of other secreted proteins of similar molecular weight but different isoelectric points that also appear to map within the left end of the vaccinia genome. The recombinant virus was attenuated as judged by the increased intracranial LD50 for mice but nevertheless induced antibody and cytotoxic responses after intradermal and intraperitoneal injections. Relative to the parental virus, the recombinant was also more attenuated for immunodeficient nude mice, based on their survival time after infection.


Development of an approach to lameness examination in cattle.
In the preceding pages, the various components of an examination have been described. Each clinician will formulate his own method of utilizing these components. The following are suggested steps in a practical approach to the diagnosis of lameness. Evaluate the conditions under which restraint of the animal is possible (corral, free stall, stanchion barn). Assess the temperament, physical strength, and general amenability of the animal to be examined. If the animal is at liberty, its movements can be studied. If it is standing quietly observe it. If the animal is tied up and it seems feasible to examine the animal while it is walking, observe the animal and take its history while it is still tied up. Aggressive and wild animals should be observed, but little beneficial information will be gained from a regional approach to evaluating movement at this stage. Even if the seat of lameness seems to be in the proximal limb, it is mandatory in every case to examine the hooves and interdigital space at some stage during the evaluation. However, if the probability of proximal lameness is high, the clinician may proceed to make a complete evaluation, which would include studying the quality of movement before checking that the distal extremity is normal in every way. If the clinician decides to proceed with an examination of the digital region and is unable to locate a causative lesion in the extremity, he would then evaluate movement and manipulate and palpate the proximal limb. If no cause of the lameness becomes apparent, the nerve supply to the extremity would then be blocked.(ABSTRACT TRUNCATED AT 250 WORDS)


Using the Rasch model to quantify the causal effect of test instructions.
This experimental study was designed to quantify, by means of the Rasch model (RM), the effects of three instruction/scoring conditions on student measures and on the reliability of an achievement multiple-choice test in a field context. Examinees performed the test in one of three conditions which differed only in the instructions provided. Predictions regarding performance indicators were fulfilled, and the expected differences in reliability favoring omission-inducing instructions did appear. This difference in reliability was found for both Rasch and raw data and thus it can be concluded that the fact that results from previous studies failed to corroborate this prediction must have been due to the lack of important consequences of test scores for the students. The RM has served to neatly quantify the differences between instructions promoting guessing and instructions promoting omission under uncertainty, showing that the recommendation to omit is not only educationally but also psychometrically sound.


Wave induced coastal flooding along the southwest coast of India during tropical cyclone Tauktae.
The coastal flood during the tropical cyclone Tauktae, 2021, at Chellanam coast, Kerala, India, has invited wide attention as the wave overtopping severely affected coastal properties and livelihood. We used a combination of WAVEWATCHIII and XBeach to study the coastal inundation during high waves. The effect of low-frequency waves and the rise in the coastal water level due to wave setup caused the inundation at Chellanam, even during low tide with negligible surge height. Wave setup raised the water level at the coast with steep slopes to more than 0.6 m and peaked during low tide, facilitating wave breaking at the nearshore region. The coastal regions adjacent to these steep slopes were subjected to severe inundation. The combined effect of long and short waves over wave setup formed extreme wave runups that flooded inland areas. At gently sloping beaches, the longwave component dominated and overtopped the seawalls and damaged households along the shoreline. The study emphasizes the importance of longwave and wave setup and its interaction with nearshore bathymetry during the high wave. The present study shall lead to the development of a coastal inundation prediction system for the low-lying hot spots using the combination of WAVEWATCHIII and XBeach models.


Liquid chromatographic determination of homovanillic acid, 5-hydroxyindoleacetic acid and probenecid levels in human cerebrospinal fluid during probenecid test.
Homovanillic acid and 5-hydroxyindoleacetic acid, respectively the major metabolites of the central neurotransmitters dopamine and serotonin, are present in human lumbar cerebrospinal fluid, and their determination during the probenecid test is used to study brain monoamine abnormalities in man. We developed a high performance liquid chromatographic technique coupled with electrochemical detection which allows the simultaneous determination of homovanillic acid and 5-hydroxyindoleacetic acid on 10 microliter of human lumbar cerebrospinal fluid. Another liquid chromatographic technique coupled with ultra-violet detection was applied to the measurement of probenecid after injection of 10 microliter of native cerebrospinal fluid. The main advantages of these two techniques are their simplicity, sensitivity, rapidity (five cerebrospinal fluid samples are analysed within one hour) and their good day-to-day reproducibility . These methods were applied to probenecid tests performed in several neurological patients.


Overexpression of zmm28 increases maize grain yield in the field.
Increasing maize grain yield has been a major focus of both plant breeding and genetic engineering to meet the global demand for food, feed, and industrial uses. We report that increasing and extending expression of a maize MADS-box transcription factor gene, zmm28, under the control of a moderate-constitutive maize promoter, results in maize plants with increased plant growth, photosynthesis capacity, and nitrogen utilization. Molecular and biochemical characterization of zmm28 transgenic plants demonstrated that their enhanced agronomic traits are associated with elevated plant carbon assimilation, nitrogen utilization, and plant growth. Overall, these positive attributes are associated with a significant increase in grain yield relative to wild-type controls that is consistent across years, environments, and elite germplasm backgrounds.


Bacterial adherence and contamination during radiographic processing.
Oral fluids are potential contaminants of radiographic processors. This investigation measured bacterial contamination in a radiographic processing room during times of high and low clinical activity and processing effects on five types of microorganisms. Cultures in the clinical setting, during high and low activity, were taken by brain-heart infusion agar plates placed near automatic processors. Site samples were taken of entrance, developer, fixer, water, and exit surfaces. Measurements of processing effects were accomplished by intentional contamination of films run in series through an automatic processor. Site samples were again taken of the processor. In the clinical setting colony-forming units increased with activity. Radiographic processing after intentional contamination decreased colony-forming units on films, but they increased for all processing solutions. Bacteria on radiographic film survived processing. Although processing procedures significantly reduce the number of bacteria on films, the potential for contamination and cross-contamination remains.


Sugar alters the level of serum insulin and plasma glucose and the serum cortisol:DHEAS ratio in female migraine sufferers.
Early work has highlighted that a large percentage of migraineurs may have an altered glucidic methabolis due to carbohydrate-induced hyperinsulinism. The aim of this study was to assess the effect of sucrose on biomarkers of energy metabolism and utilization in migraineous females. A total of 16 participants (8 = Migraine, 8 = Non-migraine) at the mid-point of their menstrual cycle underwent a 15-h fast prior to ingesting 75 g sucrose dissolved in 175 g water. Blood sampling for the assessment of serum insulin, serum cortisol and serum dehydroepiandrosterone sulfate (DHEAS) and plasma glucose was conducted upon arrival at 09:00 h and then at regular 15-min intervals across a 150-min experimental period. The results showed a significant alteration in serum insulin and plasma glucose following sucrose ingestion in the migraine and non-migraine groups. In addition, significant group differences were observed in the level of serum insulin, serum DHEAS, and the cortisol:DHEAS ratio with migraine participants on average recording a higher sucrose-induced serum insulin level and lower DHEAS level and cortisol:DHEAS ratio when group data was compared. It was concluded that while sucrose consumption may potentiate serum insulin in migraineurs this does not result in the development of sucrose-induced hypoglycemia in migraine or non-migraine participants.


[Use of psoralen plus UV-A therapy in the autonomous community of Valencia, Spain].
Photochemotherapy with 8-methoxypsoralen and long-wavelength UV-A (PUVA) has been extensively used for the treatment of various skin diseases since its approval in 1982 by the US Food and Drug Administration. A retrospective study was performed of patients treated with PUVA, including topical and systemic treatment, over a period of 14 years. All patients were treated using a standard PUVA therapy regimen. A total of 877 patients were analyzed for the period 1982 to 1996. Forty-one skin diseases were treated, including 341 cases of psoriasis and 71 cutaneous T-cell lymphomas. The aim of the study was to describe the characteristics of the patients treated with PUVA therapy during that period and compare the results with those observed in other regions. Although PUVA therapy is widely used in a large number of countries for the treatment of various skin diseases, few studies have described the characteristics of the patients and the differences in the parameters of PUVA according to the disease.


Characteristics and source apportionment of atmospheric volatile organic compounds in Beijing, China.
This paper focused on VOCs and their source apportionment in urban Beijing. Our monitoring measured 52 VOCs in July 2014 and January 2015. The concentration of VOCs was in the range of 14.5~95.2 ppb in July and 2.1~93.1 ppb in January, with the top five compounds of toluene (10.7%), ethane (6.9%), ethylene (6.3%), n-butane (5.7%), and propane (5.6%) in July and ethylene (14.7%), n-butane (14.2%), ethane (9.6%), propylene (8.0%), toluene (7.9%), and benzene (6.9%) in January. The ratio of VOCs to CO reached 0.059 in July and 0.022 in January on average. These differences implied a potential seasonal difference in the VOC source contribution. Then, we conducted a source apportionment study based on 21 major VOCs and CO by using probabilistic matrix factorization (PMF) receptor model. According to the similarity between the PMF analysis profiles and the known source profiles, combustion sources, petrochemical industry sources, solvent utilization sources, and gasoline evaporation sources were identified. The correlation coefficient (R) between the PMF analysis profile and the source profile reached 0.68~0.87 in July and 0.53~0.92 in January. The better apportionment performance in July was mainly due to the use of intensive VOC observations at a 3-h resolution. When we conducted another PMF source apportionment for July based on 12-h resolved concentration input, the R values decreased to 0.47~0.73. Thus, the PMF model depends heavily on the sample number of concentration inputs, and intensive observation is more propitious. Our PMF apportionment results showed that combustion sources, petrochemical industry, solvent utilization, gasoline evaporation, and other sources contributed ambient VOCs in Beijing urban areas of 13.7 ppb, 5.1 ppb, 7.7 ppb, 12.8 ppb, and 3.3 ppb in July and 13.2 ppb, 2.0 ppb, 5.7 ppb, 6.6 ppb, and 1.0 ppb in January, respectively, on a monthly average. These apportionment results match well with the 2013 VOC emission inventory calculated by this study, but also presented significant seasonal differences in the petrochemical industry and gasoline evaporation, in which VOC emissions strongly respond to environmental temperature.


Transcription Factor EGR1 Regulates the Expression of the Clock Gene PER2 under IL-4 Stimulation in Human Keratinocytes.
PER2 is a core circadian clock gene that regulates circadian rhythms. IL-4 plays a critical role in the pathogenesis of skin inflammation, including atopic dermatitis. IL-4 enhances PER2 expression, suggesting a relationship between inflammation and the circadian clock. However, little is known about the molecular and cellular mechanisms regulating PER2 expression by inflammatory cytokines. This study showed that transcription factor EGR1 interacted with the PER2 promoter and promoted IL-4‒induced transcriptional activation of the PER2, as revealed by promoter‒reporter assay, electrophoretic mobility shift assay, DNA affinity precipitation assay, and chromatin immunoprecipitation analysis. We also found that IL-4 can use both MAPK and Jak signaling pathways to induce EGR1-mediated PER2 expression, and c-Jun N-terminal kinase 1/2 can augment IL-4‒induced activation of the Jak‒signal transducer and activator of transcription 3 pathway. Consistently, Per2 expression was reduced in dinitrochlorobenzene-induced atopic dermatitis‒like skin lesions in Egr1‒/‒ mice compared with that in Egr1+/+ mice. In addition, using a real-time bioluminescence assay, we observed that EGR1 is required for rhythmic oscillation of PER2 expression under IL-4 exposure. These findings provide further insight into the role of EGR1 in regulating PER2 expression in impaired circadian rhythm in skin inflammation.


Spontaneous passage of glass beads from the canine gallbladder: facilitation by sphincterotomy.
To investigate the mechanism by which ablation of the sphincter of Oddi prevents gallstone formation, we assessed passage of glass beads out of the gallbladders of dogs with sphincterotomy and sham sphincterotomy. One month after bead implantation, dogs with an intact sphincter passed 52%, 26%, 22%, 10%, 0%, and 0% of beads with diameters of 2, 3, 4, 5, 6, and 8 mm, respectively. For the same respective bead diameters, dogs with a sphincterotomy passed 90%, 90%, 88%, 75%, 75%, and 42% of beads (p less than 0.05 for all bead diameters). No beads were in the common bile duct of any dog. In separate dogs studied by cholescintigraphy, sphincterotomy significantly increased gallbladder ejection fraction from 0.46 to 0.76 (p less than 0.01). In addition, sphincterotomy significantly lowered resting gallbladder volume from 24.4 to 15.8 ml (p less than 0.025) and lowered cholecystokinin-stimulated gallbladder volume from 13.3 to 5.9 ml (p less than 0.025). These data indicate that even with an intact sphincter, small solids can pass from the gallbladder and into the duodenum. Sphincterotomy facilitates passage of solids, apparently by general improvement in gallbladder emptying. Facilitated passage of crystals, microliths, or small stones seems the most likely explanation for prevention of gallstone formation by sphincterotomy.


Preferential adsorption of pentachlorophenol from chlorophenols-containing wastewater using N-doped ordered mesoporous carbon.
Preferential removal of pentachlorophenol (PCP) from chlorophenols-containing wastewater has been attracted more attentions in wastewater treatment, since it is one of the most toxic pollutants. The adsorbent of N-doped ordered mesoporous carbon (M-OMC) with high BET surface area of 1901 m(2)/g, large pore volume of 1.64 cm(3)/g and uniform pore size of 3.45 nm has been successfully synthesized via evaporation-induced self-assembly (EISA) method. The effects of solution pH, pore structure of adsorbent and their surface chemical properties on PCP adsorption by M-OMC were investigated in comparison with ordered mesoporous carbon (OMC), and much higher PCP adsorption capacities of M-OMC were obtained. The significantly preferential adsorption of PCP was achieved in the treatment of tri-component wastewater including PCP, p-chlorophenol (CP) and 2.4.6-trichlorophenol (TCP), and its adsorption process well fitted the pseudo-second-order kinetics model and the Langmuir isotherm. The initial sorption rate of PCP was 103.5 μmol/(g/min), which was 2.97 times of TCP. It may be attributed to the intensification of π-π interaction between PCP and M-OMC with the nitrogen functional groups. Therefore, M-OMC is promising for removal of PCP in the adsorption pretreatment of chlorophenols-containing wastewater.


Technique for applying porcelain to pontics of fixed partial dentures.
A technique has been presented to be used to make pontics for metal-ceramic fixed partial dentures. Use of the porcelain/wax system is advantageous, especially when multiple pontics are involved.


[Anicety Renier (1804-1877) and his significance for development of Druskienkai].
The aim of his paper is to present the silhouette of Anicety Renier (1804-1877), an eminent physician, patriot, deportee. His professional achievements as a physician of health resort in Druskienkai are presented widely. He was the author of publications on this health resort.


Fabricating Amorphous g-C3N4/ZrO2 Photocatalysts by One-Step Pyrolysis for Solar-Driven Ambient Ammonia Synthesis.
Solar-driven nitrogen fixation remains a significant challenge. Graphitic carbon nitride (g-C3N4) is considered as a promising visible light photocatalyst. However, the photocatalytic performance of g-C3N4 is unsatisfactory because of the random transfer of charge carriers in the plane and the low activation efficiency of the reactants. Herein, amorphous ZrO2 was used as a robust cocatalyst of g-C3N4 to increase the NH3 production activity. The g-C3N4/ZrO2 lamellar composites were constructed by a simple one-step pyrolysis of the deep eutectic solvent ZrOCl2·8H2O/urea. The optimum NH4+ yield could reach as high as 1446 μmol·L-1·h-1 at 30 wt % ZrO2 in the g-C3N4/ZrO2 composites, with an apparent quantum efficiency over 2.14% at 400 nm. It is 7.9 times that of pristine g-C3N4 and 27.5 times that of ZrO2. The introduction of amorphous ZrO2 restrained the hydrogen generation, and the amorphous ZrO2 and g-C3N4 together contribute to the rapid photoproduced electron transfer of less electron-hole pair recombination.


Metallothionein-mediated cisplatin resistance in human ovarian carcinoma cells.
We have determined the ability of two human ovarian carcinoma cells to over-express metallothioneins (MTs) and the subsequent effect this elevation has on DDP cytotoxicity. Cells of 2008 and COLO 316 human ovarian carcinomas that were resistant to CdCl2 were obtained by stepwise selection and chronic culture in CdCl2 and ZnCl2. The 2008/MT cells were 3.2-fold resistant to CdCl2 and 4.1-fold resistant to DDP; they had 23-fold elevated MTs. The COLO/MT cells were 1.2-fold resistant to CdCl2 and 3.3-fold resistant to DDP, and they had 9-fold elevated MTs. Glutathione (GSH) was also elevated in the Cd-resistant sublines. However, four times more intracellular thiols were contributed by the MTs than by the GSH. 2008 and 2008/MT cells were examined in more detail to elucidate the mechanism of DDP resistance. Depletion of GSH with D,L-buthionine-S,R-sulfoximine (BSO) had no effect on the sensitivity of these cells to either CdCl2 or DDP. Uptake of [195m Pt]DDP in 2008 and 2008/MT cells was identical. Fractionation of the cytosol from [195mPt]DDP-exposed cells on Sephadex G-75 revealed that 17% of the total cellular Pt in 2008/MT cells was associated with the MT fraction, as against 4% in the parent 2008 cells. This increase corresponded to a concomitant loss of Pt from the particulate fraction. Fractionation of 2008 cells selected with DDP (2008/DDP cells) indicated that elevated MTs did not contribute to the DDP resistance of these cells. Only 2% of the total cellular Pt was in the MT fraction in 2008/DDP cells. These results showed that elevation of MTs may be one mechanism of DDP resistance in ovarian carcinoma; however, in vitro selection with DDP does not trigger this mechanism.


A provincial program of blood conservation: The Ontario Transfusion Coordinators (ONTraC).
Although often life-saving, blood transfusions are associated with significant risk to the patient and escalating costs to the blood system and hospital. Transfusions are often given unnecessarily. Blood conservation represents the use of alternatives to transfusion. The ONTraC program attempts to enhance transfusion practice outside the blood transfusion laboratory, promote blood conservation in surgery patients, and reduce allogeneic red cell use. In the first such large scale program, funding was obtained from the Ontario MOHLTC for a Transfusion Coordinator in 23 Ontario hospitals selected based on blood utilization and geography. At specific time periods, detailed anonymized information was collected in a defined number of all consecutive patients admitted for the three designated surgical procedures: knee arthroplasty (N=approximately 1200 at each time point), abdominal aortic aneurysm (AAA; N=300 at each time) and coronary artery bypass graft (CABG) surgery (N=300 at each time point). Considerable inter-institutional variation was observed in the proportion of patients and amount of blood transfused. At the 12 month analysis, most, although not all, hospitals had decreased use of allogeneic blood and there was an overall 24% reduction in blood use in patients undergoing knee surgery, 14% in AAA and 23% in CABG. In addition to reduction in proportion of patients transfused, transfused patients received fewer units of allogeneic blood. Patients who did not receive allogeneic transfusions had significantly lower postoperative infection rates (p<0.05) and length of stay (p<0.0001); multivariate analysis showed that allogeneic transfusion was an independent predictor of increased length of stay. Eighteen-month analysis indicates even greater reduction in allogeneic transfusion. The main measures of blood conservation employed were preoperative autologous donation and education, with recent increasing use of erythropoietin and the cell saver. These measures have been demonstrated to be very effective in avoiding allogeneic transfusion. The ONTraC have become leaders locally, nationally and internationally in blood conservation. The reduction in allogeneic transfusion associated with the implementation of the ONTraC program represents important savings in costs associated with blood components, hospital stay and work in transfusion laboratories and nursing units, as well as enhancing patient satisfaction and safety.


Evaluation of five methods for determining low-density lipoprotein cholesterol (LDL-C) in hemodialysis patients(1).
Current recommendations for the management of dyslipidemia are largely based on the concentration of LDL-C. Most clinical laboratories estimate the concentration of LDL-C by the recommended routine method, the equation of Friedewald, in specimens from fasting subjects and with TG concentrations < 4.52 mmol/L. Because of the limitations of the Friedewald calculation, direct methods for an accurate quantification of LDL-C are needed. In the present study we evaluated the accuracy of the following 5 different procedures for LDL-C in 98 patients on hemodialysis: the Friedewald equation, where LDL-C is calculated from HDL-C, measured either by the precipitation procedure with dextran sulfate-Mg(2+) (Method 1), or by a direct HDL-C assay (Method 2), the Direct LDL assay (Method 3), the homogeneous N-geneous LDL assay (Method 4) and the calculated LDL-C values deriving from the ApoB based equation: 0.41TC - 0.32TG + 1.70ApoB - 0.27, (Clin Chem 1997;43:808-815) (Method 5). All five LDL-C methods were found to be in good agreement with ultracentrifugation/dextran sulfate-Mg(2+) precipitation with the coefficients of correlation of the assays to ranging between 0.93-0.95. However, significant differences in the mean values and biases vs. the reference method were observed. The Friedewald equation and the Direct assay were less affected by high LDL-C levels, and they presented higher sensitivity and higher negative predictive value. The N-geneous assay and the ApoB derived calculation were less affected by high triglyceride levels, and they presented higher specificity and higher positive predictive value. At the diagnostic LDL-C level of 3.37 mmol/L, both Friedewald calculations correctly classified 82/92 patients; Direct assay 86/98; N-geneous assay 88/98; and ApoB derived calculation 88/98. At the diagnostic LDL-C level of 2.98 mmol/L, Friedewald calculations (Method 1 and Method 2) correctly classified 82/92 and 81/92 patients, respectively; Direct assay (LDL-3) 87/98; N-geneous assay (LDL-4) 91/98; and ApoB derived calculation (LDL-5) 91/98. Among hemodialysis patients, who commonly present "average" LDL-C concentrations and high TG levels, the N-geneous assay and the apoB derived calculation seem to yield more acceptable results for the estimation of LDL-C.


Antitumor effect of a calmodulin antagonist on the growth of solid sarcoma-180.
A calmodulin antagonist, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), proved to have antitumor activity against solid Sarcoma-180. In particular, intraperitoneal injection of W-7 strongly inhibited the growth of solid Sarcoma-180 in mice given a daily dose, 10 mg/kg for 10 consecutive days. A chlorine deficient analogue, N-(6-aminohexyl)-1-naphthalenesulfonamide (W-5), which interacts weakly with calmodulin, had no antitumor activity against solid Sarcoma-180, in a dose similar to that of W-7. Thus, W-7, a calmodulin antagonist may inhibit the growth of solid Sarcoma-180 by modulation of the Ca2+-calmodulin-dependent processes.


Metastases after mastectomy for ductal carcinoma in situ of the male breast; A case report.
and importance: Male breast cancer is a rare entity. Ductal carcinoma in situ (DCIS), constituting 10% of all male breast cancer, is confined within the breast ducts and lobules, rarely metastasizing and even less so after mastectomy. A 71 years old male with no history of trauma presented with pain, swelling, and deformity of the left arm. He had continuous back pain for 6 months and a history of mastectomy of the right breast. Fracture of shaft of the left humerus was detected on X-ray. Computed tomography (CT) showed multiple vertebral metastases later confirmed to be metastasized from the breast by biopsy. Tumor cells were progesterone receptor (PR) positive, estrogen receptor (ER) negative, and human epidermal growth factor receptor 2 (HER2) negative. The fracture was treated and the patient was kept on Tamoxifen. On follow-up after four months, the patient is doing well with relief of back pain. Despite mastectomy and the histopathological diagnosis of pure DCIS, distant metastases can occur even in absence of locoregional recurrence. Therefore, the aggressive phenotype of DCIS rather than diagnostic or treatment variables can be thought to bring worse outcome in the form of metastases. Early hormonal status identification and hormone therapy could result in a better outcome. Skeletal metastases should be strongly suspected in patients presenting with bone pain and having a history of DCIS of the breast, even after mastectomy. Even though distant metastasis after mastectomy is rare, regular follow-up and surveillance is necessary.


Endowing homodimeric carbamoyltransferase GdmN with iterative functions through structural characterization and mechanistic studies.
Iterative enzymes, which catalyze sequential reactions, have the potential to improve the atom economy and diversity of industrial enzymatic processes. Redesigning one-step enzymes to be iterative biocatalysts could further enhance these processes. Carbamoyltransferases (CTases) catalyze carbamoylation, an important modification for the bioactivity of many secondary metabolites with pharmaceutical applications. To generate an iterative CTase, we determine the X-ray structure of GdmN, a one-step CTase involved in ansamycin biosynthesis. GdmN forms a face-to-face homodimer through unusual C-terminal domains, a previously unknown functional form for CTases. Structural determination of GdmN complexed with multiple intermediates elucidates the carbamoylation process and identifies key binding residues within a spacious substrate-binding pocket. Further structural and computational analyses enable multi-site enzyme engineering, resulting in an iterative CTase with the capacity for successive 7-O and 3-O carbamoylations. Our findings reveal a subclade of the CTase family and exemplify the potential of protein engineering for generating iterative enzymes.


Inhibition of IRF3 expression reduces TGF-β1-induced proliferation of hepatic stellate cells.
Therapeutic management of liver fibrosis remains an unresolved clinical problem. Activation of hepatic stellate cell (HSC) is a pivotal event in the progression of liver fibrosis. Recent reports have showed that inhibition of activated HSC proliferation contributes to the reversal of liver fibrosis. Interferon regulatory factor 3 (IRF3), one member of the interferon regulatory factor (IRF) family, is recently proven to be a critical modulator in cardiac fibrosis. And accumulating evidence demonstrated that IRF3 plays a crucial role in liver diseases, such as hepatic steatosis, liver inflammation, and alcoholic liver injury. However, the understanding of the function of IRF3 in liver fibrosis remains limited. Our results identified the role of IRF3 in regulating human HSC (LX-2 cell) cell proliferation and apoptosis. The present study indicated that the expression of IRF3 was significantly increased in HSCs in response to TGF-β1 stimulation. Moreover, a stable and unlimited source of human HSC, the LX-2 cell line, transfected with IRF3-siRNA significantly decreases the expression level of type I collagen (Col1a1) and α-smooth muscle actin (α-SMA) in activated LX-2 cells. On the contrary, overexpression of IRF3 gives rise to an upregulation of Col1a1 and α-SMA in LX-2 cells, and further promoted HSC proliferation. Moreover, the inhibition of IRF3 significantly suppressed TGF-β1-induced HSC proliferation and increased its apoptosis. Of note, the present study indicated IRF3 may regulate LX-2 cell proliferation by via AKT signaling pathway. In summary, these observations suggest IRF3 may function as a novel regulator to modulate TGF-β1-induced LX-2 proliferation, at least in part, via AKT signaling pathway.


Ultradian cycles in mice: definitions and links with REMS and NREMS.
Sleep can be organized in two quite different ways across homeothermic species: either in one block (monophasic), or in several bouts across the 24 h (polyphasic). Yet, the main relationships between variables, as well as regulating mechanisms, are likely to be similar. Correlations and theories on sleep regulation should thus be examined on both types of sleepers. In previous studies on monophasic humans, we have shown preferential links between the number of ultradian cycles and the rapid eye movement sleep (REMS) time, rather than with its counterpart non-rapid eye movement sleep (NREMS). Here, the sleep of 26 polyphasic mice was examined, both to better describe the NREMS distribution, which is far more complex than in humans, and to replicate the analyses performed on humans. As in humans, the strongest links with the number of cycles were with REMS. Links were not significant with NREMS taken as a whole, although positive correlations were found with the NREMS immediately preceding REMS episodes and inversely significant with the residue. This convergence between monophasic and polyphasic patterns supports the central role played by REMS in sleep alternation.


Variations in heavy metal contamination of stream water and groundwater affected by an abandoned lead-zinc mine in Korea.
This study evaluated variations in heavy metal contamination of stream waters and groundwaters affected by an abandoned lead-zinc mine, where a rockfill dam for water storage will be built 11 km downstream. For these purposes, a total of 10 rounds of stream and groundwater samplings and subsequent chemical analyses were performed during 2002-2003. Results of an exploratory investigation of stream waters in 2000 indicated substantial contamination with heavy metals including zinc (Zn), iron (Fe) and arsenic (As) for at least 6 km downstream from the mine. Stream waters near the mine showed metal contamination as high as arsenic (As) 8,923 microg L(-1), copper (Cu) 616 microg L(-1), cadmium (Cd) 223 microg L(-1) and lead (Pb) 10,590 microg L(-1), which greatly exceeded the Korean stream water guidelines. Remediation focused on the mine tailing piles largely improved the stream water qualities. However, there have still been quality problems for the waters containing relatively high concentrations of As (6-174 microg L(-1)), Cd (1-46 microg L(-1)) and Pb (2-26 microg L(-1)). Rainfall infiltration into the mine tailing piles resulted in an increase of heavy metals in the stream waters due to direct discharge of waste effluent, while dilution of the contaminated stream waters improved the water quality due to mixing with metal free rain waters. Levels of As, Cu and chromium (Cr) largely decreased after heavy rain but that of Pb was rather elevated. The stream waters were characterized by high concentrations of calcium (Ca) and sulfate (SO(4)), which were derived from dissolution and leaching of carbonate and sulfide minerals. It was observed that the proportions of Ca and SO(4) increased while those of bicarbonate (HCO(3)) and sodium and potassium (Na+K) decreased after a light rainfall event. Most interestingly, the reverse was generally detected for the groundwaters. The zinc, being the metal mined, was the most dominant heavy metal in the groundwaters (1758-10,550 microg L(-1)) near the mine, which far exceeded the Korean standard of 1000 microg L(-1) for drinking water. The decreases in the heavy metals contents in the groundwaters associated with reduced rainfall were quite different from the increases observed for the stream waters, which is not clearly understood at this time and warrants further investigation.


Central Catheter-related Bloodstream Infection Rates in Children on Home Parenteral Nutrition.
Reliable venous access is prerequisite for patients receiving long-term parenteral nutrition (PN). Central venous catheters are, however, an important risk factor for the development of potentially lethal complications, including catheter-related bloodstream infection (CRBSI). We have retrospectively assessed the incidence of CRBSIs in children on long-term PN who were treated at the Children's Hospital Zagreb from January 2011 until January 2019 and the cost effectiveness of the use of taurolidine line locks in children at home PN (HPN). During this period 48 children received long-term PN and 24 children were discharged to HPN. The rate of CRBSI 1.15/1000 catheter days in total; 2.35/1000 days in the hospital; and 0.48/1000 days at home. If taurolidine line lock was used every day of PN for children on HPN total costs would exceed existing CRBSI treatment costs >5 times.


Welcome to the era of CKD and the eGFR.
Estimating glomerular filtration rate using a simplified formula will lead to a vast increase in detection of chronic kidney disease in Australia.


Phosphatidic acid-producing enzymes regulating the synaptic vesicle cycle: Role for PLD?
In cortical and hippocampal neurons of the mammalian brain, the synaptic vesicle cycle is a series of steps that tightly regulate exo- and endocytosis of vesicles. Many proteins contribute to this regulation, but lipids have recently emerged as critical regulators as well. Of all the many lipid signaling molecules, phosphatidic acid is important to the physical processes of membrane fusion. Therefore, the lipid-metabolizing enzymes that produce phosphatidic acid are vital to the regulation of the cycle. Our lab is particularly interested in the potential regulatory mechanisms and neuronal roles of two phosphatidic acid-producing enzymes: diacylglycerol kinase theta (DGKθ) and phospholipase D (PLD). We recently discovered a regulatory role of DGKθ on evoked endocytosis (Goldschmidt et al., 2016). In addition to this enzyme, studies implicate PLD1 in neurotransmission, although its precise role is of some debate. Altogether, the production of phosphatidic acid by these enzymes offer an interesting and novel pathway for the regulation of the synaptic vesicle cycle.


Virtual anthropology: Forensic applications to cranial skeletal remains from the Spanish Civil War.
Biological and forensic anthropologists face limitations while studying skeletal remains altered by taphonomic alterations and perimortem trauma, such as in remains from the Spanish Civil War. However, virtual anthropology techniques can optimize the information inferred from fragmented and deformed remains by generating and restoring three-dimensional bone models. We applied a low-cost 3D modelling methodology based on photogrammetry to develop novel forensic applications of virtual 3D skull reconstruction, assembly, restoration and ancestry estimation. Crania and mandible fragments from five Spanish Civil War victims were reconstructed with high accuracy, and only one cranium could not be assembled due to extensive bone loss. Virtual mirroring successfully restored reconstructed crania, producing 3D models with reduced deformation and perimortem trauma. High correlation between traditional and virtual craniofacial measurements confirmed that 3D models are suitable for forensic applications. Craniometric databases of world-wide and Spanish populations were used to assess the potential of discriminant analysis to estimate population ancestry. Our protocol correctly estimated the continental origin of 86.7 % of 15 crania of known origin, and despite low morphological differentiation within European populations, correctly identified 54.5 % as Spanish and 27.3 % of them with high posterior probabilities. Two restored crania from the Civil War were estimated as Spanish, and one as a non-Spanish European. Results were not conclusive for one cranium and did not confirm previous archeological hypotheses. Overall, our research shows the potential to assess the presence of foreign volunteers in the Spanish Civil War and highlights the added value of 3D-virtual techniques in forensic anthropology.


Role of cyclic AMP in cardiac beta-adrenoceptor desensitization: studies using prenalterol and inhibitors of phosphodiesterase.
We examined the effects of prolonged exposure of cardiac cells in primary culture to the partial beta-adrenoceptor agonist prenalterol and inhibitors of phosphodiesterase on their subsequent ability to increase intracellular cyclic AMP during a 5-min exposure to 50 microM isoprenaline (receptor responsiveness). Although prenalterol possesses only 7% of the agonist activity of isoprenaline on adenylate cyclase, it induces extensive beta-adrenoceptor desensitization. Three hours after exposing the cells to 1 microM prenalterol, beta-adrenoceptor responsiveness was reduced by 40% (p less than 0.05), whereas after 12 h the reduction averaged 55%. Prolonging the incubation time to 48 h had no further effect on the magnitude of receptor desensitization. The magnitude of the desensitization was concentration dependent. On exposure of cells to 10(-8) M prenalterol for 16 h, receptor responsiveness was reduced by 19%, and at concentrations of 1 microM and higher responsiveness was reduced by 60% (p less than 0.01). Receptor desensitization appeared to be due to an inability of receptors to activate adenylate cyclase as well as to receptor loss. To investigate if beta-adrenoceptor desensitization as well as receptor loss could be mediated by cyclic AMP, the cells were exposed for 16 h to inhibitors of phosphodiesterase. Exposure of cells to the phosphodiesterase inhibitor isobutylmethylxanthine (0.1 mM) (which increased intracellular cyclic AMP by between 50 and 150%) also induced receptor desensitization. The reduction in receptor responsiveness averaged 62% (p less than 0.01). The loss in responsiveness could be accounted for by an inability of receptors to activate adenylate cyclase as well as by receptor loss.(ABSTRACT TRUNCATED AT 250 WORDS)


The estimation of recruitment from stage frequency data.
A general relationship is derived for the area of the sample curve for a given stage in the life-cycle, in terms of the mean survival rate and duration and the number entering the stage. The influence of recruitment, development and mortality patterns on the area of the sample curve is investigated. The results indicate a simple method of analysing stage frequency data to produce a best estimate of the number entering each stage. A test example, using simulated data, is provided with a discussion of the method in relation to other techniques based on the area of the sample curve.


Comparative evaluation of the impact of the COVID-19 lockdown on perinatal experience: A prospective multicentre study.
To compare in the early postpartum the perinatal experience during a COVID-19 related lockdown ('lockdown' group) and a pandemic control group subject to looser restrictions. This national multicentre prospective cohort study took place in four French maternity units. Women were recruited during the postpartum stay for the lockdown and pandemic control groups, according to their enrolment period. Both faced the same labour and delivery restrictions but only the pandemic control group could have a postpartum visitor. The primary outcome was the perinatal experience during childbirth, assessed by the Labour Agentry Scale (LAS) self-administered questionnaire, completed before discharge. The study included 596 women and analysed 571 of them: 260 in the lockdown group and 311 in the pandemic control group. The mean LAS score was lower in the lockdown group (161.1 ± 26.8, 95% confidence interval [CI] 157.8-164.3 versus 163.3 ± 24.0, 95% CI 160.6-166.0; P = 0.289). In multivariable analysis, the LAS score was lower in the lockdown group (-6.2 points, P = 0.009), in women with caesarean (-21.6 points, P < 0.001) versus spontaneous deliveries, and among women financially impacted by the lockdown (-6.4 points, P = 0.007) or who experienced restrictions during childbirth (-8.1 points, P < 0.001). The LAS score rose with the prenatal care quality score (P < 0.001). The perinatal experience was more negatively affected by lockdown restrictions than by the looser pandemic restrictions for controls, but mode of delivery remained the main factor influencing this experience.


Cyclic nucleotide distribution in identified layers of suprafused rabbit retinas.
To assess the effects of pharmacologic agents on metabolite levels in identified retinal layers we have devised an apparatus that allows suprafusion of everted eye cups (vitreal surface exposed) or isolated retinas (photoreceptor surface exposed), recording of electroretinograms (ERG), and rapid freezing in a configuration which permits obtaining frozen tangential sections of the retina. Samples from identified layers of the freeze-dried sections can be subsequently weighed and processed for biochemistry by Lowry methods (Lowry and Passonneau , 1972), or cyclic nucleotides can be extracted from weighed samples and assayed. To test the usefulness of the apparatus, we have compared levels of guanosine 3',5'-monophosphate (cGMP) and adenosine 3',5'-monophosphate (cAMP), measured in defined layers of suprafused rabbit retinas, to concentrations reported for retinal layers from eyes removed from light- and dark-adapted rabbits and quickly frozen. For most layers the concentrations of cyclic nucleotides were similar in both the suprafused and whole eye preparations. In experimental applications we suprafused dark-adapted eye cups and retinas with a physiological saline whose free calcium level was sharply reduced by chelation, and observed that cGMP levels were elevated only in those retinal layers containing photoreceptor portions. Ethyleneglycol bis-(beta-amino-ethyl ether)N,N'-tetraacetic acid (EGTA) (3 mM) yielded a threefold increase in cGMP in 10 min when applied to the photoreceptor surface of isolated retinas. However, when the vitreal surfaces of everted eye cups were suprafused with calcium-chelated medium for 10 min, a threefold increase in cGMP required 20 mM EGTA, 3 mM EGTA being ineffective. In another study, vitreal surfaces of eye cups or photoreceptor surfaces of retinas were suprafused with physiological saline containing 3 mM 3-isobutyl-1-methylxanthine (IBMX) in order to alter cAMP retinal levels. In both cases, eightfold increases in cAMP were measured in the inner nuclear and inner plexiform layers, while three- to fourfold increases occurred in the outer nuclear and outer plexiform layers. This agent also caused a large increase in cAMP in the pigment epithelium.


Comprehensive multi-omics analysis reveals m7G-related signature for evaluating prognosis and immunotherapy efficacy in osteosarcoma.
Osteosarcoma is one of the most prevalent bone malignancies with a poor prognosis. The N7-methylguanosine (m7G) modification facilitates the modification of RNA structure and function tightly associated with cancer. Nonetheless, there is a lack of joint exploration of the relationship between m7G methylation and immune status in osteosarcoma. With the support of TARGET and GEO databases, we performed consensus clustering to characterize molecular subtypes based on m7G regulators in all osteosarcoma patients. The least absolute shrinkage and selection operator (LASSO) method, Cox regression, and receiver operating characteristic (ROC) curves were employed to construct and validate m7G-related prognostic features and derived risk scores. In addition, GSVA, ssGSEA, CIBERSORT, ESTIMATE, and gene set enrichment analysis were conducted to characterize biological pathways and immune landscapes. We explored the relationship between risk scores and drug sensitivity, immune checkpoints, and human leukocyte antigens by correlation analysis. Finally, the roles of EIF4E3 in cell function were verified through external experiments. Two molecular isoforms based on regulator genes were identified, which presented significant discrepancies in terms of survival and activated pathways. Moreover, the six m7G regulators most associated with prognosis in osteosarcoma patients were identified as independent predictors for the construction of prognostic signature. The model was well stabilized and outperformed traditional clinicopathological features to reliably predict 3-year (AUC = 0.787) and 5-year (AUC = 0.790) survival in osteosarcoma cohorts. Patients with increased risk scores had a poorer prognosis, higher tumor purity, lower checkpoint gene expression, and were in an immunosuppressive microenvironment. Furthermore, enhanced expression of EIF4E3 indicated a favorable prognosis and affected the biological behavior of osteosarcoma cells. We identified six prognostic relevant m7G modulators that may provide valuable indicators for the estimation of overall survival and the corresponding immune landscape in patients with osteosarcoma.


Structure-activity relationship study of hypoxia-activated prodrugs for proteoglycan-targeted chemotherapy in chondrosarcoma.
Due to an abundant chondrogenic, poorly vascularized and particularly hypoxic extracellular matrix, chondrosarcoma, a malignant cartilaginous tumour, is chemo- and radio-resistant. Surgical resection with wide margins remains the mainstay of treatment. To address the lack of therapy, our strategy aims to increase anticancer drugs targeting and delivery in the tumour, by leveraging specific chondrosarcoma hallmarks: an extensive cartilaginous extracellular matrix, namely the high negative fixed charge density and severe chronic hypoxia. A dual targeted therapy for chondrosarcoma was investigated by conjugation of a hypoxia-activated prodrug (HAP) to quaternary ammonium (QA) functions which exhibit a high affinity for polyanionic sites of proteoglycans (PGs), the major components of the chondrosarcoma extracellular matrix. Based on preclinical results, an imidazole prodrug, ICF05016, was identified and provided the basis for a lead optimization study. A series of 27 QA-phosphoramide mustard conjugates, differing by the type of QA function and the length of the alkyl linker, was yielded by a common multi-step sequence involving phosphorylation of a key 2-nitroimidazole alcohol. Then, a screening was realized by surface plasmon resonance technology to assess biomolecular interactions between QA derivatives and aggrecan, the most abundant PG in chondrosarcoma. Results revealed that affinity depends more on the type of QA function, than on the linker length. Moreover, the presence of a benzyl group enhanced affinity to aggrecan. Twelve compounds were shortlisted and evaluated for antiproliferative activity (i.e., growth inhibiting concentration 50), under normoxic and hypoxic conditions using the human extraskeletal myeloid chondrosarcoma cell line (HEMC-SS). For all prodrugs, hypoxic selectivity was maintained and even increased, compared with the lead. From this study, compound 31f emerged as the most effective PG-targeted HAPs with a dissociation constant of 2.10 μM in the SPR experiment, a hypoxia cytotoxicity ratio of 24 and an efficient reductive cleavage under chemical and enzymatic conditions.


A minimally invasive method to prevent postlaryngectomy major pharyngocutaneous fistula using infrahyoid myofascial flap.
To prevent postoperative pharyngocutaneous fistula (PCF) after total (pharyngo)laryngectomy, simultaneous coverage of pharyngeal anastomosis with vascularised flaps such as pectoralis major muscle, anterolateral thigh or radial forearm, has been reported to be effective. As an alternative to the invasive methods using distant flaps, we used the infrahyoid myofascial flap (IHMFF), which was harvested from the same operation field of (pharyngo)laryngectomy, for covering the site of pharyngeal anastomosis. Herein, we describe the safety and effectiveness of our minimally invasive method for preventing PCF. Eleven patients who were at a high risk of developing PCF due to previous chemoradiotherapy underwent simultaneous coverage of pharyngeal anastomosis with IHMFF after total (pharyngo)laryngectomy. The incidence of PCF and the rate of major fistula requiring surgical closure were determined, and the results were compared with the control group (23 patients without IHMFF cover after laryngectomy). PCF developed in 2 of the 11 patients (18.2%). The fistulae of these two patients were closed conservatively and did not require additional surgery. PCF developed in 6 of 23 patients (26.1%) in patients without IHMFF cover. All the six patients with fistula required additional closure surgery. The incidence of PCF did not differ in patients with or without IHMFF cover (Fisher's exact probability test; p=0.939, NS). However, the rate of major PCF requiring surgical closure was significantly lower in patients with IHMFF cover (Fisher's exact probability test; p=0.036<0.05). For (pharyngo)laryngectomy patients, IHMFF cover is a minimally invasive method that can prevent major PCF.


Pd(II)-mediated triad multilayers with zinc tetrapyridylporphyrin and pyridine-functionalized nano-TiO2 as linkers: assembly, characterization, and photocatalytic properties.
Triad hybrid multilayers containing the light sensitizers of zinc tetrapyridylporphyrin (ZnTPyP) and pyridine-functionalized TiO(2) (TiO(2)-Py) nanoparticles were constructed on substrate surfaces with the use of Pd(II) ions as the connectors using the layer-by-layer (LBL) method. The assembly process was monitored using ultraviolet-visible (UV-vis) absorption and X-ray photoelectron spectra as well as scanning electron microscopy and atomic force microscopy. The content of the pyridine substituents in the TiO(2)-Py nanocomposites was about 2% (w/w). The Soret absorption band of ZnTPyP was 24 nm red-shifted in the hybrid multilayers due to a strong intermolecular electronic coupling interaction among porphyrin macrocycles or porphyrin macrocycle/TiO(2)-Py nanoparticles. The average surface density of each ZnTPyP layer was about 1.4 × 10(-10) mol/cm(2). Aggregation of the small TiO(2)-Py nanoparticles to larger domains with sizes up to hundreds of nanometers occurred in the hybrid multilayers; however, such an aggregation behavior was weaker than that in the solutions. The quartz substrate modified with the as-prepared Pd/ZnTPyP/Pd/TiO(2)-Py triad hybrid multilayers was used as a heterogeneous photocatalyst for the degradation of methyl orange (MO) under irradiation (λ > 420 nm) at room temperature with a catalytic efficiency of about 1.3 × 10(-3) MO/ZnTPyP·s. Without the use of the filter, the catalytic efficiency increased because both ZnTPyP and TiO(2)-Py nanocomposites acted as the light sensitizers. It is suggested that the present heterogeneous catalyst has the advantages of facile separation, high stability, structural controllability on the molecular and nanoscale level, and good recyclability.


Gender differences in murine pulmonary responses elicited by cellulose nanocrystals.
Cellulose-based materials have been used for centuries to manufacture different goods derived from forestry and agricultural sources. In the growing field of nanocellulose applications, its uniquely engineered properties are instrumental for inventive products coming to competitive markets. Due to their high aspect ratio and stiffness, it is speculated that cellulose nanocrystals (CNC) may cause similar pulmonary toxicity as carbon nanotubes and asbestos, thus posing a potential negative impact on public health and the environment. The present study was undertaken to investigate the pulmonary outcomes induced by repeated exposure to respirable CNC. C57BL/6 female and male mice were exposed by pharyngeal aspiration to CNC (40 μg/mouse) 2 times a week for 3 weeks. Several biochemical endpoints and pathophysiological outcomes along with gene expression changes were evaluated and compared in the lungs of male and female mice. Exposure to respirable CNC caused pulmonary inflammation and damage, induced oxidative stress, elevated TGF-β and collagen levels in lung, and impaired pulmonary functions. Notably, these effects were markedly more pronounced in females compared to male mice. Moreover, sex differences in responses to pulmonary exposure to CNC were also detected at the level of global mRNA expression as well as in inflammatory cytokine/chemokine activity. Overall, our results indicate that there are considerable differences in responses to respirable CNC based on gender with a higher pulmonary toxicity observed in female mice.


Alteration of the structure and function of photosystem I by Pb2+.
The toxic effects of Pb(2+) on photosynthetic electron transport were studied in photosystem I (PSI) submembrane fractions isolated from spinach. Structural and spectroscopic analysis using FTIR, fluorescence and X-ray photoelectron spectroscopy (XPS) showed that Pb(2+) binds with proteins via oxygen and nitrogen atoms with an overall binding constant of KPb-PSI=4.9×10(3) (±0.2) M(-1) and the number of bound Pb(2+) cation was 0.9 per PSI complex. Pb(2+) binding altered the protein conformation indicating a partial protein destabilization. Electron transport and P700 photooxidation/reduction measurements showed that the interaction of Pb(2+) cations with PSI produced a donor side limitation of electron transport presumably due to Pb(2+) binding to or in the vicinity of plastocyanin.


Fine and Coarse Particulate Matter Exposures and Associations with Acute Cardiac Events among Participants in a Telemedicine Service: A Case-Crossover Study.
Subclinical cardiovascular changes have been associated with ambient particulate matter (PM) exposures within hours. Although the U.S. Environmental Protection Agency continues to look for additional evidence of effects associated with sub-daily PM exposure, this information is still limited because most studies of clinical events have lacked data on the onset time of symptoms to assess rapid increased risk. Our objective was to investigate associations between sub-daily exposures to PM and acute cardiac events using telemedicine data. We conducted a case-crossover study among telemedicine participants [Formula: see text] of age who called a service center for cardiac-related symptoms and were transferred to a hospital in Tel Aviv and Haifa, Israel (2002-2013). Ambient [Formula: see text] and [Formula: see text] measured by monitors located in each city during the hours before the patient called with symptoms were compared with matched control periods. We investigated the sensitivity of these associations to more accurate symptom onset time and greater certainty of diagnosis. We captured 12,661 calls from 7,617 subscribers experiencing ischemic (19%), arrhythmic (31%), or nonspecific (49%) cardiac events. PM concentrations were associated with small increases in the odds of cardiac events. For example, odds ratios for any cardiac event in association with a [Formula: see text] increase in 6-h and 24-h average [Formula: see text] were 1.008 [95% confidence interval (CI): 0.998, 1.018] and 1.006 (95% CI: 0.995, 1.018), respectively, and for [Formula: see text] were 1.003 (95% CI: 1.001, 1.006) and 1.003 (95% CI: 1.000, 1.007), respectively. Associations were stronger when using exposures matched to the call time rather than calendar date and for events with higher certainty of the diagnosis. Our analysis of telemedicine data suggests that risks of cardiac events in telemedicine participants [Formula: see text] of age may increase within hours of PM exposures. https://doi.org/10.1289/EHP2596.