Promising small molecule anti-fibrotic agents: Newly developed or repositioned drugs targeting myofibroblast transdifferentiation.
Fibrosis occurs in all organs and tissues except the brain, and its progression leads to dysfunction of affected organs. Fibrosis-induced organ dysfunction results from the loss of elasticity, strength, and functionality of tissues due to the extracellular matrix secreted by myofibroblasts that express smooth muscle-type actin as a marker. Myofibroblasts, which play a major role in fibrosis, were once thought to originate exclusively from activated fibroblasts; however, it is now clear that myofibroblasts are diverse in origin, from epithelial cells, endothelial cells, adipocytes, macrophages, and other cells. Fibrosis of vital organs, such as the heart, lungs, kidneys, and liver, is a serious chronic disease that ultimately leads to death. Currently, anti-cancer drugs have made remarkable progress, as evidenced by the development of many molecular-targeted drugs, and are making a significant contribution to improving the prognosis of cancer treatment. However, the development of anti-fibrotic agents, which also play an important role in prognosis, has lagged. In this review, the current knowledge regarding myofibroblasts is summarized, with particular attention given to their origin and transdifferentiation signaling pathways (e.g., TGF-β, Wnt/β-catenin, YAP/TAZ and AMPK signaling pathways). The development of new small molecule anti-fibrotic agents and the repositioning of existing drugs targeting myofibroblast transdifferentiation are discussed.


Design and discovery of novel quinazolinedione-based redox modulators as therapies for pancreatic cancer.
Altered cellular bioenergetics and oxidative stress are emerging hallmarks of most cancers including pancreatic cancer. Elevated levels of intrinsic reactive oxygen species (ROS) in tumors make them more susceptible to exogenously induced oxidative stress. Excessive oxidative insults overwhelm their adaptive antioxidant capacity and trigger ROS-mediated cell death. Recently, we have discovered a novel class of quinazolinediones that exert their cytotoxic effects by modulating ROS-mediated signaling. Cytotoxic potential was determined by colorimetric and colony formation assays. An XF24 Extracellular Flux Analyzer, and colorimetric and fluorescent techniques were used to assess the bioenergetics and oxidative stress effects, respectively. Mechanism was determined by Western blots. Compound 3a (6-[(2-acetylphenyl)amino]quinazoline-5,8-dione) was identified through a medium throughput screen of ~1000 highly diverse in-house compounds and chemotherapeutic agents for their ability to alter cellular bioenergetics. Further structural optimizations led to the discovery of a more potent analog, 3b (6-[(3-acetylphenyl)amino]quinazoline-5,8-dione) that displayed anti-proliferative activities in low micromolar range in both drug-sensitive and drug-resistant cancer cells. Treatment with 3b causes Akt activation resulting in increased cellular oxygen consumption and oxidative stress in pancreatic cancer cells. Moreover, oxidative stress induced by 3b promoted activation of stress kinases (p38/JNK) resulting in cancer cell death. Treatment with antioxidants was able to reduce cell death confirming ROS-mediated cytotoxicity. In conclusion, our novel quinazolinediones are promising lead compounds that selectively induce ROS-mediated cell death in cancer cells and warrant further preclinical studies. Since 3b (6-[(3-acetylphenyl)amino]quinazoline-5,8-dione) exerts Akt-dependent ROS-mediated cell death, it might provide potential therapeutic options for chemoresistant and Akt-overexpressing cancers.


Use of an inexpensive magnifier with light source in the diagnosis of trichiasis among community-based case finders in Nigeria.
Trichiasis case finding is challenging in many trachoma endemic countries. We sought to determine how the use of an inexpensive hand-held magnifier with light source aids in the diagnosis of people with trichiasis. We undertook a study on the use of the "Wilson", an inexpensive hand-held magnifier with light source between two groups of trained case finders: community drug distributors (CDD) volunteers and non-health volunteers. Each group was randomly allocated to be provided a Wilson or not. We compared the number of people with suspected trichiasis and the proportion of true cases referred by each group. Focus group discussions explored the perceptions of the case finders that used the Wilson on its utility in the diagnosis of trichiasis and its ease of use. Among the 121 case finders, 61 were CDD volunteers (31 were provided the Wilson) and 60 were non-health worker volunteers (30 were provided the Wilson). Case finders that used the Wilson were 4.57 times (95% CI 2.46-8.47) more likely to have patients presenting confirmed as trichiasis compared to those not using the Wilson. Case finders that used the Wilson felt the instrument was easy to use, enlarged the eyelashes and made diagnoses of trichiasis easier. In Nigeria it appears that the Wilson is easy to handle and use among trichiasis case finders enhances the diagnosis of trichiasis.


Structural and dynamical aspects of HIV-1 protease and its role in drug resistance.
Acquired immunodeficiency syndrome (AIDS) caused by the retrovirus human immunodeficiency virus (HIV) has become a major epidemic afflicting mankind. The Joint United Nations Program on HIV/AIDS (UNAIDS) projection shows the existence of millions of AIDS patients at the end of 2012. All the Food and Drug Administration (FDA)-approved drugs are getting ineffective due to resistance offered by the mutation-prone HIV. Hence, there is an urgent need for developing new drugs with greater potential. HIV life cycle is controlled by the activities of its essential proteins like glycoproteins (gp41 and gp120), HIV reverse transcriptase (HIV-RT), HIV integrase (HIV-IN), and HIV-1 protease (HIV-pr). This chapter focuses on the protein HIV-pr, which is important for the cleavage of Gag and Gag-Pol polyproteins to form mature, structural, and functional virions. The conformation and dynamics of the protein HIV-pr play a pivotal role in ligand binding and the catalytic process, which is affected by the rapid point mutations and various physiological parameters. The effect of the mutations and the varied simulation protocols on conformational dynamics and drug resistance of HIV-pr is discussed.


Immune Functions Alterations Due to Racing Stress in Thoroughbred Horses.
Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed in thirty Thoroughbred racehorses, which were divided in two groups. The samples were taken right after races (moment 0 d), during rest periods (-11 d, +1 d, +3 d), and after training (-8, +2, +5). In both groups, the phagocytosis showed a decrease in percentage and intensity immediately after the race when comparing samples collected during rest or training periods. In the mean values of oxidative burst on samples collected immediately after the race, group I animals demonstrated a decrease (524.2 ± 248.9) when compared with those samples collected in other moments. No significant differences were found between the results of different moments regarding the apoptotic cells and lymphoproliferation assays. The mean values of serum cortisol levels were increased immediately after racing. There was an increase in the percentage of neutrophils found immediately after the race. It was possible to conclude that, although a transient reduction was found in the number of neutrophils, the horses' adaptive function was not affected.


On circuit functionality in Boolean networks.
It has been proved, for several classes of continuous and discrete dynamical systems, that the presence of a positive (resp. negative) circuit in the interaction graph of a system is a necessary condition for the presence of multiple stable states (resp. a cyclic attractor). A positive (resp. negative) circuit is said to be functional when it "generates" several stable states (resp. a cyclic attractor). However, there are no definite mathematical frameworks translating the underlying meaning of "generates." Focusing on Boolean networks, we recall and propose some definitions concerning the notion of functionality along with associated mathematical results.


DAMPs are able to skew CD4+ T cell subsets and increase the inflammatory profile in pregnant women with preeclampsia.
Preeclampsia (PE) is characterized by abnormal activation of the immune system. The intense systemic inflammatory reaction, could be related to the presence of molecules released after cell stress or death, that are capable of inducing inflammation and are known as damage-associated molecular patterns (DAMP). This study evaluated the profile of T cells through the analysis of transcription factors and the cytokines produced after culture with or without DAMPs: heat shock protein 70 (Hsp70), hyaluronan (HA) and monosodium urate (MSU). Twenty pregnant women with PE, 20 normotensive (NT) pregnant women and 20 non-pregnant (NP) women were studied. The results showed polarization toward Th1/Th17 and a decrease in Th2/Treg profiles in preeclamptic women associated with elevated levels of TNF, IFN-γ, and IL-17A and diminished levels of TGF-β1 and IL-10 when compared to the normotensive group. In addition, preeclamptic women had a higher percentage of cells co-expressing T-bet/GATA-3 and T-bet/RORγt and fewer T-bet/FoxP3 cells when compared to normotensive group. MSU induced an increase in IFN-γ and IL-22 in all studied groups. MSU, HA, and Hsp70 induced significant higher production of TNF in the PE and NP groups. The PE group showed elevated levels of TGF-β1 after incubation with MSU, HA, and Hsp70, whereas HA and Hsp70 decreased TGF-β1 production in NT group. The results suggest that these alarmins may play a role in the activation of innate and adaptive immune systems by skewing CD4 + T cells and increasing the release of inflammatory cytokines, thereby contributing to the pathogenesis of this important syndrome.


Patients with Low National Institutes of Health Stroke Scale Scores Have Longer Door-to-Needle Times: Analysis of a Telestroke Network.
The benefits of intravenous tissue-type plasminogen activator (IV-tPA) in acute ischemic stroke (AIS) are time dependent. Because emergency rooms quickly initiate a stroke alert with more severe symptoms, we hypothesized that patients with lower National Institutes of Health Stroke Scale (NIHSS) scores, indicating a less severe stroke, would have longer door-to-needle (DTN) times compared to patients with higher NIHSS scores. Data obtained from the 19-hospital Providence Stroke Registry were used to identify AIS patients who received IV-tPA within 4.5 hours of last-known-well. NIHSS scores were obtained prior to tPA administration at the time of emergency department presentation and categorized as low-NIHSS (score = 0-5) or high-NIHSS (score = 6-42) strokes. Median DTN times were collected for both groups as the primary outcome variable. Linear mixed-effects regression models were used to assess the effect of NIHSS scores on DTN and its 2 components: door-to-CT (DCT) and CT-to-needle (CTN) times. We identified 692 AIS patients who received IV-tPA within 4.5 hours of last-known-well, with 198 patients presenting with low-NIHSS strokes and 494 patients with high-NIHSS strokes. In multivariable analysis, median DTN time was estimated to be 18% higher for low-NIHSS strokes than high-NIHSS strokes after adjusting for covariates (P < .001). Median DCT times were also higher for low-NIHSS (19 minutes) compared to high-NIHSS (11 minutes) strokes after adjusting for covariates (P < .001), whereas CTN times were unchanged (P = .055). In AIS patients receiving IV-tPA in a telestroke network, lower NIHSS scores were associated with longer DTN and DCT times.


Glycogen synthase kinase-3β inhibitor ameliorates imbalance of connexin 43 in an acute kidney injury model.
This study was designed to evaluate whether glycogen synthase kinase-3β (GSK-3β) inhibitor, 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8) induced the the expression of connexin 43 (Cx43) to protect against renal ischemia-reperfusion (I/R) injury (RI/RI) in rats. Rats were subjected to 45 min ischemia followed 2 h reperfusion with TDZD-8 (1 mg/kg) for 5 min prior to reperfusion. The results indicated that TDZD-8 improved the recovery of renal function, reduced oxidative stress and inflammation injury, and upregulated the expression of (Cx43) as compared to I/R group. Therefore, our study demonstrated that TDZD-8 provided a protection to the kidney against I/R injury in rats through inducing the expression of (Cx43).


Hippocampal dopamine receptors modulate cFos expression in the rat nucleus accumbens evoked by chemical stimulation of the ventral hippocampus.
Recently, we have shown that D1 and D2 receptors in the ventral hippocampus (VH) modulate both the locomotor activation and the increase in dopamine (DA) levels in the rat nucleus accumbens (NAc) induced by NMDA stimulation of the VH. In the present study we analyze the possible role of VH D1 and D2 receptors in the modulation of the cFos expression in NAc (core and shell subregions) and in dorsal striatum. This was assessed by immunohistochemical analysis of cFos expression in the rat brains after retro-dialysis application of NMDA (50mM, 10 min) into VH, in absence and in presence of either the D1/D5 receptor antagonist SCH 23390 (100 and 250 microM, 60 min) or the D2 receptor antagonist raclopride (100 and 250 microM, 60 min). NMDA induced a robust increase in the cFos expression in the NAc shell, both in the ipsilateral and contralateral side. No statistically significant increases were observed in the NAc core and in the dorsal striatum. Simultaneous application of SCH 23390 and NMDA into the VH attenuated the NMDA-evoked cFos expression in NAc shell. In contrast, raclopride had no significant effect. Our present results show that the NMDA receptor mediated effects in the VH require D1 receptors and suggest that DA in VH strongly modulates the excitatory outputs from this brain area.


Vascular response to 1 week of sleep restriction in healthy subjects. A metabolic response?
Sleep loss may induce endothelial dysfunction, a key factor in cardiovascular risk. We examined the endothelial function during one week of sleep restriction and a recovery period (from 3-to-13 days) in healthy subjects, and its link to autonomic, inflammatory and/or endocrine responses. 12 men were followed at baseline (B1, 8-h sleep), after 2 (SR2) and 6 (SR6) days of SR (4-h sleep: 02:00-06:00) and after 1 (R1) and 12 (R12) recovery nights (8h sleep). At 10:00, we assessed changes in: arm cutaneous vascular conductance (CVC) induced by local application of methacholine (MCh), cathodal current (CIV) and heat (44°C), finger CVC and skin temperature (Tfi) during local cold exposure (5°C, 20-min) and passive recovery (22°C, 20-min). Blood samples were collected at 08:00. Compared with baseline (B1), MCh and heat-induced maximal CVC values (CVC peak) were decreased at SR6 and R1. No effect of SR was observed for Tfi and CVC during immersion whereas these values were lower during passive recovery on SR6 and R1. From SR2 to R12, plasma concentrations of insulin, IGF-1 (total and free) and MCP-1 were significantly increased while those of testosterone and prolactin were decreased. Whole-blood blood mRNA concentrations of TNF-α and IL-1β were higher than B1. No changes in noradrenaline concentrations, heart rate and blood pressure were observed. These results demonstrate that SR reduces endothelial-dependent vasodilatation and local tolerance to cold. This endothelial dysfunction is independent of blood pressure and sympathetic activity but associated with inflammatory and metabolic pathway responses (ClinicalTrials-NCT01989741).


Effect of biochar addition on the dynamics of antibiotic resistant bacteria during the pig manure composting.
In present study, the taxonomic variation of antibiotic resistant bacteria (ARB) in pig manure (PM) composting with coconut shell biochar (CSB) and bamboo biochar (BB) addition was investigated. The experiment was divided into three treatments: T1 (as control or without biochar amendment), T2 was added 10% coconut shell biochar and T3 supplemented with 10% bamboo biochar. The initial feed stock were properly homogenized using a mechanical crusher. PM and wheat straw (WS) were mixed in a 5: 1 dry weight ratio to adjust the initial carbon/nitrogen ratio 25:1, bulk density to ~0.5 (kg/L) and ~60% moisture content, respectively. This experiment was lasted for 42 days. The results indicated the bacterial communities in the three treatments were more different in terms of relative abundance and diversity of dominant bacteria. The control group had the highest abundance of Kingdome bacteria. The changes in ARB was noticed by variation in the relative abundances of Actinobacteria, Proteobacteria, Firmicutes and Bacteroidota. At the end of composting (on day 42), the total RAs of ARB at the class, order, and family levels were considerably reduced in T2 and T3 by ~35.78-38.75%, 36.42-40.63% and 45.82-47.70%, respectively. But in T1 was decreased by 6.16-8.62%, 7.93-8.72% and 8.70-10.15%, as compared with the day 0 sample. However, the CSB was much more effective to reduce 55 to 60% of ARB than T3 or BB applied treatment has 40 to 42% ARB reduction, while control has certainly very less RAa of ARB reduction. Finally, the biochar amendment was significant approach to mitigate the total ARB abundance in compost and it's further used for organic farming purposes.


Lack of an association between angiotensin-converting enzyme gene insertion/deletion polymorphism and ischaemic stroke.
Numerous factors have been reported to influence the pathogenesis of stroke. The angiotensin I-converting enzyme (ACE) gene is a candidate gene for atherosclerotic-related diseases. In the present study, the association between the polymorphism of the ACE gene and ischaemic stroke was investigated. Using polymerase chain reaction techniques, 100 patients (48 males, age 69.3 +/- 9.7 years) with cerebral infarction and 100 age- and sex-matched controls were divided into the following three ACE genotypes [deletion (D) and insertion (I)]: II, ID and DD. There was no evidence of any association between the ACE gene polymorphism and the presence of ischaemic stroke (odds ratio 0.874, 95% confidence interval 0.386-1.973). The DD genotype in the human ACE gene does not appear to be a risk factor for ischaemic stroke. Further evaluation in a larger population study is required to examine the possibility of an increased risk of ischaemic stroke in DD homozygotes.


Corrigendum: Global tendency and frontiers of research on myopia from 1900 to 2020: A bibliometrics analysis.
[This corrects the article DOI: 10.3389/fpubh.2022.846601.].


Effects of a sunscreen formulation on albino hairless mice: a morphological approach.
The purpose of the study was to evaluate the effects of a sunscreen formulation on the skin of albino hairless mice subjected to simulated solar light (SSL) in terms of morphological changes. Young adult albino hairless mice HRS/J (n = 36) were used as an experimental model for determining skin photoaging changes. Mice were irradiated with SSL, and the sunscreen (estimated SPF 30, PF-UVA) was obtained from the Pharmacy College/UFRJ, Brazil. The animals were divided into four groups: non-treated (G1), radiation only (G2), sunscreen-treated (G3) and vehicle + radiation (G4). Animals from groups G2, G3 and G4 were irradiated weekly (5 weeks), with no immobilization. One week after the final exposure, the dorsal skin was observed using a dermatoscopic camera. Biopsies were analyzed in order to quantify neovascularization and to evaluate histological aspects of the skin. Neovascularization was also evaluated with immunohistochemical reactions for the Von Willebrand factor. Animals from G2 displayed classical morphological changes denoting skin photoaging: thickening of the epidermis, increased dermal cellularity, follicular keratosis, sebaceous gland hyperplasia, and angiogenesis. Animals from groups G3 and G1 displayed similar morphological profiles, without these changes. Animals from group G4 showed more morphological changes than group G2, emphasizing the relative importance of the putative photosensitizing components present in the vehicle formulation. The extent of the morphological skin changes suggested that the sunscreen formulation was effective against SSL, and showed the importance of assessing the phototoxicity of vehicle formulations.


Parametric hazard rate models for long-term sickness absence.
In research on the time to onset of sickness absence and the duration of sickness absence episodes, Cox proportional hazard models are in common use. However, parametric models are to be preferred when time in itself is considered as independent variable. This study compares parametric hazard rate models for the onset of long-term sickness absence and return to work. Prospective cohort study on sickness absence with four follow-up years of 53,830 employees working in the private sector in the Netherlands. The time to onset of long-term (>6 weeks) sickness absence and return to work were modelled by parametric hazard rate models. The exponential parametric model with a constant hazard rate most accurately described the time to onset of long-term sickness absence. Gompertz-Makeham models with monotonically declining hazard rates best described return to work. Parametric models offer more possibilities than commonly used models for time-dependent processes as sickness absence and return to work. However, the advantages of parametric models above Cox models apply mainly for return to work and less for onset of long-term sickness absence.


Microbial Aerosols: New Diagnostic Specimens for Pulmonary Infections.
Pulmonary infections are important causes of global morbidity and mortality, but diagnostics are often limited by the ability to collect specimens easily, safely, and in a cost-effective manner. We review recent advances in the collection of infectious aerosols from patients with TB and with influenza. Although this research has been focused on assessing the infectious potential of such patients, we propose that these methods have the potential to lead to the use of patient-generated microbial aerosols as noninvasive diagnostic tests of disease and tests of infectiousness.


A novel PD-1/PD-L1 pathway molecular typing-related signature for predicting prognosis and the tumor microenvironment in breast cancer.
Currently, the development of breast cancer immunotherapy based on the PD-1/PD-L1 pathway is relatively slow, and the specific mechanism affecting the immunotherapy efficacy in breast cancer is still unclear. Weighted correlation network analysis (WGCNA) and the negative matrix factorization (NMF) were used to distinguish subtypes related to the PD-1/PD-L1 pathway in breast cancer. Then univariate Cox, least absolute shrinkage and selection operator (LASSO), and multivariate Cox regression were used to construct the prognostic signature. A nomogram was established based on the signature. The relationship between the signature gene IFNG and breast cancer tumor microenvironment was analyzed. Four PD-1/PD-L1 pathway-related subtypes were distinguished. A prognostic signature related to PD-1/PD-L1 pathway typing was constructed to evaluate breast cancer's clinical characteristics and tumor microenvironment. The nomogram based on the RiskScore could be used to accurately predict breast cancer patients' 1-year, 3-year, and 5-year survival probability. The expression of IFNG was positively correlated with CD8+ T cell infiltration in the breast cancer tumor microenvironment. A prognostic signature is constructed based on the PD-1/PD-L1 pathway typing in breast cancer, which can guide the precise treatment of breast cancer. The signature gene IFNG is positively related to CD8+ T cell infiltration in breast cancer.


Four synaptic vesicle-specific proteins: identification by monoclonal antibodies and distribution in the nervous tissue and the adrenal medulla.
Synaptic vesicles from the guinea-pig cerebrum were isolated and administered to mice for the production of monoclonal antibodies (MAb). Four vesicle-associated proteins in the guinea-pig nervous tissue were specifically and differentially recognized by MAbs thus obtained. These proteins had molecular weights of 30,000, 36,000, 38,000 and 65,000 Da and were named SVPs (synaptic vesicle proteins) 30, 36, 38 and 65, respectively. Immunohistochemistry demonstrated that all SVPs were localized in the synaptic regions throughout the central nervous system and in the adrenal medulla. Nerve terminals in skeletal muscle, smooth muscle and sympathetic ganglion contained SVPs 36 and 38. Immunoelectron microscopy of the cerebellar cortex confirmed the localization of SVPs in the synaptic vesicles and the adjacent membranes of the presynaptic nerve terminals. Fractionation of the cerebral tissue and treatment with various agents showed that SVPs were localized in the synaptic vesicles and the synaptic plasma membrane and that SVPs were integrated within the membrane and liberated only after solubilization of the membrane.


Primary lens extraction for glaucoma management: A review article.
Recently, primary lens extraction alone gained more acceptance as an alternative surgical approach for glaucoma management. This view was supported by the advances in phacoemulsification and intraocular lenses with greater safety and visual recovery, in addition to a substantial reduction of intraocular pressure and deepening of the anterior chamber and filtration angle. The decrease in IOP after cataract surgery in primary open-angle glaucoma (POAG) is mild, less predictable, related to baseline levels, and may return to presurgical values after an initial period of reduction. Therefore, the IOP-lowering effect of primary cataract extraction in POAG may be insufficient to achieve adequate IOP control. The IOP reduction after lens extraction is consistently greater in eyes with primary angle closure glaucoma (PACG) than in eyes with POAG. Primary lens extraction in acute PACG eliminates, or at least, reduces the risk of recurrence of acute attacks and deepens the anterior chamber and widens the angle which reduces the risk of progression of peripheral anterior synechiae and development of chronic PACG. Primary lens extraction may be more preferable to glaucoma incisional surgery in mild to moderate PACG eyes with appositional angle closure. The decision to do lens extraction as a primary treatment for glaucoma should be individualized based upon several factors other than the effect on IOP. These factors include patients' characteristics, surgeons' skills and preferences, status of glaucoma control, type of cataract and intraocular lens implanted, and potential harm of laser treatment for late capsular opacification and fibrosis.


Integrative taxonomy of anisakid nematodes in stranded cetaceans from Brazilian waters: an update on parasite's hosts and geographical records.
Anisakidae are represented mainly by species of Anisakis Dujardin, 1845, Pseudoterranova Krabbe, 1878, and Contracaecum Railliet and Henry, 1913. Integrative taxonomy, based on morphological and genetic approaches, can provide a precise identification of species, increasing the knowledge of their systematics, biology, and evolution. In this study, integrative taxonomy was applied in order to identify, at generic and/or specific taxonomic level, 245 anisakids from 63 cetaceans of 12 different species, which stranded in the Brazilian coast. Parasites from nine hosts were preserved by freezing and, from 54 hosts, were fixed in 70% ethanol (n = 43) or alcohol-formalin-acetic (n = 11) for a period varying from 6 to 19 years. The morphological analysis allowed the identification of 171 specimens at genus level: 120 as Anisakis sp. clade I, 47 as Anisakis sp. clade II, and four as Pseudoterranova sp. From those 15, specimens were identified at species level, seven as Anisakis typica, seven as Anisakis paggiae and one as Anisakis brevispiculata. Moreover, 74 specimens were identified as belonging to Anisakidae family. The genetic analysis based on cox2 gene allowed the identification/confirmation of Anisakis typica (n = 33), Anisakis paggiae (n = 1), and for the first time in the Brazilian coast, Anisakis brevispiculata (n = 4) and Anisakis ziphidarum (n = 1). Additionally, a probably new Anisakis species, genetically close to A. paggiae, is reported infecting a dwarf sperm whale. The results clearly indicated that the preservation of nematodes influenced significantly on the level of taxonomic definition, in both morphological and, mainly, genetic analyses. Pseudoterranova sp. was identified for the first time in the spinner dolphin, Stenella longirostris. The detection of nine A. paggiae specimens, in three Kogia breviceps animals, substantiates the austral occurrence of this species. The identification of A. brevispiculata in Kogia sima and K. breviceps confirms Kogiidae whales as their most common hosts. Anisakis typica was corroborated as a generalist species, the most widely distributed in the Brazilian coast, infecting a broad variety of Delphinidae and Kogiidae, with two new geographical records in Stenella frontalis and Lagenodelphis hosei. The study showed a new scenery of parasite diversity of marine anisakids, updating the Anisakis species distribution.


Trimethyl chitosan coated palladium nanoparticles as a photothermal agent and its in vitro evaluation in 2D and 3D model of breast cancer cells.
The potential of palladium has been scantily explored in biomedical applications. In the present study, palladium nanoparticles (PdNPs) were synthesized and were successfully coated with trimethyl-chitosan (TMC) to improve their biocompatibility. Coating with TMC improved the nanoparticle accumulation in MDAMB231 breast cancer cells, compared to nanoparticles coated with native chitosan. The TMC coated palladium nanoparticles (TMC/PdNPs) exhibited good biocompatibility and physiological stability, as compared to the plain(uncoated) PdNPs. TMC coated PdNPs resulted in photothermal therapeutic effect, when irradiated with a near-infrared (NIR) laser having the wavelength of 808-nm. The TMC/PdNPs resulted in good cytotoxic effect upon laser treatment in both, 2D monolayers and 3D spheroids of MDAMB231 cells, the latter mimicking the tumor microenvironment. These results clearly indicated that TMC/PdNPs acted as ideal photothermal agents for anti-cancer therapy in combination with a non-invasive near-infrared laser.


Prevalence of asymptomatic and electrically undetectable intracardiac inside-out abrasion in silicon-coated Riata® and Riata® ST implantable cardioverter-defibrillator leads.
Recently, several cases of symptomatic and/or electrically detectable intracardiac inside-out abrasions in silicon-coated Riata® and Riata® ST leads have been described. However, the prevalence in asymptomatic patients with unremarkable implantable cardioverter defibrillator (ICD) interrogation is unknown. The aim of this study was to determine the prevalence of asymptomatic and electrically undetectable intracardiac inside-out abrasion in silicon-coated Riata® and Riata® ST leads. All 52 patients with an active silicone-coated Riata® and Riata® ST lead followed up in our outpatient clinic were scheduled for a premature ICD interrogation and a biplane chest radiograph. When an intracardiac inside-out abrasion was suspected, this finding was confirmed by fluoroscopy. Mean time since implantation was 71 ± 18 months. An intracardiac inside-out abrasion was confirmed by fluoroscopy in 6 patients (11.5%). Mean time from lead implantation to detection of intracardiac inside-out abrasion was 79 ± 14 months. In all patients with an intracardiac inside-out abrasion, ICD interrogation showed normal and stable electrical parameters. Retrospectively, in 4 of these 6 patients, a coronary angiography performed 25 ± 18 months before diagnosis of intracardiac inside-out abrasion already showed the defect. Despite undetected intracardiac inside-out abrasion, 2 of these 4 patients experienced adequate antitachycardia pacing and ICD-shocks. ICD leads were replaced in all 6 patients. The prevalence of asymptomatic intracardiac inside-out abrasion in silicon-coated Riata® and Riata® ST leads is higher than 10% when assessed by fluoroscopy, and most intracardiac inside-out abrasions are not detectable by ICD interrogation.


Mytilus edulis hemolymph contains pro-opiomelanocortin: LPS and morphine stimulate differential processing.
Mytilus edulis hemolymph contains mammalian-like proopiomelanocortin (POMC). The 20 kDa protein was purified by high pressure gel permeation chromatography, anti-adrenocorticotropin (ACTH)-affinity column and reverse-phase HPLC. The amino acid sequence determination was by Edman degradation, enzymatic treatments and Western blot analysis. Of the six peptides found in this opioid precursor, methionine-enkephalin, gamma-melanocyte stimulating hormone (MSH), alpha-MSH and ACTH exhibited 100, 80, 85 and 74% sequence identity, respectively, with the mammalian counterparts. beta-Endorphin and gamma-LPH exhibited only 25 and 10% sequence identity. Dibasic amino acid residues were found at the C-terminus of MSH and ACTH, indicating cleavage sites. The alpha-MSH is flanked at the C-terminus by Gly-Lys-Lys, representing an amidation signal. ACTH and CLIP (80% sequence identity) are also C-terminally flanked by dibasic amino acid residues. Furthermore, morphine, in a dose-dependent manner, increased the hemolymph levels of alpha-MSH and ACTH (1-39) in a naloxone and phosphoramidon antagonizable manner, indicating a neutral endopeptidase (24.11; NEP) mediated cleavage. Lipopolysaccharide (10 microg/animal) stimulated the processing of ACTH (1-39) yielding ACTH (1-24) in a cleavage that is independent of NEP, but dependent on aspartyl proteases, demonstrating differential enzymatic cleavage of ACTH (1-39). Taken together, POMC is present in invertebrates and its processing can be altered depending on the signal.


Effects of ONO-3708, an antagonist of the thromboxane A2/prostaglandin endoperoxide receptor, on platelet aggregation and thrombosis.
The beneficial effects of an antagonist of the thromboxane A2/prostaglandin endoperoxide receptor, 7-[2 alpha,4 alpha-(dimethylmethano)-6 beta-(2-cyclopentyl-2 beta- hydroxyacetamido)-1 alpha-cyclohexyl]-5(Z)-heptenoic acid (ONO-3708) on thrombosis were examined. ONO-3708 at 0.1-3 microM inhibited the human platelet aggregation induced by thromboxane A2, prostaglandin H2, collagen, ADP (secondary phase) and epinephrine (secondary phase) without affecting prostanoid synthesis and the content of cyclic AMP in platelets. The in vivo effects, on coronary thrombosis in this case, were examined in two canine models. ONO-3708, 3 to 300 micrograms/kg i.v., prevented dose dependently the coronary thrombosis induced by partial obstruction of the coronary artery. ONO-3708, 3 micrograms/kg per min i.v., significantly prevented electrically stimulated coronary thrombosis without affecting systemic blood pressure and heart rate. These results indicate that the thromboxane A2/prostaglandin endoperoxide receptor could play an important role in the pathogenesis of thrombosis and that ONO-3708 may have therapeutic advantages in preventing thrombosis.


Abnormal FISH in patients with immunoglobulin light chain amyloidosis is a risk factor for cardiac involvement and for death.
Importance of interphase fluorescent in situ hybridization (FISH) with cytoplasmic staining of immunoglobulin FISH (cIg-FISH) on bone marrow is not well understood in light chain amyloidosis (AL). This is in contrast with multiple myeloma where prognostic and treatment related decisions are dependent on cytogenetic testing. This retrospective study reviewed 401 AL patients with cIg-FISH testing performed at our institution between 2004 and 2012. Eighty-one percent of patients had an abnormal cIg-FISH. Common abnormalities involved translocations of chromosome 14q32 (52%), specifically: t(11;14) (43%), t(14;16) (3%) and t(4;14) (2%). Other common abnormalities include monosomy 13/deletion 13q (30%), trisomies 9 (20%), 15 (14%), 11 (10%) and 3 (10%). Median overall survival for this cohort of patients is 3.5 years. When plasma cell burden was greater than 10% trisomies predicted for worse survival (44 vs 19 months), and when it was ⩽10% t(11;14) predicted for worse survival (53 months vs not reached). Abnormal cIg-FISH was significantly associated with advanced cardiac involvement, and remained a prognostic factor on multivariate analysis. This large AL cohort demonstrates that abnormal FISH at diagnosis is prognostic for survival and advanced cardiac disease. Particularly, trisomies and t(11;14) affect survival when degree of plasma cell burden is considered.


Progesterone induces endothelium-independent relaxation of rabbit coronary artery in vitro.
The effect of progesterone on isolated rabbit coronary arteries and its possible mechanism was investigated by measuring changes of isometric tension. Progesterone (1, 3, 10 and 30 microM) induced significant coronary relaxation in K+ (30 mM)-, prostaglandin F2 alpha (3 microM)- or Bay K 8644 (1 microM plus 15 mM K+)- precontracted arteries. There was no difference between endothelium-intact and -denuded coronary arteries from both male and female rabbits, precontracted with these three agents. Haemoglobin, indomethacin, methylene blue, glibenclamide or barium chloride did not affect the relaxation. In endothelium-denuded rabbit coronary arteries, progesterone shifted calcium concentration-dependent constrictor-response curves to the right, the maximal contraction was also reduced. The -log ED50s were 3.6 in control, and 3.3 and 2.9 after incubation with progesterone (3 and 30 microM), respectively. Similar results were obtained in rat aorta. We conclude that progesterone induces significant endothelium-independent relaxation in rabbit coronary arteries in vitro, possibly by affecting calcium influx.


TGF-β1 contributes to the hepatic inflammation in animal models with nonalcoholic steatohepatitis by Smad3/TLR2 signaling pathway.
Non-alcoholic fatty liver disease (NAFLD) is increasingly affecting human health and the economy worldwide due to various factors. Here, we found that the expression of TGF-β1 and TLR2 was significantly up-regulated in liver samples from both rats and mice nonalcoholic steatohepatitis (NASH) models. By constructing corresponding cell model, we found that TGF-β1 challenge can positively regulate the expression of TLR2 and p-Smad2/3, and the dual luciferase reporter gene system and EMSA assay confirmed the existence of Smad3 binding site (-916 ∼ -906) in the promoter region of TLR2. The overexpression and interference changes of Smad2/3 further verified the above experimental results. Taken together, these findings suggest that TGF-β1 promotes TLR2 transcription and its target gene expression via Smad3, leading to malignant exacerbation of liver inflammation in NASH, which provides new insights into the treatment of NASH.


Sleep restriction by sleep timing late night or early wake: The impact on physical activity and dietary intake in adults.
To date, few studies have assessed whether the timing of sleep restriction impacts physical activity and energy intake patterns. Thus, we aimed to quantify physical activity and energy intake during an early wake (EW) and late sleep (LS) period. Fourteen participants who met the inclusion criteria (sleep 7-9 h/night and a BMI of <40 kg/m2) participated in 3 crossover free-living conditions: normal sleep (NS, 7-9 h), EW (2-h early wake-time), and LS (2-h late to sleep) for 4 nights. Sleep duration (via Actiwatch), energy intake (via food diaries), and physical activity (via hip accelerometry) were recorded for 4 days/4 nights throughout each condition. Sleep duration was reduced in both sleep restriction conditions compared to NS (p < 0.001) with no difference between sleep restriction conditions. Daily energy intake tended to increase in the LS condition (p = 0.056) but was unchanged during EW (p = 0.56). Fat (p = 0.031) and sodium (p = 0.039) intake were increased in the LS condition only compared to NS. During the EW condition, fat (p = 0.24) and sodium (p = 0.18) intake were not altered. No changes in carbohydrate or protein intake occurred between conditions. Daily steps tended to increase in the EW condition compared to NS (p = 0.058), while steps during the LS condition were unchanged (p = 0.28), with no differences between sleep restriction conditions. The timing of sleep curtailment differentially influences physical activity and EI the following day, such that EW results in increased physical activity, while LS leads to poorer dietary choices.


Wilhelm Ebstein and the history of surgery for Ebstein's Disease.
In the mirror of the life-story of Wilhelm Ebstein, the discovery of "Ebstein's Disease" is presented. The readers are guided through the individual stages of the development of surgical treatment of the condition.


Contrast-enhanced ultrasonography with Sonazoid™ for the evaluation of bowel ischemia.
To determine the usefulness of contrast-enhanced ultrasonography with the contrast agent Sonazoid™ for the detection of bowel ischemia. From March 2007 to February 2009, 65 patients (35 men and 30 women, mean age 70.4 ± 16.1 years) were enrolled. Fifty-three patients complained of acute abdominal pain with small bowel dilatation (n = 40) or reduced bowel peristalsis (n = 13). Twelve patients were clinically suspected of having bowel ischemia. After Sonazoid™ injection, bowel segments were scanned using harmonic imaging, and the signal intensities were classified as normal or diminished. The definitive diagnosis was confirmed by surgery in 30 patients, autopsy in 6, endoscopy in 3, angiography in 1, and clinical follow-up in 25. All 50 patients with normal signal intensities were confirmed not to have bowel ischemia. In the 15 patients with diminished signal intensities, 14 patients were confirmed to have bowel ischemia, resulting in an overall sensitivity of 100% [95% confidence interval (CI) 80.7-100%], a specificity of 98% [95% CI 89.5-99.9%], a positive predictive value of 93% (95% CI 68.1-99.8%), and a negative predictive value of 100% (95% CI 94.1-100%). Contrast-enhanced ultrasonography with Sonazoid™ is a highly sensitive and specific method for the diagnosis of bowel ischemia.


Preoperative Frailty Assessment Predicts Postoperative Mortality, Delirium and Pneumonia in Elderly Lung Cancer Patients: A Retrospective Cohort Study.
The purpose of this study was to investigate the predictive value of the 5-factor modified frailty index (mFI-5) for postoperative mortality, delirium and pneumonia in patients over 65 years of age undergoing elective lung cancer surgery. Data were collected from a single-center retrospective cohort study conducted in a general tertiary hospital from January 2017 to August 2019. In total, the study included 1372 elderly patients aged over 65 who underwent elective lung cancer surgery. They were divided into frail group (mFI-5, 2-5), prefrail group (mFI-5, 1) and robust group (mFI-5, 0) on the basis of mFI-5 classification. The primary outcome was postoperative 1-year all-cause mortality. Secondary outcomes were postoperative pneumonia and postoperative delirium. Frailty group had the highest incidence of postoperative delirium (frailty 31.2% versus prefrailty 1.6% versus robust 1.5%, p < 0.001), postoperative pneumonia (frailty 23.5% versus prefrailty 7.2% versus robust 7.7%, p < 0.001), and postoperative 1-year mortality (frailty 7.0% versus prefrailty 2.2% versus robust 1.9%. p < 0.001). Frail patients have significantly longer length of hospitalization than those in the robust group and prefrail patients (p < 0.001). Multivariate analysis showed a clear link between frailty and increased risk of postoperative delirium (aOR 2.775, 95% CI 1.776-5.417, p < 0.001), postoperative pneumonia (aOR 3.291, 95% CI 2.169-4.993, p < 0.001) and postoperative 1-year mortality (aOR 3.364, 95% CI, 1.516-7.464, p = 0.003). mFI-5 has potential clinical utility in predicting postoperative death, delirium and pneumonia incidence in elderly patients undergoing radical lung cancer surgery. Frailty screening of patients (mFI-5) may provide benefits in risk stratification, targeted intervention efforts, and assist physicians in clinical decision-making.


Proteomic Signatures During Treatment in Different Stages of Heart Failure.
Proteomics have already provided novel insights into the pathophysiology of heart failure (HF) with reduced ejection fraction. Previous studies have evaluated cross-sectional protein signatures of HF, but few have characterized proteomic changes following HF with reduced ejection fraction treatment with ARNI (angiotensin receptor/neprilysin inhibitor) therapy or left ventricular assist devices. In this retrospective omics study, we performed targeted proteomics (N=625) of whole blood sera from patients with American College of Cardiology/American Heart Association stage D (N=29) and stage C (N=12) HF using proximity extension assays. Samples were obtained before and after (median=82 days) left ventricular assist device implantation (stage D; primary analysis) and ARNI therapy initiation (stage C; matched reference). Oblique principal component analysis and point biserial correlations were used for feature extraction and selection; standardized mean differences were used to assess within and between-group differences; and enrichment analysis was used to generate and cluster Gene Ontology terms. Core sets of proteins were identified for stage C (N=9 proteins) and stage D (N=18) HF; additionally, a core set of 5 shared HF proteins (NT-proBNP [N-terminal pro-B type natriuretic peptide], ESM [endothelial cell-specific molecule]-1, cathepsin L1, osteopontin, and MCSF-1) was also identified. For patients with stage D HF, moderate (δ, 0.40-0.60) and moderate-to-large (δ, 0.60-0.80) sized differences were observed in 8 of their 18 core proteins after left ventricular assist devices implantation. Additionally, specific protein groups reached concentration levels equivalent (g<0.10) to stage C HF after initiation on ARNI therapy. HF with reduced ejection fraction severity associates with distinct proteomic signatures that reflect underlying disease attributes; these core signatures may be useful for monitoring changes in cardiac function following initiation on ARNI or left ventricular assist device implantation.


Deep blue autofluorescence reflects the oxidation state of human transthyretin.
Human transthyretin (TTR) is a tetrameric protein transporting thyroid hormones and retinol. TTR is a neuroprotective factor and sensor of oxidative stress which stability is diminished due to mutations and aging, leading to amyloid deposition. Adverse environmental conditions, such as redox and metal ion imbalances, induce destabilization of the TTR structure. We have previously shown that the stability of TTR was disturbed by Ca2+ and other factors, including DTT, and led to the formation of an intrinsic fluorophore(s) emitting blue light, termed deep blue autofluorescence (dbAF). Here, we show that the redox state of TTR affects the formation dynamics and properties of dbAF. Free thiols lead to highly unstable subpopulations of TTR and the frequent ocurrence of dbAF. Oxidative conditions counteracted the destabilizing effects of free thiols to some extent. However, strong oxidative conditions led to modifications of TTR, which altered the stability of TTR and resulted in unique dbAF spectra. Riboflavin and/or riboflavin photoproducts bound to TTR and crosslinked TTR subunits. Riboflavin-sensitized photooxidation increased TTR unfolding, while photooxidation, either in the absence or presence of riboflavin, increased proteolysis and resulted in multiple oxidative modifications and dityrosine formation in TTR molecules. Therefore, oxidation can switch the role of TTR from a protective to pathogenic factor.


Minor Loops in Major Folds: Enhancer-Promoter Looping, Chromatin Restructuring, and Their Association with Transcriptional Regulation and Disease.
The organization and folding of chromatin within the nucleus can determine the outcome of gene expression. Recent technological advancements have enabled us to study chromatin interactions in a genome-wide manner at high resolution. These studies have increased our understanding of the hierarchy and dynamics of chromatin domains that facilitate cognate enhancer-promoter looping, defining the transcriptional program of different cell types. In this review, we focus on vertebrate chromatin long-range interactions as they relate to transcriptional regulation. In addition, we describe how the alteration of boundaries that mark discrete regions in the genome with high interaction frequencies within them, called topological associated domains (TADs), could lead to various phenotypes, including human diseases, which we term as "TADopathies."


Brain alarm by self-extracellular nucleic acids: from neuroinflammation to neurodegeneration.
Neurological disorders such as stroke, multiple sclerosis, as well as the neurodegenerative diseases Parkinson's or Alzheimer's disease are accompanied or even powered by danger associated molecular patterns (DAMPs), defined as endogenous molecules released from stressed or damaged tissue. Besides protein-related DAMPs or "alarmins", numerous nucleic acid DAMPs exist in body fluids, such as cell-free nuclear and mitochondrial DNA as well as different species of extracellular RNA, collectively termed as self-extracellular nucleic acids (SENAs). Among these, microRNA, long non-coding RNAs, circular RNAs and extracellular ribosomal RNA constitute the majority of RNA-based DAMPs. Upon tissue injury, necrosis or apoptosis, such SENAs are released from neuronal, immune and other cells predominantly in association with extracellular vesicles and may be translocated to target cells where they can induce intracellular regulatory pathways in gene transcription and translation. The majority of SENA-induced signaling reactions in the brain appear to be related to neuroinflammatory processes, often causally associated with the onset or progression of the respective disease. In this review, the impact of the diverse types of SENAs on neuroinflammatory and neurodegenerative diseases will be discussed. Based on the accumulating knowledge in this field, several specific antagonistic approaches are presented that could serve as therapeutic interventions to lower the pathological outcome of the indicated brain disorders.


Coherent delta-band oscillations between cortical areas correlate with decision making.
Coherent oscillations in the theta-to-gamma frequency range have been proposed as a mechanism that coordinates neural activity in large-scale cortical networks in sensory, motor, and cognitive tasks. Whether this mechanism also involves coherent oscillations at delta frequencies (1-4 Hz) is not known. Rather, delta oscillations have been associated with slow-wave sleep. Here, we show coherent oscillations in the delta frequency band between parietal and frontal cortices during the decision-making component of a somatosensory discrimination task. Importantly, the magnitude of this delta-band coherence is modulated by the different decision alternatives. Furthermore, during control conditions not requiring decision making, delta-band coherences are typically much reduced. Our work indicates an important role for synchronous activity in the delta frequency band when large-scale, distant cortical networks coordinate their neural activity during decision making.


Comprehensive metabolome analyses reveal N-acetylcysteine-responsive accumulation of kynurenine in systemic lupus erythematosus: implications for activation of the mechanistic target of rapamycin.
Systemic lupus erythematosus (SLE) patients exhibit depletion of the intracellular antioxidant glutathione and downstream activation of the metabolic sensor, mechanistic target of rapamycin (mTOR). Since reversal of glutathione depletion by the amino acid precursor, N-acetylcysteine (NAC), is therapeutic in SLE, its mechanism of impact on the metabolome was examined within the context of a double-blind placebo-controlled trial. Quantitative metabolome profiling of peripheral blood lymphocytes (PBL) was performed in 36 SLE patients and 42 healthy controls matched for age, gender, and ethnicity of patients using mass spectrometry that covers all major metabolic pathways. mTOR activity was assessed by western blot and flow cytometry. Metabolome changes in lupus PBL affected 27 of 80 KEGG pathways at FDR p < 0.05 with most prominent impact on the pentose phosphate pathway (PPP). While cysteine was depleted, cystine, kynurenine, cytosine, and dCTP were the most increased metabolites. Area under the receiver operating characteristic curve (AUC) logistic regression approach identified kynurenine (AUC = 0.859), dCTP (AUC = 0.762), and methionine sulfoxide (AUC = 0.708), as top predictors of SLE. Kynurenine was the top predictor of NAC effect in SLE (AUC = 0.851). NAC treatment significantly reduced kynurenine levels relative to placebo in vivo (raw p = 2.8 × 10-7, FDR corrected p = 6.6 × 10-5). Kynurenine stimulated mTOR activity in healthy control PBL in vitro. Metabolome changes in lupus PBL reveal a dominant impact on the PPP that reflect greater demand for nucleotides and oxidative stress. The PPP-connected and NAC-responsive accumulation of kynurenine and its stimulation of mTOR are identified as novel metabolic checkpoints in lupus pathogenesis.


Investor Outlook: Rising from the Ashes; GSK's European Approval of Strimvelis for ADA-SCID.
GlaxoSmithKline's (GSK) and partner San Raffaele Telethon Institute for Gene Therapy's recent positive European approval for Strimvelis for treatment of severe combined immunodeficiency due to adenosine deaminase deficiency (ADA-SCID) represents the second EU-approved gene therapy and the first γ-retrovirus and first ex vivo gene therapy. In this article we discuss the significance and implications of this historic approval for the broader gene therapy field.


Interferon-gamma release assays for the tuberculosis serial testing of health care workers: a systematic review.
Interferon-gamma release assays (IGRAs) are increasingly used in the tuberculosis (TB) screening of health care workers (HCWs). However, comparatively high rates of conversions and reversion as well as growing evidence of substantial within-subject variability of interferon-gamma responses complicate their interpretation in the serial testing of HCWs. We conducted a systematic review on the repeat use of the two commercial IGRAs, the QuantiFERON-TB Gold or In-Tube version (QFT) and the T-SPOT.TB (T-SPOT), in the serial testing and its with-subject variability among HCWs in order to provide guidance on how to interpret serial testing results in the context of the periodic screening of subjects with an increased occupational risk of latent TB infection (LTBI) in countries with low and intermediate TB incidence rates. The Medline, Embase, and Cochrane databases were searched without restrictions. Retrieved articles were complemented by additional hand searched records. Only studies that used commercial IGRAs among HCWs apart from contact and outbreak investigations and those fulfilling further predefined criteria were included. Overall, 20 studies, five using the T-SPOT and 19 using the QFT assay, were included. Fifteen studies met eligibility criteria for serial testing and five studies for within-subject variability. Irrespective of TB incidence rates in the study's country of origin, reversion rates were consistently higher than conversion rates (range 22-71% vs. 1-14%). Subjects with baseline results around the diagnostic threshold were more likely to show inconsistent results on retesting. The within-subject variability of interferon-gamma responses was considerable across all studies systematically assessing it. On the basis of reviewed studies we advocate using a borderline zone from 0.2-0.7 IU/ml for the interpretation of repeat QFT results in the routine screening of HCWs with an increased LTBI risk. Subjects with QFT results within this borderline zone, with suspected fresh infection, and those who are considered for preventive chemotherapy should be retested with the QFT within a period of about four weeks before preventive chemotherapy is recommended. However, the available data regarding the use of the T-SPOT in the serial testing of HCWs is remarkably limited and warrants further research.


Analysis of Mitochondrial DNA in Induced Pluripotent and Embryonic Stem Cells.
The mitochondrial genome has a major role to play in establishing and maintaining pluripotency. Furthermore, mitochondrial DNA (mtDNA) copy is strictly regulated during differentiation. Undifferentiated, pluripotent cells possess fewer than 300 copies of mtDNA, which establishes the mtDNA set point and promotes cell proliferation and, as a result, these cells rely on glycolysis with some support from oxidative phosphorylation (OXPHOS) for the generation of ATP. The mtDNA set point provides the starting point from which cells increase their mtDNA copy number as they differentiate into mature functional cells. Dependent on cell types, mtDNA copy number ranges from ~10 copies in sperm to several thousand in cardiomyocytes. Consequently, differentiating cell types can acquire the appropriate numbers of mtDNA copy to meet their specific requirements for ATP generated through OXPHOS. However, as reprogrammed somatic cells do not always achieve this, it is essential to analyze them for their OXPHOS potential and ability to regulate mtDNA copy number. Here, we describe how to assess mtDNA copy number in pluripotent and differentiating cells using real-time PCR protocols; assess expression of the mtDNA specific replication factors through real-time RT-PCR; identify mtDNA variants in embryonic and induced pluripotent stem cells; determine DNA methylation patterns of the mtDNA-specific replication factors; and assess mitochondrial OXPHOS capacity.


Nurse practitioner preceptor education to increase role preparedness.
The public has become aware of the need for nurses and nurse practitioners in primary care with the widespread public health crisis. As the need for nurses and nurse practitioners grows, there is a need for the clinical educator and preceptor. However, preceptors have noted barriers to precepting such as time constraints, lack of preceptor role education, and role preparedness. Currently, neither preceptor role development nor education has been implemented within a local institution. Preceptor vocalization led the charge for preceptor role development education within the institution. This quality improvement project focused on the preceptor role preparedness and education. A convenience sample was recruited from a large, diverse academic institution. The modified Clinical Supervision Self-Assessment Tool-Skills was utilized to determine the effectiveness of online education in preceptor role preparedness. An online, self-directed, evidence-based preceptor education webpage with easily reproducible information related to collaborating with students and faculty situations. Literature, such as the One Minute Preceptor, Ask, Tell, Ask, and engaging student scenarios, was included from the literature review. A voice-recorded PowerPoint summarizing the webpage was included with a transcript for accessibility. A paired t-test was used to assess the mean differences between the preintervention and postintervention periods. The results indicated the online preceptor education was effective. Preparing nurse practitioner preceptors for their roles benefits the health care system. An online, self-directed, evidence-based preceptor education webpage can be beneficial in future preceptor education planning.


Tumor necrosis predicts poor clinical outcomes in patients with node-negative upper urinary tract urothelial carcinoma.
Tumor necrosis has been indicated as a factor for the poor clinical outcome in human cancers. We aim to disclose the association between tumor necrosis and overall survival and recurrence-free survival in node-negative upper urinary tract urothelial carcinoma patients treated with radical nephroureterectomy. A retrospective cohort of 100 patients with upper urinary tract urothelial carcinoma from January 1990 to June 2011 was enrolled in this study. Univariate analysis with Log-rank test and multivariate analysis with Cox proportional hazards regression models were conducted to determine the correlations of tumor necrosis with overall survival and recurrence-free survival. Tumor necrosis was presented in 48 patients with upper urinary tract urothelial carcinoma and was significantly associated with the advanced pathological stage (P < 0.001), high tumor grade (P < 0.001), subsequent bladder tumor (P = 0.018), vascular invasion (P < 0.001) and lymph node metastasis (P = 0.026). Multivariate analysis revealed tumor necrosis as an independent unfavorable predictor of overall survival in node-negative upper urinary tract urothelial carcinoma patients by multivariate analysis (hazard ratio = 9.23, 95% confidence interval = 1.05-80.89, P = 0.045). Tumor necrosis was an independent factor of adverse clinical outcomes in node-negative upper urinary tract urothelial carcinoma patients who received radical nephroureterectomy. Evaluation of tumor necrosis might be of clinical significance to determine whether patients with node-negative upper urinary tract urothelial carcinoma should be given further therapy after radical nephroureterectomy.


Confocal scanning microscopy: three-dimensional biological imaging.
Confocal scanning optical microscopy, arguably the most significant in biological light microscopy in this decade, enables one to obtain quantitative non-invasive optical sections through labelled biological specimens, virtually free from out-of-focus blur. A set of these optical sections collected at a series of focal levels through an object constitutes a three-dimensional image which may then be processed digitally for display as a computer reconstruction, a stereo pair or an animation sequence.


Levels of circulating intercellular adhesion molecule-1 in patients with Behçet's disease.
The purpose of this study was to investigate the possible relationship between circulating intercellular adhesion molecule-1 (cICAM-1) plasma levels and Behçet's disease. Plasma levels of cICAM-1 were measured in 44 patients with Behçet's disease and in 30 healthy controls. cICAM-1 levels were higher in patients with Behçet's disease than in the controls (P=0.001). The difference was more marked between patients with active Behçet's disease and controls (P < 0.001). cICAM-1 levels were also higher in patients with active Behçet's disease than in those with inactive disease (P<0.001). cICAM-1 seems to be related to disease activity in Behçet's disease. It may be useful in assessing disease activity in Behçet's disease, as well as in monitoring response to treatment.


Induction of activation of the antioxidant response element and stabilization of Nrf2 by 3-(3-pyridylmethylidene)-2-indolinone (PMID) confers protection against oxidative stress-induced cell death.
The antioxidant response elements (ARE) are a cis-acting enhancer sequence located in regulatory regions of antioxidant and detoxifying genes. Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a member of the Cap 'n' Collar family of transcription factors that binds to the ARE and regulates the transcription of specific ARE-containing genes. Under oxidative stress, Nrf2/ARE induction is fundamental to defense against reactive oxygen species (ROS) and serves as a key factor in the protection against toxic xenobiotics. 3-(3-Pyridylmethylidene)-2-Indolinone (PMID) is a derivative of 2-indolinone compounds which act as protein kinase inhibitors and show anti-tumor activity. However, the role of PMID in the oxidative stress remains unknown. In the present study, we showed that PMID induced the activation of ARE-mediated transcription, increased the DNA-binding activity of Nrf2 and then up-regulated the expression of antioxidant genes such as HO-1, SOD, and NQO1. The level of Nrf2 protein was increased in cells treated with PMID by a post-transcriptional mechanism. Under CHX treatment, the stability of Nrf2 protein was enhanced by PMID with decreased turnover rate. We showed that PMID reduced the ubiquitination of Nrf2 and disrupted the Cullin3 (Cul3)-Keap1 interaction. Furthermore, cells treated with PMID showed resistance to cytotoxicity by H(2)O(2) and pro-oxidant 6-OHDA. PMID also up-regulated the antioxidant level in BALB/c mice. Taken together, the compound PMID induces the ARE-mediated gene expression through stabilization of Nrf2 protein and activation of Nrf2/ARE pathway and protects against oxidative stress-mediated cell death.


Diagnostic value of the combination of DAPK methylation in urinary sediment and B ultrasound for recurrent urinary bladder cancer.
Urinary bladder cancer (UBC) is the most common malignancy affecting the urinary system. This study aimed to investigate the diagnostic value of combining DAPK methylation in urinary sediment and B ultrasound in the detection of recurrent UBC. A total of 1021 cases with primary UBC who underwent electrocision of bladder tumor through urethra were included in this study and followed up. Various parameters including B ultrasound, DAPK methylation in urinary sediment, examination of exfoliated cells in the urine, and cystoscopy were performed. The data collected was analyzed using the Kappa test, and receiver operating characteristic (ROC) curve was constructed to assess the diagnostic role in recurrent UBC. Among the 1021 patients, 115 patients experienced recurrence confirmed by cystoscopy and biopsy within two years and were excluded from the study, resulting in an effective sample size of 906 primary UBC cases. The results of cystoscopy showed agreement with B ultrasound (Kappa = 0.785, P < 0.05), as well as with DAPK methylation in urinary sediment, and the combination of B ultrasound and DAPK methylation (Kappa = 0.517, P < 0.05, Kappa = 0.593, P < 0.05). The combination of B ultrasound with DAPK methylation yielded an area under the curve of 0.922, with a sensitivity of 92.86%, specificity of 91.63%, and a negative predictive value of 99.4%, suggesting that a negative result indicates a low risk of recurrence. The combination of DAPK methylation in urinary sediment with B ultrasound demonstrates high diagnostic performance for recurrent UBC.


Hemolysis following treatment of theophylline overdose with coated charcoal hemoperfusion.
Life-threatening theophylline toxicity is an indication for coated charcoal hemoperfusion. This can usually be accomplished without serious side effects. We describe here a case of acute severe intravascular hemolysis following coated charcoal hemoperfusion.


Complication Avoidance in Spine Surgery.
The outcomes of spine surgery are closely related to postoperative morbidity. Therefore, an experienced surgeon must be aware of various complications and should apply all necessary preventive measures to avoid them. It is widely considered that complications of spine surgery are underreported and that their real incidence is much higher than expected. This review highlights methods to prevent various types of morbidity that may be encountered during different spinal procedures, considering general complications, approach-related complications, fusion- and implant-related complications, and systemic complications.


A shrinking strategy for creating dynamic SERS hot spots on the surface of thermosensitive polymer nanospheres.
This study reports a strategy that allows one to finely tune the distance between Ag nanoparticles decorated on thermoresponsive microgels to create vast 'hot spots' for electromagnetic enhancement. The dynamic gap (i.e. the interparticle distance) can be tuned easily because the microgel template undergoes a phase transition upon changing the temperature. This change could increase the SERS intensity for three orders of magnitude.


Association between Gestational Diabetes Mellitus and Postpartum Depression among Women in Eastern India: A Cohort Study.
The study was planned to evaluate the association between Gestational Diabetes Mellitus (GDM) and Postpartum Depression (PPD) in a rural population of Odisha, Eastern India. Pregnant women in the first trimester were recruited and followed up till six weeks of postpartum. Gestational Diabetes Mellitus was assessed with 75 grams glucose challenge test and PPD was assessed at six weeks after delivery with Edinburgh Postnatal Depression Scale. Statistical difference between variables was assessed using Chi-square test, Fischer's exact test, and unpaired T-test. Covariates were adjusted using bivariate and multivariate logistic regression to estimate the association between GDM and PPD. Out of 436 pregnant women recruited, 347 (89.6%) remained in the study. Prevalence of GDM was 13.9% (95% CI: 10.7-17.3) and PPD was 9.8% (95% CI: 6.6-12.9). Incidence of PPD in the GDM group was 14.58% (95% CI: 4.2-24.9) and in women without GDM was 9.06% (95% CI: 5.76-12.3). However, the association was not significant on multivariate logistic regression (Risk Ratio (RR) = 1.56, 95% C.I: 0.61-6.16; P-value = 0.35). This study demonstrated that women with GDM were at higher risk of developing PPD suggesting that an "at risk" approach should be implemented for screening.


Lacrimal sac rhinosporidiosis: case report and review of literature with a new grading system to optimize treatment.
Rhinosporidiosis is a chronic granulomatous disease affecting the mucous membrane primarily and is caused by Rhinosporidium seeberi, an aquatic protistan parasite. The nose is the most common site of involvement and is seen in 83.3% cases, followed by ocular involvement in 11.2% cases and other sites like larynx, trachea and bronchus in 5.5% cases. In various oculosporidiosis case series, lacrimal drainage system involvement was seen to vary from 14.3% to 59.6% cases. Isolated lacrimal sac involvement in rhinosporidiosis was found in 45.8% (72 out of 157) cases of the lacrimal drainage system in a review of 31 studies. A variety of surgical procedures have been used to treat rhinosporidiosis of lacrimal sac like dacryocystorhinostomy, Dacryocystectomy, lateral rhinotomy and local lesion excision with a success rate varying from 28.5% to 92.3%. This wide variation in the success rate was due to the fact that a uniform surgical procedure was performed in all the cases of a particular series irrespective of the extent of disease. Grading the lacrimal sac rhinosporidiosis to decide the extent of surgical excision may help achieve better results. We present a grading system based on our own experience in a case of extensive rhinospodiosis of lacrimal sac and review of 31 studies published in the literature. A 24-year-old male from Nepal presented with the complaints of watering from his right eye of 13 years duration, swelling in the right medial canthal area with an extension to the inferior part of the orbit for 12 years and nasal blockage for 1.5 years. The patient had a history of previous intervention in which biopsy was taken from the nose and sent for histopathology that confirmed rhinosporidiosis. An extended intranasal endoscopic dacryocystectomy was done along with debridement and coblation of the lesion over the septum and nasopharynx. Intraoperatively a large rhinosporidiosis mass was seen filling the sac and was removed in toto along with the sac and nasolacrimal duct. Recurrence of a tiny lesion after 6 months in our case despite wide excision with the drilling of bony nasolacrimal duct and coblation, made us review the literature.


Use of the Hage framework for theory construction: Factors affecting glucose control in the college-aged student with type 1 diabetes.
This article describes the Hage framework for theory construction and its application to the clinical problem of glycemic control in college-aged students with type 1 diabetes. College-aged students with type 1 diabetes struggle to self-manage their condition. Glycated hemoglobin (HbA1c), if controlled within acceptable limits (6-8%), is associated with the prevention or delay of serious diabetic complications such as kidney and cardiovascular disease. Diabetes educators provide knowledge and skills, but young adults must self-manage their condition on a daily basis, independent of parents. The Hage framework includes five tasks of theory construction: narrowing and naming the concepts, specifying the definitions, creating the theoretical statements, specifying the linkages, and ordering components in preparation for model building. During the process, concepts within the theory were revised as the literature was reviewed, and measures and hypotheses, foundational to research, were generated. We were successful in applying the framework and creating a model of factors affecting glycemic control, emphasizing that physical activity, thought of as a normal part of wellness, can be a two-edged sword producing positive effect but also serious negative effects in some college-aged students with type 1 diabetes. Contextual factors important to self-management in college-aged students are emphasized. The Hage framework, already used to a small extent in nursing curricula, deserves more attention and, because of its generic nature, may be used as a template for theory construction to examine a wide variety of nursing topics.


Strengthening Home Visitor Practice Through an Embedded Model of Infant and Early Childhood Mental Health Consultation.
Home visitors face the difficult task of engaging and supporting families who experience economic, social, and mental health challenges. Infant and Early Childhood Mental Health Consultation (IECMHC) is one strategy to improve early childhood professionals' abilities to promote maternal and children's mental health through relationship building and collaboration. To date, most of the evidence showing positive effects of IECMHC comes from evaluations conducted in center-based early childhood programs. Only a few have examined the implementation and outcomes of IECMHC in home visiting programs. Using a longitudinal, matched-comparison group design, a three-year pilot study of a cross-system, embedded model of IECMHC assessed implementation and outcomes for teachers and home visitors. Here, we focus on intervention effects on home visitor-family relationships based on an analysis of 42 recordings of home visits. We used the Home Visit Rating Scales Adapted & Extended to Excellence (HOVRS-A+ v.2.1; Roggman et al., in 2010) and created another scale, the IECMHC scale, of items aligned with the expected outcomes of the model. Home visitors in the intervention group scored significantly higher on the HOVRS-A+ home visitor responsiveness to family scale over time than those in the comparison group. Additionally, scores of home visitors in the intervention group increased at a greater rate, on average, on the newly developed IECMHC scale than scores of home visitors in the comparison group. We conclude that the Illinois model of IECMHC successfully improved home visitors' responsiveness to families and improved the quality of home visits. Due to the small sample, we recommend further study of the model in home visiting and further development of the IECMHC scale as part of the HOVRS-A+ .


The expression of DCC protein in female breast cancer.
The deleted in colorectal cancer (DCC) gene has been shown to be frequently deleted or its expression reduced or absent in glioblastomas, colorectal, gastro-intestinal, pancreatic and prostatic tumors. In the present study, we investigated the expression of DCC in surgical specimen from 75 patients with primary breast cancer. The expression of the DCC, estrogen receptors (ER), and progesterone receptors (PR) was studied in 75 surgical specimens of primary breast cancer using an immunohistochemical method. To evaluate the outcomes of the breast cancer patients, we followed up the patients during minimum of 10 years. Reduced or loss of expression of DCC was identified in 45 out of 75 samples. There were significant differences between cases without metastasis or local recurrences versus these with metastasis or local recurrences (p = 0.006), and between patients alive with no evidence of malignancy versus those with recurrence or dead of disease (p = 0.005). There were no significant differences between the DCC status and age, sex, tumor location, stage, grade, or proportion of patients who received adjuvant therapy. These findings suggest that a decrease in DCC expression may influence the prognosis of breast carcinoma patients.


Galactose clearance measurements and liver blood flow.
Galactose clearance, measured during low galactose infusion and calculated as infusion rate divided by peripheral galactose concentration (systemic clearance), has been proposed as a measure of liver blood flow. This requires nearly complete hepatic extraction as well as negligible extrahepatic elimination. The purpose of the study was to examine if these assumptions are fulfilled in subjects with no liver disease, and to compare the galactose clearance measurement with an independent measurement of liver blood flow. Liver blood flow was measured in 6 subjects by means of a constant indocyanine green infusion, indocyanine green concentration measurements in a peripheral artery and a hepatic vein, and calculation according to Fick's principle. The mean (+/- SEM) blood flow rate was 1.2 +/- 0.1 L/min. Galactose was given at a constant infusion rate of 142 +/- 10 mumol/min, and steady-state concentrations were measured in the peripheral artery (A) and the hepatic vein (V). The hepatic extraction fraction [(A - V)/A] was 0.91 +/- 0.03. The hepatic galactose elimination rate [(A - V) X flow] was 101 +/- 12 mumol/min; this is about two-thirds of the total elimination rate (viz., infusion rate). Urinary excretion was negligible. This indicates an extrahepatic galactose elimination of approximately 41 mumol/min. Systemic galactose clearance, calculated as mentioned above, was 1.5 +/- 0.1 L blood/min. It was significantly higher than the liver blood flow in each subject (paired t-test, each p less than 0.02), on average 133% of the flow. Thus the systemic galactose clearance value overestimates liver blood flow, probably due to a small, but in this context quantitatively important, extrahepatic galactose elimination.


Immunogenicity and safety of an inactivated quadrivalent influenza vaccine co-administered with a 23-valent pneumococcal polysaccharide vaccine versus separate administration, in adults ≥50years of age: Results from a phase III, randomized, non-inferiority trial.
We compared co-administration versus separate administration of an inactivated quadrivalent influenza vaccine (IIV4) with a 23-valent pneumococcal polysaccharide vaccine (PPV23) in adults at high risk of complications of influenza and pneumococcal infection. This phase III, placebo-controlled, observer-blind trial (NCT02218697) was conducted in France and Belgium during the 2014-2015 influenza season. Adults≥50years of age meeting their country's vaccination recommendations were randomized 1:1 to co-administration or separate administration. Immunogenicity was assessed by hemagglutination inhibition (HI) titers for IIV4 and 22F-inhibition ELISA for PPV23. Co-primary objectives were to demonstrate non-inferiority of co-administration versus separate administration in terms of geometric mean titer (GMT) ratio for each influenza strain in the IIV4 and geometric mean concentration (GMC) ratio for six pneumococcal serotypes (1, 3, 4, 7F, 14, 19A) in the PPV23 in the per-protocol cohort (N=334). The study met its co-primary objectives, with the upper limit of the 95% confidence interval of the GMT and GMC ratios (separate administration over co-administration) being ≤2.0 for all four antigens of the IIV4 and the six pre-selected serotypes of the PPV23, respectively. Immunogenicity of the IIV4 and PPV23 was similar regardless of administration schedule. In a post hoc analysis pooling participants ≥60years of age from the co-administration and separate administration groups, IIV4 immunogenicity was similar in higher risk adults with comorbidities (diabetes; respiratory, heart, kidney, liver, or neurological diseases; morbid obesity) versus those without. Both vaccines had an acceptable safety and reactogenicity profile; pain was the most common symptom, occurring more often with co-administration than separate administration. The IIV4 and PPV23 can be co-administered without reducing antibody responses reflecting protection against influenza or pneumococcal disease. Co-administration of PPV23 at the annual influenza vaccination visit may improve uptake. Comorbidities had no impact on IIV4 immunogenicity, supporting its value in older adults with chronic medical conditions. Clinical Trial Registry Number: NCT02218697.


Cardiac tamponade related to a coronary injury by a pericardial calcification: an unusual complication.
Cardiac tamponade is a rare but severe complication of pericardial effusion with a poor prognosis. Prompt diagnosis using transthoracic echocardiography allows guiding initial therapeutic management. Although etiologies are numerous, cardiac tamponade is more often due to a hemopericardium. Rarely, a coronary injury may result in such a hemopericardium with cardiac tamponade. Coronary artery aneurysm are the main etiologies but blunt, open chest trauma or complication of endovascular procedures have also been described. A 83-year-old hypertensive man presented for dizziness and hypotension. The patient had oliguria and mottled skin. Transthoracic echocardiography disclosed a circumferential pericardial effusion with a compressed right atrium, confirmed by contrast-enhanced thoracic CT scan. A pig-tail catheter allowed to withdraw 500 mL of blood, resulting in a transient improvement of hemodynamics. Rapidly, recurrent hypotension prompted a reoperation. An active bleeding was identified at the level of the retroventricular coronary artery. The pericardium was thickened with several "sharping" calcified plaques in the vicinity of the bleeding areas. On day 2, vasopressors were stopped and the patient was successfully extubated. Final diagnosis was a spontaneous cardiac tamponade secondary to a coronary artery injury attributed to a "sharping"calcified pericardial plaque. Cardiac tamponade secondary to the development of a hemopericardium may develop as the result of a myocardial and coronary artery injury induced by a calcified pericardial plaque.


ACT-107, a novel variant of AmpC β-lactamase from Enterobacter huaxiensis isolated from Neotropical leaf frog (Phyllomedusa distincta) inhabiting the Brazilian Atlantic Forest.
The aim of this study was to characterise a broad-spectrum cephalosporin-resistant AmpC-positive Enterobacter huaxiensis colonising the skin of a Neotropical frog (Phyllomedusa distincta) inhabiting the Brazilian Atlantic Forest. During a genomic surveillance study of antimicrobial resistance, we screened skin samples from P. distincta. Gram-negative bacteria growing on MacConkey agar plates supplemented with 2 µg/mL ceftriaxone were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. A cephalosporin-resistant E. huaxiensis was sequenced using the Illumina NextSeq platform. Genomic data were analysed using bioinformatics tools, whereas AmpC β-lactamase was characterised in depth by comparative analysis of amino acids, in silico modelling, and analysis of susceptibility to β-lactam antibiotics and combinations of β-lactamase inhibitors. Whole-genome sequencing analysis revealed a novel variant of AmpC β-lactamase belonging to the ACT family, designated ACT-107 by NCBI. This variant contains 12 novel amino acid mutations within the ACT family, 5 in the signal peptide sequence (Ile2, Met14, Tyr16, Gly18 and Thr20), and 7 in the mature protein (Gln22, His43, Cys60, Thr157, Glu225, Ala252 and Asn310). In silico modelling showed that substitutions occurring in the mature chain are localised in the solvent-accessible surface of the protein, where they are not expected to affect the β-lactamase activity, as observed in the resistance profile. Strikingly, 'not designated' ACT variants from E. huaxiensis were clustered (> 96% identity) with ACT-107. Since E. huaxiensis has been isolated from human infection, ACT-107 requires surveillance and the attention of clinicians.


Preparation of new surface coating based on modified oil-based polymers blended with ZnO and CuZnO NPs for steel protection.
In our paper, we have synthesized modified PEA and alkyd resin by replacing the new source of polyol (SDEA) which was confirmed by different analyses such as IR, and 1HNMR spectra. A series of conformal, novel, low-cost, and eco-friendly hyperbranched modified alkyd and PEA resins were fabricated with bio ZnO, CuO/ZnO) NPs through an ex-situ method for mechanical and anticorrosive coatings. The synthesized biometal oxides NPs and its composite modified alkyd and PEA were confirmed by FTIR, SEM with EDEX, TEM, and TGA, and can be stably dispersed into modified alkyd and PEA resins at a low weight fraction of 1%. The nanocomposite coating was also subjected to various tests to determine their surface adhesion, which ranged from (4B-5B), physico-mechanical characteristics such as scratch hardness, which improved from < 1.5 to > 2 kg, gloss (100-135) Specific gravity (0.92-0.96) and also chemical resistance test which passed for water, acid, and solvent except alkali, was poor because of the hydrolyzable ester group in the alkyd and PEA resins. The anti-corrosive features of the nanocomposites were investigated through salt spray tests in 5 wt % NaCl. The results indicate that well-dispersed bio ZnO and CuO/ZnO) NPs (1.0%) in the interior of the hyperbranched alkyd and PEA matrix improve the durability and anticorrosive attributes of the composites, such as degree of rusting, which ranged from 5 to 9, blistering size ranged from 6 to 9, and finally, scribe failure, which ranged from 6 to 9 mm. Thus, they exhibit potential applications in eco- friendly surface coatings. The anticorrosion mechanisms of the nanocomposite alkyd and PEA coating were attributed to the synergistic effect of bio ZnO and (CuO/ZnO) NPs and the prepared modified resins are highly rich in nitrogen elements, which might be regarded as a physical barrier layer for steel substrates.


Feasibility of using humic substances from compost to remove heavy metals (Cd, Cu, Ni, Pb, Zn) from contaminated soil aged for different periods of time.
There is a need for inexpensive, readily-available and environmentally-friendly soil washing agents to remediate polluted soils. Thus, batch washing experiments were performed to evaluate the feasibility of using a solution of humic substances (HS) extracted from compost as a washing agent for simultaneous removal of Cu, Cd, Zn, Pb and Ni from artificially contaminated soils aged for 1 month, 12 months and 24 months. The efficiency of metal removal in single and multiple washings and kinetic constants (equilibrium metal concentration qe and rate constant k from the second-order kinetic equation) were determined. On average, triple washing removed twice as much metal as that removed with a single washing. At pH 7 and a HS concentration of 2.2 g C L(-1), metal removal from all soils decreased in this order: Cd (79.1-82.6%) > Cu (51.5-71.8%) > Pb (44.8-47.6%) > Ni (35.4-46.1%) > Zn (27.9-35.8%). However, based on qe (mg kg(-1)), metal removal was in this order: Pb > Zn ≈ Cu > Ni > Cd. This difference was due to different concentrations of metals, which is typical for multi-metal contaminated soils. Regardless of washing mode, removal of Cd and Pb was not affected by soil age, whereas removal of Cu, Ni and Zn was higher in soils that had been aged for a shorter time. These results indicate that HS are suitable for remediating soil contaminated with multiple heavy metals in extremely high concentrations.


Narrative ability in high-functioning children with autism or Asperger's syndrome.
This study examines the narrative abilities of 28 high-functioning children with autism or Asperger's Syndrome and 22 typically developing children across two different discourse contexts. As compared with the typically developing children, the high-functioning group performed relatively well in the storybook context but exhibited difficulty imbuing their narratives of personal experience with the more sophisticated characteristics typically employed by the comparison group. Furthermore, children with autism or Asperger's Syndrome demonstrated impairments inferring and building on the underlying causal relationships both within and across story episodes in both narrative contexts. Findings further revealed that the narrative abilities of children with autism or Asperger's Syndrome were associated with performance on measures of emotional understanding, but not theory of mind or verbal IQ. Findings are discussed in relation to the social and emotional underpinnings of narrative discourse.


Excision of a submandibular gland: a safe day case procedure?
There are considerable benefits, both for patients and hospitals, if operations are done as day case procedures. Excision of a submandibular gland is a relatively common operation and it is usual practice for surgeons to be cautious, admit the patient for an overnight stay, and leave a drain in place. To assess the amount of postoperative bleeding into the wound (and hence potential risk to the airway) we have studied prospectively the amount of drainage that occurs. Sixty consecutive patients admitted for overnight postoperative monitoring after excision of a submandibular gland had a suction drain placed as part of the procedure. Drainage was measured by departmental staff at regular intervals during the following 24h. Nearly all the patients drained 40ml or less (mean 18ml) and in all cases there was a clear decrease in the volumes drained over the first 6-8h postoperatively. Drainage then became negligible. The plateau in drainage was evident regardless of the initial volume drained. Surgeons should be confident that drainage will cease after 6-8h in most patients, and residual drainage is negligible.


Bioconcentration, bioaccumulation and biomagnification of mercury in plankton of the Mediterranean Sea.
Plankton plays a prominent role in the bioaccumulation of mercury (Hg). The MERITE-HIPPOCAMPE campaign was carried out in spring 2019 along a north-south transect including coastal and offshore areas of the Mediterranean Sea. Sampling of sea water and plankton by pumping and nets was carried out in the chlorophyll maximum layer. Two size-fractions of phytoplankton (0.7-2.7 and 2.7-20 μm) and five of zooplankton (between 60 and >2000 μm) were separated, and their total mercury (THg) and monomethylmercury (MMHg) contents were measured. Bioconcentration of THg was significantly higher in the smallest phytoplankton size-fraction dominated by Synechococcus spp. The bioaccumulation and biomagnification of MMHg in zooplankton was influenced by size, food sources, biochemical composition and trophic level. MMHg was biomagnified in the plankton food web, while THg decreased toward higher trophic levels. Higher MMHg concentrations were measured in oligotrophic areas. Plankton communities in the Southern Mediterranean Sea had lower MMHg concentrations than those in the Northern Mediterranean Sea. These results highlighted the influence of environmental conditions and trophodynamics on the transfer of Hg in Mediterranean plankton food webs, with implications for higher trophic level consumers.


Hyponatremia as a risk factor for microvascular spasm following subarachnoid hemorrhage.
Hyponatremia is a water-electrolyte balance disorder diagnosed in about 30% of patients after subarachnoid hemorrhage (SAH). The main factors responsible for hyponatremia in these patients are increased plasma concentrations of either vasopressin (leading to water retention and dilutional hyponatremia) or natriuretic peptides (leading to plasma sodium ions deficiency). Data demonstrates that the leading causes of post-SAH disability - delayed cerebrovascular spasm (CVS) and delayed cerebral ischemia (DCI) - are more often diagnosed in patients who develop hyponatremia than in normonatremic patients with SAH. Data also indicates that reducing sodium ion concentration in the blood/perfusate affects the tone and regulation of cerebral blood vessels in a manner that depends on the vessel's location in a vascular tree (intraparenchymal arterioles vs. large vessels on the brain surface) and environmental conditions. In the present article, we review possible mechanisms underlying the effects of hyponatremia on cerebral blood vessels and discuss the potential role of hyponatremia in the development of large vessels and microvascular spasm, taking into consideration the presence of vasopressin and natriuretic peptides.


A CRISPR-enhanced metagenomic NGS test to improve pandemic preparedness.
The lack of preparedness for detecting and responding to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pathogen (i.e., COVID-19) has caused enormous harm to public health and the economy. Testing strategies deployed on a population scale at day zero, i.e., the time of the first reported case, would be of significant value. Next-generation sequencing (NGS) has such capabilities; however, it has limited detection sensitivity for low-copy-number pathogens. Here, we leverage the CRISPR-Cas9 system to effectively remove abundant sequences not contributing to pathogen detection and show that NGS detection sensitivity of SARS-CoV-2 approaches that of RT-qPCR. The resulting sequence data can also be used for variant strain typing, co-infection detection, and individual human host response assessment, all in a single molecular and analysis workflow. This NGS work flow is pathogen agnostic and, therefore, has the potential to transform how large-scale pandemic response and focused clinical infectious disease testing are pursued in the future.


Inhibition of macrophage phagocytosis by methylation inhibitors. Lack of correlation of protein carboxymethylation and phospholipid methylation with phagocytosis.
Adenosine (Ado), deoxyadenosine (dAdo), and adenine arabinoside (AraA) inhibit the phagocytosis of IgG-coated erythrocytes and zymosan by resident and thioglycollate-elicited macrophages (thio-macrophages) in a dose-dependent and reversible manner. 3-Deazaadenosine (3cAdo) and adenine (Ade) also inhibit the phagocytosis by resident macrophages. Homocysteine thiolactonate (Hcy) potentiates the inhibition by Ado and 3cAdo while erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) potentiates the inhibition by Ado, dAdo and AraA. This inhibition has a very rapid onset and the drugs do not interfere with the binding of IgG-coated erythrocytes to macrophages. The combination of Ado, Hcy and EHNA does not appreciably affect the intracellular level of ATP and S-adenosyl-L-methionine (AdoMet) in thio-macrophages but causes accumulations of Ado and S-adenosyl-L-homocysteine (AdoHcy) up to 135 and 145 nmol/mg of protein, respectively. During phagocytosis reversal, Ado is metabolized within 15 min while AdoHcy decreases log-arithmically with a half-life of 50 min. Carboxymethylation and phospholipid methylation, however, resume about 60-90 min after phagocytosis has recovered, and thus cannot function as transmembrane signals for phagocytosis. Other evidence showing the lack of correlation between phagocytosis and carboxymethylation inhibition include 1) Ado + Hcy inhibit carboxymethylation much better than Ado + EHNA (91 versus 75%) in thio-macrophage, but the two combinations show comparable phagocytosis inhibition potency; 2) Ado + Hcy inhibit carboxymethylation almost as well as Ado + Hcy + EHNA, but the latter is a much more effective drug combination for phagocytosis inhibition; 3) Ade and 3cAdo, although inhibiting resident macrophage phagocytosis as well as Ado + EHNA + Hcy, are much weaker carboxymethylation inhibitors; 4) dAdo and AraA potently inhibit phagocytosis but not carboxymethylation. The difference in the apparent methylation levels is not due to changes in the specific activities of AdoMet, which decrease with a half-life of 88 min. Interestingly, after the initial lag phase of about 90 min after the initiation of inhibition reversal, carboxymethylation and phagocytosis increase in parallel. In a log-log plot of carboxymethylation, phospholipid methylation, or phagocytosis versus the intracellular AdoHcy accumulation, a linear relationship is obtained. It is possible that AdoHcy accumulation is responsible for phagocytosis inhibition but inhibits by a mechanism other than interfering with protein and lipid methylations.


zDHHC9 Regulates Cardiomyocyte Rab3a Activity and Atrial Natriuretic Peptide Secretion Through Palmitoylation of Rab3gap1.
Production and release of natriuretic peptides by the stressed heart reduce cardiac workload by promoting vasodilation, natriuresis, and diuresis, which has been leveraged in the recent development of novel heart-failure pharmacotherapies, yet the mechanisms regulating cardiomyocyte exocytosis and natriuretic peptide release remain ill defined. We found that the Golgi S-acyltransferase zDHHC9 palmitoylates Rab3gap1 resulting in its spatial segregation from Rab3a, elevation of Rab3a-GTP levels, formation of Rab3a-positive peripheral vesicles, and impairment of exocytosis that limits atrial natriuretic peptide release. This novel pathway potentially can be exploited for targeting natriuretic peptide signaling in the treatment of heart failure.