Molecular Docking and Dynamics Investigations for Identifying Potential Inhibitors of the 3-Chymotrypsin-like Protease of SARS-CoV-2: Repurposing of Approved Pyrimidonic Pharmaceuticals for COVID-19 Treatment.
This study demonstrates the inhibitory effect of 42 pyrimidonic pharmaceuticals (PPs) on the 3-chymotrypsin-like protease of SARS-CoV-2 (3CLpro) through molecular docking, molecular dynamics simulations, and free binding energies by means of molecular mechanics-Poisson Boltzmann surface area (MM-PBSA) and molecular mechanics-generalized Born surface area (MM-GBSA). Of these tested PPs, 11 drugs approved by the US Food and Drug Administration showed an excellent binding affinity to the catalytic residues of 3CLpro of His41 and Cys145: uracil mustard, cytarabine, floxuridine, trifluridine, stavudine, lamivudine, zalcitabine, telbivudine, tipiracil, citicoline, and uridine triacetate. Their percentage of residues involved in binding at the active sites ranged from 56 to 100, and their binding affinities were in the range from -4.6 ± 0.14 to -7.0 ± 0.19 kcal/mol. The molecular dynamics as determined by a 200 ns simulation run of solvated docked complexes confirmed the stability of PP conformations that bound to the catalytic dyad and the active sites of 3CLpro. The free energy of binding also demonstrates the stability of the PP-3CLpro complexes. Citicoline and uridine triacetate showed free binding energies of -25.53 and -7.07 kcal/mol, respectively. Therefore, I recommend that they be repurposed for the fight against COVID-19, following proper experimental and clinical validation.


Target-cell specificity of fusogenic liposomes: membrane fusion-mediated macromolecule delivery into human blood mononuclear cells.
Fusogenic liposome, a unique vector prepared by fusing ultraviolet-inactivated Sendai virus and liposome, is known to efficiently deliver content into various animal cells through membrane fusion. In this study, we examined the target-cell specificity of fusogenic liposome (FL)-mediated macromolecule delivery into human blood cells using diphtheria toxin fragment A (DTA) as a probe. Among the peripheral blood mononuclear cells (PBMC), FL was able to deliver its encapsulates into CD14+ monocytes and CD4-/CD8- T-cells, but not into CD19+ B-lymphocytes, CD4+ T-cells or CD8+ T-cells. The susceptibility of human leukemia cell lines to FL was similar to that of PBMC; the order of the reactivity was U937 (monoblastic leukemia)>MOLT4, Jurkat (T-lymphoma)>Daudi, BALL1 (B-lymphoma)>K562 (erythroblastic leukemia). Interestingly, FL showed similar binding activity to all of these leukemia cell lines. These findings indicate that, among blood cells, monocytes, monoblastic leukemia cells, CD4-/CD8- T-cells and T-lymphoma cells are preferable targets for FL-mediated macromolecule delivery. This is the first demonstration of the existence of non-permissive cells against FL. Our results also suggest that some molecules on target-cells other than the binding targets of SV-derived protein may participate in fusion between FL and cells.


Feasibility of image-based simulation to estimate ablation target in human ventricular arrhythmia.
Previous studies suggest that magnetic resonance imaging with late gadolinium enhancement (LGE) may identify slowly conducting tissues in scar-related ventricular tachycardia (VT). To test the feasibility of image-based simulation based on LGE to estimate ablation targets in VT. We conducted a retrospective study in 13 patients who had preablation magnetic resonance imaging for scar-related VT ablation. We used image-based simulation to induce VT and estimate target regions according to the simulated VT circuit. The estimated target regions were coregistered with the LGE scar map and the ablation sites from the electroanatomical map in the standard ablation approach. In image-based simulation, VT was inducible in 12 (92.3%) patients. All VTs showed macroreentrant propagation patterns, and the narrowest width of estimated target region that an ablation line should span to prevent VT recurrence was 5.0 ± 3.4 mm. Of 11 patients who underwent ablation, the results of image-based simulation and the standard approach were consistent in 9 (82%) patients, where ablation within the estimated target region was associated with acute success (n = 8) and ablation outside the estimated target region was associated with failure (n = 1). In 1 (9%) case, the results of image-based simulation and the standard approach were inconsistent, where ablation outside the estimated target region was associated with acute success. The image-based simulation can be used to estimate potential ablation targets of scar-related VT. The image-based simulation may be a powerful noninvasive tool for preprocedural planning of ablation procedures to potentially reduce the procedure time and complication rates.


Bis(1,10-phenanthroline-5,6-dione-κN,N')silver(I) 2-hy-droxy-3,5-dinitro-benzoate.
In the cation of the title salt, [Ag(C(12)H(6)N(2)O(2))(2)](C(7)H(3)N(2)O(7)), the Ag(I) atom is coordinated in a distorted tetra-hedral geometry by four N atoms from two 1,10-phenanthroline-5,6-dione ligands, while the 3,5-dinitro-salicylate anion has only a short contact [2.847 (6) Å] between one of its O atoms and the Ag(I) atom. The dihedral angle between the two 1,10-phenanthroline-5,6-dione ligands is 58.4 (1)°. There is an intra-molecular O-H⋯O hydrogen bond in the 3,5-dinitro-salicylate anion.


Emissive Supramolecular Systems Based on Reversible Bond Formation and Noncovalent Interactions.
Noncovalent interactions and reversible bond formations are widely seen in natural systems for the construction of sophisticated molecular systems that perform various biological processes. Inspired by the natural systems, luminescent supramolecular systems constructed by coordination-driven self-assembly and homometallic metal-metal interations have been studied increasingly. These supramolecular systems show fascinating luminescent behaviors that are not observed from single components. This review summarizes our progress in the development of two types of unique luminescent supramolecular systems. The mononuclear Pt(II) complex units can sandwich coinage metal ions to form heteropolynuclear complexes involving heterometallic metal-metal interactions. A close proximity of the two or three different metal ions by the noncovalent forces lead to orbital overlapping among the coinage metal ions and the Pt(II) complex units, showing emission color change accompanied with structural transformation and reversible metal binding behaviors. Emissive host-guest systems consisting of mononuclear metal complexes and a hydrogen-bonded capsule are also developed, that show a unique encapsulation-induced emission enhancement (EIEE) behavior.


[Protection of corneal endothelium from apoptosis by gene and cell therapy].
Protection of corneal endothelium from apoptosis using gene and cell therapy is in a translational phase. This approach offers advantages for eye banking and after transplantation. Safe vehicles for gene or cell therapeutic transduction of corneal endothelium with nucleic acids are available. This strategy will be further developed in consultation with the Paul Ehrlich Institute and European regulatory authorities.


Myenteric neurons of the mouse small intestine undergo significant electrophysiological and morphological changes during postnatal development.
Organized motility patterns in the gut depend on circuitry within the enteric nervous system (ENS), but little is known about the development of electrophysiological properties and synapses within the ENS. We examined the electrophysiology and morphology of myenteric neurons in the mouse duodenum at three developmental stages: postnatal day (P)0, P10–11, and adult. Like adults, two main classes of neurons could be identified at P0 and P10–11 based on morphology: neurons with multiple long processes that projected circumferentially (Dogiel type II morphology) and neurons with a single long process. However, postnatal Dogiel type II neurons differed in several electrophysiological properties from adult Dogiel type II neurons. P0 and P10–11 Dogiel type II neurons exhibited very prominent Ca(2+)-mediated after depolarizing potentials (ADPs) following action potentials compared to adult neurons. Adult Dogiel type II neurons are characterized by the presence of a prolonged after hyperpolarizing potential (AHP), but AHPs were very rarely observed at P0. The projection lengths of the long processes of Dogiel type II neurons were mature by P10–11. Uniaxonal neurons in adults typically have fast excitatory postsynaptic potentials (fEPSPs, 'S-type' electrophysiology) mainly mediated by nicotinic receptors. Nicotinic-fEPSPs were also recorded from neurons with a single long process at P0 and P10–11. However, these neurons underwent major developmental changes in morphology, from predominantly filamentous neurites at birth to lamellar dendrites in mature mice. Unlike Dogiel type II neurons, the projection lengths of neurons with a single long process matured after P10–11. Slow EPSPs were rarely observed in P0/P10–11 neurons. This work shows that, although functional synapses are present and two classes of neurons can be distinguished electrophysiologically and morphologically at P0, major changes in electrophysiological properties and morphology occur during the postnatal development of the ENS.


Characterization of the participation of sodium channels on the rise in Na+ induced by 4-aminopyridine (4-AP) in synaptosomes.
The participation of voltage-sensitive Na+ channels (VSSC) on the changes on internal (i) Na+, K+, Ca2+, and on DA, Glu, and GABA release caused by different concentrations of 4-AP was investigated in striatum synaptosomes. TTX, which abolished the increase in Na(i) (as determined with SBFI), induced by 0.1 mM 4-AP only inhibited by 30% the rise in Na(i) induced by 1 mM 4-AP. One millimolar 4-AP markedly decreased the fluorescence of the K+ indicator dye PBFI but 0.1 mM 4-AP did not. Like 1 mM 4-AP, ouabain decreased PBFI fluorescence and increased a considerable fraction of Na(i) in a TTX-insensitive manner. In contrast with the different TTX sensitivity of the rise in Na(i) induced by 0.1 and 1 mM 4-AP, the rise in Ca(i) (as determined with fura-2) induced by the two concentrations of 4-AP was markedly inhibited by TTX, as well as by omega-agatoxin in combination with omega-conotoxin GVIA, indicating that only the TTX-sensitive fraction of the rise in Na(i) induced by 4-AP is linked with the activation of presynaptic Ca2+ channels. It is concluded that the TTX-sensitive fraction of neurotransmitter release evoked by 4-AP is released by exocytosis, and the TTX insensitive fraction involves reversal of the neurotransmitters transporters. This contrasts with the exocytosis evoked by high K+ that is unchanged by TTX and with the neurotransmitter-transporter-mediated release evoked by veratridine, which is highly TTX sensitive and does not require activation of Ca2+ channels.


Clinical evaluation of autoantibodies to p53 protein in patients with chronic liver disease and hepatocellular carcinoma.
In hepatocellular carcinoma (HCC) of patients from the Western hemisphere, mutations in the p53 tumour suppressor gene are present in up to 37% of cases. Conformational change and cellular accumulation can initiate an immune response with generation of circulating autoantibodies to p53 protein. In the present study, we investigated 711 consecutive patients with chronic liver disease to evaluate the sensitivity and specificity of autoantibodies to p53 protein as a serological marker for HCC. Detection of p53 autoantibodies was performed using an enzyme-linked immunosorbent assay with immobilised recombinant p53 protein. Liver cirrhosis was present in 259 patients (36.4%) and a HCC was diagnosed in 75 patients (10.6%). Autoantibodies to p53 protein were detectable in 15 of 377 patients with chronic liver disease (4.0%) and in 10 of 259 patients presenting with liver cirrhosis (3.9%). All 25 p53 autoantibody-positive/HCC-negative patients were carefully investigated and no underlying malignancy was clinically detected, suggesting that elevated p53 antibody levels may not exclusively be detectable in patients with malignant disease. In patients with clinically manifest HCC, p53 autoantibodies were detected in 17 of 75 cases, thus resulting in a sensitivity of 22.7% and a specificity of 96.1%. In contrast, assessment of serum alpha-fetoprotein (AFP) resulted in a sensitivity and specificity of 69.3 and 91.8% (AFP > 20 ng/ml) and 53.3 and 99.1% (AFP > 100 ng/ml) for the detection of HCC, respectively. The data of the present study reveal that the presence of p53 autoantibodies in patients with chronic liver disease is not completely specific for HCC. Moreover, we obtained no direct evidence that p53 autoantibody formation precedes the clinical diagnosis of HCC. However, serological screening for HCC might be improved by combining AFP and p53 autoantibody assays.


Prevention of cardiac dysfunction, kidney fibrosis and lipid metabolic alterations in l-NAME hypertensive rats by sinapic acid--Role of HMG-CoA reductase.
The present study was designed to evaluate the effect of sinapic acid, a bioactive phenolic acid on high blood pressure associated cardiac dysfunction, kidney fibrosis and lipid alterations in N(ω)-nitro-l-arginine methyl ester hydrochloride (l-NAME) induced hypertensive rats. Sinapic acid was administered to rats orally at a dosage of 40 mg/kg everyday for a period of 4 weeks. Sinapic acid treatment significantly decreased mean arterial pressure, left ventricular end diastolic pressure, organ weights (liver and kidney), lipid peroxidation products in tissues (liver and kidney), activities of hepatic marker enzymes and the levels of renal function markers in serum of l-NAME rats. Sinapic acid treatment also significantly increased the level of plasma nitric oxide metabolites, and enzymatic and non-enzymatic antioxidants in tissues of l-NAME rats. Tissue damage was assessed by histopathological examination. Alterations in plasma angiotensin-converting enzyme activity, level of plasma lipoproteins and tissue lipids were corrected by sinapic acid treatment in l-NAME rats. Sinapic acid treatment significantly decreased the activity of 3-hydroxy-3-methylglutaryl-Coenzyme A (HMG-CoA) reductase in plasma and liver, whereas the activity of lecithin cholesterol acyl transferase was significantly increased in the plasma of hypertensive rats. Docking result showed the interaction between sinapic acid and HMG-CoA reductase. Sinapic acid has shown best ligand binding energy of -5.5 kcal/M. Moreover, in chick embryo model, sinapic acid improved vessel density on chorioallantoic membrane. These results of the present study concludes that sinapic acid acts as a protective agent against hypertension associated cardiac dysfunction, kidney fibrosis and lipid alterations.


The pathogenicity and immunologic relationship of a virulent and a tissue-culture-adapted Babesia bovis.
Babesia bovis grown in tissue culture was used to inoculate 12, 2-year-old Holstein steers. All 12 developed serological evidence of infection but only six had a febrile response of greater than or equal to 40 degrees C, and only one had a demonstrable B. bovis parasitemia. An average modest drop of 19% was observed in packed cell volume (PCV) during the period of reaction. All 12 steers were subsequently challenged with virulent B. bovis: seven on day 78 post inoculation (p.i.), two on day 106 p.i., and three on day 251 p.i. No demonstrable clinical response was observed in any of the 12 steers previously exposed to the tissue-culture organism, whereas severe signs of babesiosis occurred in seven 2-year-old, non-vaccinated control steers given a comparably virulent B. bovis challenge. All seven controls showed a febrile response, B. bovis parasitemias, with an average drop of 55% in PCV and a 28% mortality.


Designing split reporter proteins for analytical tools.
A current focus of biological research is to quantify and image cellular processes in living cells and animals. To detect such cellular processes, genetically-encoded reporters have been extensively used. The most common reporters include firefly luciferase, renilla luciferase, green fluorescent protein (GFP) and its variants with various spectral properties. This review describes novel design of split-GFP and luciferase reporters based on protein splicing, and highlights some potential applications with the reporters to study protein-protein interactions, protein localization, intracellular protein dynamics, and protein activity in living cells and animals.


Geophysical evidence from the MELT area for compositional controls on oceanic plates.
Magnetotelluric and seismic data, collected during the MELT experiment at the southern East Pacific Rise, constrain the distribution of melt beneath this mid-ocean-ridge spreading centre and also the evolution of the oceanic lithosphere during its early cooling history. Here we focus on structures imaged at distances approximately 100 to 350 km east of the ridge crest, corresponding to seafloor ages of approximately 1.3 to 4.5 million years (Myr), where the seismic and electrical conductivity structure is nearly constant and independent of age. Beginning at a depth of about 60 km, we image a large increase in electrical conductivity and a change from isotropic to transversely anisotropic electrical structure, with higher conductivity in the direction of fast propagation for seismic waves. Conductive cooling models predict structure that increases in depth with age, extending to about 30 km at 4.5 Myr ago. We infer, however, that the structure of young oceanic plates is instead controlled by a decrease in water content above a depth of 60 km induced by the melting process beneath the spreading centre.


Current state-of-the-art systemic therapy for pediatric soft tissue sarcomas.
Pediatric soft tissue sarcomas (STS) are a heterogeneous group of tumors. Rhabdomyosarcomas (RMS) are the most common histologic subtype, while synovial sarcomas and undifferentiated sarcomas are among the more common non-rhabdomyosarcomatous soft tissue sarcomas (NRSTS) encountered. While the survival outcome for certain groups of RMS patients is quite good, the prognosis for those with alveolar histology or those with metastatic or relapsed disease remains dismal. Also, the response rate for some NRSTS to conventional chemotherapy is suboptimal. Thus increased understanding of involved molecular pathways, such as the insulin growth factor and mammalian target of rapamycin pathways, may indicate potential targets for therapy. In addition, immunotherapy-based approaches that include both non-specific activation with interleukins as well as targeted tumor antigen specific T lymphocytes are emerging avenues in the treatment of children with soft tissue sarcomas.


Na/Ca exchange in the basolateral membrane of the A6 cell monolayer: role in Cai homeostasis.
The presence of a Na/Ca exchanger in A6 cells was investigated by measuring intracellular calcium (Cai) fluctuations and the 45Ca fluxes through the basolateral membranes (blm) of the cell monolayer. Removal of Na+ from the medium produced a transient increase in Cai followed by a regulatory phase returning Cai to control levels in 3-4 min, this phase being greatly accelerated (< 60 s) by NaCl addition (apparent Km of approximately 5 mM Na+). The Cai increase was only found with the Na(+)-free medium on the basolateral side of the cell monolayer. A twofold increase in the 45Ca influx was observed under these conditions. In Ca(2+)- depleted cells, the initial Cai increase after Ca2+ addition to the medium was greater when the putative Na/Ca exchanger was not functioning (i.e. in a Na(+)-free medium). 45Ca effluxes through the blm of the monolayer were greatly and transiently increased by a Na(+)-free medium on the serosal side and blocked by orthovanadate (1 mM). The Cai increased induced by a hypo-osmotic shock was greater in cells bathed in a Na(+)-medium, conditions expected to block the activity of the Na/Ca exchanger. These findings support the hypothesis that a Na/Ca exchanger is present on the blm of A6 cells and affirm its role in Cai homeostasis in steady-state conditions and following osmotic shock. In addition, a Ca2+ pump also located on the blm and Ca2+ stores sensitive to inositol 1,4,5-trisphosphate were found to be implicated in Cai homeostasis.


Diffusion imaging in dementia with Lewy bodies: Associations with amyloid burden, atrophy, vascular factors and clinical features.
White matter disruption in dementia has been linked to a variety of factors including vascular disease and cortical pathology. We aimed to examine the relationship between white matter changes on diffusion tensor imaging (DTI) in DLB and factors including vascular disease, structural atrophy and amyloid burden. Participants with DLB (n = 29), Alzheimer's disease (AD, n = 17) and healthy controls (n = 20) had clinical and neuropsychological assessments followed by structural and diffusion tensor 3T MRI and 18F-Florbetapir PET-CT imaging. Voxelwise statistical analysis of white matter fractional anisotropy (FA) and mean diffusivity (MD) was carried out using Tract-Based Spatial Statistics with family-wise error correction (p < 0.05). DLB and AD groups demonstrated widespread increased MD and decreased FA when compared with controls. There were no differences between the DLB and AD groups. In DLB, increased MD and decreased FA correlated with decreased grey matter and hippocampal volumes as well as vascular disease. There was no correlation with cortical florbetapir SUVR. The relationship between DTI changes and grey matter/hippocampal volumes remained after including Cumulative Illness Rating Scale-Geriatric vascular score as a covariate. Widespread disruption of white matter tracts is present in DLB and is associated with vascular disease, reduced hippocampal volume and reduced grey matter volume, but not with cortical amyloid deposition. The mechanism behind the correlation observed between hippocampal volume and white matter tract disruption should be investigated in future cohorts using tau imaging, as hippocampal atrophy has been shown to correlate with tau deposition in DLB.


Investigation of the effect of training attachment behaviors to pregnant mothers on some physical indicators of their infants from birth to three months based on the separation of male and female infants.
Maternal attachment to the fetus during pregnancy improves infant's growth and development and is an important criterion for public health. The aim is to investigate the effect of training attachment behaviors to pregnant women on some growth health indicators of infants. This is a clinical trial with convenience sampling which was conducted on 190 pregnant women in Hafez Hospital in Shiraz who randomly classified into intervention and control groups. For the intervention group, 6 sessions of 90-min classes were held. After delivery, both groups were compared in terms of physical growth (weight, height, head circumference) at birth, one and three months after birth. The data were analyzed using repeated Measurement. Weight increase in female infants in the intervention group compared to the control group was significant at birth (p = 0.016), one month (p = 0.010) and three months after birth (p = 0.014). Height increase in female infants in the intervention group compared to the control group was significant at birth (p = 0.025), one month (p < 0.001) and three months after birth (p = 0.009). Male gender was not statistically significant in any age group but it increased the size of the head circumference of three-month-old infants. Training of attachment skills to mothers improves height and weight of female infants in three steps. The average index for boys in the intervention group was higher than that of control group, but it was not significant.


Enhance wastewater biological treatment through the bacteria induced graphene oxide hydrogel.
The interaction between bacteria and graphene-family materials like pristine graphene, graphene oxide (GO) and reduced graphene oxide (rGO) is such an elusive issue that its implication in environmental biotechnology is unclear. Herein, two kinds of self-assembled bio-rGO-hydrogels (BGHs) were prepared by cultivating specific Shewanella sp. strains with GO solution for the first time. The microscopic examination by SEM, TEM and CLSM indicated a porous 3D structure of BGHs, in which live bacteria firmly anchored and extracellular polymeric substances (EPS) abundantly distributed. Spectra of XRD, FTIR, XPS and Raman further proved that GO was reduced to rGO by bacteria along with the gelation process, which suggests a potential green technique to produce graphene. Based on the characterization results, four mechanisms for the BGH formation were proposed, i.e., stacking, bridging, rolling and cross-linking of rGO sheets, through the synergistic effect of activities and EPS from special bacteria. More importantly, the BGHs obtained in this study were found able to achieve unique cleanup performance that the counterpart free bacteria could not fulfill, as exemplified in Congo red decolorization and Cr(VI) bioreduction. These findings therefore enlighten a prospective application of graphene materials for the biological treatment of wastewaters in the future.


Balancing of energy-consuming processes of rat hepatocytes.
A method for the quantification of energy consuming processes described by Siems et al. for reticulocytes and by Müller et al. for ascites tumour cells was applied to balance the ATP-consumption of isolated rat hepatocytes. On the basis of decreased coupled respiration rates following the specific inhibition of energy-requiring reactions, the energy demands of protein turnover, nucleic acid synthesis, Na+/K(+)-ATPase and Ca2(+)-transport of hepatocytes in different incubation media were assessed. These processes together with urea synthesis account for about 60 per cent of the total energy consumption in a glucose and amino acid-enriched Eagle/Borsook medium. The metabolic flux rates of total ATP-consumption and ATP-consumption of single energy-requiring processes in hepatocytes are compared with those in reticulocytes and different tumour cell types.


Genetic characterization of Sézary's syndrome by conventional cytogenetics and cross-species color banding fluorescent in situhybridization.
Sezary's syndrome is a peripheral T-cell neoplasm characterized by a pruritic exfoliative or infiltrated erythroderma, lymphadenopathies, and atypical T lymphocytes in the peripheral blood. Cytogenetic studies are scarce. This study was designed to increase cytogenetic information on this disorder. Peripheral blood samples were collected from 21 patients with Sezary's syndrome (10 men, 11 women, mean age 64 years) and analyzed by conventional cytogenetics (72-hr cultures with phytohemagglutinin). For a better characterization of multiple chromosomal rearrangements, cross-species color banding (RxFISH) was used in four cases. Fifteen (71.4%) of the 21 cases showed cytogenetic aberrations, with the karyotype being complex in 14. Among the 15 patients with an abnormal karyotype, 8 presented a diploid/near-diploid karyotype and 7 a near-tetraploid karyotype. The chromosomes most frequently involved were 1, 6, 8, 9, 10, 11, and 17. The most common structural rearrangements affected 1q, 2q, 6q23-27, and 8q22. Monosomies of chromosomes 9 and 10 and trisomies of chromosome 18 were recurrently observed. A statistical trend between abnormal and complex karyotypes, the presence of monosomy 10, the number of Sezary cells, and a decreased overall survival was observed. RxFISH technology allowed the description of 27 previously undetected chromosomal abnormalities. Abnormal karyotypes, particularly complex karyotypes, were frequently detected in patients with Sezary's syndrome. Monosomy 10 was the most frequent recurrent cytogenetic marker (73% in abnormal cases). There was a high diversity of chromosomal breakpoints. RxFISH is a useful novel technology for redefining complex karyotypes.


Drug interaction and pharmacokinetic modeling of oxcarbazepine in korean patients with epilepsy.
To determine first whether there was a clear relationship between concentrations of the active metabolite of oxcarbazepine (OXC), 10-hydroxycarbamazepine (OHC), and dose adjusted for weight, and second, whether the clearance of OHC was influenced by comedication with enzyme-inducing antiepileptic drugs (EIAED). We analyzed 254 cases (patients 3-80 years of age) of OXC therapeutic drug monitoring, retrospectively. The cases were categorized into OXC monotherapy (n = 78), OXC in combination with EIAED (n = 73), and OXC in combination with non-EIAED (n = 103). The serum OHC concentrations of predose samples were measured by high-performance liquid chromatography. A population pharmacokinetic model was developed using NONMEM. The mean ± SD serum concentration of OHC was 14.47 ± 8.28 μg/mL at a mean daily dose of 16.22 ± 7.99 mg/kg. The serum concentration of OHC was correlated with the OXC dose per body weight (r = 0.6005; P < 0.0001). No association was found between OHC concentration and patient age, weight, sex, or seizure type. The concentration-to-dose ratio on OXC in combination with EIAED was significantly lower than that on OXC monotherapy (P = 0.002) or OXC in combination with non-EIAED (P < 0.0001). In population pharmacokinetic modeling, the apparent clearance of OHC was higher by 31.2% in combination with EIAED than in other groups. The serum concentration of OHC was statistically significantly correlated with the dose of OXC and negatively correlated with comedication of EIAED. Population pharmacokinetic analysis showed that the apparent clearance of OHC increased with comedication with EIAEDs.


Patient and Caregiver Reciprocal Support: Impact on Decision Making in Specialist Palliative Care.
Patients and informal caregivers in palliative care can reciprocate in supporting one another. However, how reciprocal support among patients and informal caregivers in palliative care impacts on their decision making for care is not well understood. To identify how reciprocal support among patients with advanced illness and their informal caregivers in specialist palliative care impacts on their decision making for care. Between July 2021 and May 2022, 30 qualitative interviews were conducted with 14 patient and caregiver dyads, seven nondyad caregiver participants and one nondyad patient participant (total n = 36), recruited from a large regional specialist palliative care service. Data were analyzed using Corbin and Strauss grounded theory method. Reciprocal support among patients and informal caregivers was underpinned by obligation and choice. Caregivers who felt obliged to care had difficulty communicating with the patient about the patient's preferences for care and their own wishes for patient care. Patients who felt obliged to accept support from their caregiver tended to minimize caregiver participation in decision making which made caregivers feel disempowered in discussions about patient care. Caregivers tended to be more involved in decision making when caregivers assumed caregiving duties by choice and when the patient did not feel restricted by their reliance on their caregiver. Open communication between patients and caregivers made patients more trusting of their caregiver. Patient and caregiver dyadic interventions in specialist palliative care involving decision making need to account for how obligation and choice manifest and function between the patient and caregiver.


Quality and Certification of Electronic Health Records: An overview of current approaches from the US and Europe.
Numerous projects, initiatives, and programs are dedicated to the development of Electronic Health Records (EHR) worldwide. Increasingly more of these plans have recently been brought from a scientific environment to real life applications. In this context, quality is a crucial factor with regard to the acceptance and utility of Electronic Health Records. However, the dissemination of the existing quality approaches is often rather limited. The present paper aims at the description and comparison of the current major quality certification approaches to EHRs. A literature analysis was carried out in order to identify the relevant publications with regard to EHR quality certification. PubMed, ACM Digital Library, IEEExplore, CiteSeer, and Google (Scholar) were used to collect relevant sources. The documents that were obtained were analyzed using techniques of qualitative content analysis. The analysis discusses and compares the quality approaches of CCHIT, EuroRec, IHE, openEHR, and EN13606. These approaches differ with regard to their focus, support of service-oriented EHRs, process of (re-)certification and testing, number of systems certified and tested, supporting organizations, and regional relevance. The analyzed approaches show differences with regard to their structure and processes. System vendors can exploit these approaches in order to improve and certify their information systems. Health care organizations can use these approaches to support selection processes or to assess the quality of their own information systems.


New implant surgical guide.
This article describes the process of making and using a surgical guide stent. It is intended to enable the surgeon to maintain the same horizontal and vertical axes of cylindrical endosseous implants during the surgical phase of implant placement as those determined at the treatment planning stage. Furthermore, the stent can be used with the entire series of surgical drills, thereby minimizing the chance of inadvertently enlarging the implant site as a result of freehand use of the handpiece.


Plasma unconjugated estriol values in high-risk pregnancy.
The usefulness of plasma estriol concentrations in the management of high-risk pregnancy is controversial. In this study, serial plasma unconjugated estriol (E3) concentrations determined by radioimmunoassay were evaluated in 321 patients with high-risk pregnancy for correlation with fetal and neonatal performance. A plasma E3 concentration of 4 ng. per milliliter or less in late pregnancy correlated significantly with low one-and five-minute Apgar scores and selected neonatal problems in pregnancies complicated by chronic hypertension and intrauterine growth retardation, with a similar trend for pregnancies complicated by moderate and severe pre-eclampsia. Most perinatal deaths in the study were associated with acute problems which began during labor and delivery, or fetal death occurred before early intervention was practical. Therefore, the perinatal mortality rate is not likely to be influenced by the use of E3 values, and the perinatal mortality rate is not a realistic measure of the usefulness of plasma E3 values in management of high-risk pregnancy. Low plasma E3 values were not always ominous, and for this reason it is suggested that clinical indications and the results of other tests for fetal-placental function be considered along with plasma E3 values in the selection of patients for early delivery to reduce the possibility of inappropriate intervention.


In vitro assays of the antibacterial and antioxidant activity of aqueous leaf extracts from different Prunus salicina Lindl. cultivars.
The growing interest in the substitution of synthetic food antioxidants and antimicrobial additives by natural ones has fostered research on vegetable sources and on the screening of raw materials, for identifying new antioxidants and antimicrobial natural agents. The aim of the present study was to assess total phenolic contents and determine polyphenolic composition, related antioxidative and antimicrobial properties of plum leaves extracts from six cultivars. It was observed that the content of total phenolic compounds was cultivar dependent. High antioxidant capacity has been observed and related to the relative amounts of polyphenolic compounds with good antioxidant properties. The antimicrobial activity was confirmed against Listeria innocua and Escherichia coli, and it has found to be related with the high phenolic contents. Our results suggest that the use of plum leaf extracts is a feasible alternative as antibacterial and antioxidant agents to prevent the deterioration of stored foods by bacteria and oxidation.


Identification of cytochrome P450 2D6 and 2C9 substrates and inhibitors by QSAR analysis.
This paper presents four new QSAR models for CYP2C9 and CYP2D6 substrate recognition and inhibitor identification based on human clinical data. The models were used to screen a large data set of environmental chemicals for CYP activity, and to analyze the frequency of CYP activity among these compounds. A large fraction of these chemicals were found to be CYP active, and thus potentially capable of affecting human physiology. 20% of the compounds within applicability domain of the models were predicted to be CYP2C9 substrates, and 17% to be inhibitors. The corresponding numbers for CYP2D6 were 9% and 21%. Where the majority of CYP2C9 active compounds were predicted to be both a substrate and an inhibitor at the same time, the CYP2D6 active compounds were primarily predicted to be only inhibitors. It was demonstrated that the models could identify compound classes with a high occurrence of specific CYP activity. An overrepresentation was seen for poly-aromatic hydrocarbons (group of procarcinogens) among CYP2C9 active and mutagenic compounds compared to CYP2C9 inactive and mutagenic compounds. The mutagenicity was predicted with a QSAR model based on Ames in vitro test data.


Strike-slip 23 January 2018 MW 7.9 Gulf of Alaska rare intraplate earthquake: Complex rupture of a fracture zone system.
Large intraplate earthquakes in oceanic lithosphere are rare and usually related to regions of diffuse deformation within the oceanic plate. The 23 January 2018 MW 7.9 strike-slip Gulf of Alaska earthquake ruptured an oceanic fracture zone system offshore Kodiak Island. Bathymetric compilations show a muted topographic expression of the fracture zone due to the thick sediment that covers oceanic basement but the fracture zone system can be identified by offset N-S magnetic anomalies and E-W linear zones in the vertical gravity gradient. Back-projection from global seismic stations reveals that the initial rupture at first propagated from the epicenter to the north, likely rupturing along a weak zone parallel to the ocean crustal fabric. The rupture then changed direction to eastward directed with most energy emitted on Aka fracture zone resulting in an unusual multi-fault earthquake. Similarly, the aftershocks show complex behavior and are related to two different tectonic structures: (1) events along N-S trending oceanic fabric, which ruptured mainly strike-slip and additionally, in normal and oblique slip mechanisms and (2) strike-slip events along E-W oriented fracture zones. To explain the complex faulting behavior we adopt the classical stress and strain partitioning concept and propose a generalized model for large intra-oceanic strike-slip earthquakes of trench-oblique oriented fracture zones/ocean plate fabric near subduction zones. Taking the Kodiak asperity position of 1964 maximum afterslip and outer-rise Coulomb stress distribution into account, we propose that the unusual 2018 Gulf of Alaska moment release was stress transferred to the incoming oceanic plate from co- and post-processes of the nearby great 1964 MW 9.2 megathrust earthquake.


Successful cryopreservation of purified autologous CD34+ cells: influence of freezing parameters on cell recovery and engraftment.
Conventional hematopoietic stem cell cryopreservation methods use a DMSO concentration of 10%. However, cells manipulated ex vivo may require more refined freezing protocols adapted to the specific cell suspension. In this retrospective study, we evaluated the results obtained with CD34+ cells purified from peripheral blood of 39 patients on the CEPRATE SC System and frozen in 7.5% DMSO with a view to transplantation. The post-freezing recovery of progenitor cells was 89.4 +/- 27.87% for CD34+ cells, 59.13 +/- 36.93% for CFU-GM, and 53.49 +/- 40.71 for BFU-E. Neither the purity of the suspension nor the nucleated cell density during freezing was predictive of cell recovery. No difference was observed between cells stored in vials and bags. Thirty-seven patients transplanted with the concentrated CD34+ fraction received 4.46 x 10(6) CD34+ cells/kg and 33.04 x 10(4) CFU-GM/kg. The median time to granulocyte (>0.5 x 10(9)/l) and platelet (>50 x 10(9)/l) engraftment was 11 and 13 days, respectively. Only cell density and the infused number of CD34+ cells and CFU-GM were significantly related to hematological recovery. Our data suggest that purified CD34+ cells can be successfully cryopreserved in 7.5% DMSO and may represent a first step in establishing freezing parameters for selected CD34+ cells.


Potential Antiosteoporotic Natural Product Lead Compounds That Inhibit 17β-Hydroxysteroid Dehydrogenase Type 2.
17β-Hydroxysteroid dehydrogenase type 2 (17β-HSD2) converts the active steroid hormones estradiol, testosterone, and 5α-dihydrotestosterone into their weakly active forms estrone, Δ4-androstene-3,17-dione, and 5α-androstane-3,17-dione, respectively, thereby regulating cell- and tissue-specific steroid action. As reduced levels of active steroids are associated with compromised bone health and onset of osteoporosis, 17β-HSD2 is considered a target for antiosteoporotic treatment. In this study, a pharmacophore model based on 17β-HSD2 inhibitors was applied to a virtual screening of various databases containing natural products in order to discover new lead structures from nature. In total, 36 hit molecules were selected for biological evaluation. Of these compounds, 12 inhibited 17β-HSD2 with nanomolar to low micromolar IC50 values. The most potent compounds, nordihydroguaiaretic acid (1), IC50 0.38 ± 0.04 μM, (-)-dihydroguaiaretic acid (4), IC50 0.94 ± 0.02 μM, isoliquiritigenin (6), IC50 0.36 ± 0.08 μM, and ethyl vanillate (12), IC50 1.28 ± 0.26 μM, showed 8-fold or higher selectivity over 17β-HSD1. As some of the identified compounds belong to the same structural class, structure-activity relationships were derived for these molecules. Thus, this study describes new 17β-HSD2 inhibitors from nature and provides insights into the binding pocket of 17β-HSD2, offering a promising starting point for further research in this area.


Antibiotic consumption trends among acute care hospitals in Catalonia (2008-2016): impact of different adjustments on the results.
Hospital antibiotic consumption is measured using defined-daily-doses (DDD) divided by bed days. However,other denominators as discharges could provide a more accurate interpretation of consumption. The main objective was to analyze trends of antibiotic consumption among hospitals in Catalonia during the period 2008-2016, using both DDD/100 bed days and DDD/100 discharges. Retrospective, descriptive, and longitudinal study performed among acute care hospitals affiliated to VINCat Program. Antibiotic consumption was expressed using the Anatomical Therapeutic Chemical/DDD classification and trends with a mixed linear model. Trends after using both DDD/100 bed days and DDD/100 discharges were determined and compared. Overall antibiotic consumption from 2008 to 2016 increased by 10.24% (P < 0.001) DDD/100 bed days, but remained stable (-0.87%, P = 0.051) in DDD/100 discharges. Although DDD and discharges remained unchanged, a significant reduction in bed days (-9.63%) and length of stay (-8.19%) was observed. A worrisome increase in the consumption of carbapenems and anti-MRSA drugs was noticed. Whereas a significant upward trend in antibiotic consumption in DDD/100 bed days was noticed, DDD/100 discharges remained stable. The description of both indicators seems therefore essential for a correct interpretation of data.


RNA LEGO: magnesium-dependent formation of specific RNA assemblies through kissing interactions.
The high affinity and specificity of nucleic acid base complementarity has been proven to be a powerful method for constructing specific molecular assemblies. On the other hand, recent structural studies of RNA have revealed the wide range of tertiary interactions utilized in RNA folding, which may potentially be used as tools for the design of specific macromolecular assemblies. Here, RNA building blocks containing two hairpin loops, based on the dimerization initiation site (DIS) of HIV RNA, connected by a short linker were used to construct large RNA assemblies through hairpin loop-loop ("kissing") interactions. We show that specific linear and circular assemblies can be constructed in a magnesium-dependent manner using several non-self-complementary loop-loop interactions designed in this study. These results show that the use of RNA tertiary interactions may broaden the repertoire of nucleic acid-based nanostructures.


Plant health status effects on arbuscular mycorrhizal fungi associated with Lavandula angustifolia and Lavandula intermedia infected by Phytoplasma in France.
We investigated root communities of arbuscular mycorrhizal fungi (AMF) in relation to lavender (Lavandula angustifolia) and lavandin (Lavandula intermedia) health status from organic and conventional fields affected by Phytoplasma infection. The intensity of root mycorrhizal colonization was significantly different between diseased and healthy plants and was higher in the latter regardless of agricultural practice. This difference was more pronounced in lavender. The root AMF diversity was influenced by the plant health status solely in lavender and only under the conventional practice resulting in an increase in the AMF abundance and richness. The plant health status did not influence the distribution of root AMF communities in lavandin unlike its strong impact in lavender in both agricultural practices. Finally, among the most abundant molecular operational taxonomic units (MOTUs), four different MOTUs for each plant species were significantly abundant in the roots of healthy lavender and lavandin in either agricultural practice. Our study demonstrated that the plant health status influences root colonization and can influence the diversity and distribution of root AMF communities. Its effects vary according to plant species, can be modified by agricultural practices and allow plants to establish symbiosis with specific AMF species.


A Preliminary Study Investigating the Influence of Auditory Stimulation on the Occurrence of Nocturnal Equine Sleep-Related Behavior in Stabled Horses.
The physical environment is known to influence nocturnal behavioral time budgets of the stabled horse, but less evidence exists to suggest how this might be affected by including additional sensory stimuli. This study aimed to establish the impact of novel auditory stimuli on the frequency of equine sleep-related behavior. Seven horses stabled for 24 hours per day on the same yard receiving the same daily management routine were observed from 2030 to 0630 over nine nights. Frequency of nocturnal behavior was recorded using focal intermittent sampling against a predetermined ethogram and an infrared CCTV camera system. Data were recorded under the following conditions: without music for two nights (phase A1), exposure to music for five nights (Beethoven's ninth Symphony) played at an average of 62.3 decibels (phases B1 [nights 3-4] and B2 [nights 6-7]), and two further nonconsecutive nights (phase A2) when music was no longer played. A general linear model was used to determine differences in the frequency of parametric behavioral data with a significantly higher occurrence of "ingestion" (F [3,18] = 7.910, P = .001) during phases in B compared with A, and a significant decrease in the occurrence of "other" behavior (F [3,18] = 10.25, P = .000) comparing phase A1 with all other phases. The Wilcoxon signed rank test highlighted significant differences in the frequency of "lateral recumbency" between specific phases (P < .05). The addition of music appears to have a significant effect on the equine nocturnal time budget that might be beneficial from an equine sleep perspective.


Effects of inulin on gilthead seabream (Sparus aurata L.) innate immune parameters.
Inulin, a fructooligossacharide, is a prebiotic that plays an important role in the immune function in mammals, but it has never been assayed in other vertebrate groups. Thus, we have studied the inulin effects on the gilthead seabream (Sparus aurata L.) innate immune response both in vitro and in vivo. For the in vitro study, head-kidney leucocytes were incubated with inulin (ranging from 0 to 1000 microg ml(-1)) for 30, 90, 180 and 300 min and 24h and any effect was observed on leucocyte viability or the main innate cellular immune responses (leucocyte peroxidase, phagocytic, respiratory burst and natural cytotoxic activities). For the in vivo study, seabream specimens were fed for 1 or 2 weeks with a commercial diet supplemented with inulin: 0 (control), 5 or 10 g inulin kg(-1) diet (0.5 and 1%, respectively). Inulin produced a significant inhibition in phagocytosis and respiratory burst in leucocytes from specimens fed diets containing 0.5% or 1% of inulin for 1 week. Based on the present results, inulin does not seem to be a good immunostimulant for seabream, though its effects in other species and combined with other immunostimulans (i.e. probiotics) might be of great interest.


Immunogenicity of four sequential polypeptides in inbred guinea-pigs and their recognition at the T lymphocyte and antibody levels.
Four sequential polypeptides containing equimolar amounts of tyrosine, glutamic acid, alanine and glycine were shown to be T lymphocyte-dependent immunogens in inbred guinea-pigs. Poly (Glu-Ala-Tyr-Gly) was immunogenic only in strain 2 guinea-pigs; poly (Glu-Tyr-Ala-Gly) and poly (Ala-Tyr-Glu-Gly) were immunogenic only in strain 13 guinea-pigs; and poly (Ala-Glu-Tyr-Gly) was immunogenic in both inbred strains. The specificity of immune recognition was probed at the T lymphocyte and humoral levels with the heterologous polypeptides. Only a few cases of heterologous cross-stimulation or cross-reaction were observed, indicating the great selectivity of immune recognition. The results showed considerable variability in immune recognition from animal to animal. Nevertheless, at the T-cell level, cross-stimulation appears to necessitate that the antigen is itself immunogenic in that strain, whereas at the antibody level, cross-reaction is not similarly restricted. The structural basis for mutual recognition of immunogen and antigen at these levels is discussed.


Experimental study on the explosion characteristics of hydrogen-methane premixed gas in complex pipe networks.
To explore the overpressure evolution laws and flame propagation characteristics in complex pipe networks after the addition of hydrogen to methane, we experimentally studied the explosive pressure wave and flame wave propagation laws for three different premixed gas mixtures with hydrogen-methane concentrations of 0, 10% and 20% when the equivalence ratio was 1. Experimental results indicate that the maximum explosion overpressure of the premixed gas increases with increasing distance from the explosion source, and it shows a gradually decreasing trend. In the complex pipe network, an overpressure zone is formed in the B-E-H and D-E sections of the network. The flame temperature is superimposed with the superimposition of the pressure, showing a trend of first increasing, then decreasing, then increasing, and finally decreasing in the complex pipe network. The flame arrival time increases with increasing distance, and the maximum flame speed shows a decreasing trend. The peak overpressure and maximum flame velocity of the premixed gas under a hydrogen volume fraction of 20% are 1.266 MPa and 168 m/s. The experimental research results could provide important theoretical guidelines for the prevention and control of fuel gas explosions in urban pipe networks.


Two neuronal, nuclear-localized RNA binding proteins involved in synaptic transmission.
While there is evidence that distinct protein isoforms resulting from alternative pre-mRNA splicing play critical roles in neuronal development and function, little is known about molecules regulating alternative splicing in the nervous system. Using Caenorhabditis elegans as a model for studying neuron/target communication, we report that unc-75 mutant animals display neuroanatomical and behavioral defects indicative of a role in modulating GABAergic and cholinergic neurotransmission but not neuronal development. We show that unc-75 encodes an RRM domain-containing RNA binding protein that is exclusively expressed in the nervous system and neurosecretory gland cells. UNC-75 protein, as well as a subset of related C. elegans RRM proteins, localizes to dynamic nuclear speckles; this localization pattern supports a role for the protein in pre-mRNA splicing. We found that human orthologs of UNC-75, whose splicing activity has recently been documented in vitro, are expressed nearly exclusively in brain and when expressed in C. elegans, rescue unc-75 mutant phenotypes and localize to subnuclear puncta. Furthermore, we report that the subnuclear-localized EXC-7 protein, the C. elegans ortholog of the neuron-restricted Drosophila ELAV splicing factor, acts in parallel to UNC-75 to also affect cholinergic synaptic transmission. In conclusion, we identified a new neuronal, putative pre-mRNA splicing factor, UNC-75, and show that UNC-75, as well as the C. elegans homolog of ELAV, is required for the fine tuning of synaptic transmission. These findings thus provide a novel molecular link between pre-mRNA splicing and presynaptic function.


PuMYB21/PuMYB54 coordinate to activate PuPLDβ1 transcription during peel browning of cold-stored "Nanguo" pears.
Refrigeration is commonly used to extend the storage life of "Nanguo" pears, but fruit in long-term refrigeration is prone to peel browning, which is related to membrane lipid degradation. To determine the mechanism of membrane lipid degradation, we identified two R2R3-MYB transcription factors (TFs), PuMYB21 and PuMYB54, from "Nanguo" pears, which were notably expressed in response to cold stress and during the peel-browning process. The results from yeast one-hybrid, electrophoretic mobility shift, and transient expression assays indicated that both PuMYB21 and PuMYB54 directly bind to the promoter of PuPLDβ1 (a key enzyme catalyzing the hydrolysis of membrane phospholipids) and activate its expression, which probably enhances the degradation of membrane phospholipids and eventually results in peel browning. Moreover, the overexpression of PuMYB21 and PuMYB54 can greatly activate the transcription of endogenous PuPLDβ1 in both "Nanguo" pear fruits and calli, and their silencing can inhibit its transcription. Furthermore, yeast two-hybrid, bimolecular fluorescence complementation, and pull-down assays verified that PuMYB21 interacts with PuMYB54 to enhance the expression of PuPLDβ1. In summary, we demonstrate that PuMYB21 and PuMYB54 may have roles in membrane lipid metabolism by directly binding to the downstream structural gene PuPLDβ1 during the low temperature-induced peel browning of "Nanguo" pears.


Optic neuropathy associated with vitamin B12 deficiency.
A 17-year-old boy with vitamin B12 deficiency that occurred after a small bowel resection developed bilateral centrocecal scotomas during folic acid therapy and improved on therapy including vitamin B complex.


Rehabilitation of people with rheumatoid arthritis.
Rehabilitation, as an adjunct to pharmacological and surgical therapies in patients with rheumatoid arthritis (RA), aims to minimize the consequences of the disease. For a systematic assessment of the consequences of disease, an appropriate definition and evaluation of the goals of therapy and interventions, and an active partnership with the patient, a structured approach to rehabilitation management is needed.Despite widespread positive clinical experience with rehabilitative interventions, the scientific evidence of their effectiveness is, in general, scanty, owing to a lack of studies with sufficient methodological quality. Further well-designed clinical studies are warranted with respect to several interventions where evidence is falling short.


Ionic strength, pH and temperature effects upon selectivity for transition and heavy metal ions when using chelation ion chromatography with an iminodiacetic acid bonded silica gel column and simple inorganic eluents.
An investigation into the selectivity of an iminodiacetic acid (IDA) modified silica gel column for transition and heavy metal ions using non-chelating inorganic eluents has been carried out. A number of eluent parameters were investigated to determine the exact retention mechanism taking place and to control selectivity. The parameters studied were eluent ionic strength and the nature of the inorganic salt used, eluent pH and eluent temperature. The results obtained showed how despite certain metal ions exhibiting similar stability constants with the bonded IDA groups, careful control of each of the above parameters, in particular eluent chloride ion concentration and eluent temperature, could result in large changes in selectivity. Optimal conditions for the isocratic and gradient separation of Mg(II), Ca(II), Mn(II), Cd(II), Co(II), Zn(II) and Pb(II) were determined. An isocratic method using a 0.035 M KCl, 0.065 M KNO3 (pH 2.5) eluent was successfully applied to the determination of Mn(II), Cd(II), Co(II) and Zn(II) at concentrations between 20 and 121 microg/l in a freshwater certified reference material (NIST 1640).


MCP-1 facilitates VEGF production by removing miR-374b-5p blocking of VEGF mRNA translation.
Monocyte chemotactic protein-1 (MCP-1) is known to be able to facilitate vascular endothelial growth factor (VEGF) gene expression, hence promoting vascular hyperpermeability and neovascularization. We show here that a microRNA molecule, miR-374b-5p can target the 3'-untranslated region of the VEGF mRNA, thus preventing VEGF production. Additionally, MCP-1 promotes the acetylation of transcription factor stat3 at Lys685, which facilitates the formation of an ac-stat3-DNA methyltransferase-histone methyltransferase complex (ac-stat3/DNMT1/EZH2) that binds to the promoter of the miR-374b-5p gene. This results in diminished miR-374b-5p expression and enhanced VEGF production. Moreover, treatment of appropriate animal models either with a miR-374b-5p mimicry or with inhibitors of either stat3 acetylation, DNMT1, or EZH2, leads to marked inhibition of MCP-1-promoted neovascularization and tumor growth. These findings indicate that MCP-1 facilitated inhibition of miR-374b-5p gene expression leads to the removal of a block of VEGF mRNA translation by miR-374b-5p. This mechanism could be of importance in the modulation of inflammatory conditions.


Pseudo-malignant paranasal invasive aspergillosis showing bone destruction and FDG uptake on PET/CT: A case report.
Aspergillosis is a rare disease that often occurs in patients with diabetes mellitus or compromised hosts. This study describes a 60-years-old woman with unusual pseudo-malignant paranasal invasive aspergillosis with 18F-fluorodeoxyglucose (18F-FDG) uptake treated by endoscopic sinus surgery. To the best of our knowledge, there are few reported cases of paranasal fungal infection with 18F-FDG uptake. A 60-years-old woman was presented with headache and nasal obstruction. Computed tomography (CT) showed a shadow with bone destruction in the sinus cavity and accumulation of 18F-FDG uptake. The patient was diagnosed with a malignant tumor or pseudo-malignant paranasal invasive aspergillosis. The patient underwent endoscopic sinus surgery; no neoplastic lesions were detected in the areas with CT shadows. All the observed fungal mass reservoirs were removed. The patient remained hospitalized for 1 week after the surgery with no significant postoperative abnormalities. There was no recurrence of the disease for 6 months, and the patient's symptoms resolved, indicating a good course of follow-up. Invasive aspergillosis should be considered a differential disease when positron emission tomography (PET)/CT scans show FDG uptake, CT shows bone destruction, and T2-weighted MRI scans show a low signal.


Selective and Stepwise Functionalization of the Pyridazine Scaffold by Using Thio-Substituted Pyridazine Building Blocks.
We described a regioselective tri- and tetra-functionalization of the pyridazine scaffold using two readily available building blocks: 3-alkylthio-6-chloropyridazine and 3,4-bis(methylthio)-6-chloropyridazine by performing selective metalations with TMPMgCl ⋅ LiCl and catalyst-tuned cross-coupling reactions with arylzinc halides. Several of the resulting pyridazines were converted into more elaborated N-heterocycles such as thieno[2,3-c]pyridazines and 1H-pyrazolo[3,4-c]pyridazines.


Expanding the Role of Continuous Glucose Monitoring in Modern Diabetes Care Beyond Type 1 Disease.
Application of continuous glucose monitoring (CGM) has moved diabetes care from a reactive to a proactive process, in which a person with diabetes can prevent episodes of hypoglycemia or hyperglycemia, rather than taking action only once low and high glucose are detected. Consequently, CGM devices are now seen as the standard of care for people with type 1 diabetes mellitus (T1DM). Evidence now supports the use of CGM in people with type 2 diabetes mellitus (T2DM) on any treatment regimen, not just for those on insulin therapy. Expanding the application of CGM to include all people with T1DM or T2DM can support effective intensification of therapies to reduce glucose exposure and lower the risk of complications and hospital admissions, which are associated with high healthcare costs. All of this can be achieved while minimizing the risk of hypoglycemia and improving quality of life for people with diabetes. Wider application of CGM can also bring considerable benefits for women with diabetes during pregnancy and their children, as well as providing support for acute care of hospital inpatients who experience the adverse effects of hyperglycemia following admission and surgical procedures, as a consequence of treatment-related insulin resistance or reduced insulin secretion. By tailoring the application of CGM for daily or intermittent use, depending on the patient profile and their needs, one can ensure the cost-effectiveness of CGM in each setting. In this article we discuss the evidence-based benefits of expanding the use of CGM technology to include all people with diabetes, along with a diverse population of people with non-diabetic glycemic dysregulation.


Postprandial accumulation of chylomicrons and chylomicron remnants is determined by the clearance capacity.
To better understand the postprandial clearance of triglyceride-rich lipoproteins (TRLs) and its relation to the fasting kinetics of TRLs. Two studies were performed on 30 male subjects: a fasting kinetic study to determine the fasting secretion and clearance rates of apolipoprotein B (apoB) 100 and triglycerides in the very low-density lipoprotein 1 and 2 (VLDL(1) and VLDL(2)) fractions; and a postprandial study to determine the postprandial accumulation of apoB48, apoB100 and triglycerides in the chylomicron, VLDL(1) and VLDL(2) fractions. Results from these two studies were combined to characterize the postprandial clearance of TRLs in a physiologically relevant setting. Our results show that postprandial accumulation of the apoB48-carrying chylomicrons can be predicted from the clearance capacity of the lipolytic pathway, determined in the fasting state. Furthermore, we show that chylomicrons and VLDL(1) particles are not cleared equally by the lipoprotein lipase pathway, and that chylomicrons seem to be the preferred substrate. Subjects with a rapid fasting lipid metabolism accumulate lower levels of postprandial triglycerides with less accumulation of apoB100 in the VLDL(1) fraction and a faster transfer of apoB100 into the VLDL(2) fraction. In contrast, fasting VLDL(1) secretion does not predict postprandial triglyceride accumulation. Non-fasting triglyceride levels have recently been identified as a major predictor of future cardiovascular events. Here we show that the capacity of the lipolytic pathway is a common determinant of both the fasting and non-fasting triglyceride levels and may thus play an important role in the development of dyslipemia and atherosclerosis.


Cytomegalovirus infection with multiple colonic perforations in a renal transplant recipient.
Cytomegalovirus (CMV) continues to be potentially the most important pathogen affecting organ transplant recipients. Severe gastrointestinal complications have been reported to occur in about 10% of renal transplant recipients, sometimes with dramatic presentations. We report the case of a 57-year-old CMV-seropositive woman with end-stage renal failure who developed CMV-related colonic multiple perforation 30 days after cadaveric CMV-positive renal transplantation. CMV pp65 antigenemia test and CMV-PCR had always been negative on all the weekly controls routinely performed in the postoperative period. Only after the sudden onset of this complication did the antigenemia and PCR become positive. The relationship between infection and perforation has been established beyond any doubt, as the histology of the resected colonic segment revealed florid CMV infection with evidence of typical inclusions in both macrophages and endothelial cells. Colonic perforations are often fatal in transplant recipients because of inability to contain the perforation, and only a rapid diagnosis and an aggressive surgical treatment can improve the prognosis.


The Use of Ratings of Perceived Exertion in Children and Adolescents: A Scoping Review.
In adults, ratings of perceived exertion (RPE) can be used to predict maximal oxygen uptake, estimate time to exhaustion, assess internal training load and regulate exercise intensity. However, the utility of RPE in children is less researched and therefore, warrants investigation. The purpose of this scoping review is to map out the literature around the application of RPE specifically during aerobic exercise in paediatric populations. Seven bibliographic databases were systematically searched. Grey literature searching and pearling of references were also conducted. To be included for the review, studies were required to comply with the following: (1) participants aged ≤ 18 years asymptomatic of any injuries, disabilities or illnesses; (2) applied RPE in aerobic exercise, testing and/or training; (3) included at least one measure of exercise intensity; and (4) be available in English. The search identified 22 eligible studies that examined the application of RPE in children. These studies involved a total of 718 participants across ten different countries. Nine different types of RPE scales were employed. Overall, the application of RPE in paediatric populations can be classified into three distinct themes: prediction of cardiorespiratory fitness/performance, monitoring internal training loads, and regulation of exercise intensity. The utility of RPE in paediatric populations remains unclear due to the small body of available research and inconsistencies between studies. However, findings from the included studies in this scoping review may show promise. Further research focussing on child-specific RPE scales across various sports, subgroups, and in field-based settings is needed.


Acceptability of a self-sampling technique to collect vaginal smears for gram stain diagnosis of bacterial vaginosis.
To diagnose asymptomatic bacterial vaginosis (BV), self-sampled vaginal smears were collected during a study of risk factors for preterm birth in African American women. More than 90% of those women who were willing to participate in the interview portion of the study were also willing to provide a self-sampled vaginal smear. The smears are an acceptable and efficient way of detecting BV in an urban minority population.


Deep diving with Clio.
An autonomous underwater vehicle, named Clio, can sample ocean basin-scale biogeochemistry at depths up to 6000 meters.


Association of dietary components with dyslipidemia and low-grade inflammation biomarkers in adults with heterozygous familial hypercholesterolemia from different countries.
The association of components of a low saturated fat (SFA) and of a Mediterranean diet was tested with atherosclerosis biomarkers in 190 familial hypercholesterolemia adults (FH) from Brazil (BR) and Spain (SP). Median blood LDL-C, Apolipoprotein B (apoB), and C reactive protein (hs-CRP) concentrations were higher in BR than in SP: 179.0 vs.161 mg/dL; 141 vs. 103 mg/dL; and 1.6 vs. 0.8 mg/L respectively (all p < 0.001). In BR there was lower median total fat (22.3 vs. 38.3%) and SFA (8.1 vs. 12.5%) but higher cholesterol (283.3 mg vs.188.9 mg) and carbohydrate (57.1 vs. 42.5%) consumption (all p < 0.001). Inverse associations were encountered between fibers, mono, and polyunsaturated fats and their ratios to SFA with LDL-C and ApoB (all p < 0.001). There was a direct association respectively of cholesterol with lipid biomarkers and of carbohydrates and trans-fatty acids with hs-CRP while other fats showed inverse relations with the latter (p < 0.001).


Surfactant phospholipid metabolism in 3-day and 3-day postmature rabbits in vivo.
Three-day-old normal and 3-day postmature rabbits (prepared by human chorionic gonadotropin treatment of the does) were given injections of the radiolabeled phospholipid precursors palmitic acid, choline, and glycerol. The specific activities as cpm/mumol phospholipid phosphate were measured for phosphatidylcholine, disaturated phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, and phosphatidylethanolamine from lung and alveolar wash fraction lipid extracts of 3-day-old rabbits. Specific activities similarly were measured for phosphatidylcholine and disaturated phosphatidylcholine from postmature rabbits. Curves describing the appearance of labeled phospholipids into the alveolar wash fraction demonstrated that in both 3-day and postmature rabbits, phospholipid secretion was delayed for 3 to 4 hr. Thereafter, there was a rapid accumulation of labeled phospholipids until 12 to 16 hr after labeled precursor injection. The curves were similar for all measured phospholipids and all labeled precursors studied. The phospholipid compositions of lung and alveolar wash fraction lipid extracts were determined by analysis of the various phospholipids separated by two-dimensional thin-layer chromatography. Forty percent of lung samples and 35% of alveolar wash samples from postmature rabbits contained no detectable phosphatidylglycerol, whereas all 22 lung and surfactant samples from 3-day-old rabbits contained this phospholipid.


Hypoxia related long non-coding RNAs in ischemic stroke.
With high rates of mortality and disability, stroke has caused huge social burden, and 85% of which is ischemic stroke. In recent years, it is a progressive discovery of long non-coding RNA (lncRNA) playing an important regulatory role throughout ischemic stroke. Hypoxia, generated from reduction or interruption of cerebral blood flow, leads to changes in lncRNA expression, which then influence disease progression. Therefore, we reviewed studies on expression of hypoxia-related lncRNAs and relevant molecular mechanism in ischemic stroke. Considering that hypoxia-inducible factor (HIF) is a crucial regulator in hypoxic progress, we mainly focus on the HIF-related lncRNA which regulates the expression of HIF or is regulated by HIF, further reveal their pathogenesis and adaption after brain ischemia and hypoxia, so as to find effective biomarker and therapeutic targets.


Different Resistance Mechanisms of Medicago truncatula Ecotypes Against the Rust Fungus Uromyces striatus.
ABSTRACT A pathosystem consisting of the model plant Medicago truncatula and the rust fungus Uromyces striatus was characterized. From a collection of 113 mostly European accessions of M. truncatula, the vast majority were found to be susceptible to U. striatus, whereas 5 accessions showed strong resistance reactions. Stomatal surface characteristics, even if partly occluded, did not interfere with the ability of U. striatus germ tubes to infect. After penetration, the resistant ecotypes reacted with various degrees of cell death during different stages of haustorial establishment. Whereas four ecotypes showed a typical hypersensitive reaction by developing necrotic lesions, one ecotype (F11.008) exhibited a prehaustorial type of defense without hypersensitive response. This ecotype may be used as a source of nonhost-type of resistance against U. striatus.


The Influence of Presentation Format of Story on Narrative Production in Chinese Children Learning English-as-a-Second-Language: A Comparison Between Graphic Novel, Illustration Book and Text.
Past studies have shown that multimodal presentation of story can improve story-retelling performance in the first language. The purpose of the present study was to investigate whether similar multimedia effects can be observed in second language learning and graphic novel reading. A total of 51 Chinese elementary school children, aged 7-8, who were learning English as a second language were recruited. They were randomly assigned to one of the three experimental conditions that differed in the format of story presentation: English text, English text with pictorial illustrations or graphic novel. After reading the same story, the children retold the story in English. The narratives produced were then rated by two independent raters. The results of group comparison showed that children from the three experimental groups had similar performance, indicating that multimedia presentation may not always facilitate narrative production in English as a second language. Within-subject comparison further showed that the children were relatively strong in language skills and capturing the main ideas of the story, while showing weakness in story structure awareness, elaboration, as well as local and global cohesion. Suggestions for the application of multimodal presentation of narrative texts are discussed.


Students in Turkey During the Early Days of the COVID-19 Pandemic.
This study aims to analyse the subjective well-being of university students during the early days of the COVID-19 pandemic process and to examine how students have been affected economically, psychologically and socially. Other subjects included in the scope of the study are the suitability of the physical conditions during the process of online education, the students' motivation, and their expectations towards the future. The study also analyses the connection between these factors and the subjective well-being concepts of happiness and life satisfaction. The study focuses on students of the Faculty of Economics at Marmara University in Turkey. In total, 428 students took part in the survey prepared for the study. Descriptive analyses indicate that students have been negatively affected in terms of psychological and socio-economic factors along with subjective well-being. Ordered probit models show a statistically significant relationship between a major part of these changes and subjective well-being variables.


A new method for the assay of factor XI in plasma and the preparation and use of a new artificial substrate.
An improved method for the one-stage assay of factor XI in plasma has been proposed; it is reproducible and compares well with an old, established method.A short investigation into the preparation and use of artificially prepared factor XI-deficient plasma has been carried out: this substrate can be used as an alternative to the congenitally deficient substrate.


Retrospective and prospective analyses of the treatment of overanticoagulated patients.
To compare withholding warfarin therapy with low-dose (2.5 mg) oral vitamin K therapy in excessively anticoagulated patients without bleeding complications. Prospective and retrospective studies. Anticoagulation clinic at a Veterans Affairs institution. Twenty-eight men were matched according to initial international normalized ratio (INR) and INR goal ranges. The retrospective arm of the study consisted of chart reviews of overanticoagulated patients whose warfarin doses were held until therapeutic INR values were reached. The prospective arm included overanticoagulated patients who were administered a single 2.5-mg dose of oral vitamin K. Mean days to therapeutic INR values were 2.3+/-0.6 and 1.4+/-0.6 (p=0.001), and mean reduction in INR 1 day after treatment intervention was 1.32+/-0.79 and 3.46+/-1.31 (p<001) U for the withholding and vitamin K groups, respectively. Compared with withholding the warfarin dose, administration of 2.5 mg of oral vitamin K to excessively anticoagulated patients receiving warfarin significantly reduced the time required to reach a therapeutic INR as well as final INR.


FOXO1 reduces tumorsphere formation capacity and has crosstalk with LGR5 signaling in gastric cancer cells.
Gastric cancer (GC) is a major of cause of cancer-related death and is characterized by its heterogeneity and molecular complexity. FOXO1 is a transcription factor that plays a key role in GC growth and metastasis. However, the implication of FOXO1 in GC cell stemness has been elusive. This study, for the first time, demonstrates that FOXO1 regulates GC cell stemness in association with LGR5. FOXO1 expression was significantly lower in GC tumorsphere cells than in adherent GC cells. FOXO1 silencing and overexpression promoted and inhibited the tumorsphere formation capacity of GC cells, respectively. Additionally, there was an inverse correlation between FOXO1 and GC stem cell marker LGR5 in human GC specimens. Further in vitro and in vivo experiments showed that negative crosstalk between these two molecules exists and that LGR5 silencing reversed the FOXO1 shRNA-induced tumorsphere formation even without FOXO1 restoration. Taken together, our results suggest that FOXO1 inhibits the self-renewal capacity of GC cells through interaction with LGR5. Thus, FOXO1/LGR5 signaling pathway may provide a novel targeted therapy for GC.


Platinum drug adduct formation in the nucleosome core alters nucleosome mobility but not positioning.
Nucleosome positioning and reorganization regulate DNA site exposure in chromatin. Platinum anticancer agents form DNA adducts that disrupt nuclear activities, triggering apoptosis. Mechanistic insight would aid in the development of improved therapies to circumvent drug toxicity and resistance. We show that platinum adducts formed by reaction of cisplatin or oxaliplatin with the nucleosome core inhibit histone octamer-DNA sliding but do not cause significant alteration of positioning. Thus, adduct formation reinforces positional preferences intrinsic to the DNA sequence, which indicates that modulation of platinum drug site selectivity by histone octamer association may relate to nucleosome-specific properties of DNA. This sheds light on platinum drug-mediated inhibition of chromatin remodeling in vivo and suggests that adducts can shield their own repair and interfere with genomic activities by directly altering nucleosome dynamics.


Age-related trends in neuromagnetic transient beta burst characteristics during a sensorimotor task and rest in the Cam-CAN open-access dataset.
Non-invasive neurophysiological recordings, such as those measured by magnetoencelography (MEG), provide insight into the behaviour of neural networks and how these networks change with factors such as task performance, disease state, and age. Recently, there has been a trend in describing neurophysiological recordings as a series of transient bursts of neural activity rather than averaged sustained oscillations as burst characteristics may be more directly correlated with the neurological generators of brain activity. In this work, we investigate how beta burst characteristics change with age in a large open access dataset. The objectives are (1) to detect and characterize transient beta bursts over the ipsilateral and contralateral primary sensorimotor cortices during a unilateral motor task performance and during wakeful resting, and (2) to identify age-related changes in beta burst characteristics, in the context of earlier reports of age-related changes in beta suppression and the post-movement beta rebound. MEG data, acquired at the Cambridge Centre for Ageing and Neuroscience, of roughly 600 participants with a nearly uniform distribution of ages between 18 and 88 years old was used for analysis. We found that burst rate is the predominant factor related to age-related changes in the amplitude of the induced beta rhythm responses associated with a button press task. Furthermore, we present a cross-validation of burst parameters detected at the sensor- (peak sensor and sensor ROI) and source-level (beamformer spatial filter). This work is as an important step in characterizing transient bursts in neuromagnetic signals in the temporal domain, towards a better understanding of the healthy aging human brain.


The Trypanosoma rangeli trypomastigote surfaceome reveals novel proteins and targets for specific diagnosis.
Sympatric distribution and sharing of hosts and antigens by Trypanosoma rangeli and Trypanosoma cruzi, the etiological agent of Chagas' disease, often incur in misdiagnosis and improper epidemiological inferences. Many secreted and surface proteins (SP) have been described as important antigens shared by these species. This work describes the T. rangeli surfaceome obtained by gel-free (LC-ESI-MS/MS) and gel-based (GeLC-ESI-MS/MS) proteomic approaches, and immunoblotting analyses and the comparison of these SP with T. cruzi. A total of 138 T. rangeli proteins and 343 T. cruzi proteins were obtained, among which, 42 and 157 proteins were exclusively identified in T. rangeli or T. cruzi trypomastigotes, respectively. Immunoblotting assays using sera from experimentally infected mice revealed a distinct band pattern for each species. MS/MS analysis of T. rangeli exclusive bands revealed two unique GP63-related proteins and flagellar calcium-binding protein. Also, a ~32kDa band composed of 12 distinct proteins was exclusively recognized by anti-T. cruzi serum. This highly sensitive proteomic assessment of surface proteins characterized the T. rangeli surfaceome, revealing several differences and similarities between these two parasites. The study reports new T. rangeli-specific proteins with promising use in differential diagnosis from T. cruzi. In this manuscript, we report the first proteomic analysis of the T. rangeli surface (surfaceome), a non-pathogenic parasite occurring in sympatry with T. cruzi, the etiological agent of Chagas disease. This comparative proteomic analysis was performed using high-throughput in-gel and gel-free proteomic approaches combined with immunoblotting, allowing us to identify new T. rangeli-specific proteins with promising use in differential serodiagnosis, among several other protein not previously reported for this taxon. Additionally, cross-recognition assays showed that T. cruzi surface proteins were recognized by heterologous serum (anti-T. rangeli) that strengthens the possibility of misdiagnosis of Chagas disease in humans and other mammals. Thus, this work provides new insights to understand the serological cross-reactivity between T. cruzi and T. rangeli, as well as, the identification of targets for specific T. rangeli diagnosis as revealed by the comparative surfaceome analysis. We strongly believe that this research is of importance to the readers of Journal of Proteomics since it provides new potential markers for diagnosis of both T. cruzi and T. rangeli parasites increasing the spectrum of specific targets for unambiguous diagnosis of T. rangeli and T. cruzi infections, besides describing new approaches to assess the trypanosomatids proteome.


Location of the globular region in chicken erythrocyte histone H5.
Chicken erythrocyte histone H5 has been cleaved by acetic acid hydrolysis at the two aspartic acid residues 65 and 99 and the 4 peptides (1-65), (66-185) (1-99) (100-185) recovered in a pure form. 270 MHz magnetic resonance and circular dichroic studies show that the two C-terminal peptides are unable to form secondary or tertiary structure. The N-terminal peptides however, form both secondary and tertiary structure. In particular, the peptide (1-99) at high ionic strength possesses a similar number of helical residues to intact histone H5 and also had a closely related nuclear magnetic resonance spectrum. It is concluded that the peptide (1-99) contains most, but not quite all of the residues that are included in the globular segment of histone H5.


Vertical transmission of the hepatitis C virus to infants of anti-human immunodeficiency virus-negative mothers: molecular evolution of hypervariable region 1 in prenatal and perinatal or postnatal infections.
In a prospective study of 33 infants born to hepatitis C virus (HCV)-positive human immunodeficiency virus-negative mothers the vertical transmission of HCV occurred in 6.8%. The evolution of HCV infection in two babies was studied from birth up to 5 or 6 years of age, and the sequencing of the hypervariable region (HVR) of the putative envelope-encoding E2 region of the HCV genome was performed. The HVR1 sequence variability and the different serological profiles during follow-up could reflect the differences in HCV transmission routes, HCV genotypes, and clinical evolution of infection.


Unique functional responses of fungal communities to various environments in the mangroves of the Maowei Sea in Guangxi, China.
Fungi are important compartments of microbial communities of mangroves. Their diversity might be influenced by their habitat environment. This study analyzed the distribution and function of fungal communities in the sediments and plant samples from mangrove ecosystem of the Maowei Sea area in Guangxi, China. The results showed that phytopathogenic fungi Cladosporium (17.00%) was mainly observed in the sediments from the protected zone, while endophytic fungi Alternaria (9.22%) and Acremonium (6.09%) were only observed in the sediments from wharf. The fungi in the sediments from village and park were mainly consisted of high-activity endophytes and fungi related to lignin-degrading, respectively. Acaulospora and Aspergillus with higher relative abundance discovered in plant tissues could help plant growth. Cirrenalia (37.66%) and Lignincola (26.73%) with high-activity for lignin-degrading were discovered in decayed leaves. The distribution and function of fungi were highly dependent on the environment settings, thus the fungi can be used as indicators for monitoring the environmental change of mangrove ecosystems.


Identification of a druggable pocket of the calcium-activated chloride channel TMEM16A in its open state.
The calcium-activated chloride channel TMEM16A is a potential drug target to treat hypertension, secretory diarrhea, and several cancers. However, all reported TMEM16A structures are either closed or desensitized, and direct inhibition of the open state by drug molecules lacks a reliable structural basis. Therefore, revealing the druggable pocket of TMEM16A exposed in the open state is important for understanding protein-ligand interactions and facilitating rational drug design. Here, we reconstructed the calcium-activated open conformation of TMEM16A using an enhanced sampling algorithm and segmental modeling. Furthermore, we identified an open-state druggable pocket and screened a potent TMEM16A inhibitor, etoposide, which is a derivative of a traditional herbal monomer. Molecular simulations and site-directed mutagenesis showed that etoposide binds to the open state of TMEM16A, thereby blocking the ion conductance pore of the channel. Finally, we demonstrated that etoposide can target TMEM16A to inhibit the proliferation of prostate cancer PC-3 cells. Together, these findings provide a deep understanding of the TMEM16A open state at an atomic level and identify pockets for the design of novel inhibitors with broad applications in chloride channel biology, biophysics, and medicinal chemistry.


Pretreatment body mass index predicts survival among patients administered nivolumab monotherapy for pretreated non-small cell lung cancer.
Biomarker assessments for nivolumab monotherapy efficacy in previously treated patients with non-small cell lung cancer (NSCLC) remain unclear. We evaluated whether body mass index (BMI) and Glasgow prognostic score (GPS) are useful for assessing the efficacy of nivolumab alone as a second-line treatment in patients with pretreated NSCLC. Data of 99 patients treated with second-line nivolumab monotherapy for NSCLC between January 2016 and December 2019 were evaluated for prognostic values of BMI and GPS to assess their usefulness in predicting progression-free survival (PFS) and overall survival (OS). The Eastern Cooperative Oncology Group-performance status (PS) independently predicted the second-line nivolumab monotherapeutic effect; good PS (0-1) correlated with significantly longer PFS (4.3 vs. 1.9 months, log-rank; p = 0.0004) and OS (17.7 vs. 4.6 months, log-rank; p < 0.0001) than poor PS. BMI independently predicted survival, with high BMI (≥22.1 kg/m2 ) associated with significantly longer OS (19.1 vs. 8.5 months, log-rank; p = 0.0023) than low BMI (<22.1 kg/m2 ). However, GPS showed no significant difference for PFS or OS. Among patients with NSCLC treated with nivolumab monotherapy as second-line treatment, PS was significantly correlated with both PFS and OS and BMI with OS. Thus, BMI could be a useful predictor of survival in these patients.


Prevention of peroxynitrite-dependent damage by carnosine and related sulphonamido pseudodipeptides.
The naturally occurring dipeptides carnosine and anserine have been proposed to act as antioxidants in vivo. We investigated whether these compounds can act as protective agents able to counteract peroxynitrite-dependent reactions. The results showed that the dipeptides efficiently protect tyrosine against nitration, alpha1-antiproteinase against inactivation and human low-density lipoprotein against modification by peroxynitrite. Carnosine exerts its protective effect at concentrations similar to those found in human tissues. In addition, some synthetic pseudodipeptides, stucturally related to carnosine but stable to hydrolytic enzymes, possess protective properties against peroxynitrite-dependent damage similar to the natural dipeptides. These pseudodipeptides may represent stable mimics of the biologically active carnosine suitable for pharmacological applications.


Prevalence and significance of ischemic cerebrovascular events in giant cell arteritis.
To determine the prevalence of cerebrovascular events (CVE) in giant cell arteritis (GCA) and to alert clinicians to the importance of early detection of CVE in this disease. Retrospective observational study involving a cohort of GCA patients. Demographic, clinical and laboratory data were collected. All patients fulfilled the American College of Rheumatology (ACR) 1990 GCA classification criteria and had a positive ultrasound test for GCA in agreement with the EULAR recommendations. Demographic and clinical parameters were recorded with special attention paid to ischemic cranial events. We studied 123 consecutive GCA patients, 74 (60.2%) women with a mean age of 79 years. Twelve patients (9.75%) suffered from neurologic symptoms other than AION, of whom 9 (7.3%) experienced ischemic events related to GCA and 3 (2.44%) likely experienced CVE due to other common causes. Of the 9 patients with CVE caused by GCA, 5 were diagnosed with transient ischemic attacks (TIAs), 2 with ischemic stroke, and 2 were cases involving cranial nerve palsies. High rates of mortality were found in patients with a TIA or stroke, while polymyalgia rheumatica (PMR) appeared to confer some protection against ischemic pathologies in GCA patients. Stroke and TIA are common presentation patterns associated with GCA and should be suspected in all CVE-related cases with high acute-phase reactants commonly present in the elderly. This ischemic subgroup exhibited a higher mortality rate.


Cytogenetic abnormalities and genomic copy number variations in EPO (7q22) and SEC-61(7p11) genes in primary myelodysplastic syndromes.
Myelodysplastic syndromes (MDSs) are heterogeneous clonal haematopoeitic stem cell disorders characterized by ineffective haematopoeisis, cytopenias and risk of progression to AML. We studied 150 MDS patients for cytogenetic aberrations and 60 patients with normal karyotype and 40 patients harboring cytogenetic abnormalities for copy number variations (CNVs). Cytogenetic abnormalities were detected in 46% of patients with a majority of patients harboring abnormalities of chromosome 7 and del (20q) at frequencies of 16% and 12% respectively. We explored the potential of quantitative multiplex PCR assay of short fluorescent fragments (QMPSF) to identify CNVs and correlated the findings with cytogenetic data and disease prognosis. CNVs (n=31) were detected in 28.3% of karyotypically normal and 23% patients with abnormal karyotype. Genetic losses or deletions (n=26) were more frequent than duplications (n=5). EPO (7q22) and SEC-61(7p11) emerged as new candidate genes susceptible to genetic losses with 57.7% deletions identified in regions on chromosome 7. The CNVs correlated with International Prognostic Scoring System (IPSS) intermediate disease risk group. Our integrative cytogenetic and copy number variation study suggests that abnormalities of chromosome 7 are predominant in Indian population and that they may play a secondary role in disease progression and should be evaluated further for asserting their clinical significance and influence on disease prognosis.


The burden of lung disease in Hong Kong: a report from the Hong Kong Thoracic Society.
The burden of lung disease in Hong Kong is not known. This study determined the mortality and hospitalization rates of respiratory diseases in Hong Kong in 2005, their trend in the past decade and their incidence/prevalence. Mortality data were obtained from the Department of Health and hospitalization data from the Hospital Authority, Hong Kong. Incidence/prevalence data were obtained from local registries or local studies. Trends of mortality and hospitalization rates of various respiratory diseases from 1997 and 2005 were calculated after age standardization and were tested for significance using negative binomial regression analysis. Age standardized mortality rates in Hong Kong were compared with those of the UK and globally. Respiratory disease was the most common cause of mortality and hospitalization in Hong Kong in 2005. Globally and in the UK, cardiovascular disease ranked first in mortality. Respiratory infections ranked first in respiratory mortality, followed by respiratory tract cancer and chronic obstructive lung disease. Respiratory infections also ranked first followed by chronic obstructive lung disease in the utilization of respiratory inpatient bed-days. While mortality rates from all respiratory diseases decreased in the past decade, hospitalization rates remained unchanged. Unlike other respiratory diseases, mortality from respiratory infections have increased since 2001. Smoking is the most important risk factor in non-communicable respiratory diseases. Respiratory disease is responsible for the highest health-care burden locally. Increased efforts in improving management and prevention of these diseases, including tobacco control, improving air quality and vaccination against influenza and pneumococci, are necessary.


An Experimental and Theoretical Investigation of the Electronic Structures and Photoelectrical Properties of Ethyl Red and Carminic Acid for DSSC Application.
The photoelectrical properties of two dyes-ethyl red and carminic acid-as sensitizers of dye-sensitized solar cells were investigated in experiments herein described. In order to reveal the reason for the difference between the photoelectrical properties of the two dyes, the ground state and excited state properties of the dyes before and after adsorbed on TiO₂ were calculated via density functional theory (DFT) and time-dependent DFT (TDDFT). The key parameters including the light harvesting efficiency (LHE), the driving force of electron injection ( Δ G inject ) and dye regeneration ( Δ G regen ), the total dipole moment ( μ normal ), the conduction band of edge of the semiconductor ( Δ E CB ), and the excited state lifetime (τ) were investigated, which are closely related to the short-circuit current density ( J sc ) and open circuit voltage ( V oc ). It was found that the experimental carminic acid has a larger J sc and V oc , which are interpreted by a larger amount of dye adsorbed on a TiO₂ photoanode and a larger Δ G regen , excited state lifetime (τ), μ normal , and Δ E CB . At the same time, chemical reactivity parameters illustrate that the lower chemical hardness (h) and higher electron accepting power (ω⁺) of carminic acid have an influence on the short-circuit current density. Therefore, carminic acid shows excellent photoelectric conversion efficiency in comparison with ethyl red.