Aberrant expression of two miRNAs promotes proliferation, hepatitis B virus amplification, migration and invasion of hepatocellular carcinoma cells: evidence from bioinformatic analysis and experimental validation.
As key negative regulators of gene expression, microRNAs (miRNAs) play an important role in the onset and progression of hepatocellular carcinoma (HCC). This study aimed to identify the miRNAs involved in HCC carcinogenesis and their regulated genes. The Gene Expression Omnibus (GEO) dataset (GSE108724) was chosen and explored to identify differentially expressed miRNAs using GEO2R. For the prediction of potential miRNA target genes, the miRTarBase was explored. Enrichment analysis of Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed by the DAVID online tool. The hub genes were screened out using the CytoHubba plug-in ranked by degrees. The networks between miRNAs and hub genes were constructed by Cytoscape software. MiRNA mimics and negative control were transfected into HCC cell lines and their effects on proliferation, hepatitis B virus DNA (HBV-DNA) replication, TP53 expression, migration, and invasion were investigated. The following methods were employed: MTT assay, quantitative PCR (qPCR) assay, western blotting, wound healing assay, and transwell assay. A total of 50 differentially expressed miRNAs were identified, including 20 upregulated and 30 downregulated miRNAs, in HCC tumor tissues compared to matched adjacent tumor-free tissues. The top three upregulated (miR-221-3p, miR-222-3p, and miR-18-5p) and downregulated (miR-375, miR-214-3p and miR-378d) miRNAs, ranked by |log2 fold change (log2FC)|, were chosen and their potential target genes were predicted. Two gene sets, targeted by the upregulated and the downregulated miRNAs, were identified respectively. GO and KEGG pathway analysis showed that the predicted target genes of upregulated and downregulated miRNAs were mainly enriched in the cell cycle and cancer-related pathways. The top ten hub nodes of gene sets ranked by degrees were identified as hub genes. Analysis of miRNA-hub gene network showed that miR-221-3p and miR-375 modulated most of the hub genes, especially involving regulation of TP53. The q-PCR results showed that miR-221-3p and miR-375 were markedly upregulated and downregulated, respectively, in HCC cells and HCC clinical tissue samples compared to non-tumoral tissues. Furthermore, miR-221-3p overexpression significantly enhanced proliferation, HBV-DNA replication, as well as the migration and invasion of HCC cells, whereas miR-375 overexpression resulted in opposite effects. Western blotting analysis showed that the overexpression of miR-221-3p and miR-375 reduced and increased TP53 expression, respectively. The present study revealed that miR-211-3p and miR-375 may exert vital effects on cell proliferation, HBV-DNA replication, cell migration, and invasion through the regulation of TP53 expression in HCC.


KIDS CARE: A Behavioral Model To Strengthen Patient and Family Partnerships.
Patient- and Family-Centered Care is a core value of The Children's Hospital of Philadelphia (CHOP). Satisfaction/ experience data are closely tracked to assist in determining if hospital staff are partnering effectively with patients and families. When opportunities for improvement were identified within the Nursing Department, an existing institutional model, KIDS CARE, was used to promote change. KIDS CARE was developed to teach and reinforce respectful behaviors for nurses initiating partnerships with patients and families. The Patient Satisfaction Committee partnered with the Family Advisory Council and Shared Governance Council to revise this model to help achieve the goals of improving quality of care. Next steps involved educating patients, families, and staff using innovative multimodal strategies. By engaging in this renewed commitment to Patient- and Family-Centered Care, systems and structures were developed to keep KIDS CARE relevant and make strides toward improved outcomes for patients and families.


Efficacy and Safety of Peroral Endoscopic Myotomy in Achalasia Patients with Failed Previous Intervention: A Systematic Review and Meta-Analysis.
Peroral endoscopic myotomy (POEM) has emerged as a rescue treatment for recurrent or persistent achalasia after failed initial management. Therefore, we aimed to investigate the efficacy and safety of POEM in achalasia patients with failed previous intervention. We searched the MEDLINE, Embase, Cochrane, and PubMed databases using the queries "achalasia," "peroral endoscopic myotomy," and related terms in March 2019. Data on technical and clinical success, adverse events, Eckardt score and lower esophageal sphincter (LES) pressure were collected. The pooled event rates, mean differences (MDs) and risk ratios (RR) were calculated. A total of 15 studies with 2,276 achalasia patients were included. Overall, the pooled technical success, clinical success and adverse events rate of rescue POEM were 98.0% (95% confidence interval [CI], 96.6% to 98.8%), 90.8% (95% CI, 88.8% to 92.4%) and 10.3% (95% CI, 6.6% to 15.8%), respectively. Seven studies compared the clinical outcomes of POEM between previous failed treatment and the treatment naïve patients. The RR for technical success, clinical success, and adverse events were 1.00 (95% CI, 0.98 to 1.01), 0.98 (95% CI, 0.92 to 1.04), and 1.17 (95% CI, 0.78 to 1.76), respectively. Overall, there was significant reduction in the pre- and post-Eckardt score (MD, 5.77; p<0.001) and LES pressure (MD, 18.3 mm Hg; p<0.001) for achalasia patients with failed previous intervention after POEM. POEM appears to be a safe, effective and feasible treatment for individuals who have undergone previous failed intervention. It has similar outcomes in previously treated and treatment-naïve achalasia patients.


Hamstring injuries in Australian football.
To identify risk factors for hamstring injury in community-level Australian football players. Cohort study. The preseason and 2000 season of 4 adult Victorian Amateur Football Association clubs. All players in the clubs who were training at the time of the baseline assessment were invited to participate (n = 148). Players who were injured and unable to participate in the baseline assessment were excluded (22 players; 15%). During 3 weeks before preseason practice, each player had a series of musculoskeletal tests and completed a questionnaire. The musculoskeletal screening included flexibility tests of the hamstrings, lower extremities, quadriceps, and iliopsoas; range of motion assessments of the lumbar spine and hip rotation; and the active slump test of neural mobility. Testing was done after a brief warm up by 4 trained screeners. A questionnaire with pre-established validity and reliability included information on playing experience, injury history, and training habits. The main outcome measure was a first hamstring injury that resulted in missed participation time and/or treatment from a health professional in relation to predictive factors. Criteria defining a hamstring injury were sudden onset of posterior thigh pain, and tenderness on palpation with or without pain on stretching or contracting the hamstring muscle group. Through the season, the clubs' coaching staff collected information on exposure (hours of match and training participation). A total of 26 hamstring injuries occurred (incidence, 4 injuries per 1000 player hours). More injuries were sustained during competition (77%) than training. Rapid acceleration during running or sprinting was the primary mechanism of injury (81%), with the remainder occurring as the player kicked the ball. After adjustment for exposure, younger age was associated with a lower relative risk (RR) of injury (RR for age > or =23 years, 3.8; 95% CI, 1.1-14.0), as were quadriceps flexibility (RR for >51 degree knee flexion, 0.3; CI, 0.1-0.8) and active knee extension range of motion (RR for >27 degree knee flexion, 2.8; CI, 0.9-8.5; not significant). Frequency of off-season running, body height, and neural mobility were not significantly associated with hamstring injury. Hamstring injuries in amateur Australian football players most commonly occurred with sprinting and were more frequent in players older than 23 years or with lesser quadriceps flexibility.


[Morphological analysis of transgenic tobacco plants expressing the PnEXPA3 gene of black poplar (Populus nigra)].
Transgenic tobacco plants overexpressing the PnEXPA3 gene of black poplar (Populus nigra), which encodes alpha-expansin, were obtained. The transgenic plants were characterized by increased size of epidermic and mesophyll cells of leaves. However, the size of leaves remained normal. Overexpression of the PnEXPA3 gene provided stimulatory effect only on the stem length. Other morphological traits of the transgenic plants remained unchanged.


Enhancing electrical energy storage capability of dielectric polymer nanocomposites via the room temperature Coulomb blockade effect of ultra-small platinum nanoparticles.
Introducing a high dielectric constant (high-k) nanofiller into a dielectric polymer is the most common way to achieve flexible nanocomposites for electrostatic energy storage devices. However, the significant decrease of breakdown strength and large increase of dielectric loss has long been known as the bottleneck restricting the enhancement of practical energy storage capability of the nanocomposites. In this study, by introducing ultra-small platinum (<2 nm) nanoparticles, high-k polymer nanocomposites with high breakdown strength and low dielectric loss were prepared successfully. Core-shell structured polydopamine@BaTiO3 (PDA@BT) and core-satellite ultra-small platinum decorated PDA@BT (Pt@PDA@BT) were used as nanofillers. Compared with PDA@BT nanocomposites, the maximum discharged energy density of the Pt@PDA@BT nanocomposites is increased by nearly 70% because of the improved energy storage efficiency. This research provides a simple, promising and unique way to enhance energy storage capability of high-k polymer nanocomposites.


A rat brain bicistronic gene with an internal ribosome entry site codes for a phencyclidine-binding protein with cytotoxic activity.
The cloning and characterization of the gene for the fourth subunit of a glutamate-binding protein complex in rat brain synaptic membranes are described. The cloned rat brain cDNA contained two open reading frames (ORFs) encoding 8.9- (PRO1) and 9.5-kDa (PRO2) proteins. The cDNA sequence matched contiguous genomic DNA sequences in rat chromosome 17. Both ORFs were expressed within the structure of a single brain mRNA and antibodies against unique sequences in PRO1- and PRO2-labeled brain neurons in situ, indicative of bicistronic gene expression. Dicistronic vectors in which ORF1 and ORF2 were substituted by either two different fluorescent proteins or two luciferases indicated concurrent, yet independent translation of the two ORFs. Transfection with noncapped mRNA led to cap-independent translation of only ORF2 through an internal ribosome entry sequence preceding ORF2. In vitro or cell expression of the cloned cDNA led to the formation of multimeric protein complexes containing both PRO1 and PRO2. These complexes had low affinity (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801)-sensitive phencyclidine-binding sites. Overexpression of PRO1 and PRO2 in CHO cells, but not neuroblastoma cells, caused cell death within 24-48 h. The cytotoxicity was blocked by concurrent treatment with MK-801 or by two tetrahydroisoquinolines that bind to phencyclidine sites in neuronal membranes. Co-expression of two of the other subunits of the protein complex together with PRO1/PRO2 abrogated the cytotoxic effect without altering PRO1/PRO2 protein levels. Thus, this rare mammalian bicistronic gene coded for two tightly interacting brain proteins forming a low affinity phencyclidine-binding entity in a synaptic membrane complex.


Effects of chronic ethanol consumption on the metabolism and carcinogenicity of N'-nitrosonornicotine in F344 rats.
The effects of chronic ethanol consumption on the carcinogenicity and metabolism of N'-nitrosonornicotine (NNN) in male F344 rats have been investigated. Groups of 26 to 30 rats were maintained on either a control liquid diet (Groups 1, 3, and 5) or an ethanol-containing liquid diet (Groups 2, 4, and 6) for 4 weeks prior to and during treatment with NNN. The carcinogen was injected s.c. (10 mg/kg, Groups 3 and 4) three times weekly or added to the liquid diet (17.5 mg/liter, Groups 5 and 6). The total dose was 1 mmol of NNN per rat. Control rats (Groups 1 and 2) received s.c. injections of 0.9% NaCl solution. The nasal mucosa was the main target tissue of NNN in Groups 3 and 4, but both the nasal mucosa and esophagus were major target tissues in Groups 5 and 6. In rats treated s.c. with NNN (Groups 3 and 4), ethanol consumption had no effect on the distribution and incidence of nasal cavity tumors. In rats treated with NNN added to the control liquid diet or to the ethanol-containing liquid diet, the number of tumors of the nasal cavity was 18 in Group 5 and 26 in Group 6 (p less than 0.05). In contrast, the number of rats with esophageal tumors was 25 in Group 5 and 20 in Group 6 (p less than 0.05). The effects of ethanol on the enzyme system which activates NNN were studied in rats which had been maintained on an ethanol-containing liquid diet for 4 weeks. Explants of nasal mucosae, lingual mucosae, esophagi , and livers were cultured in vitro with NNN. Nasal mucosae of ethanol-consuming rats had a 1.5-fold higher (p less than 0.05) alpha-carbon-hydroxylating activity than did those of control rats. Activating enzymes in the lingual mucosae, esophagi , and livers were not induced by ethanol. The results show that the increased susceptibility of the rat nasal mucosa to the carcinogenic effects of NNN added to an ethanol-containing diet could be due in part to an induction of activating enzymes by ethanol. However, since chronic ethanol consumption had no apparent effect on the incidence of nasal cavity tumors in rats treated by s.c. injection of NNN, factors other than enzyme induction are important in determining the effects of ethanol on NNN carcinogenicity.


The changing age and seasonal profile of pertussis in Canada.
During the postvaccine era in Canada, most cases of pertussis have been reported in children <5 years of age, with the highest incidence, morbidity, and mortality in infants <1 year old. Population-based data, with very high laboratory confirmation rates and hospital separation and mortality statistics, chronicle the changing age and seasonal profile associated with pertussis over recent successive outbreaks in British Columbia, Canada. A large outbreak during 2000 highlights 2 important changes to the postvaccine profile. For the first time in Canada, the incidence of pertussis among preteens and teens surpassed that of all other age groups. At the same time, a decreasing incidence of pertussis among infants and preschool children highlights reduced susceptibility in the very young. Recent changes in the childhood immunization program (including introduction of an acellular pertussis vaccine), waning immunity, and changes in laboratory methods are considered in explaining these 2 simultaneous but divergent trends in the pertussis profile.


Acousto-optic modulation and opto-acoustic gating in piezo-optomechanical circuits.
Acoustic wave devices provide a promising chip-scale platform for efficiently coupling radio frequency (RF) and optical fields. Here, we use an integrated piezo-optomechanical circuit platform that exploits both the piezoelectric and photoelastic coupling mechanisms to link 2.4 GHz RF waves to 194 THz (1550 nm) optical waves, through coupling to propagating and localized 2.4 GHz acoustic waves. We demonstrate acousto-optic modulation, resonant in both the optical and mechanical domains, in which waveforms encoded on the RF carrier are mapped to the optical field. We also show opto-acoustic gating, in which the application of modulated optical pulses interferometrically gates the transmission of propagating acoustic pulses. The time-domain characteristics of this system under both pulsed RF and pulsed optical excitation are considered in the context of the different physical pathways involved in driving the acoustic waves, and modelled through the coupled mode equations of cavity optomechanics.


Value-based recruitment in midwifery: do the values align with what women say is important to them?
The aim of this study was to discuss theoretical conceptualization and definition of values and value-based recruitment in the context of women's views about what they would like from their midwife. Value-based recruitment received headline status in the UK government's response to pervasive deficiencies in compassionate care identified in the health service. Core values which aim to inform service user's experience are defined in the National Health Service Constitution but clarity about whether these encompass all that women say is important to them is needed. Discussion paper. A literature search included published papers written in English relating to values, VBR and women's views of a 'good' midwife with no date limiters. Definitions of values and value-based recruitment are examined. Congruence is explored between what women say is important to them and key government and professional regulatory documentation. The importance of a 'sustainable emotional' dimension in the midwife-mother relationship is suggested. Inconsistencies are identified between women's views, government, professional documentation and what women say they want. An omission of any reference to emotions or emotionality in value-based recruitment policy, professional recruitment and selection guidance documentation is identified. A review of key professional documentation, in relation to selection for 'values', is proposed. We argue for clarity and revision so that values embedded in value-based recruitment are consistent with health service users' views. An enhancement of the 'values' in the value-based recruitment framework is recommended to include the emotionality that women state is a fundamental part of their relationship with their midwife.


A sequential screning test based on the running component of audiogenic seizures in mice, including reference compound PD50 values.
1. The running component of audiogenic seizures in mice has been used as the basis of a sequential screening test for the detection of a variety of centrally acting drugs.2. For acceptance by the test, an active compound must completely suppress the running component in a total of sixteen mice at a dose of 1/5 LD50 intraperitoneally.3. Considerable economies in the numbers of animals required for screening have been achieved, the mean number of mice required to reject an inactive compound being 2.0.4. The running component is highly sensitive to anticonvulsants and general central depressants, and insensitive to phenothiazine tranquillizers and morphine. Reserpine caused an increase in the severity of the running component.5. The statistical model used in this test is of general application to screening test situations which use quantal data.


Design of resonant mirrors with negative group delay.
Resonant mirrors introduce large spectral gradients in reflected phase while maintaining high reflectivity, allowing synthesis of optimized reflected phase for many practical applications. In this paper we show theoretically that asymmetry is required for negative group delay in lossless mirrors and explore the limits of reflected phase in resonant mirrors through the use of coupled mode theory and rigorous couple wave analysis. Our coupled mode theory shows that the phase response of resonant mirrors is determined by interacting resonances and gives insight into tradeoffs in design of mirrors with desired phase response.


A Compact Ultra-Wideband Monocone Antenna with Folded Shorting Wires for Vehicle-to-Everything (V2X) Applications.
In this paper, a capacitively-fed, ultra-wideband (UWB), and low-profile monocone antenna is proposed for vehicle-to-everything (V2X) applications. The proposed antenna consists of a monocone design with an inner set of vias. Additionally, an outer ring is added with a small gap from the monocone and shorted with six folded wires of different lengths to extend the operating band. The proposed antenna covers the frequency range from 0.75 GHz to 7.6 GHz and has a 164% fractional bandwidth, with a gain value varying between 2 and 10 dBi. The dimensions of the antenna are 0.37λL × 0.37λL × 0.067λL. The antenna was fabricated using a 3D printer with low-cost polylactic acid plastic (PLA) material and then sprayed with aerosol copper nanoparticles. The efficiency was approximately 90% throughout the frequency bands of interest. Finally, the proposed antenna was installed on a vehicle and tested with an OBU (onboard unit) and a RSU (roadside unit) in the field. The results show a longer wireless communication range for V2X applications.


Fluorescence Quenching of a Conjugated Polymer by Synergistic Amine-Carboxylic Acid and π-π Interactions for Selective Detection of Aromatic Amines in Aqueous Solution.
Fluorescence sensing of amine in aqueous solution is challenging. The various basicity and chemical structures of amines may lead to poor selectivity in aqueous solution, and selective fluorescence detection of primary aromatic amine is rarely reported. This paper presents design and synthesis of a fluorescent conjugated polymer for rapid and selective sensing of aromatic amines in aqueous solution. The fluorescent conjugated polymer, poly[fluorenyl-alt-p-phenyleneethynylene] with pendant carboxylic acid groups and long alky chains, is synthesized via palladium-catalyzed Sonogashira coupling reaction. The fluorescence of the polymer is selectively quenched by the aromatic amines in aqueous solution, whereas the aliphatic amines enhance the fluorescence of the polymer. The high selectivity to the aromatic amines, particularly to the environmentally important p-phenylenediamine, likely originates from the amplified π-π fluorescence quenching synergized by amine and carboxylic acid interaction. Our results demonstrate an effective material design strategy that may be extended to fluorescence sensing of other aromatic compounds.


Why do women choose to undergo oocyte aspiration without sedation or analgesia?
Oocyte pick-up (OPU) is a painful but essential part of in-vitro fertilization (IVF) that is usually performed under sedation and analgesia (SaA). Our aim was to study that why some women decide to undergo OPU without SaA? This was a prospective study using patient questionnaires and the standardized 7-item generalized anxiety disorder (GAD-7) score. The patients were asked to assess the pain experienced during OPU using a visual analog scale (VAS). The study sample was a convenience sample of 100 healthy women undergoing OPU at our unit with or without SaA. Women who chose to undergo OPU without SaA were significantly more likely to express the fear of anesthesia. A high pain score (VAS ≥ 6) was reported by significantly more patients who underwent OPU without SaA than with SaA. Yet, 98% of the patients who underwent OPU without SaA stated that in future IVF cycles, they would still choose to undergo OPU without SaA. More patients had high anxiety scores among those who underwent OPU with than without SaA. Women who chose to undergo OPU without SaA reported more often fear of anesthesia. Although these women experienced significantly more pain during OPU, almost all of them suggested that they would still choose to undergo OPU without SaA. Increased anxiety, as expressed by higher GAD-7 scores, was not associated with a tendency to choose SaA during OPU. The option of OPU without SaA seems to be an acceptable option for selected women. Egg retrieval from the ovaries is a painful part of in vitro fertilization (IVF). It is, therefore, usually performed under sedation and pain relief (analgesia). The aim of this study was to investigate: Why some women decide to undergo egg retrieval without sedation? We prospectively studied 100 women using patient questionnaires and standardized scores in order to measure patient's pain and anxiety levels. We found that women who chose to undergo egg retrieval without sedation were significantly more likely to express fear of anesthesia. As expected, women who decided to forgo sedation experienced more pain during egg retrieval, yet, 98% of them decided that in future IVF cycles, they would still choose to undergo egg retrieval without sedation. Surprisingly, women who had high anxiety scores were not more likely to ask for sedation during egg retrieval. The option to undergo egg retrieval without sedation during IVF seems to be acceptable for some women.


Enhanced photo-catalytic activity of ordered mesoporous indium oxide nanocrystals in the conversion of CO2 into methanol.
Ordered mesoporous indium oxide nanocrystal (m-In2O3) was synthesized by nanocasting technique, in which highly ordered mesoporous silca (SBA-15) was used as structural matrix. X-ray diffraction (XRD), Field Emission Scanning Electron Microscopy (FESEM) Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halanda (BJH) studies were carried out on m-In2O3 and the results revealed that this material has a highly ordered mesoporous surface with reduced grain size, increased surface area and surface volume compared to the non porous indium oxide. The diffuse reluctance spectrum exhibited substantially improved light absorption efficiency in m-In2O3 compared to normal indium oxide, however, no considerable change in the band gap energies of these materials was observed. When m-In2O3 was used as a photo-catalyst in the photo-catalytic process of converting carbon dioxide (CO2) into methanol under the pulsed laser radiation of 266-nm wavelengths, an enhanced photo-catalytic activity with the quantum efficiency of 4.5% and conversion efficiency of 46.3% were observed. It was found that the methanol production yield in this chemical process is as high as 485 µlg-1 h-1 after 150 min of irradiation, which is substantially higher than the yields reported in the literature. It is quite clear from the results that the introduction of mesoporosity in indium oxide, and the consequent enhancement of positive attributes required for a photo-catalyst, transformed photo-catalytically weak indium oxide into an effective photo-catalyst for the conversion of CO2 into methanol.


Recurrent intracranial sarcoma mimicking chronic subdural haematoma.
Primary sarcomas of the central nervous system and their coincidence with a subdural haematoma are each rare. We describe an unusual case of unsuspected dural spreading of a recurrent spindle cell sarcoma concealed in a chronic subdural haematoma, which occurred months after microsurgical resection and external radiation of a temporal parenchymatous sarcoma.


Improved computational model (AQUIFAS) for activated sludge, integrated fixed-film activated sludge, and moving-bed biofilm reactor systems, Part I: Semi-empirical model development.
Research was undertaken to develop a model for activated sludge, integrated fixed-film activated sludge (IFAS), and moving-bed biofilm reactor (MBBR) systems. The model can operate with up to 12 cells (reactors) in series, with biofilm media incorporated to one or more cells, except the anaerobic cells. The process configuration can be any combination of anaerobic, anoxic, aerobic, post-anoxic with or without supplemental carbon, and reaeration; it can also include any combination of step feed and recycles, including recycles for mixed liquor, return activated sludge, nitrates, and membrane bioreactors. This paper presents the structure of the model. The model embeds a biofilm model into a multicell activated sludge model. The biofilm flux rates for organics, nutrients, and biomass can be computed by two methods--a semi-empirical model of the biofilm that is relatively simpler, or a diffusional model that is computationally intensive. The values of the kinetic parameters for the model were measured using pilot-scale activated sludge, IFAS, and MBBR systems. For the semiempirical version, a series of Monod equations were developed for chemical oxygen demand, ammonium-nitrogen, and oxidized-nitrogen fluxes to the biofilm. Within the equations, a second Monod expression is used to simulate the effect of changes in biofilm thickness and fraction nitrifiers in the biofilm. The biofilm flux model is then linked to the activated sludge model. The diffusional model and the verification of the models are presented in subsequent papers (Sen and Randall, 2008a, 2008b). The model can be used to quantify the amount of media and surface area required to achieve nitrification, identify the best locations for the media, and optimize the dissolved oxygen levels and nitrate recycle rates. Some of the advanced features include the ability to apply different media types and fill fractions in cells; quantify nitrification, denitrification, and biomass production in the biofilm and mixed liquor suspended solids; and perform dynamic simulations.


Conjugation of β-Glucan with the Hydrazone and Disulfide Linkers Markedly Improves the Immunogenicity of Zika Virus E Protein.
The diseases caused by Zika virus (ZIKV) have received widespread concerns. As a key viral element of ZIKV, E protein was an ideal antigen for vaccine development. However, the poor immunogenicity of E protein necessitated the formulation with adjuvants. Formulation of E protein by conjugation with β-glucan was a strategy to improve the immunogenicity of E protein, where β-glucan was a polysaccharide adjuvant that could activate macrophages and trigger intracellular processes. However, the antigenic epitopes of E protein and the immunomodulatory sites of β-glucan were shielded in the conjugate. Moreover, the conjugate might elicit the undesired immune response to β-glucan. Thus, the acidic-labile hydrazone and the thiol-sensitive disulfide bonds were used as the linkers between E protein and β-glucan. Hydrazone hydrolysis and disulfide reduction could sufficiently detach the two components in the immune cells to overcome the two disadvantages. As compared with the conjugate without the two linkers, the conjugate with the two linkers (E-PS-4) elicited high E protein-specific IgG titers and low β-glucan-specific IgG titers. E-PS-4 elicited high levels of IFN-γ, TNF-α, IL-2, and IL-10. Moreover, E-PS-4 greatly facilitated the activation of dendritic cells without significant toxicity to the organs. A pharmacokinetic study revealed that the serum duration of E-PS-4 was longer than that of E protein. Accordingly, conjugation of E protein with β-glucan by the hydrazone and disulfide linkers could promote a potent cellular and humoral immune response to E protein. Thus, our study could facilitate the development of an effective vaccine against ZIKV.


Genetic dissection of cell wall defects and the strigolactone pathway in Arabidopsis.
Defects in the biosynthesis and/or deposition of secondary plant cell wall polymers result in the collapse of xylem vessels causing a dwarfed plant stature and an altered plant architecture termed irregular xylem (irx) syndrome. For example, reduced xylan O-acetylation causes strong developmental defects and increased freezing tolerance. Recently, we demonstrated that the irx syndrome in the trichome birefringence-like 29/eskimo1 (tbl29/esk1) mutant is dependent on MORE AXILLARY GROWTH 4 (MAX4), a key enzyme in the biosynthesis of the phytohormone strigolactone (SL). In this report, we show that other xylan- and cellulose-deficient secondary wall mutants exhibit increased freezing tolerance correlated with the irx syndrome. In addition, these phenotypes are also dependent on MAX4, suggesting a more general interaction between secondary wall defects and SL biosynthesis. In contrast, MAX4 does not play a role in developmental defects triggered by primary wall deficiencies, suggesting that the interaction is restricted to vascular tissue. Through a reverse genetics approach, the requirement of different components of the SL pathway impacting the irx syndrome in tbl29 was evaluated. Our results show that the tbl29-associated irx phenotypes are dependent on the MAX3 and MAX4 enzymes, involved in the early steps of SL biosynthesis. In contrast, this signaling is independent on downstream enzymes in the biosynthesis and perception of SL such as MAX1 and MAX2.


Clonidine promotes the accumulation of 45Ca in pancreatic beta-cell organelles.
Glucose-stimulated insulin release from pancreatic islets of ob/ob-mice was inhibited by 10(-9) M of the alpha 2-adrenergic agonist clonidine. This inhibitory effect was abolished by 10(-7) M of the antagonist yohimbine. Loading the islets with 45Ca during the clonidine exposure followed by isolation of subcellular fractions under conditions known to minimize the 45Ca redistribution resulted in increased accumulation of the isotope in the mitochondrial and microsomal fractions. It is suggested that clonidine inhibits glucose-stimulated insulin release by increasing the organelle sequestration of Ca2+.


New developments in the diagnosis, prognosis and treatment of chronic lymphocytic leukemia.
The remarkable progress witnessed over the past few years in the diagnosis, prognosis, and therapy of chronic lymphocytic leukemia has profoundly changed the clinical approach to this disease. This review focuses on the most recent advances in the diagnostic and prognostic examination of patients with chronic lymphocytic leukemia, with particular emphasis on their implications in clinical management, taking into account the broadening of the therapeutic possibilities available today. Through the biologic improvements achieved during the past few years it is now possible to effectively stratify chronic lymphocytic leukemia patients prognostically at presentation on the basis of several laboratory parameters. Furthermore, the availability of purine analogs and monoclonal antibodies and the extension of autografting and allografting procedures have allowed the achievement of higher response rates, including molecular remissions. With the aim of investigating whether early and aggressive treatment intervention may improve the survival of patients with a poor prognosis, new therapeutic trials have been specifically designed. A complete biologic and clinical examination now allows the establishment of a correct diagnostic characterization of patients with chronic lymphocytic leukemia and to identify patients with early disease with a different prognostic likelihood. Multicenter prospective trials, in which the enrolled patients are stratified and treated according to their prognostic risk, will determine the best treatment for the different categories of patients. It is likely that in the near future each chronic lymphocytic leukemia patient can be offered a targeted treatment algorithm based on the clinical and biologic characteristics at presentation.


Mff-Dependent Mitochondrial Fission Contributes to the Pathogenesis of Cardiac Microvasculature Ischemia/Reperfusion Injury via Induction of mROS-Mediated Cardiolipin Oxidation and HK2/VDAC1 Disassociation-Involved mPTP Opening.
The cardiac microvascular system ischemia/reperfusion injury following percutaneous coronary intervention is a clinical thorny problem. This study explores the mechanisms by which ischemia/reperfusion injury induces cardiac microcirculation collapse. In wild-type mice, mitochondrial fission factor (Mff) expression increased in response to acute microvascular ischemia/reperfusion injury. Compared with wild-type mice, homozygous Mff-deficient (Mffgt) mice exhibited a smaller infarcted area, restored cardiac function, improved blood flow, and reduced microcirculation perfusion defects. Histopathology analysis demonstrated that cardiac microcirculation endothelial cells (CMECs) in Mffgt mice had an intact endothelial barrier, recovered phospho-endothelial nitric oxide synthase production, opened lumen, undivided mitochondrial structures, and less CMEC death. In vitro, Mff-deficient CMECs (derived from Mffgt mice or Mff small interfering RNA-treated) demonstrated less mitochondrial fission and mitochondrial-dependent apoptosis compared with cells derived from wild-type mice. The loss of Mff inhibited mitochondrial permeability transition pore opening via blocking the oligomerization of voltage-dependent anion channel 1 and subsequent hexokinase 2 separation from mitochondria. Moreover, Mff deficiency reduced the cyt-c leakage into the cytoplasm by alleviating cardiolipin oxidation resulting from damage to the electron transport chain complexes and mitochondrial reactive oxygen species overproduction. This evidence clearly illustrates that microcirculatory ischemia/reperfusion injury can be attributed to Mff-dependent mitochondrial fission via voltage-dependent anion channel 1/hexokinase 2-mediated mitochondrial permeability transition pore opening and mitochondrial reactive oxygen species/cardiolipin involved cyt-c release.


Distribution of 11-nor-9-carboxy-Delta9-tetrahydrocannabinol in traffic fatality cases.
11-nor-9-carboxy-Delta9-tetrahydrocannabinol (THC-COOH) distributions in postmortem specimens are rarely reported. Fifty New Jersey State Medical Examiner's cases in which automobile accident deaths suspected of involving marijuana intake were studied for the distributions of THC-COOH in postmortem urine, blood, vitreous humor, and bile specimens. Cases were selected based on immunoassay (TDx) urine test results. If the preliminary urine test indicated the presence of THC-COOH (apparent THC-COOH concentration > or = 20 ng/mL), urine, heart blood, vitreous humor, and bile specimens from the case were analyzed for THC-COOH concentrations by gas chromatography-mass spectrometry. The mean, standard deviation, and range of THC-COOH in heart blood, urine, and bile found in these 50 cases were 0.081, 0.082, and 0.016-0.33 microg/mL; 0.314, 0.415, and 0.044-2.33 microg/mL; and 12.9, 11.4, and 1.03-43.7 microg/mL, respectively. THC-COOH was absent (detection limit, 1 ng/mL) or at low concentration (< 10 ng/mL) in vitreous humor specimens. The mean, standard deviation, and range of the bile-to-blood and urine-to-blood ratios were 242, 196, and 17.2-888 and 4.70, 4.05, and 1.14-19.2, respectively. The highest concentrations of THC-COOH were found in bile and the lowest in vitreous humor. These findings are consistent with the high hydrophobicity nature of THC-COOH and further suggest that bile is the specimen of choice for detecting low level of THC-COOH in postmortem cases.


Rapid detection of melamine adulteration in dairy milk by SB-ATR-Fourier transform infrared spectroscopy.
Melamine is a nitrogenous chemical substance used principally as a starting material for the manufacture of synthetic resins. Due to its very high proportion of nitrogen melamine has been added illegitimately to foods and feeds to increase the measured protein content, which determines the value of the product. These issues prompted private as well as governmental laboratories to develop methods for the analysis of melamine in a wide variety of food products and ingredients. Owing to this fact present study is aimed to use single bounce attenuated total reflectance (SB-ATR) Fourier transform infrared spectroscopy (FTIR) method as an effective rapid tool for the detection and quantification of melamine in milk (liquid and powder). Partial least-squares (PLS) models were established for correlating spectral data to melamine concentration with R(2)>0.99, and RMSEC 0.370. Linear calibration curves were obtained over the calibration range of 25-0.0625%. The LOD and LOQ of the method was 0.00025% (2.5 ppm) and 0.0015% (15 ppm) respectively. Proposed SB-ATR-FTIR method requires little or no sample preparation with an assay time of 1-2 min.


Review article: glycyrrhizin as a potential treatment for chronic hepatitis C.
Chronic hepatitis C is a slowly progressive liver disease that may evolve into cirrhosis with its potential complications of liver failure or hepatocellular carcinoma. Current therapy with alpha-interferon is directed at viral clearance, but sustained response is only achieved in 20-40% of patients without cirrhosis, and less than 20% in patients with cirrhosis who have the greatest need for therapy. Treatment for those who do not respond to anti-viral therapy is highly desirable. In Japan glycyrrhizin has been used for more than 20 years as treatment for chronic hepatitis. In randomized controlled trials, glycyrrhizin induced a significant reduction of serum aminotransferases and an improvement in liver histology compared to placebo. Recently, these short-term effects have been amplified by a well-conducted retrospective study suggesting that long-term usage of glycyrrhizin prevents development of hepatocellular carcinoma in chronic hepatitis C. The mechanism by which glycyrrhizin improves liver biochemistry and histology are undefined. Metabolism, pharmacokinetics, side-effects, and anti-viral and hepatoprotective effects of glycyrrhizin are discussed.


Induction of basic helix-loop-helix protein-containing complexes during erythroid differentiation.
The involvement of basic helix-loop-helix (bHLH) transcription factors in erythroid differentiation and development has been established by forced expression of the proteins TAL1 and Id1 in cultured cell lines and by targeted disruption of the mouse TAL1 gene. To better understand the mechanism by which bHLH proteins regulate erythropoiesis, we have investigated HLH protein-DNA interactions in mouse erythroleukemia (MEL) cells before and during chemically induced differentiation. Three bHLH (E-box) binding activities were found to be induced in nuclei from differentiating MEL cells. Using specific antisera, we have demonstrated that these complexes are dimers of TAL1 and ubiquitous E proteins. Similar complexes were detected in nuclear extracts from a human erythroid cell line, K562, and from mouse fetal liver. All three bHLH complexes were disrupted in vitro by Id1, a dominant-negative HLH protein that we and others have previously shown to antagonize MEL cell differentiation. During differentiation of an Id1-overexpressing MEL cell line, induction of a complex containing TAL1 and E2A was not only blocked but reduced below the levels seen in undifferentiating cells. These observations are consistent with the idea that TAL1 and Id1 have opposing effects on erythroid differentiation and that the level of TAL1/E2A heterodimer and/or another E protein-containing complex may influence the decision of a cell to terminally differentiate.


Treatment of Uremic Pruritus: A Systematic Review.
Uremic pruritus is a common and burdensome symptom afflicting patients with advanced chronic kidney disease (CKD) and has been declared a priority for CKD research by patients. The optimal treatments for uremic pruritus are not well defined. Systematic review. Adult patients with advanced CKD (stage ≥ 3) or receiving any form of dialysis. PubMed, CINAHL, Embase, International Pharmaceutical Abstracts, Scopus, Cochrane Library, and ClinicalTrials.gov from their inception to March 6, 2017, were systematically searched for randomized controlled trials (RCTs) of uremic pruritus treatments in patients with advanced CKD (stage ≥ 3) or receiving any form of dialysis. 2 reviewers extracted data independently. Risk of bias was assessed using the Cochrane Collaboration risk-of-bias tool. Any intervention for the treatment of uremic pruritus was included. A quantitative change in pruritus intensity on a visual analogue, verbal rating, or numerical rating scale. 44 RCTs examining 39 different treatments were included in the review. These treatments included gabapentin, pregabalin, mast cell stabilizers, phototherapy, hemodialysis modifications, and multiple other systemic and topical treatments. The largest body of evidence was found for the effectiveness of gabapentin. Due to the limited number of trials for the other treatments included, we are unable to comment on their efficacy. Risk of bias in most studies was high. Heterogeneity in design, treatments, and outcome measures rendered comparisons difficult and precluded meta-analysis. Despite the acknowledged importance of uremic pruritus to patients, with the exception of gabapentin, the current evidence for treatments is weak. Large, simple, rigorous, multiarm RCTs of promising therapies are urgently needed.


Clinical Manifestations and Prognostic Factors of Pneumocystis jirovecii Pneumonia without HIV.
Pneumocystis jirovecii pneumonia (PCP) can occur in HIV patients but also in those without HIV (non-HIV PCP) but with other causes of immunodeficiency including malignancy or rheumatic diseases. To evaluate the clinical presentation and prognostic factors of non-HIV PCP, we retrospectively reviewed all patients diagnosed as having PCP without HIV at Kameda Medical Center, Chiba, Japan, from January 2005 until June 2012. For the purpose of examining a prognostic factor for non-HIV PCP with 30-day mortality, we compared the characteristics of patients, clinical symptoms, radiological images, Eastern Cooperative Oncology Group performance status (PS), and the time from the onset of respiratory symptoms to the start of therapy, in both survival and fatality groups. A total of 38 patients were eligible in this study. Twenty-five survived and 13 had died. The non-HIV PCP patients in the survivor group had a better PS and received anti-PCP therapy earlier than those in the nonsurvivor group. Rales upon auscultation and respiratory failure at initial visits were seen more frequently in the nonsurvivor group than in the survivor group. Lactate dehydrogenase and C-reactive protein values tended to be higher in the nonsurvivor group, but this was not statistically significant. Multivariate analyses using 5 variables showed that a poor PS of 2-4 was an independent risk factor for non-HIV PCP patients and resulted in death (odds ratio 15.24; 95% confidence interval 1.72-135.21). We suggest that poor PS is an independent risk factor in non-HIV PCP, and a patient's PS and disease activity may correlate with outcome.


Problematic internet use and suicidal behavior in adolescents: A review.
Suicide is a leading cause of death in adolescents. Problematic internet use (PIU) has been associated with suicidal behavior in this age group. To assess this association, we reviewed population-based studies that examined the relationship between PIU and suicidal behavior. We searched PubMed, PsycINFO, and Web of Science for studies published in English from 1995 through 2021. Only studies with the stated aim to examine the relationship between PIU and suicidal behavior in participants age 13 through 18 were included. Also, each study had to include a measure of internet overuse with resultant impairment/distress. To minimize selection bias, studies with convenience samples were excluded. Seven cross-sectional studies with at least 9,500 participants each met the inclusion/exclusion criteria. Five assessed both suicidal ideation and suicide attempts and 2 examined suicidal ideation only. All studies found a statistically significant association between suicidal behavior and PIU. Effect sizes were mostly in the minimal to small range. All 7 studies supported a positive relationship between PIU and suicidal ideation and suicide attempts, which could be clinically important. Longitudinal studies with improved methodology are needed to identify risk factors and determine whether PIU is causally related to suicidality.


Color perception in pediatric patient room design: healthy children vs. pediatric patients.
The purpose of this study was to investigate the value of color as a component of a healing environment for pediatric patient rooms. Color preferences from pediatric inpatient, pediatric outpatient, and healthy children groups were investigated and compared for group and gender differences. Positive environmental stimulation can promote patient well-being by reducing stress or negative feelings. If environmental colors can have positive influences, then those colors will make patients more comfortable, reducing their stress. Although previous color studies are suggestive, none has focused on pediatric healthcare environments. Patients' health status may affect their reaction to environmental stimulation. There was a gap in the body of knowledge regarding the consistency of color preferences with regard to patient health status. A simulation method was used because of its reliability and feasibility. It allowed for investigating the value of color in real contexts and controlling confounding variables. Previous color preference studies typically have been done with small colored squares of paper, which are visually different from seeing a color applied on wall surfaces. In addition, they failed to control confounding variables such as color attributes and light sources. Group differences were not significant. This refuted the hypothesis that the color preferences of pediatric patients are different from those of healthy children. However, overall color preferences showed that the healthy children group had higher mean scores of color preferences than both pediatric outpatients and inpatients groups. Gender effects were identified across all three groups. All three groups showed similar color preference patterns, except for yellow. Both pediatric outpatients and inpatients preferred yellow less than healthy children did. Healthy children's mean scores of color preferences were higher than the pediatric outpatients; the pediatric outpatients group had higher mean scores of color preferences than the pediatric inpatients group. Although this observation was not statistically significant, it raised the question of whether the sickest pediatric patients respond differently to color than healthy children do. Gender effects indicated that girls preferred red and purple more than boys do. Regardless of gender effects, healthy children and pediatric patients preferred blue and green the most and white the least. These results can help healthcare providers and professionals better understand appropriate colors for pediatric populations.


10th Anniversary Critical Review: The tissue-residue approach for toxicity assessment: concepts, issues, application, and recommendations.
The tissue-residue approach for toxicity assessment (TRA) is simply the use of tissue concentrations as the dose metric for characterizing toxicant potency. There are several advantages to using tissue residues over exposure concentrations (e.g., water, sediment, and diet) to calculate toxicity metrics. These include a large reduction in toxic response variability among all species for a given compound, an improved ability to address mixture toxicity, an increased use of information on modes and mechanisms of toxic action, a likely reduction in the number of species needed to characterize toxicant potency, the potential to improve ecological risk assessments, and the generation of more scientifically defensible tissue, water, and sediment toxicity guidelines or criteria. A keystone concept for the TRA is that the body/tissue residue reflects the target "dose" better than the traditional dose (e.g., water, air, soil/sediment, or diet) because the closer the dose surrogate is to the actual site of toxic action the less it is influenced by myriad modifying factors. Our goal for this review is to present the concepts and issues associated with the TRA and discuss some of the potential applications and expected improvements to the field of environmental toxicology that we believe will promote enhanced protection for species and ecosystems.


An essential role for IL-17 in preventing pathogen-initiated bone destruction: recruitment of neutrophils to inflamed bone requires IL-17 receptor-dependent signals.
IL-17 and its receptor are founding members of a novel family of inflammatory cytokines. IL-17 plays a pathogenic role in rheumatoid arthritis (RA)-associated bone destruction. However, IL-17 is also an important regulator of host defense through granulopoiesis and neutrophil trafficking. Therefore, the role of IL-17 in pathogen-initiated bone loss was not obvious. The most common form of infection-induced bone destruction occurs in periodontal disease (PD). In addition to causing significant morbidity, PD is a risk factor for atherosclerotic heart disease and chronic obstructive pulmonary disease (COPD). Similar to RA, bone destruction in PD is caused by the immune response. However, neutrophils provide critical antimicrobial defense against periodontal organisms. Since IL-17 is bone destructive in RA but a key regulator of neutrophils, we examined its role in inflammatory bone loss induced by the oral pathogen Porphyromonas gingivalis in IL-17RA-deficient mice. These mice showed enhanced periodontal bone destruction, suggesting a bone-protective role for IL-17, reminiscent of a neutrophil deficiency. Although IL-17RA-deficient neutrophils functioned normally ex vivo, IL-17RA knock-out (IL-17RA(KO)) mice exhibited reduced serum chemokine levels and concomitantly reduced neutrophil migration to bone. Consistently, CXCR2(KO) mice were highly susceptible to alveolar bone loss; interestingly, these mice also suggested a role for chemokines in maintaining normal bone homeostasis. These results indicate a nonredundant role for IL-17 in mediating host defense via neutrophil mobilization.


Vaccination with inactivated poliovirus vaccine and oral poliovirus vaccine in Denmark.
In Denmark a polio vaccination program including both inactivated poliovirus vaccine (IPV) and oral poliovirus vaccine ( OPV ) has been in use since 1968. Three injections of IPV are given when the children are five, six, and 15 months of age. Subsequently, three vaccinations with trivalent OPV are administered at the age of three, four, and five years. The acceptance rate is high-93%-98%-and greater than 95% of the population has antibodies to poliovirus. The geometric mean titer of serum antibodies is much greater than 10 IU for all three types. The epidemiology of poliomyelitis and the background for the development of the present vaccination schedule are reviewed.


The effect of environmental temperature on the immune response of a marine teleost (Paralichthys dentatus).
The kinetics of the primary immune response to horse erythrocytes were studied in summer flounder ( Paralichthys dentatus). This species produced antibody over a wide temperature range. The fish were divided into five groups and the experiments carried out at environmental temperatures of 27, 22, 17, 12, and 7.5 degrees C. At the lower environmental temperatures, a corresponding delay in the appearance of circulation antibody occurred, although the magnitude and duration of the response was not appreciably affected. After immunizing at 12 degrees C, lowering the environmental temperature gradually to 8 degrees C, did not appear to inhibit an ongoing primary responses. Typical secondary responses were seen in fishes kept at warmer temperatures, but when the temperature was lowered to 8 degrees C, no anamnestic response was seen. Individual variation was most noticeable at middle temperature ranges.


Randomized Phase II Trial of Gemcitabine Plus TH-302 Versus Gemcitabine in Patients With Advanced Pancreatic Cancer.
TH-302 is an investigational hypoxia-activated prodrug that releases the DNA alkylator bromo-isophosphoramide mustard in hypoxic settings. This phase II study (NCT01144455) evaluated gemcitabine plus TH-302 in patients with previously untreated, locally advanced or metastatic pancreatic cancer. Patients were randomly assigned 1:1:1 to gemcitabine (1,000 mg/m(2)), gemcitabine plus TH-302 240 mg/m(2) (G+T240), or gemcitabine plus TH-302 340 mg/m(2) (G+T340). Randomized crossover after progression on gemcitabine was allowed. The primary end point was progression-free survival (PFS). Secondary end points included overall survival (OS), tumor response, CA 19-9 response, and safety. Two hundred fourteen patients (77% with metastatic disease) were enrolled between June 2010 and July 2011. PFS was significantly longer with gemcitabine plus TH-302 (pooled combination arms) compared with gemcitabine alone (median PFS, 5.6 v 3.6 months, respectively; hazard ratio, 0.61; 95% CI, 0.43 to 0.87; P = .005; median PFS for metastatic disease, 5.1 v 3.4 months, respectively). Median PFS times for G+T240 and G+T340 were 5.6 and 6.0 months, respectively. Tumor response was 12%, 17%, and 26% in the gemcitabine, G+T240, and G+T340 arms, respectively (G+T340 v gemcitabine, P = .04). CA 19-9 decrease was greater with G+T340 versus gemcitabine (-5,398 v -549 U/mL, respectively; P = .008). Median OS times for gemcitabine, G+T240, and G+T340 were 6.9, 8.7, and 9.2 months, respectively (P = not significant). The most common adverse events (AEs) were fatigue, nausea, and peripheral edema (frequencies similar across arms). Skin and mucosal toxicities (2% grade 3) and myelosuppression (55% grade 3 or 4) were the most common TH-302-related AEs but were not associated with treatment discontinuation. PFS, tumor response, and CA 19-9 response were significantly improved with G+TH-302. G+T340 is being investigated further in the phase III MAESTRO study (NCT01746979).


Patient satisfaction with community psychiatric nursing: a prospective controlled study.
A sample of patients allocated to receive either community psychiatric nursing or out-patient psychiatrist follow-up was interviewed at 6-month intervals for a total period of 18 months. Two dimensions of patient satisfaction are discussed: a cross-sectional analysis of the quality of the perceived therapeutic relationship and an analysis over time of attitudes to aspects of the mental health care received. The satisfaction expressed with elements of the therapeutic relationship revealed that patients find nurses more approachable and more sympathetic. The satisfaction expressed by patients receiving the nursing mode of follow-up was superior to that expressed by out-patient psychiatrist patients. Particularly positive were responses made by nursing patients to domiciliary visiting. The greater levels of satisfaction with nursing follow-up increased over time.


The interaction of He with vibrating HCN: potential energy surface, bound states, and rotationally inelastic cross sections.
A four-dimensional potential energy surface representing the interaction between He and hydrogen cyanide (HCN) subjected to bending vibrational motion is presented. Ab initio calculations were carried out at the coupled-cluster level with single and double excitations and a perturbative treatment of triple excitations, using a quadruple-zeta basis set and mid-bond functions. The global minimum is found in the linear He-HCN configuration with the H atom pointing towards helium at the intermolecular separation of 7.94 a0. The corresponding well depth is 30.35 cm(-1). First, the quality of the new potential has been tested by performing two comparisons with previous theoretical and experimental works. (i) The rovibrational energy levels of the He-HCN complex for a rigid linear configuration of the HCN molecule have been calculated. The dissociation energy is 8.99 cm(-1), which is slightly smaller than the semi-empirical value of 9.42 cm(-1). The transitions frequencies are found to be in good agreement with the experimental data. (ii) We performed close coupling calculations of the rotational de-excitation of rigid linear HCN in collision with He and observed a close similarity with the theoretical data published in a recent study. Second, the effects of the vibrational bending of HCN have been investigated, both for the bound levels of the He-HCN system and for the rotationally inelastic cross sections. This was performed with an approximate method using the average of the interaction potential over the vibrational bending wavefunction. While this improves slightly the comparison of calculated transitions frequencies with experiment, the cross sections remain very close to those obtained with rigid linear HCN.


TLR4 promoted endoplasmic reticulum stress induced inflammatory bowel disease via the activation of p38 MAPK pathway.
Endoplasmic reticulum (ER) stress contribute to inflammatory bowel disease (IBD). However, the mechanistic link between toll-like receptor 4 (TLR4) and ER stress in IBD remains elusive. This study aimed to investigate the mechanism by which ER stress and TLR4 promote inflammation in IBD. IBD mouse model was established by the induction of TNBS, and Grp78 and TLR4 in intestine tissues were detected by immunohistochemistry. THP-1 cells were treated with lipopolysaccharides (LPS), ER stress inducer or inhibitor tauroursodeoxycholic acid (TUDCA), or p38 MAPK inhibitor. The activation of MAPK signaling was detected by Western blot, and the production and secretion of inflammatory factors were detected by PCR and ELISA. We found that the expression levels of TLR4 and GRP78 were significantly higher in the intestine of IBD model mice compared with control mice but were significantly lower in the intestine of IBD model mice treated with ER stress inhibitor TUDCA. ER stress inducer significantly increased while ER stress inhibitor TUDCA significantly decreased the expression and secretion of TNF-α, IL-1β and IL-8 in THP-1 cells treated by LPS. Only p38 MAPK signaling was activated in THP-1 cells treated by ER stress inducer. Furthermore, p38 inhibitor SB203580 inhibited the production and secretion of TNF-α, IL-1β and IL-8 in THP-1 cells treated with LPS. In conclusion, TLR4 promotes ER stress induced inflammation in IBD, and the effects may be mediated by p38 MAPK signaling. TLR4 and p38 MAPK signaling are novel therapeutic targets for IBD.


Nonselective enrichment for yeast adenine mutants by flow cytometry.
The expression of certain adenine biosynthetic mutations in the yeast Saccharomyces cerevisiae results in a red colony color. This phenomenon has historically provided an ideal genetic marker for the study of mutation, recombination, and aneuploidy in lower eukaryotes by classical genetic analysis. In this paper, it is reported that cells carrying ade1 and/or ade2 mutations exhibit primary fluorescence. Based on this observation, the nonselective enrichment of yeast cultures for viable adenine mutants by using the fluorescence-activated cell sorter has been achieved. The advantages of this approach over conventional genetic analysis of mutation, recombination, and mitotic chromosomal stability include speed and accuracy in acquiring data for large numbers of clones. By using appropriate strains, the cell sorter has been used for the isolation of both forward mutations and chromosomal loss events in S. cerevisiae. The resolving power of this system and its noninvasiveness can easily be extended to more complex organisms, including mammalian cells, in which analogous metabolic mutants are available.


Distinct Effects of Nalmefene on Dopamine Uptake Rates and Kappa Opioid Receptor Activity in the Nucleus Accumbens Following Chronic Intermittent Ethanol Exposure.
The development of pharmacotherapeutics that reduce relapse to alcohol drinking in patients with alcohol dependence is of considerable research interest. Preclinical data support a role for nucleus accumbens (NAc) κ opioid receptors (KOR) in chronic intermittent ethanol (CIE) exposure-induced increases in ethanol intake. Nalmefene, a high-affinity KOR partial agonist, reduces drinking in at-risk patients and relapse drinking in rodents, potentially due to its effects on NAc KORs. However, the effects of nalmefene on accumbal dopamine transmission and KOR function are poorly understood. We investigated the effects of nalmefene on dopamine transmission and KORs using fast scan cyclic voltammetry in NAc brain slices from male C57BL/6J mice following five weeks of CIE or air exposure. Nalmefene concentration-dependently reduced dopamine release similarly in air and CIE groups, suggesting that dynorphin tone may not be present in brain slices. Further, nalmefene attenuated dopamine uptake rates to a greater extent in brain slices from CIE-exposed mice, suggesting that dopamine transporter-KOR interactions may be fundamentally altered following CIE. Additionally, nalmefene reversed the dopamine-decreasing effects of a maximal concentration of a KOR agonist selectively in brain slices of CIE-exposed mice. It is possible that nalmefene may attenuate withdrawal-induced increases in ethanol consumption by modulation of dopamine transmission through KORs.


Opening up the advantages of the ruthenocenic bioprobes of ferroquine: distribution and localization in Plasmodium falciparum-infected erythrocytes.
A ferrocene-quinoline conjugate named ferroquine (FQ or SSR97193) is active against both chloroquine-susceptible and chloroquine-resistant Plasmodium falciparum and P. vivax strains and/or isolates. FQ was shown to be efficient for the treatment of uncomplicated malaria in humans (phase IIb of clinical trials). However, the molecular basis of FQ's mechanism of action is still unknown because few approaches (such as radioactive labelling or immunofluorescence) are available for that purpose. Previous reports from our laboratory suggest that the intramolecular hydrogen bond in the lateral side chain plays a crucial role in the antimalarial activity of the drug. We used two ruthenocenic bioprobes of FQ (with and without an intramolecular hydrogen bond) to study their localization and quantification in Plasmodium falciparum-infected erythrocytes. We first used Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) analysis to trace ruthenoquine (RQ, with an intramolecular hydrogen bond) and methylruthenoquine (Me-RQ, without an intramolecular hydrogen bond) in the infected red blood cells (iRBCs). We showed that RQ accumulates faster in the digestive vacuole of the iRBCs than Me-RQ. We next examined the ruthenium distribution at the ultrastructural level by transmission electron microscopy (TEM). We showed that RQ accumulates faster in the parasitic digestive vacuole (DV) close to its membranes than Me-RQ.


Use of the red fluorescent protein as a marker of Kaposi's sarcoma-associated herpesvirus lytic gene expression.
A hallmark of all herpesvirus is the ability to exist in either a latent, or lytic, state of replication, enabling the lifelong infection of its host. Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) can efficiently establish a latent infection in a variety of cell types in vitro, making it a valuable model for the study of latency and reactivation. To facilitate the identification of KSHV lytic replication, and allow subsequent experiments with live cells, a recombinant virus, rKSHV.219, was constructed using JSC-1 cells that expresses the red fluorescent protein (RFP) from the KSHV lytic PAN promoter, the green fluorescent protein (GFP) from the EF-1alpha promoter, and with the gene for puromycin resistance as a selectable marker. rKSHV.219 from JSC-1 cells was used to infect Vero cells for purification of the recombinant virus. Vero cells were also used for the production of rKSHV.219 at levels of 10(5)-10(6) infectious units (IU) of virus per milliliter using a combination of KSHV/RTA expressed from a baculovirus vector, BacK50, and butyrate. Virus produced from Vero cells was used to infect human fibroblasts (HF), 293, DU145, T24, HaCaT, and HEp-2 cells, and in all cells except 293 cells, only a latent infection was established with GFP expression, but no RFP expression. In 293 cells, 10-15% of cells showed lytic gene expression. Both primary and immortalized microvascular endothelial cells (MVEC) were also infected with rKSHV.219, and reduced spontaneous lytic replication was found in immortalized cells. In all cells used in this study, rKSHV.219 efficiently established latent infections from which the virus could be reactivated to productive lytic replication. This work also demonstrated strong synergy between KSHV/RTA and butyrate for the activation of KSHV lytic replication and the production of infectious virus.


Enhanced tumor cell growth suppression by an N-terminal truncated retinoblastoma protein.
The retinoblastoma (RB) gene encodes a nuclear phosphoprotein of 928 amino acids (pRB). Thus far, much effort in RB research has been focused on both the viral oncoprotein-binding domains and the C-terminal domain, whereas little is known about the N-terminal moiety of the protein. We report here that an N-terminal truncated RB protein of approximately 94 kDa (pRB94) exerts more potent cell growth suppression as compared to the full-length pRB protein in a diversity of tumor cell lines examined, including those having a normal endogenous RB gene. Tumor cells transfected with the pRB94-expressing plasmids displayed multiple morphological changes frequently associated with cellular senescence and/or apoptosis. They failed to enter S phase and rapidly died. The pRB94 expressed in recipient tumor cells had a longer half-life than the full-length pRB protein and tended to remain in an active un- or hypophosphorylated form. Since it has also been found that N-terminal truncated RB proteins often accumulated in growth-arrested and/or differentiated tumor cells, we suggest that N-terminal truncation of pRB may be one of the cellular mechanisms modulating the RB protein function in cell-cycle control.


Inhibitors and inactivators of protein arginine deiminase 4: functional and structural characterization.
Protein arginine deiminase 4 (PAD4) is a transcriptional coregulator that catalyzes the calcium-dependent conversion of specific arginine residues in proteins to citrulline. Recently, we reported the synthesis and characterization of F-amidine, a potent and bioavailable irreversible inactivator of PAD4. Herein, we report our efforts to identify the steric and leaving group requirements for F-amidine-induced PAD4 inactivation, the structure of the PAD4-F-amidine x calcium complex, and in vivo studies with N-alpha-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide (Cl-amidine), a PAD4 inactivator with enhanced potency. The PAD4 inactivators described herein will be useful pharmacological probes in characterizing the incompletely defined physiological role(s) of this enzyme. In addition, they represent potential lead compounds for the treatment of rheumatoid arthritis because a growing body of evidence supports a role for PAD4 in the onset and progression of this chronic autoimmune disorder.


Brownian dynamics investigation of magnetization and birefringence relaxations in ferrofluids.
Brownian dynamics simulations are used to investigate the dynamics of orientational properties of real charge-stabilized ferrofluids, i.e. stable colloidal dispersions of magnetic nanoparticles. The relaxation times of the magnetization and of the birefringence, data accessible by experimental techniques, have been computed at several volume fractions. Besides, the effect of hydrodynamic interactions has been investigated. Equilibrium simulations without field are found to be inadequate to determine the aforementioned relaxation times for the systems under study, the dipolar interaction being too weak. Thus a nonequilibrium simulation procedure that mimics the experimental operating mode has been developed. After equilibrium simulations under a magnetic field, both birefringence and magnetization decays are recorded once the field is suppressed. Birefringence and magnetization decays are markedly impeded as the volume fraction increases, whereas they are barely enhanced when the intensity of the initial magnetic field is increased at a fixed volume fraction. Eventually, hydrodynamic interactions exhibit a slight but systematic lengthening of the relaxation times.


Diabetic Ketoacidosis in Children Before and During COVID-19 Pandemic: A Cross-sectional Study.
Coronavirus disease 2019 (COVID-19) has spread quickly. Comorbidities, such as diabetes, have been determined as critical risk factors for COVID-19. This study aimed to determine the frequency and severity of diabetic ketoacidosis (DKA) in children before and during the COVID-19 pandemic. This retrospective study examined children aged less than 18 years diagnosed with DKA hospitalized in Yazd Shahid Sadoughi Hospital from February 20, 2020, to November 21, 2021. The collected information was compared to those obtained during the same period in 2019 (pre-pandemic). According to the inclusion criteria, only children with suspected symptoms of COVID-19 or an infected family member underwent PCR. The study included 70 children with confirmed DKA during the COVID-19 pandemic and 33 children hospitalized during the pre-pandemic period. The findings showed that the rate of DKA was higher during the pandemic than in the pre-pandemic period. In the DKA subgroups (during the COVID-19 pandemic vs. pre-pandemic), 35.7% vs. 21.2% were severe, 37.1% vs. 36.4% were moderate, and 27.1% vs. 42.4% were mild. Of 70 children, 30 underwent PCR tests for COVID-19, showing six positive cases. Among positive cases, five had mild symptoms, while one was hospitalized with signs of respiratory distress, polyuria, and polydipsia. All physical examinations of this patient were normal, except for the chest exam. A remarkable increase was observed in the frequency and severity of DKA in children during the pandemic.


Growth of density inhomogeneities in a flow of wave turbulence.
We consider the flow being a superposition of random waves and describe the evolution of the spectrum of the passive scalar in the leading (fourth) order with respect to the wave amplitudes. We find that wave turbulence can produce an exponential growth of the passive scalar fluctuations when either both solenoidal and potential components are present in the flow or there are potential waves with the same frequencies but different wave numbers.


Studies on the release of solubilized drugs from ointment bases. Part 8: The effect of emulsifiers on pharmaceutical availability of ethanolamine, diethanolamine and morpholine from emulsive ointments.
The effect of addition of lanolin and Span 60 to white vaseline and of their concentration on the pharmaceutical availability of monoethanolamine , diethanolamine and morpholine was investigated. The emulsifier type and its concentration exercise an influence on constant release rates. The liberation of the investigated amines proceeds at almost the same speed as therapeutic substances within the limits from 10 to 30%. The release half life periods showed the same dependence in relation to the content of investigated substances in the ointment.


Evaluating Dye-Labeled DNA Dendrimers for Potential Applications in Molecular Biosensing.
DNA nanostructures provide a reliable and predictable scaffold for precisely positioning fluorescent dyes to form energy transfer cascades. Furthermore, these structures and their attendant dye networks can be dynamically manipulated by biochemical inputs, with the changes reflected in the spectral response. However, the complexity of DNA structures that have undergone such types of manipulation for direct biosensing applications is quite limited. Here, we investigate four different modification strategies to effect such dynamic manipulations using a DNA dendrimer scaffold as a testbed, and with applications to biosensing in mind. The dendrimer has a 2:1 branching ratio that organizes the dyes into a FRET-based antenna in which excitonic energy generated on multiple initial Cy3 dyes displayed at the periphery is then transferred inward through Cy3.5 and/or Cy5 relay dyes to a Cy5.5 final acceptor at the focus. Advantages of this design included good transfer efficiency, large spectral separation between the initial donor and final acceptor emissions for signal transduction, and an inherent tolerance to defects. Of the approaches to structural rearrangement, the first two mechanisms we consider employed either toehold-mediated strand displacement or strand replacement and their impact was mainly via direct transfer efficiency, while the other two were more global in their effect using either a belting mechanism or an 8-arm star nanostructure to compress the nanostructure and thereby modulate its spectral response through an enhancement in parallelism. The performance of these mechanisms, their ability to reset, and how they might be utilized in biosensing applications are discussed.


Overexpression of the integrin-linked kinase promotes anchorage-independent cell cycle progression.
Cell adhesion to substratum has been shown to regulate cyclin A expression as well as cyclin D- and E-dependent kinases, the latter via the up-regulation of cyclin D1 and the down-regulation of cyclin-Cdk inhibitors p21 and p27, respectively. This adhesion-dependent regulation of cell cycle is thought to be mediated by integrins. Here we demonstrate that stable transfection and overexpression of the integrin-linked kinase (ILK), which interacts with the beta1 and beta3 integrin cytoplasmic domains, induces anchorage-independent cell cycle progression but not serum-independent growth of rat intestinal epithelial cells (IEC18). ILK overexpression results in increased expression of cyclin D1, activation of Cdk4 and cyclin E-associated kinases, and hyperphosphorylation of the retinoblastoma protein. In addition, ILK overexpression results in the expression of p21 and p27 Cdk inhibitors with altered electrophoretic mobilities, with the p27 from ILK-overexpressing cells having reduced inhibitory activity. The transfer of serum-exposed IEC18 cells from adherent cultures to suspension cultures results in a rapid down-regulation of expression of cyclin D1 and cyclin A proteins as well as in retinoblastoma protein dephosphorylation. In marked contrast, transfer of ILK-overexpressing cells from adherent to suspension cultures results in continued high levels of expression of cyclin D1 and cyclin A proteins, and a substantial proportion of the retinoblastoma protein remains in a hyperphosphorylated state. These results indicate that, when overexpressed, ILK induces signaling pathways resulting in the stimulation of G1/S cyclin-Cdk activities, which are normally regulated by cell adhesion and integrin engagement.


Role of VASP in reestablishment of epithelial tight junction assembly after Ca2+ switch.
Epithelial permeability is tightly regulated by intracellular messengers. Critical to maintaining barrier integrity is the formation of tight junction complexes. A number of signaling pathways have been implicated in tight junction biogenesis; however, the precise molecular mechanisms are not fully understood. A growing body of evidence suggests a role for intracellular cAMP in tight junction assembly. Using an epithelial model, we investigated the role of cAMP signal transduction in barrier recovery after Ca2+ switch. Our data demonstrate that elevation of intracellular cAMP levels significantly enhanced barrier recovery after Ca2+ switch. Parallel experiments revealed that epithelial barrier recovery is diminished by H-89, a specific and potent inhibitor of cAMP-dependent protein kinase (protein kinase A) activity. Of the possible PKA effector proteins, the vasodilator-stimulated phosphoprotein (VASP) is an attractive candidate, since it has been implicated in actin-binding and cross-linking functions. We therefore hypothesized that VASP may play a role in the cAMP-mediated regulation of epithelial junctional reassembly after Ca2+ switch. We demonstrate here that VASP is phosphorylated via a PKA-dependent process under conditions that enhance barrier recovery. Confocal laser scanning microscopy studies revealed that VASP localizes with ZO-1 at the tight junction and at cell-cell borders and that phospho-VASP appears at the junction after Ca2+ switch. Subsequent transfection studies utilizing epithelial cells expressing truncated forms of VASP abnormal in oligomerization or actin-binding activity revealed a functional diminution of barrier recovery after Ca2+ chelation. Our present studies suggest that VASP may provide a link between cAMP signal transduction and epithelial permeability.


Abstraction and addition kinetics of C2H radicals with CH4, C2H6, C3H8, C2H4, and C3H6: CVT/SCT/ISPE and hybrid meta-DFT methods.
Rate coefficients for the reactions of C2H radicals with methane (k1), ethane (k2), propane (k3), ethylene (k4), and propylene (k5) were computed using canonical variational transition state theory (CVT) coupled with hybrid-meta density functional theory (DFT) over a wide range of temperatures from 150 to 5000 K. The quantum chemical tunneling effect was corrected by the small curvature tunneling (SCT) method. The dynamic calculations are performed using the variational transition state theory (VTST) with the interpolated single-point energies (ISPE) method at the CCSD(T)/cc-pVTZ//M06-2X/6-31+G(d,p) level of theory. Intrinsic reaction coordinate (IRC) calculations were performed to verify that the transition states are connected to the reactants and products. The rate coefficients obtained over the studied temperature range yield the following Arrhenius expressions (cm(3) molecule(-1) s(-1)): k1 = 4.69 × 10(-19)T(2.44) exp[331/T], k2 = 4.29 × 10(-17)T(2.11) exp[432/T], k3 = 4.81 × 10(-17)T(1.98) exp[697/T], k4 = 7.54 × 10(-21)T(2.96) exp[1942/T], and k5 = 8.04 × 10(-23)T(3.44) exp[3011/T] cm(3) molecule(-1) s(-1). Branching ratio calculation for the reactions of C2H radicals with ethylene and propylene shows that the abstraction reactions are not important at lower temperatures. However, as the temperature increases, abstraction reactions become more important.


Fitting and interpreting occupancy models.
We show that occupancy models are more difficult to fit than is generally appreciated because the estimating equations often have multiple solutions, including boundary estimates which produce fitted probabilities of zero or one. The estimates are unstable when the data are sparse, making them difficult to interpret, and, even in ideal situations, highly variable. As a consequence, making accurate inference is difficult. When abundance varies over sites (which is the general rule in ecology because we expect spatial variance in abundance) and detection depends on abundance, the standard analysis suffers bias (attenuation in detection, biased estimates of occupancy and potentially finding misleading relationships between occupancy and other covariates), asymmetric sampling distributions, and slow convergence of the sampling distributions to normality. The key result of this paper is that the biases are of similar magnitude to those obtained when we ignore non-detection entirely. The fact that abundance is subject to detection error and hence is not directly observable, means that we cannot tell when bias is present (or, equivalently, how large it is) and we cannot adjust for it. This implies that we cannot tell which fit is better: the fit from the occupancy model or the fit ignoring the possibility of detection error. Therefore trying to adjust occupancy models for non-detection can be as misleading as ignoring non-detection completely. Ignoring non-detection can actually be better than trying to adjust for it.


Immunocytochemistry in liquid-based cervical cytology: analysis of clinical use following a cross-sectional study.
Cytological screening for cervical cancer is hampered by imperfect sensitivity and low inter-observer reproducibility. Human papillomavirus (HPV) testing lacks specificity as a primary screening method. Studies indicate that immunocytochemical detection of alterations caused by HPV in the host cells can optimise screening. Here, the potential of p16(INK4a) (cyclin-dependent kinase inhibitor p16) and MIB-1 (Ki-67 proliferation marker) as adjunct molecular markers for cervical lesions was investigated in a prospective, cross-sectional study of 500 samples in the framework of opportunistic screening in Flanders, Belgium. A consecutive series of 200 samples and 100 samples from the cytological categories ASC, LSIL and HSIL were investigated. Surepath samples were interpreted according to the Bethesda 2001 reporting system. HPV testing was done with MY09/MY11 consensus PCR. Immunocytochemistry for p16(INK4a) and MIB-1 was performed with an automated staining protocol. The number of immunoreactive cells/1,000 cervical cells was assessed. There was a higher mean number of p16(INK4A) and MIB-1 immunoreactive cells/1,000 cells in HSIL (4.06 +/- 1.93 and 11.13 +/- 2.83, respectively) compared to other cytological categories. Both markers showed a large spread in counts, for all categories. In cases of HSIL without immunoreactive cells for either marker, low cellularity and long-term storage in water were often the cause of false negativity. This study confirms that positive staining for p16(INK4a) and MIB-1 is highly correlated with presence of high-grade lesions. These markers could be used as adjuncts to increase the sensitivity of cytological screening as well as the specificity of the HPV test. However, clear methodological standards are needed for optimal performance of immunocytochemistry in a clinical setting.


A Model of Waardenburg Syndrome Using Patient-Derived iPSCs With a SOX10 Mutation Displays Compromised Maturation and Function of the Neural Crest That Involves Inner Ear Development.
Waardenburg syndrome (WS) is an autosomal dominant inherited disorder that is characterized by sensorineural hearing loss and abnormal pigmentation. SOX10 is one of its main pathogenicity genes. The generation of patient-specific induced pluripotent stem cells (iPSCs) is an efficient means to investigate the mechanisms of inherited human disease. In our work, we set up an iPSC line derived from a WS patient with SOX10 mutation and differentiated into neural crest cells (NCCs), a key cell type involved in inner ear development. Compared with control-derived iPSCs, the SOX10 mutant iPSCs showed significantly decreased efficiency of development and differentiation potential at the stage of NCCs. After that, we carried out high-throughput RNA-seq and evaluated the transcriptional misregulation at every stage. Transcriptome analysis of differentiated NCCs showed widespread gene expression alterations, and the differentially expressed genes (DEGs) were enriched in gene ontology terms of neuron migration, skeletal system development, and multicellular organism development, indicating that SOX10 has a pivotal part in the differentiation of NCCs. It's worth noting that, a significant enrichment among the nominal DEGs for genes implicated in inner ear development was found, as well as several genes connected to the inner ear morphogenesis. Based on the protein-protein interaction network, we chose four candidate genes that could be regulated by SOX10 in inner ear development, namely, BMP2, LGR5, GBX2, and GATA3. In conclusion, SOX10 deficiency in this WS subject had a significant impact on the gene expression patterns throughout NCC development in the iPSC model. The DEGs most significantly enriched in inner ear development and morphogenesis may assist in identifying the underlying basis for the inner ear malformation in subjects with WS.


Formation and mitigation of acrylamide in oven baked vegetable fries.
Investigation into oven baked sweet potato and carrot fries at various temperatures and times demonstrated the in situ formation of acrylamide in an exponential manner. High levels of acrylamide were found in these food items: up to 327 µg/kg for sweet potato baked at 190 °C for 14 min, and 99 µg/kg for carrot baked at 190 °C for 13 min. Risk assessment via Margin of Exposures estimation showed that consumption of these fries might pose adverse health effects to consumers from toddlers to adults, especially when the fries were prepared at high temperatures above 175 °C and for a long time. Raw ingredient blanching and immersion in acetic acid prior to preparation have been proven to greatly reduce acrylamide formation, up to 99%. It is recommendable to apply these techniques either at industrial or domestic cooking scales to ensure minimal health risk from dietary exposure to acrylamide.


Comparison of Physician-Modified Fenestrated/Branched Stent-Grafts and Hybrid Visceral Debranching Plus Stent-Graft Placement for Complex Thoracoabdominal Aortic Aneurysm Repair.
Purpose: To evaluate the immediate postoperative and midterm outcomes of complex thoracoabdominal aortic aneurysm (TAAA) treatment with fenestrated/branched physician-modified endovascular grafts (PMEGs) or open debranching of the visceral aorta with bypass graft revascularization plus endovascular aneurysm exclusion (hybrid repair). Materials and Methods: A retrospective analysis was conducted of 88 patients (mean age 70.0±10.6 years; 73 men) with complex TAAAs who underwent treatment with PMEGs (60, 68%) or a hybrid technique (28, 32%) between 2016 and 2019. The mean aneurysm diameter was 64.5±11.7 mm, and 37 patients (42%) were symptomatic. The Zenith TX2 and Ankura were the main stent-grafts used in the PMEG group. The hybrid technique involved visceral debranching with extra-anatomical bypass graft revascularization and subsequent stent-graft deployment (1- or 2-stage procedure). Results: In the PMEG group, 35 patients received modified stent-grafts with 4 fenestrations, 8 patients had 4 branches per device, and 17 patients had combinations (50 fenestrations and 18 branches) that successfully revascularized 228 of the 240 targets (95%). In the 28 hybrid cases, all 110 target vessels were successfully revascularized with bypass grafts. The overall 30-day mortality was 3.4% (2 PMEG and 1 hybrid), and the early rate of target vessel stenosis/occlusion was 3.3% (5 in PMEG group and 6 in the hybrid repair group). The 30-day morbidity was mainly attributed to pulmonary complications (15%), lower limb ischemia (8%), or spinal cord ischemia with paraplegia (6%). Eleven patients (13%) had deteriorated renal function with a >30% decrease in the glomerular filtration rate. The mean follow-up was 22.3±4.9 months, and mortality was 4.5% (3.3% in the PMEG group vs 7.1% in the hybrid repair group). Conclusion: PMEGs and hybrid techniques seem to be feasible treatment options for aortic aneurysms necessitating visceral vessel revascularization. PMEGs may have a lower morbidity than the hybrid technique, which nonetheless remains an important option available for complex aortic aneurysms.


Collective excitation of Rydberg-atom ensembles beyond the superatom model.
In an ensemble of laser-driven atoms involving strongly interacting Rydberg states, the steady-state excitation probability is usually substantially suppressed. In contrast, here we identify a regime in which the Rydberg excited fraction is enhanced by the interaction. This effect is associated with the buildup of many-body coherences induced by coherent multiphoton excitations between collective states. The excitation enhancement should be observable under currently existing experimental conditions and may serve as a direct probe for the presence of coherent multiphoton dynamics involving collective quantum states.


Comparison of secretome from osteoblasts derived from sclerotic versus non-sclerotic subchondral bone in OA: A pilot study.
Osteoarthritis (OA) is characterized by cartilage degradation but also by other joint tissues modifications like subchondral bone sclerosis. In this study, we used a proteomic approach to compare secretome of osteoblast isolated from sclerotic (SC) or non sclerotic (NSC) area of OA subchondral bone. Secretome was analyzed using differential quantitative and relative label free analysis on nanoUPLC G2 HDMS system. mRNA of the more differentially secreted proteins were quantified by RT-PCR in cell culture from 5 other patients. Finally, osteomodulin and fibulin-3 sequences were quantified by western blot and immunoassays in serum and culture supernatants. 175 proteins were identified in NSC osteoblast secretome. Data are available via ProteomeXchange with identifier PXD008494. Compared to NSC osteoblast secretome, 12 proteins were significantly less secreted (Osteomodulin, IGFBP5, VCAM-1, IGF2, 78 kDa glucose-regulated protein, versican, calumenin, IGFBP2, thrombospondin-4, periostin, reticulocalbin 1 and osteonectin), and 13 proteins were significantly more secreted by SC osteoblasts (CHI3L1, fibulin-3, SERPINE2, IGFBP6, SH3BGRL3, SERPINE1, reticulocalbin3, alpha-2-HS-glycoprotein, TIMP-2, IGFBP3, TIMP-1, SERPINF1, CSF-1). Similar changes in osteomodulin, IGF2, SERPINE1, fibulin-3 and CHI3L1 mRNA levels were observed. ELISAs assays confirm the decrease by half of osteomodulin protein in SC osteoblasts supernatant compared to NSC and in OA patients serum compared to healthy subjects. Fibulin-3 epitopes Fib3-1, Fib3-2 and Fib3-3 were also increased in SC osteoblasts supernatant compared to NSC. We highlighted some proteins differentially secreted by the osteoblasts coming from OA subchondral bone sclerosis. These changes contribute to explain some features observed in OA subchondral bone, like the increase of bone remodeling or abnormalities in bone matrix mineralization. Among identified proteins, osteomodulin was found decreased and fibulin-3 increased in serum of OA patients. These findings suggest that osteomodulin and fibulin-3 fragments could be biomarkers to monitor early changes in subchondral bone metabolism in OA.


Pressure selected reactivity and kinetics deduced from photoinduced dissociation of ethylene glycol.
The photon-induced reactivity of liquid ethylene glycol (EG) was investigated in a diamond anvil cell at pressures up to ∼4 GPa and ambient temperature. The near-UV radiation at λ = 350 nm was employed to photodissociate EG via the two-photon absorption processes. The reaction evolution was monitored as a function of time and the reaction products were characterized by using in situ FTIR spectroscopy. At low initial loading pressures, the IR spectra show two distinctive sets of profile evaluations indicating sequential photoinduced chemical reactions, which are designated as primary and secondary photochemical processes, respectively. By careful examination of the characteristic IR bands and possible reaction pathways, over ten species as the primary and secondary reaction products were unambiguously identified. Significantly, we found that one of the photodissociation product CO2 forms specific clathrate hydrate structures or clusters that are both time- and pressure-dependent, indicating interesting and unique sequestration behavior of CO2 at high pressures. Quantitative analysis on selective reaction products allows detailed reaction kinetics involving competitive reaction channels to be probed. In particular, the type and quantity of reaction products as well as the kinetics were found highly pressure dependent. Moreover, the pressure variation of the system along the reaction progression allows the interpretation of possible reaction mechanisms of photodissociation of EG under high pressures.


[Left renal pelvic tumor 32 years after calyco-ileo-vesiconeostomy].
This is a case report of left renal pelvic tumor found 32 years after left calico-ileo-vesiconeostomy. The patient has undergone right nephrectomy for absence of renal function at 26 years-old and left urinary reconstruction was performed using the intestine for pyelo-ureteral junction obstruction at 28 years old because of bilateral congenital hydronephrosis. Later he was treated for recurrent left renal stone with percutaneous nephrolithotripsy, nephrolithotomy and extracorporeal shock wave lithotripsy. When he was 56 years old, macrohematuria appeared. He received left nephrectomy under the diagnosis of left renal pelvic tumor.


MUC13 Cell Surface Mucin Limits Salmonella Typhimurium Infection by Protecting the Mucosal Epithelial Barrier.
MUC13 cell surface mucin is highly expressed on the mucosal surface throughout the intestine, yet its role against bacterial infection is unknown. We investigated how MUC13 impacts Salmonella typhimurium (S Tm) infection and elucidated its mechanisms of action. Muc13-/- and wild-type littermate mice were gavaged with 2 isogenic strains of S Tm after pre-conditioning with streptomycin. We assessed clinical parameters, cecal histology, local and systemic bacterial load, and proinflammatory cytokines after infection. Cecal enteroids and epithelial cell lines were used to evaluate the mechanism of MUC13 activity after infection. The interaction between bacterial SiiE and MUC13 was assessed by using siiE-deficient Salmonella. S Tm-infected Muc13-/- mice had increased disease activity, histologic damage, and higher local and systemic bacterial loads. Mechanistically, we found that S Tm binds to MUC13 through its giant SiiE adhesin and that MUC13 acts as a pathogen-binding decoy shed from the epithelial cell surface after pathogen engagement, limiting bacterial invasion. In addition, MUC13 reduces epithelial cell death and intestinal barrier breakdown by enhancing nuclear factor kappa B signaling during infection, independent of its decoy function. We show for the first time that MUC13 plays a critical role in antimicrobial defense against pathogenic S Tm at the intestinal mucosal surface by both acting as a releasable decoy limiting bacterial invasion and reducing pathogen-induced cell death. This further implicates the cell surface mucin family in mucosal defense from bacterial infection.