bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of functionally active ATP‐sensitive K‐channels in insect cells using baculovirus
Authors: Mikhailov Michael V. , Proks Peter , Ashcroft Frances M. , Ashcroft Stephen J.H.
Published: July 2002

We have expressed active ATP‐sensitive K‐channels (KATPcharmels) inSpodoptera frugiperda(Sf9) cells using a baculovirus vector. A high yield of active channels was obtained on co‐infection with SUR1 and Kir6.2 engineered to contain N‐ and/or C‐terminal tags to permit detection by Western blotting. Channel activity was sensitive to ATP, glibenclamide and diazoxide. Channel activity was also obtained on expression of a C‐terminally truncated Kir6.2 (Kir6.2ΔC26): these channels were blocked by ATP but were insensitive to sulphonylureas. In contrast toXenopusoocytes and mammalian cells the full length Kir6.2 also gave rise to active channels in Sf9 cells when expressed alone. The highest yield of active KATPchannels was obtained on infection with a fusion protein containing SUR1 linked to Kir6.2ΔC26 via a 6‐amino acid linker.
Growth factor protection against cytokine‐induced apoptosis in neonatal rat islets of Langerhans: role of Fas
Authors: Harrison M. , Dunger A.M. , Berg S. , Mabley J. , John N. , Green M.H.L. , Green I.C.
Published: July 2002

Treatment of neonatal rat islets of Langerhans with combined cytokines (interleukin‐1β 10−10 M, tumour necrosis factor‐α 10−10 M, interferon‐γ 5 U/ml) led to extensive cell death, which was potentiated by Fas activation with the anti‐Fas cytolytic antibody JO2. Pre‐treatment with insulin (25 ng/ml) or insulin‐like growth factor‐1 (10−8 M) gave only partial protection against cell killing, but prevented the Fas‐mediated component. In the absence of cytokine treatment, Fas‐mediated killing was not observed.
Discrimination of 5′‐terminal start codons by translation initiation factor 3 is mediated by ribosomal protein S1
Authors: Moll Isabella , Resch Armin , Bläsi Udo
Published: July 2002

The interrelation between ribosomal protein S1 and IF3 in recognition/discrimination of 5′‐terminal start codons by 30S ribosomes has been studied using in vitro toeprinting. The study has been performed with two naturally occurring leaderless mRNAs, λ cI and phage r1t rro mRNA, as well as with an artificial leaderless mRNA derived from the E. coli ompA gene. We show that in the absence of S1, IF3 does not discriminate against the authentic 5′‐terminal start codon of both cI and rro mRNA. Since IF3 was able to exert its proofreading function for initiator on 30S ribosomes lacking S1, this observation cannot be attributed to a lack of binding to or action of IF3 on 30S(−S1) ribosomes. In contrast to leaderless mRNAs, ternary complex formation occurs in the presence of IF3 with 30S ribosomes when the start codon is preceded by a short 20‐nucleotide 5′‐untranslated region containing a canonical Shine and Dalgarno sequence. This suggests that 5′‐terminal start codons are recognised by IF3 as non‐standard because of the lack of 16S rRNA‐mRNA contacts.
Dissimilar phorbol ester binding properties of the individual cysteine‐rich motifs of protein kinase D
Authors: Iglesias Teresa , Matthews Sharon , Rozengurt Enrique
Published: July 2002

Protein kinase D (PKD) is a serine/threonine kinase that binds phorbol esters in a phospholipid‐dependent manner via a tandemly repeated cysteine‐rich, zinc finger‐like motif (the cysteine‐rich domain, CRD). Here, we examined whether the individual cysteine‐rich motifs of the CRD of PKD (referred to as cys1 and cys2) are functionally equivalent in mediating phorbol ester binding both in vivo and in vitro. Our results demonstrate that the cys1 and cys2 motifs of the CRD of PKD are functionally dissimilar, with the cys2 motif responsible for the majority of [3H]phorbol 12,13‐dibutyrate (PDB) binding, both in vivo and in vitro.
Tissue transglutaminase expression affects hypusine metabolism in BALB/c 3T3 cells
Authors: Beninati S , Gentile V , Caraglia M , Lentini A , Tagliaferri P , Abbruzzese A
Published: July 2002

Post‐translational formation of hypusine in eukaryotic initiation factor 5A (eIF‐5A) is essential for cell viability. Recently, we showed that hypusine protein is an in vitro substrate for transglutaminases (TGases). We report the effect of tissue TGase expression on the in vivo hypusine metabolic pathway. The stable expression of tTGase in BALB/c 3T3 cells induced a 100‐fold reduction of hypusine levels and a 50% increase of γ‐glutamyl‐ϖ‐hypusine formation. Such changes were paralleled by a consistent decrease in the free polyamine pool and an enhancement of their excretion and of the formation of their γ‐glutamyl derivatives. These effects occurred together with a significant reduction of cell proliferation. In this report we suggest, for the first time, that tTGase affects hypusine metabolism, thus regulating the eIF‐5A activity and cell proliferation.
The major myosin phosphatase in skeletal muscle is a complex between the β‐isoform of protein phosphatase 1 and the MYPT2 gene product
Authors: Moorhead Greg , Johnson Deborah , Morrice Nick , Cohen Philip
Published: July 2002

Myosin is dephosphorylated by distinct forms of protein phosphatase 1 (PP1) in smooth muscle and skeletal muscle that are composed of PP1 complexed to different regulatory subunits. The smooth muscle myosin phosphatase (smPP1M) has been characterised previously and is composed of PP1β complexed to M110 and M21 subunits that enhance the dephosphorylation of smooth muscle myosin, but not skeletal muscle myosin. In contrast, the regulatory subunit(s) of skeletal muscle myosin phosphatase (skPP1M) greatly enhance(s) the dephosphorylation of skeletal muscle myosin. Here we identify a regulatory subunit of skPP1M as the product of the MYPT2 gene, a protein whose sequence is 61% identical to the M110 subunit of smPP1M. Surprisingly, the M21 subunit of smPP1M appears to be produced from the same gene that encodes MYPT2.
The structure and expression of a preprohormone of a neuropeptide, Hym‐176 in Hydra magnipapillata
Authors: Yum Seungshic , Takahashi Toshio , Hatta Masayuki , Fujisawa Toshitaka
Published: July 2002

Hym‐176 (APFIFPGPKVamide) is a novel myoactive neuropeptide which was identified in systematic screening of signaling peptides in Hydra magnipapillata. By using PCR and library screening, we cloned and sequenced a full length cDNA which encoded a preprohormone of Hym‐176. In the preprohormone, a typical signal sequence, one copy of Hym‐176 precursor peptide and one copy of precursor sequence of another novel peptide, Hym‐357 (KPAFLFKGYKPamide), were detected. In situ hybridization analysis revealed a strong signal in peduncle neurons. Signals were also detected, though weaker, in neurons in the gastric region and around the mouth. No signals were detected in the two extremities of the body, tentacles and basal disk. The expression pattern is correlated with the distribution of Hym‐176 and its myoactive function.
The Rhodococcus erythropolis DCL14 limonene‐1,2‐epoxide hydrolase gene encodes an enzyme belonging to a novel class of epoxide hydrolases
Authors: Barbirato Fabien , Verdoes Jan C , de Bont Jan A.M , van der Werf Mariët J
Published: July 2002

Recently, we reported the purification of the novel enzyme limonene‐1,2‐epoxide hydrolase involved in limonene degradation by Rhodococcus erythropolis DCL14. The N‐terminal amino acid sequence of the purified enzyme was used to design two degenerate primers at the beginning and the end of the 50 amino acids long stretch. Subsequently, the complete limonene‐1,2‐epoxide hydrolase gene (limA) was isolated from a genomic library of R. erythropolis DCL14 using a combination of PCR and colony hybridization. The limA gene encoded a 149‐residue polypeptide with a deduced molecular mass of 16.5 kDa. It was functionally expressed in Escherichia coli. The amino acid sequence of limA contains neither any of the conserved regions of the α,β‐hydrolase fold enzymes, to which most of the previously reported epoxide hydrolases belong, nor any of the conserved motifs present in leukotriene A4 hydrolase. The structural data presented in this paper confirm previous physical and biochemical findings [van der Werf et al. (1998) J. Bacteriol. 180, 5052–5057] that limonene‐1,2‐epoxide hydrolase is the first member of a new class of epoxide hydrolases.
Recombinant human amyloid precursor‐like protein 2 (APLP2) expressed in the yeast Pichia pastoris can stimulate neurite outgrowth
Authors: Cappai Roberto , Mok Su San , Galatis Denise , Tucker David F. , Henry Anna , Beyreuther Konrad , Small David H. , Masters Colin L.
Published: July 2002

The human amyloid precursor‐like protein 2 (APLP2) is a member of the Alzheimer's disease amyloid precursor protein (APP) gene family. The human APLP2 ectodomain (sAPLP2) was expressed in the yeast Pichia pastoris and the recombinant sAPLP2 was purified from the culture medium in a single step by metal‐chelating Sepharose chromatography. The neuritotrophic activity of APLP2 was compared to the APP isoforms sAPP695 and sAPP751 on chick sympathetic neurones. APLP2 had neurite outgrowth‐promoting activity similar to that of the APP isoforms. This suggests that APP and APLP2 have a similar or related role and supports the idea of a redundancy in function between the APP‐gene family proteins.
Protection against peroxynitrite
Authors: Arteel Gavin E. , Briviba Karlis , Sies Helmut
Published: July 2002

Peroxynitrite formed in vivo from superoxide and nitric oxide can mediate oxidation, nitration, or nitrosation reactions, leading to impaired function, toxicity, and alterations in signaling pathways. Protection against peroxynitrite is important for defense of normal tissue, especially during inflammation. Biological protection against peroxynitrite is organized in three categories: prevention, interception, and repair. Prevention is the control of the formation of peroxynitrite precursors, nitric oxide and superoxide. Interception is by direct reaction with peroxynitrite, leading to non‐toxic products. In this regard, organoselenium compounds, metalloporphyrin derivatives, and peroxidases (e.g. glutathione peroxidase and myeloperoxidase) exhibit high second‐order rate constants with peroxynitrite. Ebselen, like glutathione peroxidase, protects in a catalytic fashion utilizing glutathione as reductant in the peroxynitrite reductase reaction. Protection by metalloporphyrins can be maintained through glutathione or ascorbate. Repair processes remove damaged products and restitute intact biomolecules.
Hepatitis B virus genotype assignment using restriction fragment length polymorphism patterns
Authors: Mizokami Masashi , Nakano Tatsunori , Orito Etsuro , Tanaka Yasuhito , Sakugawa Hiroshi , Mukaide Motokazu , Robertson Betty H.
Published: July 2002

Hepatitis B virus (HBV) is classified into genotypes A–F, which is important for clinical and etiological investigations. To establish a simple genotyping method, 68 full‐genomic sequences and 106 S gene sequences were analyzed by the molecular evolutionary method. HBV genotyping with the S gene sequence is consistent with genetic analysis using the full‐genomic sequence. After alignment of the S sequences, genotype specific regions are identified and digested by the restriction enzymes, HphI, NciI, AlwI, EarI, and NlaIV. This HBV genotyping system using restriction fragment length polymorphism (RFLP) was confirmed to be correct when the PCR products of the S gene in 23 isolates collected from various countries were digested with this method. A restriction site for EarI in genotype B was absent in spite of its presence in all the other genotypes and genotype C has no restriction site for AlwI. Only genotype E is digested with NciI, while only genotype F has a restriction site for HphI. Genotype A can be distinguished by a single restriction enzyme site for NlaIV, while genotype D digestion with this enzyme results in two products that migrates at 265 and 186 bp. This simple and accurate HBV genotyping system using RFLP is considered to be useful for research on HBV.
Isoforms of cytochrome P450 on organic nitrate‐derived nitric oxide release in human heart vessels
Authors: Minamiyama Yukiko , Takemura Shigekazu , Akiyama Takashi , Imaoka Susumu , Inoue Masayasu , Funae Yoshihiko , Okada Shigeru
Published: July 2002

Glutathione S‐transferases and the cytochrome P450 system have been proposed for the vascular biotransformation systems in the metabolic activation of organic nitrates. The present study was designed to elucidate the role of human cytochrome P450 isoforms on nitric oxide formation from organic nitrates using lymphoblast microsomes transfected with human CYP isoforms cDNA. CYP3A4‐transfected microsomes had the most effective potential of nitric oxide formation from isosorbide dinitrate. Anti‐CYP3A2 antibody (which cross‐reacts with CYP3A4) or ketoconazole (an inhibitor of the CYP3A superfamily) inhibited nitric oxide formation from isosorbide dinitrate in rat heart microsomes. Immunohistochemistry of human heart also showed intense bindings of CYP3A4 antibody in the endothelium of the endocardium and coronary vessels. These results suggest that the CYP3A4‐NADPH‐cytochrome P450 reductase system specifically participates in nitric oxide formation from isosorbide dinitrate.
Active/de‐active state transition of the mitochondrial complex I as revealed by specific sulfhydryl group labeling
Authors: Gavrikova E.V , Vinogradov A.D
Published: July 2002

The sensitivities of NADH oxidase and/or NADH‐ubiquinone reductase activities of submitochondrial particles and purified complex I towards N‐ethylmaleimide (NEM) and other SH‐reagents were studied. Only thermally de‐activated preparations [A.D. Vinogradov (1998) Biochim. Biophys. Acta 1364, 169–185] were inhibited by SH‐reagents whereas the redox‐pulsed, activated enzyme was resistant to the inhibitors. The pH profile of the pseudo‐first order inhibition rate suggested a pK
a of about 10 for the de‐activation‐dependent, NEM‐reactive sulfhydryl group. NADH‐ubiquinone reductase of activated particles treated with an excess of NEM followed by removal of the inhibitor was still capable of slow reversible active/de‐active transition. When active, NEM‐treated particles were de‐activated and further inhibited by N‐fluorescein maleimide, specific incorporation of the fluorescence label into low molecular mass polypeptide was evident. Comparison of the specific fluorescence labeling of submitochondrial particles, crude and purified complex I showed that the active/de‐active state‐dependent SH‐group is located in a 15 kDa polypeptide (most likely in the 15 kDa IP subunit of the iron‐sulfur protein‐containing fraction of complex I).
Mutations in the Ca2+ binding site of the Paracoccus denitrificans cytochrome c oxidase
Authors: Pfitzner Ute , Kirichenko Anna , Konstantinov Alexander A. , Mertens Martina , Wittershagen Axel , Kolbesen Bernd O. , Steffens Guy C.M. , Harrenga Axel , Michel Hartmut , Ludwig Bernd
Published: July 2002

Recent structure determinations suggested a new binding site for a non‐redox active metal ion in subunit I of cytochrome c oxidase both of mitochondrial and of bacterial origin. We analyzed the relevant metal composition of the bovine and the Paracoccus denitrificans enzyme and of bacterial site‐directed mutants in several residues presumably liganding this ion. Unlike the mitochondrial enzyme where a low, substoichiometric content of Ca2+ was found, the bacterial wild‐type (WT) oxidase showed a stoichiometry of one Ca per enzyme monomer. Mutants in Asp‐477 (in immediate vicinity of this site) were clearly diminished in their Ca content and the isolated mutant enzyme revealed a spectral shift in the heme a visible absorption upon Ca addition, which was reversed by Na ions. This spectral behavior, largely comparable to that of the mitochondrial enzyme, was not observed for the bacterial WT oxidase. Further structure refinement revealed a tightly bound water molecule as an additional Ca2+ ligand.
Subcellular reorganization of mitochondria producing heavy DNA in aging wheat coleoptiles
Authors: Bakeeva L.E , Kirnos M.D , Aleksandrushkina N.I , Kazimirchyuk S.B , Shorning B.Yu , Zamyatnina V.A , Yaguzhinsky L.S , Vanyushin B.F
Published: July 2002

Unusual closed membrane vesicles containing one or more mitochondria were isolated from homogenates of aging wheat coleoptiles. Very similar (or the same) bodies were shown to exist in situ in vacuoles of undividing cells in the apical part of intact senescent coleoptiles. Vesicles isolated from coleoptile homogenate free of nuclei by 10 min centrifugation at 1700×g and traditional mitochondria (sedimented at between 4300×g and 17 400×g) are similar in respiration rate, composition and content of cytochromes and sensitivity to respiration inhibitors. However, vesicles contain about 2‐fold more Ca2+ ions than free mitochondria do. The specific feature of vesicles containing mitochondria in aging coleoptiles is an intensive synthesis of heavy (ρ=1.718 g/cm3) mitochondrial DNA (H‐mtDNA). Thus, aging in plants is accompanied by an increased selective H‐mtDNA production and change in subcellular organization of mitochondria.
Farnesol‐induced generation of reactive oxygen species dependent on mitochondrial transmembrane potential hyperpolarization mediated by F0F1‐ATPase in yeast
Authors: Machida Kiyotaka , Tanaka Toshio
Published: July 2002

An isoprenoid farnesol (FOH) inhibited cellular oxygen consumption and induced mitochondrial generation of reactive oxygen species (ROS) in cells of Saccharomyces cerevisiae in correlation with hyperpolarization of the mitochondrial transmembrane potential (mtΔΨ). The FOH‐induced events were coordinately abolished with the F1‐ATPase inhibitor sodium azide as well as the F0F1‐ATPase inhibitor oligomycin, suggesting the dependence of ROS generation on mtΔΨ hyperpolarization mediated by the proton pumping function of F0F1‐ATPase as a result of ATP hydrolysis. The role of F1‐ATPase activity in mtΔΨ hyperpolarization was supported by the intracellular depletion of ATP in FOH‐treated cells and its protection with sodium azide. An indirect mechanism was suggested to exist in the regulation of F0F1‐ATPase by FOH to accelerate its ATP‐hydrolyzing activity.
Anterior pituitary response to stress : time‐related changes and adaptation
Authors: Martí Octavi , Armario Antonio
Published: July 2002

A wide array of physical and psychological stressors alter the secretion of anterior pituitary hormones. However, both the qualitative and the quantitative features of the stressors as well as its duration markedly influence the final endocrine response. In addition, among all anterior pituitary hormones, only ACTH and prolactin levels appear to reflect the intensity of the stress experienced by the animals. Although physical stressors show a somewhat specific neuroendocrine profile, the response of the pituitary‐adrenal (PA) and sympathomedulloadrenal axes are common to almost all stressors. After an initial stimulatory effect of stress, an inhibition of all anterior pituitary hormones, except ACTH, can be found provided the stressor is intense enough. The mechanisms responsible for this biphasic response to stress are likely to be located at sites above the pituitary. When the animals are repeatedly exposed to the same stressor, some behavioural and physiological consequences of stress exposure are reduced, suggesting that the animals become adapted to the stimulus. This process has been also termed habituation. Among all the pituitary hormones, only ACTH and prolactin levels are reduced as a consequence of repeated exposure to the same (homotypic) stressor, although some negative results have been reported. However, it has been recently reported that subtle changes in the characteristics of the stressors or in their regularity can greatly influence adaptation, and these factors might explain failure to find adaptation of ACTH and prolactin in some works. Habituation of ACTH and prolactin, when observed, appears to be specific for the chronically applied stressor so that the potentiality of the PA axis and prolactin to respond to a novel (heterotypic) stressor can be preserved. In the case of the PA axis, an intact or potentiated response to a novel stressor is observed in spite of presumably negative feedback exerted by daily stress‐induced glucocorticoid release and the high resting levels of glucocorticoids. This phenomenon has been termed as facilitation and can be unmasked alternating stress. Although with the exception of the PA axis, developmental aspects of anterior pituitary response to stress have been poorly studied, available data suggest that dramatic changes occur in some hormones during weaning, with some, but less profound, changes thereafter. Responsiveness to stressors appears to mature with age, but developmental patterns differ among the various anterior pituitary hormones.
Gap junction intercellular communication in gliomas is inversely related to cell motility
Authors: McDonough Wendy S. , Johansson Angie , Joffee Hylton , Giese Alf , Berens Michael E.
Published: July 2002

Gliomas are lethal because of local invasion into brain parenchyma. Glioma cells were isolated from different regions (white matter, gray matter and tumor core) of a glioma‐bearing dog brain. Individual clonal cell lines were established from each area, and characterized for growth, migration and gap junctions. The regional clonal cell lines differed in rates and preferred substrate for migration. Cell lines generated from invaded white matter showed stimulated migration on collagen and variable migration on merosin, whereas migration of cell lines derived from invaded gray matter showed the reciprocal responses: stimulation on merosin and inhibition on collagen. Gap junctional communication showed significant degrees of variation between the different clones. A direct inverse relationship between the number of cells demonstrating gap junctional communication and migration rate of cells away from multicellular spheroids was evident. Glioma cells which have a reduced capacity to connect to each other have an accelerated migration rate onto autologous, glioma‐derived matrix. These results suggest that invasive glioma cells suppress autologous cell‐to‐cell cohesion, partly evident as reduced formation of gap junctions. In addition, glioma cells were stimulated to migrate in a dose‐dependant manner in response to epidermal growth factor (EGF) coincident with the reduction of Cx43 levels and increased serine phosphorylation. We speculate that in order for glioma cells to invade locally into brain parenchyma they must first detach from neighboring cells (“let go…let's go” paradigm of invasion).
QT interval lengthening in cardiac disease relates more to left ventricular systolic dysfunction than to autonomic function
Authors: Davey Patrick
Published: July 2002

Background:

There are multiple influences on the QTc interval, including the autonomic nervous system. Which influence is the principal determinant of the variation in QTc interval between different cardiac diseases is not yet clear, though some studies have suggested that the QTc interval primarily reflects sympatho‐vagal balance. This study investigated this claim further.

Aim:

To determine if autonomic tone was the prime determinant of variation in the QTc interval between subjects with different cardiac diseases.

Methods

Subjects with different cardiac diseases were studied, QTc interval determined and correlated with three different measures of the autonomic nervous system, that of baroreflex sensitivity, catecholamine levels (epinephrine and norepinephrine) and sympatho‐vagal balance as determined by power spectrum analysis.

Results:

47 subjects were studied, comprising 17 subjects with heart failure, 14 subjects with left ventricular hypertrophy and 16 control subjects. For the group as a whole there was no relationship between QTc interval and any measure of the autonomic nervous system function, but there was a reasonable relationship between fractional shortening and QTc interval (r = 0.47, P < 0.003). For subjects with an echocardiographic fractional shortenings less than 0.35 (which correlates with an ejection fraction of < 50%), a strong relationship between fractional shortening and QTc interval remained (r = 0.57, P < 0.002), but in addition a relationship between QTc interval and catecholamine levels developed (for epinephrine: r = 0.67, P < 0.002; and for norepinephrine: r = 0.62, P < 0.005). Multiple regression analysis showed that fractional shortening and epinephrine levels were independently related to QTc interval.

Conclusion:

In subjects with a variety of cardiac diseases, the prime determinant of QTc interval is left ventricular systolic performance rather than the autonomic nervous system, though in subjects with low normal and less fractional shortenings catecholamine levels are independently related to QTc interval.
Clinical trials update: highlights of the scientific sessions of the XXIII Congress of the European Society of Cardiology — WARIS II, ESCAMI, PAFAC, RITZ‐1 and TIME
Authors: Coletta Alison P. , Cleland John G.F.
Published: July 2002

This article continues a series of reports updating recent research developments of particular interest to personnel involved in the treatment and management of patients with heart failure. This is a summary of selected presentations made at the Scientific Sessions of the XXIII Annual Congress of the European Society of Cardiology. Summaries of the following clinical studies are included: WARIS‐II, ESCAMI, PAFAC, RITZ‐I and TIME.
Perineal Trauma: Prevention and Treatment
Authors: McCandlish Rona
Published: July 2002

This article examines two aspects of routine midwifery practice: management of the perineum at the end of the second stage of labor and management and repair of perineal injury. Although some aspects of perineal management and repair have been researched and there is reliable evidence on which to base practice, there remains a considerable and urgent collaborative clinical research agenda that midwives should actively pursue.
New drug development in the United States from 1963 to 1999
Authors: DiMasi Joseph A.
Published: July 2002

Clinical Pharmacology & Therapeutics (2001) 69, 286–296; doi: 10.1067/mcp.2001.115132
Responding to the COVID-19 pandemic: The role of occupational health services in a tertiary hospital in Singapore
Authors: Hwang Jeff , Yong Elsie , Cheong Karen , Ling Zheng Jye , Goh Lay Hoon , Lim Fong Seng , Loh Victor , Bagdasarian Natasha , Somani Jyoti , Archuleta Sophia , Sng Judy , Lim See Ming
Published: October 2020

With coronavirus disease 2019 declared a Public Health Emergency of International Concern on 30 January 2020, occupational health services in a tertiary hospital in Singapore stepped up via a three-pronged approach, namely, protection of individual staff, protection of staff workforce, and prevention of nosocomial spread so as to support business continuity plans. Despite the multiple new challenges brought by the COVID-19 pandemic, the hospital’s occupational health services were able to adapt and keep all employees and patients safe with strong support from senior management and close collaboration with various departments.
Generation of fully functional fluorescent fusion proteins to gain insights into ABCC6 biology
Authors: Szeri Flora , Niaziorimi Fatemeh , Donnelly Sylvia , Orndorff Joseph , van de Wetering Koen
Published: October 2020

ABCC6 mediates release of ATP from hepatocytes into the blood. Extracellularly, ATP is converted into the mineralization inhibitor pyrophosphate. Consequently, inactivating mutations in ABCC6 give low plasma pyrophosphate and underlie the ectopic mineralization disorder pseudoxanthoma elasticum. How ABCC6 mediates cellular ATP release is still unknown. Fluorescent ABCC6 fusion proteins would allow mechanistic studies, but fluorophores attached to the ABCC6 N‐ or C‐terminus result in intracellular retention and degradation. Here we describe that intramolecular introduction of fluorophores yields fully functional ABCC6 fusion proteins. A corresponding ABCC6 variant in which the catalytic glutamate of the second nucleotide binding domain was mutated, correctly routed to the plasma membrane but was inactive. Finally, N‐terminal His10 or FLAG tags did not affect activity of the fusion proteins, allowing their purification for biochemical characterization.
Vaccination against HPV: boosting coverage and tackling misinformation
Authors: Bigaard Janne , Franceschi Silvia
Published: October 2020

The availability of human papillomavirus (HPV) vaccines and screening tests has raised the possibility of globally eliminating cervical cancer, which is caused by HPV. Cervical cancer is a very common malignancy worldwide, especially among deprived women. High vaccination coverage is key to the containment and eventual elimination of the infection. Public HPV vaccination programmes in Italy and Denmark were swiftly established and are among the most successful worldwide. Still, in both countries, it has been challenging to achieve and maintain the recommended coverage of > 80% in girls. In a well‐studied Italian region, vaccination coverage in girls at age 15 years (World Health Organization's gold standard) reached 76% in 2015 but decreased to 69% in 2018, likely due to work overload in public immunization centres. In Denmark, doubts about safety and efficacy of the HPV vaccine generated a decline in coverage among girls age 12–17, from 80% in 2013 down to 37% in 2015, when remedial actions made it rise again. Insights from these two countries are shared to illustrate the importance of monitoring coverage in a digital vaccine registry and promptly reacting to misinformation about vaccination.
Community Mall for all generations
Authors: Mizukami Tetsuya
Published: October 2020

The renovation of a 28‐year‐old supermarket is planned as one of the main facilities of the Aburatsu Shopping Street Regeneration Project in Nichinan, Miyazaki Prefecture. The building, with its steel rigid frame structure, is partially reduced and divided into two buildings to create an outdoor space, thereby creating a complex facility that contains a meeting area and six eating and drinking establishments.
Detectability of fiducials’ positions for real‐time target tracking system equipping with a standard linac for multiple fiducial markers
Authors: Ono Shunsuke , Ueda Yoshihiro , Ohira Shingo , Isono Masaru , Sumida Iori , Inui Shoki , Morimoto Masahiro , Ashida Reiko , Miyazaki Masayoshi , Ogawa Kazuhiko , Teshima Teruki
Published: October 2020

Purpose

To investigate the detectability of fiducial markers’ positions for real‐time target tracking system equipping with a standard linac. The hypothesis is that the detectability depends on the type of fiducial marker and the gantry angle of acquired triggered images.

Methods

Three types of ball fiducials and four slim fiducials with lengths of 3 and 5 mm were prepared for this study. Triggered images with three similar fiducials were acquired at every 10° during the conformal arc irradiation to detect the target position. Although only one type of arrangement was prepared for the ball fiducials, a three‐type arrangement was prepared for the slim fiducials, such as parallel, orthogonal, and oblique with 45° to the gantry‐couch direction. To measure the detectability of the real‐time target tracking system for each fiducial and arrangement, detected marker positions were compared with expected marker positions at every angle of acquired triggered images.

Results

For the ball‐type fiducial, the maximum difference between the detected marker positions and expected marker positions was 0.3 mm in all directions. For the slim fiducial arranged parallel and oblique with 45°, the maximum difference was 0.4 mm in all directions. When each slim fiducial was arranged orthogonal to the gantry‐couch direction, the maximum difference was 1.5 mm for the length of 3 mm, and 3.2 mm for the length of 5 mm.

Conclusions

The detectability of fiducial markers’ positions for the real‐time target tracking system equipping with a standard linac depends on the form and insertion angles of the fiducials.
Novel NUDT2 variant causes intellectual disability and polyneuropathy
Authors: Diaz Frank , Khosa Shaweta , Niyazov Dmitriy , Lee Hane , Person Richard , Morrow Michelle M. , Signer Rebecca , Dorrani Naghmeh , Zheng Allison , Herzog Matthew , Freundlich Robert ,   Undiagnosed Diseases Network, Birath J. Brandon , Cervantes‐Manzo Yurivia , Martinez‐Agosto Julian A. , Palmer Christina , Nelson Stanley F. , Fogel Brent L. , Mishra Shri K.
Published: October 2020

Exome or genome sequencing was performed to identify the genetic etiology for the clinical presentation of global developmental delay, intellectual disability, and sensorimotor neuropathy with associated distal weakness in two unrelated families. A homozygous frameshift variant c.186delA (p.A63Qfs*3) in the NUDT2 gene was identified in cases 1 and 2 from one family and a third case from another family. Variants in NUDT2 were previously shown to cause intellectual disability, but here we expand the phenotype by demonstrating its association with distal upper and lower extremity weakness due to a sensorimotor polyneuropathy with demyelinating and/or axonal features.
Management of Rheumatic Diseases During the COVID‐19 Pandemic: A National Veterans Affairs Survey of Rheumatologists
Authors: Singh Jasvinder A. , Richards John S. , Chang Elizabeth , Joseph Amy , Ng Bernard
Published: October 2020

Objective

To assess the experience, views, and opinions of rheumatology providers at Veterans Affairs (VA) facilities about rheumatic disease health care issues during the COVID‐19 pandemic.

Methods

We performed an anonymized cross‐sectional survey, conducted from April 16 to May 18, 2020, of VA rheumatology providers. We assessed provider perspectives on COVID‐19 issues and resilience.

Results

Of the 153 eligible VA rheumatologists, 103 (67%) completed the survey. A significant proportion of providers reported a ≥50% increase related to COVID‐19 in visits by telephone (53%), video‐based VA video connect (VVC; 44%), and clinical video telehealth with a facilitator (29%). A majority of the responders were somewhat or very comfortable with technology for providing health care to established patients during the COVID‐19 pandemic using telephone (87%), VVC (64%), and in‐person visits (54%). A smaller proportion were comfortable with technology providing health care to new patients. At least 65% of rheumatologists considered telephone visits appropriate for established patients with gout, osteoporosis, polymyalgia rheumatica, stable rheumatoid arthritis, stable spondyloarthritis, or osteoarthritis; 32% reported a rheumatology medication shortage. Adjusted for age, sex, and ethnicity, high provider resilience was associated with significantly higher odds ratios (ORs) of comfort with technology for telephone (OR 3.1 [95% confidence interval (95% CI) 1.1–9.7]) and VVC visits for new patients (OR 4.7 [95% CI 1.4–15.7]).

Conclusion

A better understanding of COVID‐19 rheumatic disease health care issues using a health‐system approach can better inform providers, improve provider satisfaction, and have positive effects on the care of veterans with rheumatic disease.
Thermoelectric, Magnetic, and Mechanical Characteristics of Antiferromagnetic Manganese Telluride Reinforced with Graphene Nanoplates
Authors: Hooshmand Zaferani Sadeq , Ghomashchi Reza , Vashaee Daryoosh
Published: October 2020

Mechanical and thermal stability are the two challenging aspects of thermoelectric compounds and modules. Microcrack formation during material synthesis and mechanical failure under thermo‐mechanical loading is commonly observed in thermoelectric materials made from brittle semiconductors. Herein, the results of graphene‐nanoplates (GNPs) reinforcement on the mechanical and thermoelectric properties of MnTe compound are reported. The binary antiferromagnetic MnTe shown promising thermoelectric characteristics due to the paramagnon–hole drag above the Néel temperature. In this study, different bulk MnTe samples are synthesized with the addition of GNPs in a small quantity (0.25–1 wt%) by powder metallurgy and spark plasma sintering. The thermoelectric factors, magnetic behavior, microstructure, and mechanical properties of the samples are evaluated and analyzed. Nearly 33% improvement is observed in the fracture toughness of MnTe reinforced with 0.25 wt% GNPs compared to the pristine structure. The Néel temperature remains approximately unaffected with the GNP inclusion; however, the low‐temperature ferromagnetic phase impurity is significantly suppressed. The thermal conductivity and power factor decrease almost equally by ≈34% at 600 K; hence, the thermoelectric figure‐of‐merit is not affected by GNP reinforcement in the optimized sample.
An Assistive Magnetic Skin System: Enabling Technology for Quadriplegics
Authors: Almansouri Abdullah S. , Upadhyaya Lakshmeesha , Nunes Suzana P. , Salama Khaled N. , Kosel Jurgen
Published: October 2020

People with quadriplegia no longer have control over their legs, neither the hands and cannot continue living their life independently. On top of that, severely injured quadriplegics (i.e., C1 and C2 injuries) suffer from speaking difficulties and minimal head and neck movements. With the advancement in wearable artificial skins and the Internet‐of‐Things, realizing comfortable and practical solutions for quadriplegics is more tangible than ever. Herein, a comprehensive assistive magnetic skin system is presented that allows quadriplegics, including the severely injured ones, to move around individually and control their surroundings with ease. The system tracks facial expressions by tracking the movement of magnetic tattoos attached to the face, using magnetic field sensors incorporated into eyeglasses. The magnetic tattoos are made of highly flexible, stretchable, breathable, and biocompatible magnetic skins. In combination with smart‐glasses, smart‐wheelchair, and smart‐gadgets, the users can move around and control their environment with their facial expressions. The system is also designed to allow quadriplegics to perform outdoor activities effortlessly. It supports line‐of‐sight communication and does not require pretethering to the smart‐gadgets, unlike the existing solutions. Thus, enabling the user to walk on pathways, activate pedestrian lights, control public elevators, and perform various outdoor activities independently.
Flexible and Stretchable Fiber‐Shaped Triboelectric Nanogenerators for Biomechanical Monitoring and Human‐Interactive Sensing
Authors: Ning Chuan , Dong Kai , Cheng Renwei , Yi Jia , Ye Cuiying , Peng Xiao , Sheng Feifan , Jiang Yang , Wang Zhong Lin
Published: October 2020

Wearable, flexible, and even stretchable tactile sensors, such as various types of electronic skin, have attracted extensive attention, which can adapt to complex and irregular surfaces, maximize the matching of wearable devices, and conformally apply onto human organs. However, it is a great challenge to simultaneously achieve breathability, permeability, and comfortability for their development. Herein, mitigating the problem by miniaturizing and integrating the sensors is tried. Highly flexible and stretchable coaxial structure fiber‐shaped triboelectric nanogenerators (F‐TENGs) with a diameter of 0.63 mm are created by orderly depositing conductive material of silver nanowires/carbon nanotubes and encapsulated polydimethylsiloxane onto the stretchable spandex fiber. As a self‐powered multifunctional sensor, the resulting composite fiber can convert mechanical stimuli into electrical signals without affecting the normal human body. Moreover, the F‐TENGs can be easily integrated into traditional textiles to form tactile sensor arrays. Through the tactile sensor arrays, the real‐time tactile trajectory and pressure distribution can be precisely mapped. This work may provide a new method to fabricate fiber‐based pressure sensors with high sensitivity and stretchability, which have great application prospects in personal healthcare monitoring and human–machine interactions.
Self‐Assemblies of Fe3O4 Nanocrystals: Toward Nanoscale Precision of Photothermal Effects in the Tumor Microenvironment
Authors: Nicolas‐Boluda Alba , Yang Zhijie , Guilbert Thomas , Fouassier Laura , Carn Florent , Gazeau Florence , Pileni Marie Paule
Published: October 2020

Fe3O4 nanocrystals are self‐assembled into two different conformations: colloidosome and supraball that confer them with distinct properties determining their photo‐induced heating capacities. These self‐assemblies are assessed for photothermal therapy, an adjuvant strategy for tumor therapy. The tumor microenvironment is a heterogeneous ecosystem including immune cells and the extracellular matrix. The interactions between photothermal therapy agents and the different components of the tumor microenvironment determine the outcome of this therapy. In this study, the fate of both colloidosomes and supraballs within the tumor microenvironment in comparison to their Fe3O4 nanocrystal building blocks is revealed. This study highlights how these two hybrid self‐assemblies target different compartments of the tumor microenvironment and trigger local photothermal damages that are inaccessible for isolated nanocrystals and not predicted by global temperature measurements.
Additive Manufacturing of Material Scaffolds for Bone Regeneration: Toward Application in the Clinics
Authors: Garot Charlotte , Bettega Georges , Picart Catherine
Published: October 2020

Additive manufacturing (AM) allows the fabrication of customized bone scaffolds in terms of shape, pore size, material type, and mechanical properties. Combined with the possibility to obtain a precise 3D image of the bone defects using computed tomography or magnetic resonance imaging, it is now possible to manufacture implants for patient‐specific bone regeneration. This paper reviews the state‐of‐the‐art of the different materials and AM techniques used for the fabrication of 3D‐printed scaffolds in the field of bone tissue engineering. Their advantages and drawbacks are highlighted. For materials, specific criteria, are extracted from a literature study: biomimetism to native bone, mechanical properties, biodegradability, ability to be imaged (implantation and follow‐up period), histological performances, and sterilization process. AM techniques can be classified in three major categories: extrusion‐based, powder‐based, and vat photopolymerization. Their price, ease of use, and space requirement are analyzed. Different combinations of materials/AM techniques appear to be the most relevant depending on the targeted clinical applications (implantation site, presence of mechanical constraints, temporary or permanent implant). Finally, some barriers impeding the translation to human clinics are identified, notably the sterilization process.
Proton‐Conductive Melanin‐Like Fibers through Enzymatic Oxidation of a Self‐Assembling Peptide
Authors: Reddy Samala Murali Mohan , Raßlenberg Eileen , Sloan‐Dennison Sian , Hesketh Travis , Silberbush Ohad , Tuttle Tell , Smith Ewen , Graham Duncan , Faulds Karen , Ulijn Rein V. , Ashkenasy Nurit , Lampel Ayala
Published: October 2020

Melanin pigments have various properties that are of technological interest including photo‐ and radiation protection, rich coloration, and electronic functions. Nevertheless, laboratory‐based synthesis of melanin and melanin‐like materials with morphologies and chemical structures that are specifically optimized for these applications, is currently not possible. Here, melanin‐like materials that are produced by enzymatic oxidation of a supramolecular tripeptide structures that are rich in tyrosine and have a 1D morphology are demonstrated, that are retained during the oxidation process while conducting tracks form through oxidative tyrosine crosslinking. Specifically, a minimalistic self‐assembling peptide, Lys–Tyr–Tyr (KYY) with strong propensity to form supramolecular fibers, is utilized. Analysis by Raman spectroscopy shows that the tyrosines are pre‐organized inside these fibers and, upon enzymatic oxidation, result in connected catechols. These form 1D conducting tracks along the length of the fiber, which gives rise to a level of internal disorder, but retention of the fiber morphology. This results in highly conductive structures demonstrated to be dominated by proton conduction. This work demonstrates the ability to control oxidation but retain a well‐defined fibrous morphology that does not have a known equivalent in biology, and demonstrate exceptional conductivity that is enhanced by enzymatic oxidation.
Reaction of Pyridine‐N‐Oxides with Tertiary sp2‐N‐Nucleophiles: An Efficient Synthesis of Precursors for N‐(Pyrid‐2‐yl)‐Substituted N‐Heterocyclic Carbenes
Authors: Bugaenko Dmitry I. , Yurovskaya Marina A. , Karchava Alexander V.
Published: October 2020

N‐(Pyrid‐2‐yl)‐substituted azolium and pyridinium salts, precursors for hybrid NHC‐containing ligands, were obtained with excellent regioselectivity, employing a deoxygenative CH‐functionalization of pyridine‐N‐oxides with substituted imidazoles, thiazoles, and pyridine. Unlike the traditional SNAr‐based methods, this approach provides high yields for substrates bearing substituents of different electronic nature. The utility of azolium and pyridinium salts thus prepared was also highlighted by the synthesis of pyridyl‐substituted imidazolyl‐2‐thione, benzodiazepine as well as 2‐aminopyridines.

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Digital Transformation of the Agricultural Sector: Pathways, Drivers and Policy Implications
Authors: Khanna Madhu
Published: October 2020

The digital transformation of agriculture is enabling the collection of vast amounts of geo‐referenced information about growing conditions within the field and facilitating the automated implementation of spatially varying input applications. This has the potential to increase production efficiency, reduce overapplication of inputs, lower input waste and pollution, and improve farm profitability. We discuss the pathways by which digital agricultural technologies have the potential to affect crop management, and current trends and patterns of adoption of digital technologies. We provide insights from the technology adoption literature on the factors that can be expected to influence the adoption of emerging digital technologies and the findings from the empirical literature on the determinants of adoption of these types of technologies. We conclude with a discussion of the design of policy incentives to induce the adoption of digital technologies cost‐effectively, the challenges in implementing such policies and the opportunity that digital technologies offer for addressing these challenges.
Secular changes in the anthropometric and motor characteristics of Polish male university students between 2000 and 2018
Authors: Podstawski Robert , Żurek Piotr
Published: October 2020

Objectives

To determine changes in the anthropometric and motor characteristics of young males during the first 20 years of the 21st century in Poland.

Materials and Methods

The study was conducted in 2000–2018 on 2691 randomly selected male university students aged 19÷25 (20.0 ± 1.1 years). The participants' body mass and height were measured, and the students participated in 13 motor ability tests. The analyzed traits were evaluated by testing the fit of linear and curvilinear functions to empirical data.

Results

The students evaluated in 2018 were 1.7 cm taller than their peers tested in 2000. Body mass and BMI values continued to decrease between 2000 and 2006 (by 0.46 kg and 0.15 kg/m2 per year on average), whereas a steady and significant increase in both parameters was observed between 2006 and 2018 (by 0.45 kg and 0.12 kg/m2 per year on average). The results of motor tests were strongly correlated with body mass and BMI, and they continued to improve until 2006, after which a steady decline was observed up to 2018 when the students scored lowest in motor tests.

Conclusions

The trend of increasing body height has been maintained in the studied population, but unlike body mass and BMI, the rate of increase in body height was lower than in the preceding years. A decrease in body mass and BMI is correlated with an improvement in motor tests, whereas an increase in the above parameters leads to a significant decline in all evaluated motor abilities.