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The potential clinical applications of the powerful in vitro-transcribed (IVT)-mRNAs, to restore defective protein functions, strongly depend on their successful intracellular delivery and transient translation through the development of safe and efficient delivery platforms. In this study, an innovative (international patent-pending) methodology was developed, combining the IVT-mRNAs with the protein transduction domain (PTD) technology, as an efficient delivery platform. Based on the PTD technology, which enables the intracellular delivery of various cargoes intracellularly, successful conjugation of a PTD to the IVT-mRNAs was achieved and evaluated by band-shift assay and NMR spectroscopy. In addition, the PTD-IVT-mRNAs were applied and evaluated in two protein-disease models, including the mitochondrial disorder fatal infantile cardioencephalomyopathy and cytochrome c oxidase (COX) deficiency (attributed to SCO2 gene mutations) and β-thalassemia. The PTD-IVT-mRNA of SCO2 was successfully transduced and translated to the corresponding Sco2 protein inside the primary fibroblasts of a SCO2/COX-deficient patient, whereas the PTD-IVT-mRNA of β-globin was transduced and translated in bone marrow cells, derived from three β-thalassemic patients. The transducibility and the structural stability of the PDT-IVT-mRNAs, in both cases, were confirmed at the RNA and protein levels. We propose that our novel delivery platform could be clinically applicable as a protein therapy for metabolic/genetic disorders., Graphical abstract, The corresponding author and colleagues successfully developed a universal delivery platform of therapeutic in vitro-transcribed (IVT)-mRNAs conjugated to a selected PTD through an innovative patent-pending methodology. PTD-IVT-mRNAs exhibited significant stability, intracellular transduction, and protein expression efficiency and were applied in patients’ cells of two different monogenic/metabolic disorders, revealing powerful future perspectives.

Human ocular dirofilariasis is a zoonotic disease caused by several species of filarioid helminths of the genus Dirofilaria. The aim of this study was to further re-examine five preserved specimens previously isolated from patients with ocular dirofilariasis by molecular means. Four of the examined helminths had been stored in unbuffered formaldehyde solution for more than eight years; whereas, the fifth helminth was stored in ethanol buffer for more than two years. For the four specimens stored in formaldehyde, different methods of DNA recovery and amplification were applied and investigated for their efficiency in DNA extraction and PCR amplification. However, the DNA extraction and PCR amplification were successful only for the ethanol-preserved helminth. The genetic identification of the ethanol-preserved specimen as Dirofilaria repens (D. repens) and its phylogenetic position based on the analysis of mitochondrial 12S ribosomal RNA, nuclear 18S ribosomal RNA and mitochondrial cytochrome c oxidase subunit one sequences are reported in the present paper. To our knowledge, these are the only deposited sequences related to D. repens that have been isolated in Greece. Routine laboratory diagnosis is based on phenotypic characteristics of the helminthic parasites, but more accurate diagnosis requires molecular identification. Although the specimens preserved in formalin buffers may be a potential source for the enrichment of parasite genome databases, the DNA recovery of such samples is a challenging task.

Recent research on the topic of pile base resistance has demonstrated that the guidelines provided by existing design manuals and codes are rather conservative in the case of clayey soils u...

Organometallic rhenium complexes have recently been considered in the development of novel antitumor agents due to their suitable properties. A series of rhenium(I) tricarbonyl complexes was synthesized with the quinolone antimicrobial agents enrofloxacin (Herx) and levofloxacin (Hlfx) and solvent (e.g., methanol), imidazole (im) or pyridine (py) as co-ligands. The complexes were characterized by spectroscopic methods. The interaction of the rhenium complexes with bovine serum albumin was investigated by fluorescence emission spectroscopy and the corresponding binding constants were determined. The binding of the rhenium complexes to calf-thymus DNA was monitored by UV-Vis spectroscopy, viscosity measurements and competitive studies with ethidium bromide. These studies indicated that intercalation is the most possible mode of action and the corresponding DNA-binding constants of the complexes were calculated. The cytotoxicity of the Re-complexes against human K-562 erythroleukemia cells was found to be moderate to high. These preliminary results are promising and warrant the design of new Re-complexes with improved properties in future studies.

Introduction: Nowadays, nanotechnology and genomics contribute knowledge and innovative practices, enabling pharmacological interventions as well as theraputic decisions to be applicable on...

Background α-Thalassemia, a congenital hemoglobinopathy, is characterized by deficiency and/or reduced levels of α-globin chains in serious forms of α-thalassemia (HbH disease/Hb Bart’s). This research work deals with a Protein Replacement Therapy approach in order to manage α-thalassemia manifestations, caused by the excess of β-globin chain into HbH RBCs. The main goal was to produce the recombinant human α-globin chain in fusion with TAT, a Protein Transduction Domain, to ex vivo deliver it into HbH patients RBCs, to replace the endogenous missing α-globin chain. Results Cloning of the α-globin coding sequence, fused to the nucleotide sequence of TAT peptide was conducted and the human recombinant fusion proteins, 10xHis-XaSITE-α-globin-HA and 10xHis-XaSITE-TAT-α-globin-HA were produced. The ability of human recombinant 10xHis-XaSITE-α-globin-HA to interact in vitro with the previously produced 10xHis-XaSITE-TAT-β-globin-HA and form α-/β-globin heterodimers, was assessed and confirmed by size exclusion chromatography. The recombinant 10xHis-XaSITE-TAT-α-globin-HA was successfully delivered into human proerythroid K-562 cells, during the preliminary transduction evaluation experiments. Finally, the recombinant, TAT-fused α-globin was successfully transduced into RBCs, derived from HbH patients and reduced the formation of HbH-Inclusion Bodies, known to contain harmful β4-globin chain tetramers. Conclusions Our data confirm the successful ex vivo transduction of recombinant α-globin chains in HbH RBCs to replace the missing a-globin chain and reduce the HbH-inclusion bodies, seen in α-thalassemias. These findings broaden the possibility of applying a Protein Replacement Therapy approach to module sever forms of α-thalassemia, using recombinant α-globin chains, through PTD technology.

This longitudinal study aimed to investigate the predictive factors of suicidal ideation during the second lockdown in Greece. The respondents presented a 4.32% suicidal ideation in the second lockdown, which did not differ significantly to the initial 4.81%. Anxiety, depression, suicidal ideation during the first lockdown and living with a person with frail health and vulnerable for COVID-19 severe infection emerged as significant risk factors for suicidal ideation during the second lockdown, after controlling for gender, age, and mental health history. Depression was found as the only significant prognostic factor for suicidal ideation incidence of the second lockdown. © 2021

Journal of physics / B 55(14), 144001 - (2021). doi:10.1088/1361-6455/ac6b6b, We demonstrate the application of event-driven Timepix3-based detectors in combination with a double-sided velocity-map-imaging spectrometer to record the full 3D momentum of charged particles at the free-electron-laser facility FLASH. We measured the XUV induced fragmentation of $\text{N}_2$ using 250 kHz FLASH bursts with sub-pixel spatial resolution and up to 1.7 ns temporal resolution for photoelectrons. To further demonstrate the capabilities of this camera at even higher repetition rates we measured single-shot images of He($1s$) photoelectrons for bursts with a repetition rate of 1 MHz. Overall, with the Timepix3 camera we overcome limitations of standard-camera technology for advanced-imaging experiments with requirements on high event-rates and high spatio-temporal resolution., Published by IOP Publ., Bristol

Review of scientific instruments 92(12), 123004 (2021). doi:10.1063/5.0063225, We present the design, integration, and operation of the novel vacuum ultraviolet (VUV) beamline installed at the free-electron laser (FEL) FLASH. The VUV source is based on high-order harmonic generation (HHG) in gas and is driven by an optical laser system synchronized with the timing structure of the FEL. Ultrashort pulses in the spectral range from 10 to 40 eV are coupled with the FEL in the beamline FL26, which features a reaction microscope (REMI) permanent endstation for time-resolved studies of ultrafast dynamics in atomic and molecular targets. The connection of the high-pressure gas HHG source to the ultra-high vacuum FEL beamline requires a compact and reliable system, able to encounter the challenging vacuum requirements and coupling conditions. First commissioning results show the successful operation of the beamline, reaching a VUV focused beam size of about 20 ��m at the REMI endstation. Proof-of-principle photo-electron momentum measurements in argon indicate the source capabilities for future two-color pump���probe experiments., Published by American Institute of Physics, [S.l.]

The rapid progress achieved in the development of many biopharmaceuticals had a tremendous impact on the therapy of many metabolic/genetic disorders. This type of fruitful approach, called protein replacement therapy (PRT), aimed to either replace the deficient or malfunctional protein in human tissues that act either in plasma membrane or via a specific cell surface receptor. However, there are also many metabolic/genetic disorders attributed to either deficient or malfunctional proteins acting intracellularly. The recent developments of Protein Transduction Domain (PTD) technology offer new opportunities by allowing the intracellular delivery of recombinant proteins of a given therapeutic interest into different subcellular sites and organelles, such as mitochondria and other entities. Towards this pathway, we applied successfully PTD Technology as a protein therapeutic approach, in vitro, in SCO2 deficient primary fibroblasts, derived from patient with mutations in human SCO2 gene, responsible for fatal, infantile cardioencephalomyopathy and cytochrome c oxidase deficiency. In this work, we radiolabeled the recombinant TAT-L-Sco2 fusion protein with technetium-99 m to assess its in vivo biodistribution and fate, by increasing the sensitivity of detection of even low levels of the transduced recombinant protein. The biodistribution pattern of [99mTc]Tc-TAT-L-Sco2 in mice demonstrated fast blood clearance, significant hepatobiliary and renal clearance. In addition, western blot analysis detected the recombinant TAT-L-Sco2 protein in the isolated mitochondria of several mouse tissues, including heart, muscle and brain. These results pave the way to further consider this PTD-mediated Protein Therapy Approach as a potentially alternative treatment of genetic/metabolic disorders., Graphical abstract Unlabelled Image, Highlights • Radiolabeling of human recombinant mitochondrial TAT-L-Sco2 fusion protein with 99mTc for the first time. • [99mTc]Tc-TAT-L-Sco2 can be successfully transduced into the mitochondria of peripheral tissues upon injection into animals. • Protein Replacement Therapy, through PTD technology, can be a potential therapeutic approach for mitochondrial disorders.

The link between active learning and the development of graduate attributes has been cogently advocated in pedagogical research. Despite the extensive adoption of fieldwork in environmental and social sciences, enigmatically, there are no reported applications of fieldwork in construction and related engineering disciplines. This paper employs a case study approach to address this gap and exemplify the application of fieldwork in a postgraduate construction degree programme. It presents a robust design framework for assessed residential fieldwork that aids the scaffolding of a holistic set of graduate attributes. The framework promotes undertaking field-based experiential learning in international destinations in order to support the development of adaptable leadership and graduate future readiness for the fourth industrial revolution. After discussing its theoretical underpinnings, the paper presents the context for the case study, the design framework and stages of the fieldwork and concludes with an evaluation of its implementation, limitations and implications for practice.

Giardia and Cryptosporidium are recognized as leading causes of waterborne and foodborne diarrhoeal disease with worldwide distribution. The study aimed to determine the protozoan contamination of various foods of plant origin. A total of 72 samples from 27 different varieties of fresh vegetables and fruits were collected from supermarkets and open markets in North-Western Greece and were examined using conventional diagnostic methods. Two out of 72 (2.8%) samples were found positive for Cryptosporidium oocysts, while no sample was found to be positive for Giardia cysts. The results show the presence of protozoan contamination in foods of plant origin, which may constitute a potential health hazard.

BackgroundMAS is a severe, life-threatening complication of rheumatic diseases that occurs most frequently in patients with sJIA. The mainstay of treatment for MAS is high dose glucocorticoids (GCs); however, GCs do not provide adequate control in all patients. Additional treatments are used without a standardized approach; however, morbidity and mortality remain high. Data from animal models of MAS and from observational studies in patients suggest that overproduction of IFNγ is a driver of the hyperinflammation observed in MAS; neutralization of IFNγ has been shown to revert the signs and symptoms of MAS in murine models, and high IFNγ levels are strongly correlated with laboratory parameters of disease severity in patients.ObjectivesTo assess the efficacy and safety of intravenous (IV) infusions of emapalumab, a fully human, anti-IFNγ monoclonal antibody, in patients with MAS in sJIA.MethodsOpen-label, single-arm, phase 2 study (NCT03311854) that included patients with MAS (2016 ACR/EULAR criteria) in sJIA who had failed high-dose IV GCs and other treatments. Emapalumab was initiated at a dose of 6 mg/kg on Day 0 and continued at 3 mg/kg every 3 days until Day 15, and then twice weekly until Day 28 to ensure rapid and complete IFNγ neutralization after initiating treatment. As per protocol, 10 infusions were planned over the 4 weeks; however, treatment could be shortened if MAS remission was achieved earlier, or extended if required to achieve remission. Complete response (CR) was defined as resolution of clinical signs and symptoms of MAS according to the investigator, and normalization of laboratory parameters relevant to MAS. All patients were followed up for 4 weeks after the last infusion of emapalumab and offered to enter a 1-year, follow-up study (NCT02069899).ResultsFourteen patients (10 females) were enrolled (11 in Europe, 3 in the USA). Several patients had previously received cyclosporine A and/or anakinra, in addition to high-dose GCs. Six patients received emapalumab until Day 28. Seven patients discontinued emapalumab early due to MAS remission (as per investigator’s assessment); one patient received treatment up to Day 38. Emapalumab treatment rapidly neutralized IFNγ, as documented by CXCL9 levels. A CR was achieved by 13/14 patients during the study. One patient stopped emapalumab after 3 doses because of achievement of MAS remission as per investigator’s assessment, but lactate dehydrogenase levels remained >1.5× upper limit of normal. At Week 8, 11/14 patients had a CR; 2 achieved a CR during the study, but not at Week 8, because of a single laboratory parameter abnormality in each patient. Overall, all measured laboratory parameters related to MAS activity rapidly improved with emapalumab treatment. GCs were tapered in all patients by Week 8 (≥50% reduction, n=12; GC dose ≤1 mg/kg/day, n=8). Administration of anakinra for the treatment of underlying sJIA was maintained/introduced during the study, as required. No patients discontinued treatment for safety reasons. One treatment-related serious adverse event was reported (cytomegalovirus reactivation that resolved with antiviral treatment). All patients entered the long-term, follow-up study and were alive at last visit.ConclusionEmapalumab administration led to rapid IFNγ neutralization, was efficacious in controlling MAS in all patients, and was well tolerated with a favorable safety profile. These results support the pathogenic role of IFNγ in MAS in sJIA and the therapeutic value of IFNγ neutralization in MAS patients who have failed high-dose GCs.Disclosure of InterestsFabrizio De Benedetti Consultant of: Sobi, AbbVie, Pfizer, Roche, Sanofi, Novartis, Novimmune, Grant/research support from: Sobi, AbbVie, Pfizer, Roche, Sanofi, Novartis, Novimmune, Alexei Grom Consultant of: Novartis, AB2 Bio, Paul Brogan Consultant of: Sobi, Novartis, Roche, UCB, Claudia Bracaglia: None declared, Manuela Pardeo: None declared, Giulia Marucci: None declared, Despina Eleftheriou Speakers bureau: Sobi, Charalampia Papadopoulou Speakers bureau: Sobi, Pierre Quartier Speakers bureau: Sobi, AbbVie, Chugai-Roche, Lilly, Novartis, Pfizer, Consultant of: Sobi, AbbVie, Chugai-Roche, Lilly, Novartis, Pfizer, Jordi Anton Consultant of: Sobi, Novartis, Roche, Pfizer, AbbVie, GSK, Rikke Frederiksen Employee of: Sobi, Veronica Asnaghi Employee of: Sobi, Cristina de Min Consultant of: Sobi

Protein replacement therapy (PRT) has been applied to treat severe monogenetic/metabolic disorders characterized by a protein deficiency. In disorders where an intracellular protein is missing, PRT is not easily feasible due to the inability of proteins to cross the cell membrane. Instead, gene therapy has been applied, although still with limited success. β-Thalassemias are severe congenital hemoglobinopathies, characterized by deficiency or reduced production of the adult β-globin chain. The resulting imbalance of α-/β-globin chains of adult hemoglobin (α2β2) leads to precipitation of unpaired α-globin chains and, eventually, to defective erythropoiesis. Since protein transduction domain (PTD) technology has emerged as a promising therapeutic approach, we produced a human recombinant β-globin chain in fusion with the TAT peptide and successfully transduced it into human proerythroid K-562 cells, deficient in mature β-globin chain. Notably, the produced human recombinant β-globin chain without the TAT peptide, used as internal negative control, failed to be transduced into K-562 cells under similar conditions. In silico studies complemented by SDS-PAGE, Western blotting, co-immunoprecipitation and LC-MS/MS analysis indicated that the transduced recombinant fusion TAT-β-globin protein interacts with the endogenous native α-like globins to form hemoglobin α2β2-like tetramers to a limited extent. Our findings provide evidence that recombinant TAT-β-globin is transmissible into proerythroid K-562 cells and can be potentially considered as an alternative protein therapeutic approach for β-thalassemias.

Commercial milk formulae hold a major role in the diet of many infants in developed countries and assessing their mineral content is important in order to ensure compliance with the manufacturer claims and with regulation guidelines. In this study, the concentrations of K, Ca, Fe, Zn, Br, Rb and Sr were determined by Energy-Dispersive X-Ray Fluorescence (EDXRF) spectrometry in 28 infant milk formulae widely distributed in the Greek market. The concentration ranges (in mg kg−1) measured for K (2180–7600), Ca (3240–7480), Fe (29–98) and Zn (38–84) were within the limits recommended by the European legislation, with only a few exceptions. Although measured values were generally consistent with those labelled by the manufacturers, discrepancies over 50% to 100% were observed in some cases and were analyzed by multivariate statistical methods. The findings indicate the need for close monitoring to guarantee good manufacturing practices and constant quality of infant formulae. The daily intakes of minerals from consumption of different types of milk formulae were also estimated and the overall ranges were (0.23–0.85) g d−1 for K, (348–736) mg d−1 for Ca, (3.2–10.5) mg d−1 for Fe and (4.1–9.0) mg d−1 for Zn. These values conform to currently recommended safety limits.

Overlapping jet-grout columns are often used to form a grout slab at the bottom of deep excavations to support the retaining structures. Columnar geometrical imperfections, spatially varying strength and stiffness and the post-peak behaviour of the grout, render the simulation of the jet-grout column slab a very complex issue. In practice, these aspects are not taken into account and simplified models are commonly used, in conjunction with the application of a strength reduction factor. With the aim to improve the existing design process and extend its applicability to serviceability limit state analysis, a consistent design procedure is presented.

Background Cancer is one of the leading causes of death worldwide. Over the years, a number of conventional cytotoxic approaches for neoplastic diseases has been developed. However, due to their limited effectiveness in accordance with the heterogeneity of cancer cells, there is a constant search for therapeutic approaches with improved outcome, such as immunotherapy that utilizes and enhances the normal capacity of the patient's immune system. Methods Chimeric Antigen Receptor (CAR) T-cell therapy involves genetic modification of patient's autologous T-cells to express a CAR specific for a tumor antigen, following by ex vivo cell expansion and re-infusion back to the patient. CARs are fusion proteins of a selected single-chain fragment variable from a specific monoclonal antibody and one or more T-cell receptor intracellular signaling domains. This T-cell genetic modification may occur either via viral-based gene transfer methods or nonviral methods, such as DNA-based transposons, CRISPR/Cas9 technology or direct transfer of in vitro transcribed-mRNA by electroporation. Results Clinical trials have shown very promising results in end-stage patients with a full recovery of up to 92% in Acute Lymphocytic Leukemia. Despite such results in hematological cancers, the effective translation of CAR T-cell therapy to solid tumors and the corresponding clinical experience is limited due to therapeutic barriers, like CAR T-cell expansion, persistence, trafficking, and fate within tumors. Conclusion In this review, the basic design of CARs, the main genetic modification strategies, the safety matters as well as the initial clinical experience with CAR T-cells are described.

Zusammenfassung Hintergrund Mutationen im CRB1-Gen wurden bei frühkindlicher schwerer Netzhautdegeneration (EOSRD, early-onset severe retinal dystrophy) und Stäbchen-Zapfen-Degeneration des Kindesalters identifiziert. Diese Studie beschreibt das phänotypische Spektrum bei krankheitsverursachenden CRB1-Mutationen in den ersten beiden Lebensdekaden. Material und Methoden Acht Patienten im Alter von 3 Monaten bis 20 Jahren wurden einer ophthalmologischen Untersuchung mit Prüfung des bestkorrigierten Visus (BCVA), Farbsehprüfung, Funduskopie, optischer Kohärenztomografie (SD-OCT) und Fundusautofluoreszenz (FAF) unterzogen. Die SD-OCTs wurden mit der DIOCTA-Auswertesoftware sowie manuell einer Schichtanalyse unterzogen. In Abhängigkeit von der Kooperationsfähigkeit der Patienten wurden eine Ganzfeldelektroretinografie (ffERG, ISCEV-Standard) sowie eine Goldmann-Perimetrie durchgeführt. Ergebnisse Fünf der Patienten trugen Mutationen, die zu einem Verlust des Genproduktes führten (Spleißmutationen, Nonsense-Mutationen, Leserasterverschiebungen) und zeigten bereits in den ersten Lebensmonaten ein massiv reduziertes Sehvermögen (BCVAmax: 0,04). Der 6. Patient trug eine homozygote Missense-Mutation und zeigte im Alter von 6 Jahren einen BCVAmax von 0,05. Zwei weitere Patienten mit einer hypomorphen Missense-Mutation auf mindestens einem Allel zeigten einen besseren Visuserhalt mit BCVA bis zu 0,5 im Alter von 20 Jahren. Das ffERG lag bei allen Patienten unter der Norm und war bei der Mehrheit nicht mehr ableitbar. Das Gesichtsfeld war stark eingeschränkt. Die Photorezeptorschichten waren im SD-OCT, soweit überhaupt abgrenzbar, deutlich reduziert. Die Schichtdicke der inneren Körnerschicht nahm mit zunehmender Degeneration zu. Die Analyse der A-Scans zwischen Nervenfaserschicht (RNFL) und Bruch-Membran zeigte eine besser erhaltene Netzhautschichtung bei den Patienten mit Missense-Mutationen. Schlussfolgerung Patienten mit CRB1-Mutationen zeigen in erster Linie einen schweren Phänotyp mit stark reduziertem Sehvermögen von Geburt an. Missense-Mutationen mit einer möglichen Restfunktion des Genprodukts sind mit leichteren Phänotypen assoziiert. Eine schwere und progrediente Gesichtsfeldeinschränkung war allgemein in der 1. Lebensdekade vorhanden. Die reduzierte Abgrenzbarkeit der retinalen Schichtung im SD-OCT deutet auf einen Verlust der strukturellen Integrität aller Netzhautschichten hin.

This chapter was inadvertently published with one of the contributing author's name printed as Miliotou N. Androulla, whereas it should have been A. N. Miliotou . This correction has been updated in the book.

We present a VUV beamline installed as pump-probe source at the free-electron laser FLASH. The source is based on high-order harmonic generation driven by femtosec-ond near-infrared laser pulses synchronized with the FEL burst mode.

OBJECTIVES: The aim of the study was to assess differences in the diagnosis and management of eosinophilic esophagitis (EoE) by European pediatric (PG) and adult gastroenterologists (AG), and their self-reported adherence to guidelines. METHODS: A multiple-choice questionnaire gauged the diagnostic and management strategies of gastroenterologists treating children or adults in 14 European countries and the United Arab Emirates (UAE). RESULTS: Questionnaires were completed by 465 PG and 743 AG. PG were significantly more likely to take biopsies in patients with symptoms of esophageal dysfunction (86.2% PG vs 75.4% AG, P

Understanding the governing mechanisms of atmosphere–wave–ocean​ interactions is critical for unravelling the formation and evolution mechanisms of severe weather phenomena. This study aims at investigating the effects of atmosphere–wave–ocean​ feedbacks on a Mediterranean tropical-like cyclone (medicane), occurred on 27–30 September 2018 at the central-eastern Mediterranean Sea and characterized by severe environmental and socioeconomic impact. To unveil the interactions across the air–sea interface, the medicane was simulated by an integrated modelling system consisting of the Chemical Hydrological Atmospheric Ocean wave System (CHAOS), upgraded by embedding to it the Nucleus for European Modelling of the Ocean (NEMO) as ocean circulation component. Coupled simulations revealed that air–seaheat transfer and Ekman pumping, bringing sub-surface cold waters in upper ocean layers (upwelling), caused SST cooling (∼2–3 °C). SST cooling triggered a negative feedback loop procedure tending to balance between atmospheric and ocean processes. It also attenuated the cyclone and, subsequently, reduced the atmospheric energy embedded in ocean through the upper ocean vertical stratification weakening, thus, upper ocean vertical mixing, upwelling and SST cooling. The waves adjusted this feedback loop making the system more resistant in air–sea flux variations. Waves additionally weakened the cyclone not only due to the kinetic energy loss in the lower-atmosphere but also due to the enhancement of SST cooling which is attributed to the strengthening of Ekman pumping and vertical mixing, forced by wind stress increase. Nevertheless, waves partially balanced the air–wave–sea exchanges through the slight enthalpy flux gain under high wind conditions which is explained by considering the increase of enthalpy transfer coefficient in rougher sea areas. © 2020 Elsevier Ltd

This chapter was inadvertently published with one of the contributing author's name printed as Miliotou N. Androulla, whereas it should have been A. N. Miliotou . This correction has been updated in the book.

We present a HHG-based vacuum ultraviolet (VUV) source at the free electron laser FLASH2. The source provides ultrashort pulses from 10 to 40eV, coupled to the REMI end-station (beamline FL26) for VUV-FEL pump-probe experiments.

In the era of precision medicine, the tremendous progress in next-generation sequencing technologies has allowed the identification of an ever-increasing number of genes associated with known Mendelian disorders. Neonatal diabetes mellitus is a rare, genetically heterogeneous endocrine disorder diagnosed before 6 months of age. It may occur alone or in the context of genetic syndromes. Neonatal diabetes mellitus has been linked with genetic defects in at least 26 genes to date. Novel mutations in these disease-causing genes are being reported, giving us a better knowledge of the molecular events that occur upon insulin biosynthesis and secretion from the pancreatic β-cell. Of great importance, some of the identified genes encode proteins that can be therapeutically targeted by drugs per os, leading to transitioning from insulin to sulfonylureas. In this review, we provide an overview of pancreatic β-cell physiology, present the clinical manifestations and the genetic causes of the different forms of neonatal diabetes, and discuss the application of next-generation sequencing methods in the diagnosis and therapeutic management of neonatal diabetes and on research in this area. © 2020 Informa UK Limited, trading as Taylor & Francis Group.

Chimeric antigen receptor (CAR) cancer immunotherapy uses autologous immune system's cells, genetically modified, to reinforce the immune system against cancer cells. Genetic modification is usually mediated via viral transfection, despite the risk of insertional oncogenesis and off target side effects. In vitro-transcribed (IVT)-mRNA-mediated transfection could contribute to a much safer CAR therapy, since IVT-mRNA leaves no ultimate genetic residue in recipient cells. In this chapter, the IVT-mRNA generation procedure is described, from the selection of the target of the CAR T-cells, the cloning of the template for the in vitro transcription and the development of several chemical modifications for optimizing the structure and thus the stability of the produced CAR IVT-mRNA molecules. Among various transfection methods to efficiently express the CAR molecule on T-cells' surface, the electroporation and the cationic-lipid mediated transfection of the CAR IVT-mRNAs are described.

Chimeric antigen receptor (CAR) cancer immunotherapy uses autologous immune system's cells, genetically modified, to reinforce the immune system against cancer cells. Genetic modification is usually mediated via viral transfection, despite the risk of insertional oncogenesis and off target side effects. In vitro-transcribed (IVT)-mRNA-mediated transfection could contribute to a much safer CAR therapy, since IVT-mRNA leaves no ultimate genetic residue in recipient cells. In this chapter, the IVT-mRNA generation procedure is described, from the selection of the target of the CAR T-cells, the cloning of the template for the in vitro transcription and the development of several chemical modifications for optimizing the structure and thus the stability of the produced CAR IVT-mRNA molecules. Among various transfection methods to efficiently express the CAR molecule on T-cells' surface, the electroporation and the cationic-lipid mediated transfection of the CAR IVT-mRNAs are described.

The growth of eleven Salmonella serotypes cultured in modified Rappaport-Vassiliadis medium was compared with the growth in selenite broth and in nutrient broth. The growth of other enterobacteria in the three media was also studied. The mean generation times of all the serotypes were calculated by fitting the experimental data to the exponential growth law. The results showed that Rappaport-Vassiliadis medium sustained the growth of nine Salmonella serotypes and proved more successful in limiting the growth of other enterobacteria than did the selenite or nutrient broths.

New rhenium(I) tricarbonyl complexes with the quinolone antimicrobial agents oxolinic acid (Hoxo) and enrofloxacin (Herx) and containing methanol, triphenylphosphine (PPh3) or imidazole (im) as unidentate co-ligands, were synthesized and characterized. The crystal structure of complex [Re(CO)3(oxo)(PPh3)]∙0.5MeOH was determined by X–ray crystallography. The deprotonated quinolone ligands are bound bidentately to rhenium(I) ion through the pyridone oxygen and a carboxylate oxygen. The binding of the rhenium complexes to calf-thymus DNA (CT DNA) was monitored by UV spectroscopy, viscosity measurements and competitive studies with ethidium bromide; intercalation was suggested as the most possible mode and the DNA-binding constants of the complexes were calculated. The rhenium complex [Re(CO)3(erx)(im)] was assayed for its topoisomerase IIα inhibition activity and was found to be active at 100 μM concentration. The interaction of the rhenium complexes with human or bovine serum albumin was investigated by fluorescence emission spectroscopy (through the tryptophan quenching) and the corresponding binding constants were determined. The tracer complex [99mTc(CO)3(erx)(im)] was synthesized and identified by comparative HPLC analysis with the rhenium analog. The 99mTc complex was found to be stable in solution. Upon injection in healthy mice, fast tissue clearance of the 99mTc complex was observed, while both renal and hepatobiliary excretion took place. Preliminary studies in human K-562 erythroleukemia cells showed cellular uptake of the 99mTc tracer with distribution primarily in the cytoplasm and the mitochondria and less in the nucleus. These preliminary results indicate that the quinolone 99mTc/Re complexes show promise to be further evaluated as imaging or therapeutic agents.

Since the birth of maritime archaeology in the mid-twentieth century, shipwreck archaeology has occupied a predominant role in the research undertaken in the field. Shipwreck sites have enabled further understanding of shipbuilding technologies and traditions, trade routes and traded goods, as well as life on board ancient vessels. Excavating, interpreting and preserving underwater sites is a complex endeavour that requires the analysis of multiple aspects, not least the environmental and biological characteristics affecting the site and the archaeological record. Wreck site formation processes (WSFPs) theory has been extensively employed in the approach to the excavation, documentation and interpretation of archaeological shipwreck sites, since its inception by Keith Muckelroy in 1978. WSFPs allow to better understand the preserved archaeological record and to clarify the course of events which have affected the shipwreck site in the full extent of its archaeological context. The present paper aims to present an innovative approach to better understanding these processes. Employing a multidisciplinary approach, the research has used three-dimensional recording and modelling techniques, together with archaeological and biological data, in order to model the amphora cargo mound of the 4th century BCE Mazotos shipwreck (Cyprus). 3D photogrammetric models of the shipwreck site have been paired with the 3D photogrammetric models of the lifted amphorae bearing sediment and biogenic horizons marks on their walls, in order to attempt a reconstruction of the environment and sedimentation processes at the site. While the combination of the 3D site and amphora reconstructions allow to create a model frozen in time (i.e. the time of excavation), the sedimentation and biological information adds to the model the fourth dimension (time), shedding light on the natural SFP dynamics that have affected the site. The sediment horizons and encrusting communities’ marks on the amphora surfaces account for dynamic processes of burial/exposure as well as artefacts dislocation, allowing to better characterize the initial model and better interpret the available archaeological record.

Triphenylphosphonium (TPP) cations have been used for the development of tumor or myocardial diagnostic radiopharmaceuticals. In this work, the development of new [99mTc][Tc(CO)3(N,S,N)]+ complexes of the (N,S,N) tridentate chelator benzimidazol-2-yl-methylthioethylamine (L), and its triphenylphosphonium (TPP) cation derivative L-TPP is described. The TPP-moiety was conjugated at the Nτ-benzimidazol position of L. The reaction of the chelators L and L-TPP with a suitable precursor [Re(sol)3(CO)3]+ yielded single products of ReL+ and ReL-TPP2+. The complexes were characterized by spectroscopic methods and furthermore the structure of ReL was elucidated by X-ray crystallography. The respective 99mTc-radiotracers were synthesized in high yield, their lipophilicity was measured and both exhibited high stability in cysteine, histidine solutions as well as in rat plasma. The in vitro cell studies in human erythroleukemia K-562 and glioblastoma U-87MG tumor cells showed that the tracer 99mTcL+ exhibited significantly higher cellular uptake, while 99mTcL-TPP2+ exhibited significantly higher mitochondrial accumulation. The tracers 99mTcL+ and 99mTcL-TPP2+ exhibited fast blood elimination and excretion via the hepatobiliary and the renal routes after intravenous administration in healthy mice. Tracer 99mTcL+ exhibited higher myocardial uptake and renal excretion, while 99mTcL-TPP2+ exhibited primarily hepatobiliary excretion. These data confirm the high mitochondrial accumulation of 99mTcL-TPP2+ in vitro and show its potential as a candidate for tumor imaging.

Background The clinical relevance of mismatch repair (MMR) status in patients with nonmetastatic cancer across tumour types remains unclear. Our goal was to investigate the prognostic role of MMR deficiency in patients with stage I-III colorectal and endometrial cancer. Methods Patients with nonmetastatic colorectal and endometrial cancer with tumour tissue available for analysis were identified through the Hellenic Cooperative Oncology Group (HeCOG)'s tumour repository. Patients had been referred to Departments of Medical Oncology affiliated with HeCOG. MMR protein expression was evaluated by immunohistochemistry. The primary outcome measure was overall survival (OS). Results From May 1990 to September 2012, 1158 patients with nonmetastatic colorectal (N = 991) and endometrial cancer (N = 167) were identified (median age: 64 years, men: 544). All patients with colorectal and 109 (65%) with endometrial cancer had received adjuvant treatment. MMR deficiency was observed in 114 (11.5%) of colorectal and 80 (47.9%) of endometrial tumours. More commonly deficient proteins were PMS2 (69 patients, 7%) and MLH1 (63 patients, 6.5%) in colorectal cancer and MSH2 (58 patients, 34.7%) in endometrial cancer. Colorectal MMR-deficient (dMMR) tumours were more likely to be right sided (65 % dMMR vs 27 % proficient MMR, pMMR; p < 0.001), high grade (31% vs 15%, Ï ‡ 2, p < 0.001) and with a mucinous component (64% vs 42%, p < 0.001). Endometrial dMMR tumours were more often of endometrioid histology (51.4 % endometrioid vs 20 % serous/clear cell, p = 0.020). Compared with MMR proficiency, MMR deficiency was associated with improved OS in patients with endometrial cancer (HR = 0.38, 95% CI 0.20 to 0.76, p = 0.006), but not in patients with colorectal cancer (HR = 0.73, 95% CI 0.49 to 1.09, p = 0.130). After adjusting for age, stage and grade, MMR deficiency maintained its favourable prognostic significance in patients with endometrial cancer (HR = 0.42, 95% CI 0.20 to 0.88, p = 0.021). Conclusions DMMR was associated with improved outcomes in patients with nonmetastatic endometrial cancer, but not in patients with nonmetastatic colorectal cancer who received adjuvant chemotherapy. © 2019 BMJ Publishing Group Limited. No commercial re-use. See rights and permissions. Published by BMJ.

The potential antibacterial activity of basil (Ocimum basilicum), chamomile (Matricaria chamomilla), origanum (Thymus capitatus), tea tree (Melaleuca alternifolia) and thyme (Thymus vulgaris) essential oils, was investigated against 29 Gram-positive bacterial strains isolated from wastewater treatment plants, clinical samples (n = 25) and American Type Culture Collection (ATCC) reference strains (n = 4). Wild bacterial strains included methicillin-resistant Staphylococcus aureus (n = 16) and vancomycin-resistant Enterococcus spp. (n = 9). The antimicrobial activity of the selected oils was studied using the broth macrodilution method. The Minimal Inhibitory Concentration (MIC) values for S. aureus ranged from 0.06 to 0.5% (v/v) for origanum oil, 0.06 to 1% (v/v) for thyme oil, 0.12 to 1% (v/v) for tea tree oil, 0.25 to 4% (v/v) for basil oil and 2 to &gt, 4% (v/v) for chamomile oil. For enterococci the MIC values were significantly higher ranging from 0.25 to 1% (v/v), 0.5 to 2% (v/v), 1 to 4% (v/v), 4 to &gt, 4% (v/v) and &gt, 4% (v/v) for the above-mentioned oils, respectively. The main compounds of the tested essential oils were: estragole (Ocimum basilicum), bisabolol and trans-b-farnesene (Matricaria chamomilla), carvacrol and thymol (Thymus capitatus), terpinen-4-ol and p-cymene (Melaleuca alternifolia), thymol, linalool, and p-cymene (Thymus vulgaris). Origanum essential oil yielded the best antimicrobial results followed by thyme, tea tree, and basil oil, while chamomile oil exhibited weak antibacterial properties.

Basal cell carcinoma is the most common skin cancer of the eyelid. It most frequently occurs on the lower eyelid and the medial canthus. Although metastases appear only very rarely, basal cell carcinoma is locally destructive if left untreated. Nowadays several surgical and nonsurgical treatments can be used to remove or destroy basal cell carcinomas. The selection of optimal treatment depends on the initial individual situation as well as the age and the general condition of the patient. The "gold standard" method for the treatment of basal cell carcinomas is surgical removal with subsequent histological examination. Because of the complex anatomical characteristics of the medial canthus and the lateral nasal root region, basal cell carcinomas of this area present great challenges for any treatment method. These characteristics also explain the higher rates of incomplete removal and greater risk of recurrence, with in depth extension of the tumor. Additionally, surgical removal of the tumor from the medial canthus area often results in extended tissue defects. The plastic covering of these defects can be achieved by various reconstruction procedures.

Background Nurses' caring behaviors are central in the quality of care of patients undergoing sophisticated chemotherapy protocols. However, there is a scarcity of research regarding these behaviors in non-Anglo-Saxon countries. Objective The aim of this study was to explore caring behaviors that nurses perceive as important in caring for patients in Greece receiving chemotherapy. Methods We used a mixed-methods design, including a survey in 7 oncology wards in 3 cancer hospitals in Attica, Greece, and a subsequent qualitative focus group investigation. Caring behaviors were explored through the Caring Behavior Inventory 24 and content analysis of 3 focus group interviews. Results A sample of 72 nurses (response rate, 68.5%) were surveyed, and 18 nurses participated in the focus groups. "Knowledge/skills" (5 [SD, 0.7]) was the most important caring behaviors. No significant associations with nurses' characteristics were noted, except for higher scores in caring behaviors in participants who were married (P

Background: It remains unclear whether psychotic features increase the risk of completed suicides in unipolar depression. The present systematic review coupled with a meta-analysis attempts to elucidate whether unipolar psychotic major depression (PMD) compared to non-PMD presents higher rates of suicides. Methods: A systematic search was conducted in Scopus, PubMed, and “gray literature” for all studies providing data on completed suicides in PMD compared to non-PMD, and the findings were then subjected to meta-analysis. All articles were independently extracted by two authors using predefined data fields. Results: Nine studies with 33,873 patients, among them 828 suicides, met our inclusion criteria. PMD compared to non-PMD presented a higher lifetime risk of completed suicides with fixed-effect pooled OR 1.21 (95% CI 1.04-1.40). In a sub-analysis excluding a very large study (weight = 86.62%), and comparing 681 PMD to 2106 non-PMD patients, an even higher pooled OR was found [fixed-effect OR 1.69 (95% CI 1.16-2.45)]. Our meta-analysis may provide evidence that the presence of psychosis increases the risk of suicide in patients suffering from severe depression. The data are inconclusive on the contribution of age, mood congruence, comorbidity, and suicide method on PMD’s suicide risk. The lack of accurate diagnosis at the time of suicide, PMD’s diagnostic instability, and the use of ICD-10 criteria constitute the main study limitations. Conclusions: The presence of psychosis in major depression should alert clinicians for the increased risk of completed suicide. Thus, the implementation of an effective treatment both for psychotic depression and patients’ suicidality constitutes a supreme priority. © 2018 The Author(s).

Objective: It still remains unclear whether psychotic features increase the risk of suicidal attempts in major depressive disorder. Thus, we attempted, through a systematic review coupled with a meta-analysis, to elucidate further whether unipolar psychotic depression (PMD) compared to non-PMD presents higher levels of suicidal attempts. Method: A systematic search was conducted in PubMed, EMBASE, PsycINFO as well as in various databases of the so-called gray literature for all studies providing data on suicidal attempts in PMD compared to non-PMD, and the results were then subjected to meta-analysis. Results: Twenty studies met our inclusion criteria, including in total 1,275 PMD patients and 5,761 non-PMD patients. An elevated risk for suicide attempt for PMD compared to non-PMD patients was found: The total (lifetime) fixed-effects pooled OR was 2.11 (95% CI: 1.81–2.47), and the fixed-effects pooled OR of the five studies of the acute phase of the disorder was 1.93 (95% CI: 1.33–2.80). This elevated risk of suicidal attempt for PMD patients remained stable across all age groups of adult patients. Conclusion: Despite data inconsistency and clinical heterogeneity, this systematic review and meta-analysis showed that patients with PMD are at a two-fold higher risk, both during lifetime and in acute phase, of committing a suicidal attempt than patients with non-PMD. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

Imatinib mesylate (IM/Gleevec®), a selective inhibitor of chimeric Bcr-Abl tyrosine kinase, was developed as a first line drug to treat CML and ALL Ph(+) patients. Earlier studies have shown that hemin counteracts the IM-induced cell killing in human K-562 CML cells. In this study, we investigated whether IM disrupts the heme-dependent Cytochrome c Oxidase (COX) Biosynthesis and Assembly Pathway (HDCBAP) in Bcr-Abl(+) and Bcr-Abl(-) cells by affecting the expression of key-genes. Cells were exposed to IM and evaluated at time intervals for cell growth, cell death, expression of various genes by RT-PCR analysis as well as Sco2 mature protein levels by western blot analysis and COX enzymatic activity. IM at 1 μM induced extensive cell growth inhibition and cell death as well as marked suppression of the expression of SCO2 and FRATAXIN (FXN) genes in human K-562 and KU-812 Bcr-Abl(+) CML cells. IM also reduced the protein level of mature Sco2 mitochondrial protein as well as COX activity in these cell lines. However, treatment of human MOLT-4 Bcr-Abl(-) cells with 1μM and even with higher concentrations (4×10(-5)M) of IM neither reduced the expression of SCO2 and FXN genes nor suppressed the protein level of mature Sco2 protein and COX activity. Our findings indicate that SCO2 and FXN genes, involved in HDCBAP, are repressed by IM in human Bcr-Abl(+) CML cells and may represent novel target sites in leukemia therapy.