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Observation of a single protein by ultrafast X-ray diffraction

Light(Science & Applications) 2024年第1期13卷 80-90页

作者： Tomas Ekeberg Dameli Assalauova Johan Bielecki Rebecca Boll Benedikt J.Daurer Lutz A.Eichacker Linda E.Franken Davide E.Galli Luca Gelisio Lars Gumprecht Laura H.Gunn Janos Hajdu Robert Hartmann Dirk Hasse Alexandr Ignatenko Jayanath Koliyadu Olena Kulyk Ruslan Kurta Markus Kuster Wolfgang Lugmayr Jannik Lübke Adrian P.mancuso Tommaso Mazza Carl Nettelblad Yevheniy Ovcharenko Daniel E.Rivas Max Rose Amit K.Samanta Philipp Schmidt Egor Sobolev Nicusor Timneanu Sergey Usenko Daniel Westphal Tamme Wollweber Lena Worbs Paul Lourdu Xavier Hazem Yousef Kartik Ayyer Henry N.Chapman Jonas A.Sellberg Carolin Seuring Ivan A.Vartanyants Jochen Küpper Michael Meyer Filipe R.N.C.Maia Laboratory of Molecular Biophysics Department of Cell and Molecular BiologyUppsala UniversityHusargatan 3(Box 596)SE-75124 UppsalaSweden Deutsches Electronen-Synchrotron DESY Notkestrasse 8522607 HamburgGermany European XFEL Holzkoppel 422869 SchenefeldGermany Diamond Light Source Harwell Science&Innovation CampusDidcot OX110DEUK University of Stavanger Centre Organelle ResearchRichard-Johnsensgate 44021 StavangerNorway Leibniz Institute for Experimental Virology(HPI) Centre for Structural Systems BiologyNotkestraße 8522607 HamburgGermany Dipartimento di Fisica“Aldo Pontremoli” Universitàdegli Studi di Milanovia Celoria 1620133 MilanoItaly Center for Free-Electron Laser Science DESY22607 HamburgGermany Plant Biology Section School of Integrative Plant ScienceCornell UniversityIthacaNY 14853USA PNSensor GmbH Otto-Hahn-Ring 6D-81739 MünchenGermany Biomedical and X-Ray Physics Department of Applied PhysicsAlbaNova University CenterKTH Royal Institute of TechnologySE-10691 StockholmSweden ELI Beamlines/IoP Institute of Physics AS CR v.v.i.Na Slovance 218221 Prague 8Czech Republic Multi-User CryoEM Facility Centre for Structural Systems BiologyNotkestr.8522607 HamburgGermany University Medical Center Hamburg-Eppendorf(UKE) Martinistrasse 5220246 HamburgGermany The Hamburg Center for Ultrafast Imaging Universität HamburgLuruper Chaussee 14922761 HamburgGermany Department of Physics Universität HamburgLuruper Chaussee 14922761 HamburgGermany Department of Chemistry and Physics La Trobe Institute for Molecular ScienceLa Trobe UniversityMelbourneVIC 3086Australia Division of Scientific Computing Science for Life LaboratoryDepartment of Information TechnologyUppsala UniversityBox 337SE-75105 UppsalaSweden European Molecular Biology Laboratory c/o DESYNotkestrasse 8522607 HamburgGermany Department of Physics and Astronomy Uppsala UniversityBox 516SE-75120 UppsalaSweden Max Planck Institute for the Structure and Dynamics of Matter Luruper Chaussee 14922761 HamburgGe

The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired *** was one of the arguments for building X-ray free-electron *** to theory,the extremely intense pulses provide sufficient signal to dispense with using crystals as an amplifier,and the ultrashort pulse duration permits capturing the diffraction data before the sample inevitably *** was first demonstrated on biological samples a decade ago on the giant *** then,a large collaboration has been pushing the limit of the smallest sample that can be *** ability to capture snapshots on the timescale of atomic vibrations,while keeping the sample at room temperature,may allow probing the entire conformational phase space of *** we show the first observation of an X-ray diffraction pattern from a single protein,that of Escherichia coli GroEL which at 14 nm in diameter is the smallest biological sample ever imaged by X-rays,and demonstrate that the concept of diffraction before destruction extends to single *** the pattern,it is possible to determine the approximate orientation of the *** experiment demonstrates the feasibility of ultrafast imaging of single proteins,opening the way to single-molecule time-resolved studies on the femtosecond timescale.

关键词： smallest intense ultrafast

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Aluminium toxicity targets PIN2 in Arabidopsis root apices: Effects on PIN2 endocytosis, vesicular recycling, and polar auxin transport

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Chinese Science Bulletin 2008年第16期53卷 2480-2487页

作者： SHEN Hong HOU NingYan Markus SCHLICHT WAN YingLang Stefano mancuso Frantisek BALUSKA College of Resources and Environment South China Agricultural University Guangzhou 510642 China Institute of Cellular and Molecular Botany Bonn University Bonn D-53115 Germany Electrophysiology Laboratory Department of Horticulture University of Florence Viale delle Idee 30 1-50019 Florence Italy

The most obvious symptom of Al toxicity is the inhibition of root growth. However, the mechanism of Al-inhibiting root growth remains to be elucidated. In this study, auxin transport and vesicle movement of an auxin-efflux carrier (PIN2) were investigated in Arabidopsis roots in response to Al stress. Results indicated that Al inhibited the apical transport of auxin in root tips of Arabidopsis significantly. The severe inhibition was localized in the cells of transition zone, where the concentration of auxin was only 34% that of the control. Brefeldin A (BFA), an inhibitor of vesicle transport, induced the dot-like structure of PIN2 vesicle significantly. Al decreased the size of dot-like structure of PIN2 vesicles. Re- sults of real-time RT-PCR and Western-blotting analysis showed that Al increased the transcript level of PIN2 and the accumulation of PIN2 protein in horizontal direction of plasma membrane, but decreased its distribution in endosomes, suggesting that Al inhibited the transport of PIN2 vesicles from plasma membrane to endosomes. Results of cytoskeleton-depolymering drugs indicated that it was via the pathway of disruption of actin microfilaments that Al inhibited the transport of PIN2 vesicles. Exposed to Al stress, the cells of elongation zone had less Al uptake and less transport frequency of vesicles than cells of transition zone. Taken together, our results suggested that Al inhibited root growth mainly by modulating the transport of PIN2 vesicles between plasma membrane and endosomes, thus block- ing auxin transport and root growth.

关键词：铝毒性胞吞作用生长素

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Will vaccine-derived protective immunity curtail COVID-19 variants in the US?

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Infectious Disease Modelling 2021年第1期6卷 1110-1134页

作者： Marina mancuso Steffen E.Eikenberry Abba B.Gumel Arizona State University School of Mathematical and Statistical SciencesTempeAZ85287USA Department of Mathematics and Applied Mathematics University of PretoriaPretoria0002South Africa

Multiple effective vaccines are currently being deployed to combat the COVID-19 pandemic,and are viewed as the major factor in marked reductions of disease burden in regions with moderate to high vaccination *** effectiveness of COVID-19 vaccination programs is,however,significantly threatened by the emergence of new SARS-COV-2 variants that,in addition to being more transmissible than the wild-type(original)strain,may at least partially evade existing vaccines.A two-strain(one wildtype,one variant)and two-group(vaccinated or otherwise)mechanistic mathematical model is designed and used to assess the impact of the vaccine-induced cross-protective efficacy on the spread the COVID-19 pandemic in the United *** analysis of the model shows that,in the absence of any co-circulating SARS-CoV-2 variant,the vaccine-derived herd immunity threshold needed to eliminate the wild-type strain can be achieved if 59%of the US population is fully-vaccinated with either the Pfizer or Moderna *** threshold increases to 76%if the wild-type strain is co-circulating with the Alpha variant(a SARS-CoV-2 variant that is 56%more transmissible than the wild-type strain).If the wild-type strain is co-circulating with the Delta variant(which is estimated to be 100%more transmissible than the wild-type strain),up to 82%of the US population needs to be vaccinated with either of the aforementioned vaccines to achieve the vaccine-derived herd *** sensitivity analysis of the model reveal the following four parameters as the most influential in driving the value of the reproduction number of the variant strain(hence,COVID-19 dynamics)in the US:(a)the infectiousness of the co-circulating SARS-CoV-2 variant,(b)the proportion of individuals fully vaccinated(using Pfizer or Moderna vaccine)against the wild-type strain,(c)the cross-protective efficacy the vaccines offer against the variant strain and(d)the modification parameter accounting for the reduced infectio

关键词： COVID-19 Vaccine Wild-type Variant Reproduction number Herd immunity

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Management of portopulmonary hypertension:New perspectives

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World Journal of Gastroenterology 2013年第45期19卷 8252-8257页

作者： Luigi mancuso Francesca Scordato Michela Pieri Eliana Valerio Andrea mancuso Department of Cardiologia Ospedale Cervello90100 Palermo Italy Cardiologia Clinica Candela 90100 Palermo Italy Epatologia e Gastroenterologia Ospedale Niguarda Cà Granda Milano 20162 Milano Italy Medicina Interna 1 ARNAS Civico 90100 Palermo Italy

Portopulmonary hypertension(PPHTN)is a known complication of ***-to-severe PPHTN implies an extremely poor *** occurs in 5%-10%of patients referred for liver transplantation(LT),and probably with an higher incidence in patients with large portosystemic *** with moderate-tosevere pulmonary hypertension have been previously excluded from LT because of the extremely high surgical risk and since the post-transplant outcome reported was ***,new perspectives in the management of patients with portopulmonary hypertension are *** fact,some pulmonary vasoactive drugs have become routine in the treatment of patients with idiopathic pulmonary *** drugs,particularly epoprostenol,have been recently introduced in the treatment of patients with PPHTN,and have been shown to be effective in reducing pulmonary artery pressure as well as pulmonary vascular ***,recent studies seem to demonstrate that treatment with pulmonary vasoactive drugs could allow liver transplantation with acceptable surgical risks and excellent *** there are not large series nor prospective studies addressing this topic,the clinical scenario of patients with PPHTN seems to be positively changing.

关键词： Portopulmonary hypertension Cirrhosis Liver transplantation Management Epoprostenol

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An in vitro prototype of a porcine biomimetic testis-like cell culture system： a novel tool for the study of reassembled Sertoli and Leydig cells

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Asian Journal of Andrology 2018年第2期20卷 160-165页

作者： Iva Arato Giovanni Luca Francesca mancuso Catia Bellucci Cinzia Lilli Mario Calvitti Barbara C Hansen Domenico Milardi Giuseppe Grande Riccardo Calafiore Department of Experimental Medicine University of Perugia Perugia 06100 Italy Division of Medical Andrology and Endocrinology of Reproduction University of Perugia and Saint Mary Hospital Terni 05100 Italy Department of Internal Medicine and Pediatrics Morsani College of Medicine University of South Florida Tampa 33612 FL USA Division of Endocrinology Fondazione Policlinico Universitario 'A. Gemelli' 00168 Rome Italy Department of Medicine University of Perugia Perugia 06100 Italy

At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors （testicular cross-talk）, mainly orchestrated by gonadotropins such as luteinizing hormone （LH） and follicle-stimulating hormone （FSH） that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay （ELISA） for anti-Mullerian hormone （AMH）, inhibin B, and testosterone secretion in the medium, and Real-lime PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3βHSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.

关键词： human gonadotropins Leydig cells prepubertal biomimetic testis Sertoli cells

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Volatile organic compound emission and biochemical properties of degraded Ultisols ameliorated by no tillage and liming

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Pedosphere 2020年第5期30卷 597-606页

作者： Laura GIAGNONI Cosimo TAITI Paloma LEON Corrado COSTA Paolo MENESATTI Rafael ESPEJO Clara GOMEZ-PACCARD Chiquinquira HONTORIA Eduardo VAZQUEZ Marta BENITO Stefano mancuso Giancarlo RENELLA Department of Agriculture FoodEnvironment and Forestry(DAGRI)University of FlorenceP.le delle Cascine 18Florence 50144(Italy) Departamento de Production Agraria E.T.S.I.AgrondmicaAlimentaria y de BiosistemasUniversidad Politecnica de MadridAvda.Complutense s/nMadrid 28040(Spain) Consiglio per la Ricerca in Agricoltura e I'Analisi dell'Economia Agraria(CREA)-Unita di Ricerca per ilngegneria Agraria via della Pascolare 16Monterotondo ScaloRM 00015(Italy) Department of Soil Biogeochemistry and Soil Ecology University of BayreuthDr.Hans-Frisch-Strafie 1-3Bayreuth 95448(Germany) University of Padua Department of AgronomyFoodNatural ResourcesAnimals and Environment(DAFNAE)Viale dell'Universita 16Legnaro 35020(Italy)

TTie Ultisols in the Rana de Canamero area in Southwest Spain showed aluminum(Al)phytotoxicity,and the clearance of natural vegetation and decades of intensive conventional agriculture caused the deplation of soil organic matter(SOM).Therefore,we studied the long-term effects of no tillage and liming using sugar beet foam(SF)and red gypsum(RG),alone or in combination,on the restoration of Ultisols affected by acidification,Al phytotoxicity,and SOM *** measured the main soil chemical properties,soil microbial biomass,soil enzyme activities involved in carbon,nitrogen,phosphorus,and sulfur mineralization,and the emission of volatile organic compounds(VOCs).The results indicated that liming effectively neutralized the soil acidity in the long term and,in combination with no tillage,significantly increased soil microbial biomass and enzyme ***-three VOCs were detected using the proton transfer reaction-time of flight(PTR-ToF)technique,and both liming and tillage changed the VOC emission *** greatest difference in VOC emission pattern was observed between no-tilled un-amended soils and tilled lime-amended soils,suggesting the activation of different metabolic pathways within the microbial communities of soils under different *** in VOC emission patterns could be attributed to the decomposition of carbohydrates,which were also sustained by the higher enzyme activities in the lime-amended soils.

关键词： carbohydrate decomposition degraded soils long-term field trial soil acidity soil enzyme activities soil microbial biomass soil restoration

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Management of hepatocellular carcinoma: Enlightening the gray zones

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World Journal of Hepatology 2013年第6期5卷 302-310页

作者： Andrea mancuso Epatologia e Gastroenterologia Ospedale Niguarda Cà Granda20162 Milano Italy Medicina Interna 1 Azienda di Rilievo Nazionale ad Alta Specializzazione Ospedale Civico90127 Palermo Italy

Management of hepatocellular carcinoma (HCC) has been continuously evolving during recent years. HCC is a worldwide clinical and social issue and typically a complicates cirrhosis. The incidence of HCC is increasing, not only in the general population of patients with cirrhosis, but particularly in some subgroups of patients, like those with human immunodeficiency virus infection or thalassemia. Since a 3% annual HCC incidence has been estimated in cirrhosis, a biannual screening is generally suggested. The diagnostic criteria of HCC has recently had a dramatic evolution during recent years. HCC diagnosis is now made only on radiological criteria in the majority of the cases. In the context of cirrhosis, the universally accepted criteria for HCC diagnosis is contrast enhancement in arterial phase and washout in venous/late phase at imaging, the so called "typical pattern". However, recently up- dated guidelines slightly differ in diagnostic criteria. Apart from liver transplantation, the only cure of both HCC and underlying liver cirrhosis, all the other treatments have to match with higher rate of HCC recurrence. The latter can be classified into curative (resection and percutaneous ablation) and palliative treatments. The aim of this paper was to review the currentknowledge on management of HCC and to enlighten the areas of uncertainty.

关键词： Hepatocellular carcinoma Management Trans-arterial chemoembolization Orthotopic liver transplantation Surgery

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Hepatocellular carcinoma in thalassemia:A critical review

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World Journal of Hepatology 2010年第5期2卷 171-174页

作者： Andrea mancuso Emergency Area Ospedali Riuniti di Sciacca(Ag)

Due to blood transfusions,thalassemics are often infected with either hepatitis C virus(HCV)or hepatitis B virus and often have *** carcinoma(HCC)has emerged in thalassemics only recently as a result of the improvement in thalassemia *** fact,a prospective study estimated an HCC incidence inβ-thalassemia of about 2%.Although data are scanty,HCC screening in thalassemics with risk factors for HCC should be carried *** treatments have some efficacy in HCV infected thalassemics despite partial contraindication to ribavirin and iron ***,there are no data on how HCV treatment translates into HCC *** data suggest that HCC treatment in thalassemics should generally have the same outcomes as in *** coexistence of severe comorbidities makes liver transplantation challenging,this therapeutic possibility should not be precluded for well selected HCCβ-thalassemia *** fact,2 transfusion dependent adult HCCβ-thalassemia patients have recently undergone successful liver transplantation with a good *** conclusion,HCC seems to be a developing issue in thalassemia and HCC screening should be carried *** treatment,including liver transplantation,can be performed in selected patients. A multidisciplinary effort is needed for management.

关键词： Thalassemia Hepatocellular carcinoma Hemochromatosis Screening Complication Liver trans- plantation

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To mask or not to mask:Modeling the potential for face mask use by the general public to curtail the COVID-19 pandemic

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Infectious Disease Modelling 2020年第1期5卷 293-308页

作者： Steffen E.Eikenberry Marina mancuso Enahoro Iboi Tin Phan Keenan Eikenberry Yang Kuang Eric Kostelich Abba B.Gumel School of Mathematical and Statistical Sciences Arizona State UniversityTempeAZ85287USA

Face mask use by the general public for limiting the spread of the COVID-19 pandemic is controversial,though increasingly recommended,and the potential of this intervention is not well *** develop a compartmental model for assessing the communitywide impact of mask use by the general,asymptomatic public,a portion of which may be asymptomatically *** simulations,using data relevant to COVID-19 dynamics in the US states of New York and Washington,suggest that broad adoption of even relatively ineffective face masks may meaningfully reduce community transmission of COVID-19 and decrease peak hospitalizations and ***,mask use decreases the effective transmission rate in nearly linear proportion to the product of mask effectiveness(as a fraction of potentially infectious contacts blocked)and coverage rate(as a fraction of the general population),while the impact on epidemiologic outcomes(death,hospitalizations)is highly nonlinear,indicating masks could synergize with other nonpharmaceutical ***,masks are found to be useful with respect to both preventing illness in healthy persons and preventing asymptomatic *** mask adoption scenarios,for Washington and New York state,suggest that immediate near universal(80%)adoption of moderately(50%)effective masks could prevent on the order of 17e45%of projected deaths over two months in New York,while decreasing the peak daily death rate by 34e58%,absent other changes in epidemic *** very weak masks(20%effective)can still be useful if the underlying transmission rate is relatively low or decreasing:InWashington,where baseline transmission is much less intense,80%adoption of such masks could reduce mortality by 24e65%(and peak deaths 15e69%),compared to 2e9%mortality reduction in New York(peak death reduction 9e18%).Our results suggest use of face masks by the general public is potentially of high value in curtailing community transmission and the burden

关键词： Face mask Non-pharmaceutical intervention Cloth mask N95 respirator Surgical mask SARS-CoV-2 COVID-19

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Hepatocellular Carcinoma and Liver Transplantation:State of the Art

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Journal of Clinical and Translational Hepatology 2014年第3期2卷 176-181页

作者： Andrea mancuso Giovanni Perricone Epatologia e Gastroenterologia Ospedale Niguarda Ca'GrandaMilanoItaly Medicina Interna 1 Azienda di Rilievo Nazionale ad Alta Specializzazione Civico-Di Cristina-BenfratelliPalermoItaly

Hepatocellular carcinoma (HCC) is an aggressive tumor that often occurs in chronic liver disease and *** incidence of HCC is growing *** respect to any other available treatment for liver cancer,liver transplantation (LT) has the highest potential to *** for removal at once of both the tumor ("seed")and the damaged-hepatic tissue ("soil") where cancerogenesis and chronic liver disorders have progressed *** Milan criteria (MC) have been applied worldwide to select patients with HCC for LT,yielding a 4-year survival rate of 75%.These criteria represent the benchmark for patient selection and are the basis for comparison with any other suggested ***,MC are often considered to be too restrictive,and recent data show that between 25% and 50% of patients with HCC are currently transplanted beyond conventional ***,any unrestricted expansion of selection criteria will increase the need for donor organs,lengthen waiting periods,increase drop-out rates,and impair outcomes on intention-to-treat *** of HCC recurrence after LT is *** are a few reports available regarding the safety and efficacy of sorafenib for HCC recurrence after LT,but the data are heterogeneous.A multi-center prospective randomized controlled trial comparing placebo with sorafenib is ***,a metaanalysis of patient survival with sorafenib for HCC recurrence after LT could be helpful to characterize the therapeutic benefit and safety of ***,we review the use of LT for HCC,with particular emphasis on the selection criteria for transplantation in patients with HCC and management of HCC recurrence after LT.

关键词： Hepatocellular carcinoma Liver transplantation Selection criteria Milan criteria Recurrence Sorafenib

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