BIOGRAFIA DI ITALO CALVINO

Università di Bologna地址：Via Zamboni, 33 - 40126 Bologna电话：051/2099370传真：051/2099372网址：www.unibo.it博洛尼亚大学的全称是 ALMA MATER STUDIORUM - Università di Bologna，是世界上最古老的大学，其历史可追溯到 1088 年。

当时，一些学生厌倦了正规的学习生活，联合部分教师，采用一种全新的方式进行教学，产生了现代大学的雏形。

在以后的近千年的历史中，博洛尼亚大学一直保持着独立教学和研究，不受任何政治和经济势力影响的传统，并逐渐被其它大学所接受，成为办学的宗旨。

但丁、彼特拉克、哥白尼等著名的文学艺术大师和科学家都曾在此担任教学工作。

今天的博洛尼亚大学是全意大利最有影响的综合性大学之一，共有 23 个学院，68 个学系，由多个校区组成。

5 个大学校区分别座落在博洛尼亚 (Bologna)，切塞纳 (Cesena)，弗利 (Forlì)，拉韦纳 (Ravenna) 和里米尼 (Rimini)。

博洛尼亚大学可以提供 132 个学士学位课程，95 个硕士学位课程，8 个欧洲硕士学位课程，众多的博士学位课程，以及硕士进修班课程。

除了高水平的学术活动之外，学校还与企业界保持了广泛的联系。

教学质量同研究水平紧密相连，研究领域包括健康、工程与建筑、科技、人文与法律、政治、经济等。

博洛尼亚大学同国际上许多的知名大学签署了学术合作协议，鼓励、支持并促进教授、研究人员与学生的国际交流。

建在博洛尼亚大学中的中国学院是一个真正的文化中心，2005-2006 学年起每年接待50 名中国学生。

这是为了响应意大利前总统钱皮决定增加在意大利大学正式注册的中国学生数量的号召，并缩短意大利在这方面与其它欧洲国家的差距。

参与这一计划的不仅有博洛尼亚大学（该大学也同时也在推进马可·波罗计划），而且还有博洛尼亚当地的多个部门，包括工业联合会、商会、大区、省和市政府以及 Carisbo 基金会和 Ceur 基金会。

Certificate of Analysis Takara Bio USA, Inc.1290 Terra Bella Avenue, Mountain View, CA 94043, USA U.S. Technical Support: ********************United States/Canada 800.662.2566 Asia Pacific+1.650.919.7300Europe+33.(0)1.3904.6880Japan+81.(0)77.565.6999Page 1 of 6Tet-On® 3G Vector Set (with ZsGreen1)Table of ContentsDescription (1)pCMV-Tet3G Vector Information (2)pTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector Information (4)Quality Control Data (6)Catalog No. Lot Number631159 (Not sold separately) Specified on product label.DescriptionThe Tet-On 3G Vector Set (with ZsGreen1) is used to create tightly regulated and highly responsive tetracycline (Tet)-inducible mammalian expression systems that are turned on by the addition of doxycycline to the culture medium. The Tet-On 3G Vector Set (with ZsGreen1) allows the simultaneous expression of a gene of interest and a green fluorescent protein marker.Package Contents•20 μl pCMV-Tet3G Vector (500 ng/μl)•20 μl pTRE3G-ZsGreen1 Vector(500 ng/μl)•20 μl pTRE3G-Luc Control Vector(500 ng/μl)•40 μl Linear Hygromycin Marker (50 ng/μl)•40 μl Linear Puromycin Marker (50 ng/μl)Storage Conditions•Store plasmids at –20°C.•Spin briefly to recover contents.•Avoid repeated freeze/thaw cycles.Shelf Life• 1 year from date of receipt under proper storage conditions.Storage Buffer•10 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0)Shipping Conditions• Dry ice (–70°C)Product DocumentsDocuments for our products are available for download at /manualsThe following documents apply to this product:•Tet-On 3G Expression Systems User Manual (PT5148-1)pCMV-Tet3G Vector InformationFigure 1. pCMV-Tet3G Vector Map.DescriptionThe pCMV-Tet3G Vector expresses Tet-On 3G, a tetracycline-controlled transactivator that exhibits high activity in the presence of the inducer doxycycline (Dox), and exceptionally low activity in its absence. Tet-On 3G results from the fusion of amino acids 1–207 of a mutant Tet repressor (TetR) to 39 amino acids that form three minimal "F"-type transcriptional activation domains from the herpes simplex virus VP16 protein. Tet-On 3G was derived from Tet-On Advanced (Zhou et al. 2006; Urlinger et al. 2000; Gossen and Bujard 1992; Gossen et al. 1995); as a result, it’s fully synthetic, lacks cryptic splice sites, and is codon-optimized for stable expression in mammalian cells. Compared to both of its predecessors, however, this 3rd generation Tet-On transactivator demonstrates increased sensitivity to Dox (Zhou et al. 2006). Constitutive expression of Tet-On 3G is driven by the human cytomegalovirus immediately early promoter (P CMV IE).Location of Features in pCMV-Tet3G•P CMV IE(human cytomegalovirus immediate early promoter): 2–688•Tet-On 3G (transactivator gene): 775–1521•SV40 polyA signal: 1536–1991•pUC origin of replication: 2342–2996•Amp r (ampicillin resistance gene; β-lactamase): 3144–4004 (complementary)•SV40 polyA signal: 4275–4809 (complementary)•Kan r/Neo r (kanamycin/neomycin resistance gene): 5417–6211 (complementary)•P SV40 e (SV40 early promoter): 6532–6891 (complementary)Additional InformationpCMV-Tet3G is used to develop stable Tet-On 3G cell lines, which are hosts for Tet-inducible gene expression systems. To create a Tet-inducible expression system, a vector containing a gene of interest under the control of the Tet-inducible TRE3G promoter(P TRE3G) is transfected into a Tet-On 3G cell line. The addition of Dox to the system causes Tet-On 3G to undergo a conformational change that allows it to bind to P TRE3G, activating transcription of the gene of interest in a highly dose-dependent manner. Additional information on TRE-containing vectors, and protocols describing the construction of Tet-On 3G cell lines can be found in the Tet-On 3G Expression Systems User Manual (PT5148-1).Propagation in E. coli•Suitable host strain: Stellar™ Competent Cells•Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.• E. coli replication origin: pUCpTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector InformationFigure 2. pTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector Maps.Figure 3. pTRE3G-ZsGreen Vector Multiple Cloning Site. The internal start site (ATG) at the IRES2/MCS junction is indicated in bold.DescriptionpTRE3G-ZsGreen1is a Tet-inducible, mammalian expression vector designed to coexpress a gene of interest and the green fluorescent protein ZsGreen1 under the control of the Tet-responsive promoter P TRE3G. This promoter consists of a highly optimized Tet-responsive element (TRE) just upstream of a minimal CMV promoter. P TRE3G exhibits exceptionally low basal activity; it’s induced by the binding of Tet-On 3G but is virtually silent in its absence. The vector is designed to be used as part of our Tet-On 3G Inducible Expression System (Cat. No. 631164).ZsGreen1 is a human codon-optimized variant of the reef coral Zoanthus sp. green fluorescent protein (ZsGreen) that has been engineered for brighter fluorescence (excitation and emission maxima: 493 and 505 nm, respectively; Matz et al. 1999; Haas, Park, and Seed 1996). p TRE3G-ZsGreen allows Dox-inducible coexpression of ZsGreen1 and a gene of interest from a bicistronic mRNA transcript. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES2), positioned between ZsGreen1 and the gene of interest, facilitates cap-independent translation of the gene of interest from an internal start site at the IRES2/MCS junction (Jang et al. 1988). This ensures that a high percentage of ZsGreen1-expressing clones also express the gene of interest, allowing ZsGreen1 to be used as an indicator of inducibility and transfection efficiency, as well as a marker for selection by flow cytometry. The vector also contains a pUC origin of replication and an ampicillin resistance gene (Amp r) to allow for propagation and selection in E. coli.The pTRE3G-Luc is a Tet-inducible control vector that expresses firefly luciferase under the control of P TRE3G. When used with standard luciferase detection reagents, this vector can be used as a reporter of induction efficiency (see User Manual for protocol). pTRE3G-Luc is not intended to be used as a cloning vector.Location of Features in pTRE3G-ZsGreen1•P TRE3G (3rd generation Tet-responsive promoter): 7–382•ZsGreen1: 389–1084•IRES2 (encephalomyocarditis virus internal ribosome entry site): 1091–1673•MCS (multiple cloning site): 1686–1721•SV40 polyA signal: 1776–2573•pUC origin of replication: 2838–3481•Amp r (ampicillin resistance gene; β-lactamase): 3629–4489 (complementary)Location of Features in pTRE3G-Luc•P TRE3G (3rd generation Tet-responsive promoter): 7–382•Luciferase: 432–2084•SV40 polyA signal: 2151–2948•pUC origin of replication: 3213–3856•Amp r (ampicillin resistance gene; β-lactamase): 4004–4864 (complementary)Additional InformationpTRE3G-ZsGreen1 is a mammalian expression vector that allows tightly regulated, doxycycline-controlled coexpression of a gene of interest and ZsGreen1. The gene of interest must have both a start and a stop codon. The gene of interest should be cloned in-frame with the start codon at the IRES2/MCS junction (this codon is shown in bold in the MCS sequence in Figure 3, page 3; see the User Manual for details on how to use In-Fusion® to simplify your cloning). Cotransfection of pTRE3G-ZsGreen1 constructs with Linear Hygromycin or Puromycin Markers allows antibiotic selection of stable transfectants. In order to function, the system requires the presence of the Tet-On 3G transactivator protein, supplied by a stable Tet-On 3G cell line created with our Tet-On 3G Inducible Expression System (Cat. No. 631164).Propagation in E. coli•Suitable host strain: Stellar™ Competent Cells•Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.• E. coli replication origin: pUCExcitation and Emission of pTRE3G-ZsGreen1•Excitation: 493 nm•Emission: 505 nmReferences•Gossen, M. et al. Transcriptional activation by tetracyclines in mammalian cells. Science268, 1766–9 (1995).•Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U. S. A.89, 5547–51 (1992).•Haas, J., Park, E. C. & Seed, B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr.Biol.6, 315–24 (1996).•Jang, S. K. et al. A segment of the 5’ nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J. Virol.62, 2636–43 (1988).•Matz, M. V et al. Fluorescent proteins from nonbioluminescent Anthozoa species. Nat. Biotechnol.17, 969–73 (1999).•Urlinger, S. et al. Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. U. S. A.97, 7963–8 (2000).•Zhou, X., Vink, M., Klaver, B., Berkhout, B. & Das, A. T. Optimization of the Tet-On system for regulated gene expression through viral evolution. Gene Ther.13, 1382–90 (2006).Quality Control DataPlasmid Identity & Purity•Digestion with the indicated restriction enzymes produced fragments of the indicated sizes on a 0.8% agarose/EtBr gel:Vector Enzyme(s) Fragment(s)pCMV-Tet3G EcoRI 7.1 kbEcoRI & HindIII 1.2 & 5.9 kbpTRE3G-ZsGreen1 XhoI 4.7 kbEcoRV 1.2 & 3.5 kbpTRE3G-Luc XhoI 5.1 kbEcoRI & BamHI 2.1 & 3.0 kbLinear Hygromycin Marker HindIII & XbaI0.5, 0.6 & 1.1 kbLinear Puromycin Marker HindIII & XbaI0.45, 0.6, & 0.75 kb•Vector identity was confirmed by sequencing.•A260/A280: 1.8–2.0Functional Testing of Linear Markers•HEK 293 cells were transfected with 200 ng of either the Linear Hygromycin Marker or the Linear Puromycin Marker. After 5 hr at 37°C, the transfection solution was removed, and the cells were given fresh medium. 48 hr later, the cells were plated in two 10 cm plates. 48 hr after plating, medium containing either hygromycin orpuromycin (depending on the linear marker used to transfect the cells) was added to the plates. After 2–3 weeks, >20 clones were identified.It is certified that this product meets the above specifications, as reviewed and approved by the Quality Department.CATALOG NO.631159NOTICE TO PURCHASER:Our products are to be used for research purposes only. They may not be used for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. Our products may not betransferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without prior written approval of Takara Bio USA, Inc.Your use of this product is also subject to compliance with the licensing requirements listed below and described on the product´s web page at . It is your responsibility to review, understand and adhere to any restrictions imposed by these statements.STATEMENT 24The RCFPs (including DsRedExpress, DsRedExpress2, and E2-Crimson) are covered by one or more of thefollowing U.S. Patent Nos. 7,166,444; 7,157,565; 7,217,789; 7,338,784; 7,338,783; 7,537,915; 6,969,597; 7,150,979;7,442,522 and 8,012,682.STATEMENT 72Living Colors Fluorescent Protein Products: Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such licensespecifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc.Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please ***************************************************************************************.For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at ***********************.STATEMENT 42Use of the Tetracycline controllable expression systems (the "Tet Technology") is covered by a series of patents including U.S. Patent # 7541446, # 8383364, # 9181556 , European patents EP # 1200607, # 1954811, #2352833Academic research institutions are granted an automatic license with the purchase of this product to use the Tet Technology only for internal, academic research purposes, which license specifically excludes the right to sell, or otherwise transfer, the Tet Technology or its component parts to third parties. Notwithstanding the above, academicand not-for profit research institutions whose research using the Tet Technology is sponsored by for profitorganizations, which shall receive ownership to any data and results stemming from the sponsored research, shall need a commercial license agreement from TET Systems in order to use the Tet Technology. In accepting this license, all users acknowledge that the Tet Technology is experimental in nature. TET Systems GmbH & Co. KG makes no warranties, express or implied or of any kind, and hereby disclaims any warranties, representations, or guarantees of any kind as to the Tet Technology, patents, or products. All others are invited to request a license from TET Systems GmbH & Co. KG prior to purchasing these reagents or using them for any purpose. Takara Bio USA, Inc. is required by its licensing agreement to submit a report of all purchasers of the Tet-controllable expression system to TET Systems.For license information, please contact:GSF/CEOTET Systems GmbH & Co. KG,Im Neuenheimer Feld 58269120 Heidelberg GermanyTel: +49 6221 5880400Fax: +49 6221 5880404email:*******************or use the electronic licensing request form via /ip-licensing/licensing/for-profit-research TRADEMARKS:© 2015 Takara Bio Inc. All Rights Reserved.All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions.。

ATTENTION: Read this document and the documents listed in the Additional Resources section about installation, configuration and operation of this equipment before you install, configure, operate or maintain this product. Users are required to familiarize themselves with installation and wiring instructions in addition to requirements of all applicable codes, laws, and standards.Activities including installation, adjustments, putting into service, use, assembly, disassembly, and maintenance are required to be carried out by suitably trained personnel in accordance with applicable code of practice. If this equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired.注意：在安装、配置、操作和维护本产品前，请阅读本文档以及“其他资源”部分列出的有关设备安装、配置和操作的相应文档。

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abnormal vaginal bleeding requiring intervention had no statis-tical difference between VP and WVP patients group (p=0.3074)as other complications as well(table1).Median of related days of vaginal bleeding after the procedure were 7.4days(SD8.75)in VP group and7.34days(SD8.52)in WVP group,with no statistical difference(p=0.912). Conclusions Insert a vaginal pack or not,after LEEP,do not affect the number of postoperative gynecologic intervention due to vaginal bleeding or the amount of postoperative bleed-ing days.Previous pregnancies,hormonal status,cytology or LEEP specimen characteristics did not affect the disclosure. We also could not find any risk factor associated to abnormal bleeding.Based on that,the use of vaginal pack can be omit-ted with no further complications.IGCS19-0405382LATERALLY EXTENDED ENDOPELVIC RESECTION(LEER) AND NEOVAGINE,PATIENT WITH RECTALADENOCARCINOMA AND RECURRENCE IN CERVIX,VAGINA AND PELVIC WALL:A PURPOSE OF A CASE1J Torres*,2J Saenz,3O Suescun,3M Medina,4L Trujillo.1Especialista en entrenamiento–Universidad Militar Nueva Granada–Instituto Nacional de Cancerologia,Department of Gynecologic Oncology,Bogota D.C.,Colombia;2Especialista en entrenamiento–Universidad Militar Nueva Granada–Instituto Nacional de Cancerologia,Department of Gynecologic Oncology,Bogota D.C,Colombia;3Instituto Nacional de Cancerologia, Department of Gynecologic Oncology,Bogota D.C,Colombia;4Instituto Nacional de Cancerologia,Department of Gynecologic Oncology,Bogota D.C.,Colombia10.1136/ijgc-2019-IGCS.382Objectives Exenteration is used to treat cancers of the lower and middle female genital tract in the irradiated pelvis. Höckel described laterally extended endopelvic resection (LEER)as an approach in which the resection line extends to the pelvic side wall.Methods A49-year-old patient diagnosed with rectal adenocar-cinoma10years ago,managed with chemotherapy plus radio-therapy.T umor relapse at3years,management with low abdominoperineal resection and definitive colostomy.Second relapse4years later,compromising the posterior aspect of the coccyx and right side of the pelvis with irresecability criteria, management was decided with chemotherapy with capecita-bine,oxaliplatin and bevacizumab.New relapse at2years in the cervix,vagina and pelvic wall.Images without distance disease,type LEER management with extension of pelvic floor margins and resection of muscle pubococcygeus and right lat-eral iliococcygeus with neovagina(Singapore flap)and non-continent urinary derivation with bilateral cutaneous ureteros-tomy,achieving adequate lateral margin with curative intent. During follow-up with favorable evolution.Results LEER combines at least two procedures:total mesorec-tal excision,total mesometrial resection or total mesovesical resection.It may even require resection of the pelvic wall, internal obturator muscle,pubococcygeus,iliococcygeus,coccy-geus or internal iliac vessels.In combination with neovagina, it would offer better results in non-gynecological cancer relapses.Conclusions LEER with neovagina can be offered as a new therapy to a selected subset of patients with relapse in adja-cent gynecological organs with good oncological,functional and aesthetic results.Symptom Management–Supportive Cancer CareIGCS19-0706383PHOTOBIOMODULATION AND MANUAL LYMPHDRAINAGE FOR NIPPLE NECROSIS TREATMENT INBREAST CANCER:A CASE REPORT1J Baiocchi,2L Campanholi,3G Baiocchi*.1Oncofisio,Physical Therapy,Sao Paulo,Brazil;2CESCAGE,Physical Therapy,Ponta Grossa,Brazil;3AC Camargo Cancer Center, Gynecologic Oncology,Sao Paulo,Brazil10.1136/ijgc-2019-IGCS.383Objectives Recently,breast reconstruction after mastectomywith nipple preservation became an option of breast cancer surgery.Despite its efficacy and aesthetic superiority,the nip-ple preservation is associated with several complications in the postoperative period.The photobiomodulation therapy,for-merly known as low-intensity laser therapy,demonstrated tis-sue promotion repair by cellular repair biostimulation, angiogenesis and anti-inflammatory effects.These characteris-tics suggest a potential role for repair of chronic wounds andmay be applicable in necrosis treatment.Our aim was toreport the effects of the physiotherapeutic intervention through photobiomodulation therapy in a patient with nipple necrosis after risk reducing mastectomy.Methods We report a case of a breast cancer surgery with nip-ple necrosis treated with low-level laser therapy.The patientwas a36-year-old women who developed skin nipple necrosisin the right breast after bilateral reconstructive mastectomy.She had6sessions of low-level laser therapy.Results A female subject developed a nipple necrosis of morethan40%on the right breast after mastectomy and recon-struction.She was referred to Physical Therapy(PT)and thePT sessions were composed by manual lymph drainage,man-ual therapy for de AWS,exercises of strength and flexibility, followed by LLLT with laser660nm,2joules per point atevery1cm.Therapy was implemented for12times in total,from May2016to June2016.A re-evaluation was performed monthly from July13,2016to November2017.After18 months of follow-up,the sustained effects of LLLT were found.Conclusions Low-level laser therapy is effective for the skin cicatrization after nipple necrosis.IGCS19-0446384CONTRACEPTION AND FERTILITY COUNSELING INPATIENTS RECEIVING CHEMOTHERAPY1A Elnaggar*,2A Calfee,1LB Daily,2T Hasley,1T Tillmanns.1West Cancer Center and Research Institute,Gynecologic Oncology,Memphis,USA;2University of Tennessee Health Science Center,Obstetrics and Gynecology,Mempis,USA10.1136/ijgc-2019-IGCS.384Objectives Cancer care advances allow more patients to pursue fertility.Unfortunately,treatments may have detrimental effectson fertility and fetus should pregnancy occur.This study examines physician documentation and patient perceptions of fertility and contraception counseling. on December 24, 2023 by guest. Protected by copyright./ Int J Gynecol Cancer: first published as 10.1136/ijgc-2019-IGCS.384 on 18 September 2019. Downloaded fromMethods IRB approval obtained for a cross-sectional study of men and women,ages18–50,with newly diagnosed malig-nancy between May2017and2018.Prior sterilization,secon-dary or synchronous cancer,or prior chemotherapy were exclusionary.Consented patients received a survey regarding perception on receipt and quality of,counseling.Demographic, sexual,and social information was obtained.Differences were evaluated using chi-square tests.Results Fifty-three of179patients identified participated. Majority were women(75v25%).Patients were more likely to have perceived counseling for contraception and fertility than documented.The majority perceived counseling as suffi-cient regarding contraception and fertility.Men were more likely than women to be perceive counsel-ing regarding fertility(85v43%,p=0.010).However,both felt fertility counseling to be sufficient with similar rates of documentation.Caucasians were more likely to perceive receipt of fertility counseling(68v29%)and to perceive it to be sufficient(70v40%),then African Americans,with the same rate of documentation(35%).Conclusions Significant discrepancies in perception counsel-ing regarding contraception and fertility were seen.Gen-der and race were important factors for the perception of fertility counseling,while only race was a factor to qual-ity of perceived counseling.These differences occurred despite equal rates of physician documentation,across all groups.IGCS19-0430385WHO ARE YOU CALLING OLD?PRACTICE PATTERNS AND MANAGEMENT OF NONAGENARIANS PRESENTINGTO A GYNECOLOGIC ONCOLOGIST FOR INITIALCONSULTATIONE Ryan*,B Margolis,B Pothuri.New York University Langone Health,Obstetrics and Gynecology,New York,USA10.1136/ijgc-2019-IGCS.385Objectives T o describe the practice patterns and treatment of nonagenarians who initiated care with a gynecologic oncologist.Methods Retrospective chart review of women aged90or older who presented to a gynecologic oncologist between10/ 09and12/18at an urban academic medical center.Descrip-tive statistics utilized for variables of interest.Results We identified34nonagenarians(median age92,range 90–98):10(29%)had benign disease,8(24%)pre-malignancy or suspected malignancy,and16(47%)malignancy.Of these, 79%had age and/or functional status discussed in the care plan.Of the8with suspected malignancy,5declined further workup.The cancer distribution revealed5(31%)vulvar,5 (31%)uterine,4(25%)ovarian,1(6%)vaginal and1(6%) cervical bined,37%had stage I disease;6% stage3;6%stage4;13%recurrent;and25%unstaged.All received treatment plans:7(47%)with palliative intent and8 (53%)with curative intent.In the curative group,7under-went surgery(1adjuvant chemotherapy)and1chemotherapy/radiation.In the palliative group,4underwent radiation,1 chemotherapy and2declined/unknown.Overall,13(87%) completed the proposed treatment.T reatment-related complica-tions included1superficial skin infection and1thirty-day readmission.Conclusions Nonagenarians often presented with vulvar or endometrial cancer and87%successfully completed treatmentwith minimal adverse effects or toxicity.Age and/or functionalstatus were considered in the care plan for79%of women,but it did not preclude treatments that had the potential to preserve meaningful quality of life and/or cure patients oftheir disease.IGCS19-0646386RISK FACTORS COMPREHENSIVE GERIATRICASSESSMENT FOR EARLY DEATH IN ELDERLY PATIENTSWITH GYNECOLOGICAL CANCER.A PROSPECTIVECOHORT STUDY1J Sales*,2C Azevedo,2C santos,3L sales,4M Bezerra,5G Bezerra,4Z cavalcanti,6MJ Mello.1IMIP,Geriatric Oncology,Recife,Brazil;2IMIP,Oncology,Recife,Brazil;3FPS,Medical Course,Recife,Brazil;4IMIP,geriatric,Recife,Brazil;5HMV,oncology,caruaru,Brazil;6IMIP,post graduation,Recife,Brazil10.1136/ijgc-2019-IGCS.386Objectives T o determine risk factors for early death identifiedthe Comprehensive Geriatric Assessment(CGA)in elderly patients with gynecological cancer(EPGC).Methods Prospective cohort study.Participants with a recent diagnosis of cancer were from eight community hospitals andone cancer center in Northeast Brazil and were recruited dur-ing their first medical appointment at the outpatient oncologic clinic.A basal CGA was done before the treatment decision (ADL,Charlson Comorbidity Index-CCI,Karnofsky Perform-ance status–KPS,GDS15,IPAQ,MMSE,MNA,MNA-SF,PS,PPS,Polipharmacy,TUG).During the follow up of12 months,information about the treatments performed,the tar-geted interventions and early death was collected.Overall sur-vival was estimated using the Kaplan–Meier method,and survival curves were compared using the Log rank test for cat-egorical variables.A multivariate Cox proportional hazardsmodel was used.Results From2015–2017,84EPGC,mean age69,6±7,9;range60–96),were enrolled,25%were metastatic disease.tumor site:40,4%cervical uterine,36,9%endometrial,20,2%ovary and2,3vulva.Nine(10.7%)ECP died in less than12 months of follow-up.In our multivariate model,controlled byage,site of cancer and cancer stage,the remaining significantrisk factors were malnutrition/nonutrition determined byMNA-SF(HR3.70,95%CI1.81–5.99,p<0.001),Katz index(HR 3.60,CI 1.56–3.81,p<0.001)CCI>2(HR2,74,CI1.0.74–10.20,p=0.013)and Polipharmacy(HR2.65,CI0.71–9.81,p<0.001).Conclusions The CGA at admission identified risk factors (Nutritional risk,polypharmacy,functionality for Katz indexand comorbidity index)for premature death in EPGC.They can help to plan a personalized care. on December 24, 2023 by guest. Protected by copyright./ Int J Gynecol Cancer: first published as 10.1136/ijgc-2019-IGCS.384 on 18 September 2019. Downloaded from。

Se trata de un manual de instrucciones abreviado; sus instrucciones no sustituyen a las instrucciones de funcionamiento del equipo.La información detallada sobre el equipo puede encontrarse en el manual de instrucciones del equipo y en la documentación complementaria del mismo:Disponibles para todas las versiones del equipo mediante:•Internet: /deviceviewer•Teléfono móvil inteligente/tableta: Endress+Hauser Operations AppInstrucciones de seguridad básicasRequisitos que debe cumplir el personalPara desempeñar sus tareas, el personal debe satisfacer los requisitos siguientes:‣Debe tratarse de especialistas que cuenten con una formación apropiada ycuya cualificación sea adecuada para llevar a cabo dichas funciones y tareas ‣Es necesaria la autorización correspondiente por parte de la dirección/propiedad de la planta‣El personal debe estar bien familiarizado con las normas nacionalescorrespondientes‣Antes de empezar cualquier trabajo, deben haber leído y entendido lasinstrucciones que figuran en el manual, la documentación suplementaria y los certificados (según la aplicación)‣Seguir las instrucciones y cumplir con las condiciones básicasUso previstoEl Ceraphant es un presostato para la medición y monitorización de presiones absolutas y relativas. Los materiales del equipo de medición en contacto con el producto del proceso deben disponer de un nivel adecuado de resistencia a dichos productos.El equipo de medición puede utilizarse para realizar las siguientes mediciones (variables de proceso)•en cumplimiento de los valores de alarma especificados en "Datos técnicos"•en cumplimiento de las condiciones enumeradas en la este manual.Variable de proceso medidaPresión relativa o presión absolutaFuncionamiento seguroRiesgo de lesiones‣Use el equipo solo si está en buenas condiciones técnicas y funciona de modoseguro.‣El operario es responsable del funcionamiento sin interferencias del equipo.Zona con peligro de explosiónPara eliminar riesgos para el personal o la instalación si se usa el equipo en la zona homologada (p. ej., , seguridad para equipos a presión):‣Compruebe la placas de identificación para verificar que el equipo solicitadose puede utilizar del modo previsto en la zona homologada.Identificación del productoDirección del fabricanteEndress+Hauser SE+Co. KG Hauptstraße 179689 Maulburg, AlemaniaLugar de fabricación: Véase la placa de identificación.MontajeRequisitos de montaje•Evítese la entrada de humedad en la caja durante la instalación o el manejo del equipo, o cuando se establece el conexionado eléctrico.•No limpie ni toque las membranas de proceso con objetos duros o puntiagudos.•No retire la protección de la membrana de proceso hasta el momento mismo de instalarla.•Apriete siempre firmemente la entrada de cables.•Oriente el cable y el conector hacia abajo cuando sea posible para evitar que la humedad (p. ej., agua de lluvia o condensación) penetre.•Proteja el cabezal ante los posibles golpes.•La nota siguiente es aplicable a equipos que tengan una célula de medición de presión relativa y conector M12 o de válvula:Si un equipo calentado se enfría durante el proceso de limpieza (p. ej., por el uso de agua fría), durante un tiempo breve se forma un vacío y, enconsecuencia, puede entrar humedad en la célula de medición a través del elemento de compensación de presión (1).Riesgo de destrucción del equipo‣En este caso, monte el equipo con el compensador de presiones (1) enorientación diagonal hacia abajo –cuando sea posible– o hacia un lado.Products Solutions ServicesManual de instrucciones abreviado Ceraphant PTP33BMedición de la presión de procesoKA01623P/23/ES/02.23-00716191502023-05-01*71619150*71619150KA01623P2Endress+HauserInfluencia de la posición de instalaciónSe admite la instalación con cualquier orientación. No obstante, la orientación puede provocar un desplazamiento del punto cero, es decir, el valor medido que se muestra no es cero cuando el depósito está vacío o parcialmente lleno;consulte el manual de instrucciones.Lugar de montajeMedición de presión en gasesMonte el equipo de tal forma que la válvula de corte quede por encima del punto de medición y la condensación pueda pasar así a proceso.Medición de presión en vaporesPara la medición de presión en vapores, utilice un sifón. Un sifón reduce latemperatura a casi la temperatura ambiente. Monte el equipo preferentemente con la válvula de corte y el sifón de forma que queden por debajo del punto de medición.Puede montarse también por encima del punto de medición.Respete la temperatura ambiente máxima admisible del transmisor.Tenga en cuenta los efectos de la columna de agua hidrostática.Medición de presión en líquidosMonte el equipo con el equipo de corte y el sifón por debajo o al mismo nivel que el punto de medición.Tenga en cuenta los efectos de la columna de agua hidrostática.Medición de nivel•Instale el equipo siempre por debajo del punto de medición más bajo.•No instale el aparato en ninguna de las siguientes posiciones:•En la cortina de producto •En la salida del depósito•en la zona de influencia de una bomba de succión•O en algún punto del depósito en el que puedan actuar pulsos de presión procedentes del agitador.•Puede realizar una prueba de funcionamiento más fácilmente si monta los equipos aguas abajo de una válvula de corte.Conexión eléctricaConexión de la unidad de medición Asignación de terminales L ADVERTENCIARiesgo de lesiones debido a la activación sin control de procesos.‣Desconecte la tensión de alimentación antes de conectar el equipo.‣Asegúrese de que los procesos aguas abajo no arranquen de manerainvoluntaria.L ADVERTENCIA¡Una conexión incorrecta pone en peligro la seguridad eléctrica!‣De conformidad con la norma IEC/EN 61010, se debe proporcionar para elequipo un disyuntor adecuado.‣El equipo se debe hacer funcionar con un fusible de hilo fino de630 mA(acción lenta).‣El equipo dispone de circuitos de protección contra la inversión de polaridad.AVISODaños en la entrada analógica del PLC derivados de una conexión incorrecta ‣No conecte la salida de conmutación PNP activa del equipo a la entrada de4 … 20 mA de un PLC.Conecte el equipo de la siguiente forma:pruebe que la tensión de alimentación corresponde a la especificada en la placa de identificación.2.Conecte el equipo como se indica en el diagrama siguiente.Encienda la tensión de alimentación.Para equipos con conexión por cable: no cierre el conducto de aire de referencia (véase (a) en los siguientes planos). Proteja el conducto de aire de referencia contra la entrada de agua/condensados.1 x salida de conmutación PNP R1Para consultar otras opciones de conexión, véase el manual de instrucciones.Tensión de alimentaciónTensión de alimentación: de 10 a 30 V CC en una unidad de alimentación CCConsumo de corriente y señal de alarma1)El ajuste corriente de alarma mín. de ≤ 3,6 mA puede solicitarse a través de laestructura para cursar pedidos de productos. La corriente de alarma mín. de ≤ 3,6 mA puede configurarse en el equipo o por IO-Link.。

Formulation of Body Scrub Cream From Extract ofArabika Green Coffee (Coffea arabica L.) as AntioxidantDamayanti Hilda 1,* Aprilliani Arini 2 Clarissa D. Nancy 31,2,3 Department of Pharmacy, Sekolah Tinggi Farmasi Muhammadiyah Tangerang, Tangerang, Indonesia *Corresponding author. Email:ABSTRACTArabica green coffee beans (Coffea arabica L.) have many chemical contents in the seeds, namely tannins, alkaloids, flavonoids, coumarin and quinones. Arabica green coffee beans also have antioxidant activity because they have polyphenols. Oil cream in water is very preferred for topical use because it is easy to use and easy to clean. Span 60 and Tween 60 can be used as emulsifiers to form oil-type body scrub creams in water. The purpose of this study was to determine the secondary metabolite compounds contained in 70% ethanol extract of arabica green coffee beans (Coffea arabica L.), to formulate and determine the physical properties of 70% ethanol extract body scrub cream arabica green coffee beans (Coffea arabica L.) which good, to find out the activity of 70% ethanol extract cream of arabica green coffee beans (Coffea arabica L.) against free radicals. This research was conducted by first making extracts of arabica green coffee beans (Coffea arabica L.). Extraction was carried out using 70% ethanol solvent. The formulation of body scrub cream arabica green coffee bean extract (Coffea arabica L.) use extract concentration 1%, 3% and 5%. Then, Body scrub cream evaluated by physical examination, homogeneity, pH, viscosity, spreadability, adhesion, and room temperature storage tests. Antioxidant activity of body scrub cream Assess by calculating IC50 values. The antioxidant activity of body scrub creams at concentrations of 1%, 3% and 5% has IC50 values of 177.64 ppm; 118.10 ppm; and 99.76 ppmKeywords: Arabica green coffee beans, scrub cream, antioxidant1. INTRODUCTIONEveryone definitely yearns to look beautiful, handsome and youthful, especially for women where appearance is one of the aspects that can create high self-confidence. Even though someone is no longer a teenager, they must still look beautiful and enchanting. The appearance of skin that is smooth, without wrinkles, and radiant is every woman's dream [1]Skin care cosmetics include cosmetics to clean the skin, cosmetics to moisturize the skin, protective cosmetics and cosmetics to thin the skin or what are commonly called peels. One example of cosmetic skin care is scrubs. Lulur or body scrub can moisturize the skin, leaving the skin looking and feeling soft. Just like skin cells on the face, skin cells in the body are also regularly replaced with new, healthier cells under the skin [2]. The process of replacing these cells will slow down as we age [3].Coffee is widely cultivated in Indonesia. There are generally two types of coffee cultivated in Indonesia, namely robusta coffee and arabica coffee. Arabica coffee (Coffea arabica L.) is coffee originating from Africa, namely from the mountainous areas of Ethiopia. However, Arabica coffee is known to the public after the plant was developed outside of its original area, namely Yemen in the southern part of the Arabian peninsula. Arabica coffee plants grow lush and form small shrubs. Green coffeebeans are coffee beans that have been peeled and have not been roasted [4].The content contained in coffee is chlorogenic acid and caffeine which are phenolic compounds and flavonoid compounds [5]. These flavonoids are reducing compounds that act as antioxidants. The antioxidant properties of flavonoid compounds come from their ability to transfer an electron to free radical compounds and can inhibit oxidation reactions [6]. Based on research by [7].showed that the ethanol extract of arabica coffee fruit made in the form of cream type (M / A) has antiaging activity. Based on the research [8]shows that the Arabica coffee plant contains phenolic compounds and flavonoid compounds that act as antioxidants.Based on the research [9].shows that the ethanol extract of arabica green coffee beans has an IC50 value of 0.7 g / ml which is in the very strong category.Body scrub cream is a skin care cosmetic product that is a little harsh or commonly called obrasiver cosmetics [10]. Based on research by [11] which examined body scrub formulas using Span 60 and Tween 60 as surfactants with the most stable concentration of 2%. Ease of Use2. MATERIALS AND METHODSThe research method includes the type of research, namely laboratory experiments bymaking a body scrubProceedings of the 4th International Conference on Sustainable Innovation 2020–HealthScience and Nursing (ICoSIHSN 2020)cream formulation from 70% ethanol extract of arabica green coffee beans (Coffea arabica L.) as an antioxidant. Then the evaluation of the cream preparations includes organoleptic test, homogeneity test, spreadability test, adhesion test, pH test, viscosity test and room temperature storage test.This research was conducted from December 2019 to April 2020, conducted at the Center for Biological Research and Development - LIPI Cibinong, Jl. Raya Jakarta - Bogor for plant determination, Muhammadiyah Tangerang College of Pharmacy Jl. KH Syekh Nawawi Km. 4 No.13 Tigaraksa, Tangerang Regency for the Extraction Process, Preparation of cream preparations and for testing the antioxidant activity of various formulas for the cream of green coffee bean extract arabica (Coffea arabica L.) carried out at Poltekes Pasar Minggu Rt / Rw 06/01 Jati Padang, Pasar district Sunday, South Jakarta.2.1 MaterialsIn this study, 70% ethanol extract of arabica coffee (Coffea arabica L.) was used from the plantation of Sukamarga Village, Buay Pematang Ribu Ranau Tengah District, South Ulu Komering Organ Regency, South Sumatra. Coffee beans are obtained from ripe coffee with a reddish color. The materials used in this study were 70% ethanol, stearic acid, aquadest, adeps lanae, methyl paraben, propyl paraben, liquid paraffin, cetyl alcohol, Span 60, green coffee beans arabica coffee, tween 60. 2.2 Preparation of extractsThe sample of Arabica green coffee beans that had been mashed and weighed was 1 kg and macerated with 70% ethanol solvent. This maceration process lasts 3 x 24 hours while stirring occasionally. The purpose of stirring is to even out the concentration of the solution outside the simplicia powder grain, so that the smallest degree of concentration between the solution inside the cell and outside the cell is maintained.Then, it is filtered to separate the pulp from the macerate. Maserat was remacerated for 24 hours. After that, it is filtered again and the resulting macerate is evaporated using a rotary evaporator at a temperature of 60oC2.3 Phytochemical ScreeningPhytochemical screening in this study was conducted to determine what secondary metabolites are contained in the extract of green arabica coffee beans (Coffea arabica L.). This phytochemical screening includes testing for flavonoids, alkaloids, saponins, quinones, tannins, coumarin and steroids and triterpenoids.2.4 Formulation Body ScrubThe body scrub cream formulation begins with weighing the ingredients to be used, separating the oil phase and the liquid phase at the time of melting. The oil phase (Stearic acid, Adeps lanae, Cetyl alcohol, Span 60 then added with Propyl paraben) is melted in a porcelain dish at a temperature of 70oC over a water bath while stirring until homogeneous . The water phase (dissolved methyl paraben with water that has been heated and add propylenglycol, liquid paraffin, then add tween 60) is dissolved in a proselen dish over a water bath at the same temperature while stirring until homogeneous. Cream is made by mixing the oil phase to the water phase while stirring with an electric stirrer for 3 minutes, then leaving it for 20 seconds then stirring until it is homogeneous, after being formed the cream is added to the extract and rice powder (mesh 60/40).Furthermore, the physical stability test was carried out on the cream. Then the best formulas and formulas were optimized based on the evaluation that met the requirements, then the selected formula was used to make the cream of the Arabica green coffee bean extract.2.5 Body scrub evaluationThe body scrub cream was evaluated for organoleptic, pH, consistency, spreadability, irritability, washability and grittiness2.6 Measurement of antioxidant activity with DPPH method2.6.1 Preparation of 0.05 mM DPPH Solution Weigh 0.0098 gram DPPH powder in a 250 mL measuring flask, dissolved in methanol p.a up to 250 mL (0.1 mM)2.6.2 Preparation of Vitamin C Comparative SolutionsWeigh 0.005 grams of vitamin C powder dissolved with50 ml of methanol p.a in a 5 ml volumetric flask to obtaina concentration of 100 ppm (mother liquor). Then from the mother liquor a series of concentrations of 1 ppm, 2 ppm, 3 ppm, 4 ppm and 5 were made.2.6.3 Preparation of the Test Solution Concentration SeriesWeigh 0.0025 grams of ethanol extract of arabica green coffee beans dissolved in methanol up to 25 mL, obtained a stock solution of 100 ppm. 10 mL concentration series 3, 6, 9, 12, and 15 were made.2.6.4. Determination of the Maximum WavelengthDetermination of the maximum wavelength was carried out by measuring the absorbance of the DPPH 0.05 mM solution as much as 4 mL using a spectrophotometerwith a wavelength of 500-525 nm to obtain an absorbanceof 0.2-0.8. The wavelength that produces the greatest absorbance is the maximum wavelength2.6.5 Test the Antioxidant Activity of Cream PreparationsWeigh 1 g of cream and put it in erlenmeyer, then dissolve it in methanol p.a until the volume becomes 25 mL, heat it on a water bath until the cream and ethanol become homogeneous, then cooled in ice cubes. Do this 3 times into the test tube, then centrifuged at 3000 rpm for 10 minutes. Filtered and let stand in a dark place for 30 minutes. Read the absorbance using a UV-Vis spectrophotometer at a wavelength of 516.65 nm. Calculated% antioxidant activity of the cream of ethanol extract of arabica coffee with the formula.% Inhibition = Ac – As x 100% AcWhere Ac is the absorbance of the control reaction (containing all reagents except the sample extract) and As is the absorbance of the sample extract.3. RESULTS AND DISSCUSSION3.1 Phytochemical screening resultsBased on the table of phytochemical screening results, it is stated that the arabica green coffee bean extract positively contains flavonoids, tannins, saponins, quinones and alkaloids. Based on the previous research [8]shows that the Arabica coffee plant contains flavonoid compounds that act as antioxidants.The oxidation-reduction (redox) properties of phenolic compounds, such as flavonoids and phenolic acids,for example, help neutralize/stabilize free radicals [12].Green coffee beans contain large amounts of polyphenolic antioxidants, such as chlorogenic, caffeic, ferulic, and n-coumarinic acids [13].emulsion or cream Arabica green coffee beans, antioxidant activitas, are potential product for the prevention of UV radiation- induced and physiological aging [14].Table 1. Of phytochemical screening3.2Results of Scrub Cream Characteristics Figure 1. Result of organoleptic test scrub creamIn this study, body scrub creams of arabica green coffee beans are successfully developed from previous research from [11] with modification. Organoleptic test results with three repetitions show that the body scrub cream formula changes color every time the extract is added with a different concentration.The higher the concentration of the extract given to a preparation will experience a darker color change.Evaluation results obtained showed that the scrub cream was soft, easy to spread and applies comfortly to the skin based on spreadability fisic test. scrub cream without extract was traffic white odorless, homogenous, pH 6,8 21870 cP viscosity. Green Coffee beans scrub cream 1% was pure white, odorless, homogenous, pH 6,7 18542 cP viscosity. Green Coffee beans scrub cream 2% was light ivory, odorless, homogenous, pH 6.5 18542 cP viscosity. Green Coffee beans scrub cream 3% was ivory, odorless, homogenous, pH 6,2 17538 cP viscosity. Positive control scrub cream 2% was traffic white, odorless, homogenous, pH 4,6, 18542 cP viscosity. The pH of all formulas was in the ranged of pH of cosmteic product in accordance with SNI 16-4339-1996 (4.5-8.0)[16]. The viscosity of all formulas was in accordance with [16]SNI 16-4399-1996 about cosmetic products which states that the viscosity of cosmetic products ranges from 2000-50000 Cp. The Results of viscosity test show that the addition of arabica green coffee extract reduced the thickness of the preparation. For scrub cream, the greater the decrease in the viscosity of the sample, the more intense a scrubbing effect can be [17].The results of the adhesion evaluation test on the preparations that have been mixed with the extract, the formula for making the body scrub cream base becomes slightly thick so that it affects adhesion. The longer the adhesion of a preparation, the longer the time for drug penetration to the skin so that drug absorption will be maximized [18]. Acceptability of cream by the consumer and its effectiveness require the preparations to have specific mechanical properties, easy of removal from the container, spreadability on the skin, and rheological properties such as viscosity, elasticity, flowability or adhesiveness [19]. The better the consistency and texture of the cosmetic product, the better the smearing ability produced [20].Hydrophilic creams were chosen for incorporation of arabica green coffee extract because the emulsions o/w with a hydrophilic external phase are miscible with water and skin secretions and thus are easily removed from skin or clothing [21].3.3 Antioxidant activity3.3.1 Preparation of DPPH Main SolutionIn this study, DPPH mother liquor was made by weighing 2 mg of DPPH powder and put it in a 100 mL measuring flask, and dissolving it in p.a metabolism up to 100 mL (0.1 mM) [15].3.3.2 Determination of Operating TimeThe results of determining the operating time on the antioxidant activity test from 0 to 60 minutes did not show stable absorbance, so 30 minutes were taken to refer to the journal. Then the sample preparation was carried out by making a stock solution of 1000 ppm concentration and operating time and made a concentration series of 10 ppm, 20 ppm, 30 ppm, 40 ppm and 50 ppm.3.3.3 Antioxidant Activity Test of Arabica Green Coffee Bean ExtractThe determination of wavelength aims to obtain a wavelength that provides a maximum and stable absorbance value. The measurement of wavelength is carried out at the peak of the curve because the peak has the highest sensitivity as indicated by the highest absorbance value. The highest absorbance value and stable of DPPH solution is at a wavelength of 515-520 nm [22]. ThE result of determining. The maximum wavelength obtained in this study is 517.0 nm. Antioxidant activity of the test solution for the Arabica green coffee bean extract with an average IC50 value 14.89 μg / ml and was included in the very strong antioxidant category. the antioxidant activity increased with an increasing amount of extract. Based on test results Antioxidant activity with the DPPH method that antioxidant activity in body scrub cream formula 1 without the addition of extracts with an IC50 value of 238.08 μg / ml, this shows that formula 1 without the addition of body scrub cream preparation extract 70% ethanol extract seeds Arabica green coffee has a relatively weak antioxidant activity as a body scrub cream preparation because there are only excipients. Body scrub formula 2 cream preparation with 1% extract concentration has an IC50value of 177.64 μg / ml, this shows that formula 2 with the addition of 1% extract cream body scrub preparation with 70% ethanol extract of green arabica coffee beans has strong antioxidant activity. classified as weak as a body scrub cream preparation. Body scrub formula 3 cream preparation with 3% extract concentration has an IC50value of 118.10 μg / ml, this shows that formula 3 with the addition of a concentration of 3 extracts body scrub cream preparation with 70% ethanol extract of Arabica green coffee beans has the strength of antioxidant activity classified as being a body scrub cream preparation. The gel preparation formula 4 with an extract concentration of 5% has an IC50 value of 99.76 μg / ml, this shows that formula 4 with the addition of a 5% concentration extract of the cream body scrub preparation of 70% ethanol extract of green arabica coffee beans has the strength of antioxidant activity which is classified as strong as a body scrub cream preparation. The positive control formula 5 cream preparation, namely vitamin C, has an IC50value of 48.91 μg / ml, this shows that formula 5 with the addition of positive control, namely vitamin C cream preparation for body scrub with 70% ethanol extract of arabica green coffee beans has strong antioxidant activity. classified as very strong as a cream preparation.Topical application of such preparation provides greater photoprotective effect than peroral solution, and could be useful in human skin care. Hydrophilic cream optimal delivery system for such plant preparations containing polyphenols and flavonoid [23].Figure 2. Result of antioxidant activity test4. CONCLUSIONBased on the research that has been carried out,it can be concluded that the Arabica green coffee bean extract (Coffea arabica L.) with a concentration of 1%, 3% and5% can be formulated in a body scrub cream preparation with good results. The IC 50 antioxidant test results body scrub cream Arabica green coffee bean extract with a concentration of 1% of 177.64 μg / ml, 3% of 118.10 μg / ml and 5% of 99.76 μg / ml .REFERENCES[1] Bogadenta, A. 2012. Anticipation of Early AgingSymptoms with Herbal Potion. Yogyakarta. Blue Book Publisher.[2] Tranggono, R.I and Fatha Lathifa. 2007 Handbook ofCosmetic Science. Jakarta: Gramedia Pustaka Utama. [3] Hertina TN, Dwiyanti S. 2013. The use of whitesoybean dregs and coffee grounds with different ratios in making traditional scrubs for body care. Journal. Surabaya: Surabaya State University 2 (3): 70-77[4]Rahardjo, P. 2012. Guide for Arabica and RobustaCoffee Cultivation Cultivation. Depok: Self-help spreader[5]Andline, A.A. 2013. Antimicrobial and AntioxidantActivities of Microwave-Assisted Extracts From Coffee Ground Residue in Chiang Rai Province, Thailand. Essay. Bogor. Bogor Agricultural Institute.[6]Yuhernita, Juniarti. 2011. Analysis of SecondaryMetabolite Compounds from Methanol Extract of Surian Leaves which have Potential as Antioxidants.Makara Science. YARSI University School of Medicine. Jakarta[7]Jadoon, S., Karim, S., Asad, M, H. H., Akram, M., R.,Khan, A., K., Malik, A., Chen, C., ang Murtaza, G., 2015. Anti-Aging Potential of Phytoextract Loaded-Pharmaceutical Cream Of Human Skin Cell Longetivit. Journal oxidative Medicine and Cellular Longetivy Vol. 10, hal 1-17[8]Hudakova, J., Marcincakova, D., and Legath, J.,2016. Study Of Antioxidant Effect Types Of Coffea.Journal Vol. 60. Departement Of Pharmacology and Toxicology, University Of Veterinary Medicine and Pharmacy[9]Fidrianny, I., Annisa., Ruslan, K. 2016. AntioxidantActivities Of Arabica Green Coffea From Three Regions Using ABTS and DPPH Assays. Journal Of Pharmaceutical and Clinical Research.. Volume 9.Bandung : Institut Teknologi Bandung.[10]A lam, M. 2009. Cosmetic Dermatology For Skin OfColor. Th McGraw-Hill Companies Inc. United States[11]U lfa, M. Nur, K. Fadillah. M., 2016. Formulation andPhysical Evaluation of Body Scrub Cream from The Black Extract (Camellia sinensis) Concentration Variation of Emulgator Span-Tween 60. Jurnal.Makassar: College of Pharmacy.[12]F ernández C, San Miguel E and Fernández-Briera A2009 Superoxide dismutase and catalase: tissue activities and relation with age inthe long-lived species Margaritifera Margaritifera Biol. Res. 42 57–68[13]Y ashin, A., Yashin, Y., Wang, J. Y., & Nemzer, B.(2013). Antioxidant and antiradical activity of coffee. Antioxidants, 2(4), 230-245.[14]Buzanello, E. B., Machado, G. P., Kuhnen, S.,Mazzarino, L., & Maraschin, M. (2020).Nanoemulsions containing oil and aqueous extract of green coffee beans with antioxidant and antimicrobial activities. Nano Express, 1(1), 010058.[15]Molyneux, P. 2018. The use of the stable free radicaldiphenylpicryl- hydrazyl (DPPH) for estimating antioxidant activity[16]Standar Nasional Indonesia. 1996. Sediaan TabirSurya. SNI 16-4399-1996. Badan Standar Nasional.[17]Hasan, N., Biak, D. R. A., & Kamarudin, S. (2012).Application of bacterial cellulose (BC) in natural facial scrub. International Journal on Advanced Science, Engineering and Information Technology, 2(4), 272-275.[18]Prabandari, R. (2018). Formulasi dan uji stabilitassediaan lulur dari rimpang kunyit (Curcuma longa linn). Viva Medika: Jurnal Kesehatan, Kebidanan dan Keperawatan, 11(3), 52-58.[19]Kulawik-Pióro, A., Ptaszek, A., & Kruk, J. (2019).Effective tool for assessment of the quality of barrier creams-relationships between rheological, textural and sensory properties. Regulatory Toxicology and Pharmacology, 103, 113-123.[20]Sampebarra A L 2016 Mempelajari kestabilan danefek iritasi sediaan lipstick yang diformulasi dengan lemak kakao Industri Hasil Perkebunan 11(2) 97-103 [21]Betageri G, Prabhu S (2002). Semisolid preparations.In: Swarbrick J, Boylan JC (eds) Encyclopedia of Pharmaceutical Technology, 2nd ed., vol. 3. New York, Basel: Marcel Dekker, Inc., pp. 2436-2457. [22]Molyneux, P. (2004). The use of the stable freeradical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin J.sci. technol, 26(2), 211-219.[23]Bernatoniene, J., Masteikova, R., Davalgiene, J.,Peciura, R.,Gauryliene, R., Bernatoniene, R., ... & Muselik, J. (2011). Topical application of Calendula officinalis (L.): Formulation and evaluation of hydrophilic cream with antioxidant activity. Journal of Medicinal Plants Research, 5(6), 868-877.。

copper centers(Cu A and Cu B)and two hemes—low-spin heme a and high-spin heme a3.Despite many years of research,the individual absolute absorption spectra of the two hemes in the Soret band(420–460nm)have not yet been resolved because they overlap strongly. There is but a single classical work of Vanneste[1]reporting the absolute individual spectra of the reduced hemes a and a3.We revisited the problem with new approaches as summarized below.(1)Calcium binding to mitochondrial COX induces a small red shift of the absorption spectrum of heme a.Treating the calcium-induced difference spectrum as thefirst derivative(differential)of the ab-sorption spectrum of the reduced heme a,it is possible to reconstruct the line shape of the parent absolute spectrum of a2+by integration. The Soret band absolute spectrum of the reduced heme a obtained in this way differs strongly form that in ref.[1].It is fairly symmetric and can be easily approximated by two10nm Gaussians with widely split maxima at442and451nm.In contrast to Vanneste,no evidence for the~428nm shoulder is observed for heme a2+.(2)The overall Soret band of the reduced COX reveals at least5 more Gaussians that are not affected by Ca2+.Two of them at436 and443nm can be attributed to electronic B0transitions in heme a3, and two more can represent their vibronic satellites.(3)A theoretical dipole–dipole interaction model was developed [2]for calculation of absorption and CD spectra.The model allows to optimize parameters of the B x,y electronic transitions in the hemes a and a3to obtain bestfit to the experimental spectra.The optimized parameters agree with the characteristics of the reconstructed spectra of hemes a and a3.References[1]W.H.Vanneste,The stoichiometry and absorption spectra ofcomponents a and a-3in cytochrome c oxidase,Biochemistry,5 (1966)838–48.[2]A.V.Dyuba,A.M.Arutyunyan,T.V.Vygodina,N.V.Azarkina,A.V.Kalinovich,Y.A.Sharonov,and A.A.Konstantinov,Circular dichroism of cytochrome c oxidase,Metallomics,3(2011),417–432.doi:10.1016/j.bbabio.2014.05.171S9.P8Flavodiiron enzymes as oxygen and/or nitric oxide reductases Vera Gonçalves a,b,João B.Vicente b,c,Liliana Pinto a,Célia V.Romão a, Carlos Frazão a,Paolo Sarti d,e,f,Alessandro Giuffrèf,Miguel Teixeira a a Instituto de Tecnologia Química e Biológica António Xavier,Universidade Nova de Lisboa,Av.da República,2781–901Oeiras,Portugalb Metabolism and Genetics Group,Institute for Medicines and Pharmaceutical Sciences(iMed.UL),Faculty of Pharmacy,University of Lisbon,Av.Prof.Gama Pinto,1649–003Lisboa,Portugalc Department of Biochemistry and Human Biology,Faculty of Pharmacy, University of Lisbon,Av.Prof.Gama Pinto,1649-003Lisboa,Portugald Department of Biochemical Sciences,Sapienza University of Rome,Piazzale Aldo Moro5,I-00185Rome,Italye Fondazione Cenci Bolognetti—Istituto Pasteur,Italyf Institute of Biology,Molecular Medicine and Nanobiotechnology,National Research Council of Italy(CNR),ItalyE-mail:**************.ptThe Flavodiiron proteins(FDPs)are present in all life domains, from unicellular microbes to higher eukaryotes.FDPs reduce oxygen to water and/or nitrous oxide to nitrous oxide,actively contributing to combat the toxicity of O2or NO.The catalytic ability of FDPs is comparable to that of bonafide heme–copper/iron O2/NO transmem-brane reductases.FDPs are multi-modular water soluble enzymes, exhibiting a two-domain catalytic core,whose the minimal functional unit is a‘head-to-tail’homodimer,each monomer being built by a beta-lactamase domain harbouring a diiron catalytic site,and a short-chainflavodoxin,binding FMN[1–3].Despite extensive data collected on FDPs,the molecular determi-nants defining their substrate selectivity remain unclear.To clarify this issue,two FDPs with known and opposite substrate preferences were analysed and compared:the O2-reducing FDP from the eukaryote Entamoeba histolytica(EhFdp1)and the NO reductase FlRd from Escherichia coli.While the metal ligands are strictly conserved in these two enzymes,differences near the active site were observed.Single and double mutants of the EhFdp1were produced by replacing the residues in these positions with their equivalent in the E.coli FlRd.The biochemical and biophysical features of the EhFdp1WT and mutants were studied by potentiometric-coupled spectroscopic methods(UV–visible and EPR spectroscopies).The O2/NO reactivity was analysed by amperometric methods and stopped-flow absorption spectroscopy.The reactivity of the mutants towards O2was negatively affected, while their reactivity with NO was enhanced.These observations suggest that the residues mutated have a role in defining the substrate selectivity and reaction mechanism.References[1]C.Frazao,G.Silva,C.M.Gomes,P.Matias,R.Coelho,L.Sieker,S.Macedo,M.Y.Liu,S.Oliveira,M.Teixeira,A.V.Xavier,C.Rodrigues-Pousada,M.A.Carrondo,J.Le Gall,Structure of a dioxygen reduction enzyme from Desulfovibrio gigas,Nature Structural Biology,7(2000)1041–1045.[2]J.B.Vicente,M.A.Carrondo,M.Teixeira,C.Frazão,FlavodiironProteins:Nitric Oxide and/or Oxygen Reductases,in:Encyclopedia of Inorganic and Bioinorganic Chemistry,(2011).[3]V.L.Gonçalves,J.B.Vicente,L.M.Saraiva,M.Teixeira,FlavodiironProteins and their role in cyanobacteria,in: C.Obinger,G.A.Peschek(Eds.)Bioenergetic Processes of Cyanobacteria,Springer Verlag,(2011),pp.631–656.doi:10.1016/j.bbabio.2014.05.172S9.P9CydX is a subunit of Escherichia coli cytochrome bd terminal oxidase and essential for assembly and stability of the di-heme active siteJo Hoeser a,Gerfried Gehmann a,Robert B.Gennis b,Thorsten Friedrich ca Institut für Biochemie/Uni Freiburg,Germanyb Department of Biochemistry,University of Illinois at Urbana Champaign, USAc Albert-Ludwigs-Universitat Freiburg,GermanyE-mail:*****************.uni-freiburg.deThe cytochrome bd ubiquinol oxidase is part of many prokaryotic respiratory chains.It catalyzes the oxidation of ubiquinol to ubiqui-none while reducing molecular oxygen to water.The reaction is coupled to the vectorial transfer of1H+/e−across the membrane, contributing to the proton motive force essential for energy consum-ing processes.The presence of this terminal oxidase is known to be related to the virulence of several human pathogens,making it a very attractive drug target.The three heme groups of the oxidase are presumably located in subunit CydA.Heme b558is involved in ubiquinol oxidation,while the reduction of molecular oxygen is catalyzed by a di-nuclear heme center containing hemes b595and d [1].A severe change in Escherichia coli phenotype was noticed when a 111nt gene,denoted as cydX and located at the5′end of the cyd operon,was deleted.This small gene codes for a single transmem-brane helix obviously needed for the activity of the oxidase[2].WeAbstracts e98overproduced the terminal oxidase with and without the cydX gene product.The resulting enzyme was purified by chromatographic steps and the cofactors were spectroscopically characterized.We demon-strated that CydX tightly binds to the CydAB complex and is co-purified.The identity of CydX was determined by mass spectrometry. Additionally,the di-heme active site was only detectable in the variant containing CydX.Thus,CydX is the third subunit of the E.coli bd oxidase and is essential for the assembly and stability of the di-heme site[3].References[1]V.B.Borisov,R.B.Gennis,J.Hemp,M.I.Verkhovsky,The cytochromebd respiratory oxygen reductases,Biochim.Biophys.Acta.1807 (2011)1398–1413./10.1016/j.bbabio.2011.06.016.[2]C.E.VanOrsdel,S.Bhatt,R.J.Allen,E.P.Brenner,J.J.Hobson,A.Jamil,et al.,The Escherichia coli CydX protein is a member of the CydAB cytochrome bd oxidase complex and is required for cytochrome bd oxidase activity,J.Bacteriol.195(2013)3640–3650./10.1128/JB.00324-13.[3]J.Hoeser,S.Hong,G.Gehmann,R.B.Gennis,T.Friedrich,SubunitCydX of Escherichia coli cytochrome bd ubiquinol oxidase is essential for assembly and stability of the di-heme active site,FEBS Lett.(2014)./10.1016/j.febslet.2014.03.036.doi:10.1016/j.bbabio.2014.05.173S9.P10Characterization of the two cbb3-type cytochrome c oxidase isoforms from Pseudomonas stutzeri ZoBellMartin Kohlstaedt a,Hao Xie a,Sabine Buschmann a,Anja Resemann b, Julian nger c,Hartmut Michel ca MPI of Biophysics,Germanyb Bruker Daltonik GmbH,Germanyc Max-Planck-Institute of Biophysics,Department of Molecular Membrane Biology,GermanyE-mail:*****************************.deCytochrome c oxidases(CcOs)are the terminal enzymes of the respiratory chain and are members of the heme-copper oxidase superfamily(HCO).CcOs catalyze the reduction of molecular O2to water and couple this exergonic reaction with transmembrane proton pared to family A and B CcOs,the cbb3-type CcOs which represent the C-family,feature a distinctly different subunit composition,a reduced proton pumping stoichiometry and higher catalytic activity at low oxygen concentrations[1][2].The genome of Pseudomonas stutzeri ZoBell contains two independent cbb3-operons, encoding Cbb3-1(CcoNOP)and Cbb3-2(CcoNOQP).We generated variants with a focus on ccoQ whose function is unknown.The purified variants and the wildtype Cbb3were analyzed using UV–vis spec-troscopy,BN-and SDS-PAGE,O2reductase activity(ORA)and immunoblotting with an antibody specific for CcoQ.We found that the deletion of ccoQ has an influence on a b-type heme in the binuclear center,and that both the stability and the ORA are decreased without ccoQ compared to the WT.The O2affinity(OA)of Cbb3was spec-trophotometrically determined with oxygenated leghemoglobin as an O2delivery system.The determined Km values for the recombinant Cbb3-1are similar to previously published data[2].The Km value of rec.Cbb3-2is about2-fold higher than the value of rec.Cbb3-1.In addition,the OA and ORA of different variants introduced into the O2-cavity of rec.Cbb3-1show significant differences compared to the WT. In the structure of Cbb3,an additional transmembraneαhelix was detected but so far not assigned to any protein[3].We sequenced and identified the polypeptide chain using a customized MALDI-Tandem-MS-based setup and found a putative protein.The amino acid sequence of this proteinfits the electron density of the unknown helix and we are currently investigating the functional relevance of this protein.References[1]RS.Pitcher,NJ.Watmough The bacterial cytochrome cbb3oxidaseBiochim Biophys Acta,1655(2004),pp.388–399[2]O.Preisig,R.Zufferey,L.Thöny-Meyer,C.A.Appleby,H.HenneckeA high-affinity cbb3-type cytochrome oxidase terminates thesymbiosis-specific respiratory chain of Bradyrhizobium japonicum J.Bacteriol,178(1996),pp.1532–1538[3]S.Buschmann,E.Warkentin,H.Xie,nger,U.Ermler,H.MichelThe structure of cbb3cytochrome oxidase provides insights into proton pumping Science,329(2010),pp.327–330.doi:10.1016/j.bbabio.2014.05.174S9.P11Expression of terminal oxidases under nutrient-limited conditions in Shewanella oneidensis MR-1Sébastien Le Laz a,Arlette Kpebe b,Marielle Bauzan c,Sabrina Lignon d, Marc Rousset a,Myriam Brugna aa BIP,CNRS,Marseille,Franceb BIP,CNRS/AMU,Francec CNRS,Aix-Marseille Université,Unitéde fermentation,FR3479,IMM, Franced CNRS,Aix-Marseille Université,Plate-forme Protéomique,FR3479,IMM, MaP IBiSA,FranceE-mail:***************.frShewanella species are facultative anaerobic bacteria renowned for their remarkable respiratory versatility that allows them to use,in addition to O2,a broad spectrum of compounds as electron acceptors. In the aerobic respiratory chain,terminal oxidases catalyze the last electron transfer step by reducing molecular oxygen to water.The genome of Shewanella oneidensis MR-1encodes for three terminal oxidases:a bd-type quinol oxidase and two heme-copper oxidases, a A-type cytochrome c oxidase(Cox)and a cbb3-type oxidase.In a previous study,we investigate the role of these terminal oxidases under aerobic and microaerobic conditions in rich medium using a biochemical approach[1].Our results revealed the particularity of the aerobic respiratory pathway in S.oneidensis since the cbb3-type oxidase was the predominant oxidase under aerobic conditions while the bd-type and the cbb3-type oxidases were involved in respira-tion at low-O2tensions.Against all expectation,the low-affinity Cox oxidase had no physiological significance in our experimental conditions.Do these data reflect a functional loss of Cox resulting from evolutionary mechanisms as suggested by Zhou et al.[2]?Is Cox expressed under specific conditions like the aa3oxidase in Pseudo-monas aeruginosa,maximally expressed under starvation conditions [3]?To address these questions,we investigated the expression pattern of the terminal oxidases under nutrient-limited conditions and different dissolved O2tensions by measuring oxidase activities coupled to mass-spectrometry analysis.In addition to the notable modulation of the expression of the bd-type and cbb3-type oxidases in the different tested conditions,we detected Cox oxidase under carbon-starvation conditions.This constitutes thefirst report of a condition under which the A-type oxidase is expressed in S.oneidensis. We suggest that Cox may be crucial for energy conservation in carbon-limited environments and we propose that Cox may be a component of a general protective response against oxidative stress allowing S.oneidensis to thrive under highly aerobic habitats.Abstracts e99。

3B SCIENTIFIC® PHYSICSIstruzioni per l’uso10/15 ALF1 Spinotto da 4 mm per ilcollegamento dell’anodo2 Anodo3 Supporto4 Spirale riscaldante5 Piastra catodica6 Connettore da 4 mm peril collegamento diriscaldamento e anodo I tubi catodici incandescenti sono bulbi in vetro apareti sottili, sotto vuoto. Maneggiare con cura:rischio di implosione!∙Non esporre i tubi a sollecitazionimeccaniche.∙Non esporre il cavi di collegamento asollecitazioni alla trazione.∙Il tubo può essere utilizzato esclusivamentecon il supporto D (1008507).Tensioni e correnti eccessive e temperaturecatodiche non idonee possono distruggere i tubi.∙Rispettare i parametri di funzionamento indicati.Durante il funzionamento dei tubi, possonoessere presenti tensioni e alte tensioni cherendono pericoloso il contatto.∙Eseguire i collegamenti soltanto congliapparecchi di alimentazione disinseriti.∙Montare e smontare il tubo soltanto con gliapparecchi di alimentazione disinseriti.Durante il funzionamento il collo del tubo siriscalda.∙Se necessario far raffreddare i tubi prima dismontarli.Il rispetto della Direttiva CE per la compatibilitàelettromagnetica è garantito solo con glialimentatori consigliati.Il diodo consente test fondamentali sull´effettoEdison (effetto termoionico), serve perdimostrare la dipendenza della corrente diemissione dalla potenza di accensione delcatodo incandescente, per il rilevamento dellelinee caratteristiche del diodo nonché l’uso deldiodo come raddizzatore.Il diodo è un tubo a vuoto spinto con unfilamento caldo (catodo) in tungsteno puro e unapiastra metallica circolare (anodo) in una sferadi vetro trasparente, sotto vuoto. Catodo eanodo sono disposti parallelamente tra loro.Questa forma costruttiva planare corrisponde alsimbolo del diodo tradizionale. La capacità dipotenza della grande struttura geometrica èstata migliorata fissando una piastra metallicacircolare a una delle guide del filamento caldo,in modo da determinare un campo elettrico piùuniforme tra catodo e anodo.Tensione di accensione: ≤ 7,5 V Corrente di accensione: ≤ ca. 3 A Tensione anodo: max. 500 V Corrente anodo: tip. 2,5 mA conU A= 300 V,U F = 6,3 V CC Lunghezza del tubo: ca. 300 mm Diametro: ca. 130 mm Distanza tra catodo eanodo: ca. 15 mmPer il funzionamento del diodo sono inoltre necessari i seguenti dispositivi:1 Portatubo D 1008507 1 Alimentatore CC 500 V (@230 V) 1003308 oppure1 Alimentatore CC 500 V (@115 V) 1003307In aggiunta si consiglia:Adattatore di protezione bipolare 10099614.1 Inserimento del tubo nel portatubi∙Montare e smontare il tubo soltanto con gli apparecchi di alimentazione disinseriti.∙Spingere completamente all'indietro il dispositivo di fissaggio del portavalvole.∙Inserire il tubo nei morsetti.∙Bloccare il tubo nei morsetti mediante i cursori di fissaggio.∙Se necessario, inserire un adattatore di protezione sui jack di collegamento del tubo.4.2 Rimozione del tubo dal portatubi∙Per rimuovere il tubo, spingere di nuovo all'indietro i cursori di fissaggio e rimuoverlo.5.1 Produzione di portatori di caricamediante un catodo incandescente (effetto Edison) nonché misurazione della corrente anodica in funzione della tensione di accensione del catodo incandescenteSono necessari inoltre:1 Multimetro analogico AM50 1003073 ∙Realizzare il collegamento come illustrato in figura 1. Collegare il polo negativo della tensione anodica al connettore da 4 mmcontrassegnato con il segno meno sul collo del tubo.∙Avviare il test con un riscaldamento freddo (tensione di accensione U F = 0 V).∙Variare la tensione anodica U A tra 0 e 300 V. In pratica non c’è pass aggio di corrente (< 0,1 µA) tra catodo e anodo, anche se in presenza di alte tensioni.∙Applicare una tensione di 6 V al riscaldamento finché diventa caldo.Aumentare gradualmente la tensione anodica e misurare la corrente anodica.∙Riazzerare la tensione di accensione e far raffreddare il riscaldamento. Quindi, con tensione anodica costante, aumentare gradualmente la tensione di accensione e osservare la corrente anodica I A.Con tensione di accensione costante, la corrente anodica aumenta con l’aumentare della tensione anodica.Con tensione anodica costante, la corrente anodica aumenta con l’aumentare della tensione di accensione.5.2 Rilevamento delle linee caratteristichedel diodo∙Realizzare il collegamento come illustrato in figura 1. Collegare il polo negativo della tensione anodica al connettore da 4 mm contrassegnato con il segno meno sul collo del tubo.∙Selezionare la tensione 4,5 V, 5 V e 6 V.∙Determinare la corrente anodica I A per la rispettiva tensione di accensione in funzione della tensione anodica U A. All’uopo, aumentare la tensione anodica in fasi da 40 V a 300 V.∙Riportare in un diagramma le coppie di valori I A- U A per la rispettiva tensione di accensione.Con l’aumentare della tensione anodica, la corrente anodica aumenta fino a raggiungere un valore di saturazione.Con l’aumentare della tensione di accensione, aumenta l’inte nsità della corrente anodica.5.3 Il diodo come raddrizzatoreSono necessari inoltre:1 Resistenza di 10 kΩ1 Generatore di tensione per una tensione di 16 V CA 1 Oscilloscopio∙Montaggio come illustrato in Fig. 3 con U F = 6,3 V e U A = 16 V CA.∙Sull’oscilloscopio osservare l’effetto raddizzante del diodo.Nel circuito anodico del diodo azionato con tensione alternata, è presente una corrente continua determinata dal blocco di una semifase.Fig. 1 Rapporto di dipendenza della corrente anodica dalla tensione di accensione e misurazione della correnteanodicaFig. 2 Linee caratteristiche del diodo. La corrente anodica in funzione della tensione anodicaFig. 3 Il diodo come raddrizzatore3B Scientific GmbH ▪ Rudorffweg 8 ▪ 21031 Amburgo ▪ Germania ▪ 。

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Page 1 of 2 ColiComplete ®AOAC Official Method 992.30General DescriptionColiComplete ® contains 5-bromo-4-chloro-3-indolyl-ß-Dgalactopyranoside (X-Gal) and 4-methyl umbelliferyl-ß-D-glucuronide (MUG). Discs are added to LST inoculated with selected dilutions of samples. Samples are incubated at 35–37 °C and examined after 24 and 48 ±2 h for confirmed total coliforms and after 30 ±2 h for confirmed E. coli results. ß-Galactosidase, from coliforms present in samples, cleaves X-Gal into 5-bromo-4-chloro-indoxyl intermediate which undergoes oxidation to yield water-insoluble blue dimer, visually detectable on disc or in surrounding medium as confirmed positive result for total coliform activity. ß-Glucuronidase, from E. coli present in samples, cleaves MUG into glucuronide and methyl umbelliferone which fluoresces under long wave UV light (366 nm) as confirmed positive result for E. coli presence.NOTE : As E. coli O157:H7 does not produce ß-glucuronidase, ColiComplete ® is not suitable for the detection of E. coli O157:H7.A. Sample PreparationPrepare appropriate serial dilutions as indicated in FDA Bacteriological Analytical Manual (BAM), or AOAC Official Methods of Analysis according to sample type.B. InoculationInoculate LST tubes with appropriate sample dilution series selected to determine MPN levels or presence/absence of total coliforms and E. coli in sample. Aseptically add a single ColiComplete ® disc to each tube. Incubate at 35–37 °C.C. Reading ColiComplete ®a. For total coliforms — After at least 24 h incubation, examine each tube for visually detectable blue color on disc or in surrounding medium. Presence of blue color indicates confirmed positive result for total coliforms.NOTE: A wide range of blue color intensity may be expected, depending on sample composition and microflora. All blue reactions are positive regardless of intensity of color.Reincubate at 35–37 °C. After additional 24 ±2 h re-examine. Continued absence of blue indicates negative result; presence of blue indicates confirmed positive result for total coliforms. Read and record the MPN code or presence/absence of total coliforms in the sample.b. For E.coli — After 30 ±2 h from start of initial incubation, examine tubes under long-wave UV light (366 nm). Fluorescent tubes indicate confirmed positive result for E. coli. Read and record the MPN code or presence/absence of E. coli in the sample.D. CONTROLSPositive and negative controls should be used to facilitate interpretation of MUG fluorescent reaction. Use one known positive E. coli tube and two negative controls - one non -E. coli /coliform tube (e.g., Klebsiella spp.) and one uninoculated media tube.NOTE: Use borosilicate glass tubes, flint glass gives fluorescence that may be misinterpreted for a positive result.Lit. No. MK_UG4655EN Merck KGaAFrankfurter Strasse 25064293 DarmstadtGermanyPage 2 of 2 E. Method Modification for Certain JuicesApplicable to juice products/processors which rely on treatments that do not come into direct contact with all parts of the juice, as contained in 21 CFR Part 120: Rules and Regulations. Hazard Analysis and Critical Control Point (HAACP); Procedures for the Safe and Sanitary Processing and Importing of Juice; Final Rule. Vol 66 No. 13. 6137-6202. Use the modified method “Analysis for Escherichia coli in Citrus Juices - Modifi cation of AOAC Official Method 992.30” as stated in Section 120.25 (a).F. StorageStore unused discs at 2–8 °C (36–46 °F) in a sealed container, with desiccant.G. DisposalAfter use, all tubes must be steam-sterilized at 121 °C for at least 30 min before discarding. For in-vitro diagnostic use only.Manufacturing EntityBioControl Systems, Inc, 12822 SE 32nd St, Bellevue, WA 98005, USA.BioControl Systems, Inc is an affiliate of Merck KGaA, Darmstadt, Germany.。