4 results on '"G. Ozzella"'
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2. Tipizzazione HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1, -DPB1 del potenziale donatore d'organo mediante tecnica di RT PCR-SSP
- Author
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G. Ozzella, E. Poggi, A. G. Bianculli, D. Colasante, M. R. Fazio, A. Giaffreda, S. Sinopoli, L. Spano, and A. Piazza
- Subjects
trapianto d'organo ,tecniche di biologia molecolare - Abstract
TIPIZZAZIONE HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1, -DPB1 DEL POTENZIALE DONATORE D'ORGANO MEDIANTE TECNICA RT PCR-SSP. Giuseppina Ozzella1,2, Elvira Poggi1,2, Antonio G. Bianculli1, Damiano Colasante1, Maria R. Fazio1, Andrea Giaffreda1, Annarita Manfreda1, Silvia Sinopoli1, Lucia Spano1, Antonina Piazza1,2. 1 Centro Regionale Trapianti - Lazio, Roma 2 C.N.R. IFT UOS di Roma S. Camillo, Roma Introduzione. Pazienti in lista d'attesa per trapianto d'organo, sensibilizzati da pregressi eventi immunizzanti, quali trasfusioni, gravidanze o trapianti, possono sviluppare ampi patterns anticorpali, specifici per molecole HLA di classe I e II. Le nuove tecniche citofluorimetriche in fase solida, attualmente utilizzate per la caratterizzazione di allo-anticorpi anti-HLA, evidenziano infatti che i pazienti sensibilizzati possono produrre anticorpi anti-HLA specifici non solo per le molecole HLA-A,-B e DRB1, attualmente considerate nel matching donatore/ricevente, ma anche per i loci HLA-C, -DRB3/4/5, -DQA1, -DQB1, -DPA1 e DPB1, sottolineando così, anche in accordo con le Linee-Guida proposte dalla Società Italiana di Immunogenetica e Biologia dei Trapianti (AIBT), la necessità di estendere la tipizzazione a tutti i loci HLA del potenziale donatore. L'impiego della nuova tecnica di biologia molecolare Real Time PCR SSP permette di eseguire, in tempi molto brevi, la tipizzazione HLA del potenziale donatore d'organi per tutti i loci HLA di classe I e II consentendo così, attraverso un più accurato virtual crossmatch pre-trapianto, di selezionare il ricevente più idoneo al trapianto e, quindi, di valutare il reale rischio immunologico nei riceventi selezionati. Materiali e metodi. Il protocollo attualmente in uso presso il Laboratorio di Tipizzazione Tissutale ed Immunologia dei Trapianti del CRT-Lazio per la tipizzazione dei donatori d'organo prevede l'utilizzo della tecnica di PCR-SSP a bassa risoluzione per i soli loci -A, -B, -C, -DRB1 e -DQB1. Al fine di estendere la tipizzazione anche agli altri loci HLA (-DRB3/4/5, -DQA1, -DQB1, -DPA1 e DPB1), finora tipizzati pre-trapianto solamente in caso di selezione di potenziali riceventi con anticorpi specifici per dette molecole, abbiamo eseguito la validazione della tecnica di tipizzazione genomica RT PCR-SSP con l'impiego di prodotti della linea LinkSeq (Linkage Bioscience Inc., Voden Medical Instrument Spa) e dello strumento QuantStudio 5 (Thermo Fisher Scientific), con software applicativo SureTyperTM. Questa tecnica combina l'amplificazione con primers allele/gruppo specifici con una rilevazione diretta dei prodotti amplificati attraverso l'uso del colorante SYBR Green, in grado di legare il DNA a doppia elica. Lo strumento di RT PCR misura, in tempo reale, la fluorescenza emessa durante la fase esponenziale dell'amplificazione e registra il decadimento della fluorescenza nel ciclo finale di dissociazione. Il Software dedicato processa i dati di dissociazione, generando, per ogni pozzetto della piastra, una curva di melting che è esaminata come deriva negativa dei valori di fluorescenza (-dF) rispetto ad un range di temperatura (dT), rappresentata, in un sistema di assi cartesiani (-dF/dT), come "picchi di melting", definiti positivi/negativi rispetto a valori attesi di reazioni note. L'analisi dei patterns di reazioni permette l'identificazione degli alleli HLA presenti nel campione. Mediante tale tecnica sono stati tipizzati retrospettivamente, o in contemporanea alla tecnica in uso, 10 potenziali donatori d'organo e le tipizzazioni ottenute sono state comparate con le tipizzazioni eseguite mediante tecnica di PCR-SSP a bassa risoluzione. Risultati. L'analisi dei risultati ha evidenziato che la tecnica RT PCR-SSP (Tabella 1) è in grado di fornire, in tempi brevi (circa 90 minuti), la tipizzazione di 11 loci HLA (-A, -B, -C, -DRB1, -DRB3,4,5, -DQA1, Tab. 1RT PCR-SSP N. TipABCDRB1DQA1DQB1DRB3/4/5DPA1DPB1 10484*01*30*42*53*04*17*11*16*01*05*03*053*01/035*01/02*01*02*02:01*04:01 12862*24*24*18*35*04*12*11-*05-*03-3*02-*01 *04:02 12866*02*24*08*35*04*07*07*11*02:01*05:01*02*033*024*01*01:03*02:01*03:01*63:01 13363*03*24*35*51*04*15*04*11*03*05*03-3*024*01*01 *03:01*04:01 14177*02*26*49*51*01*07*01-*01-*05---*01*02*03:01*10:01 14184*03*30*35*44*02*16*03*13:02*01*04:01*04*063*013*02*02 *01:01*10:01 14195*03*26*38*51*12*15*13*16*01-*05*063*01:015*02:02*02:01 *05:01*13:01 14226*01*02*35*44*04*16*04*11*03*05*03*033*024*01*01 *02:01*04:01 14227*30*30*13*51*06*15*03*07*02:01*05*02 3*014*01*01 *04:01*04:02P 14256*03*26*35*55*03*04*10*11*01*05*03*053*02-*01*02*04:01*14:01 -DQB1, DPA e DPB) ad un livello intermedio di risoluzione e consente di eliminare diverse ambiguità alleliche delle tecniche genomiche di PCR-SSP a bassa risoluzione (Tabella 2). Da un punto di vista tecnico, con l'uso di tale metodica, sono completamente eliminati numerosi passaggi manuali, come la preparazione del gel di agarosio e la manipolazione del bromuro di etidio. Tab. 2PCR-SSP N. TipABCDRB1DQA1DQB1DRB3/4/5DPA1DPB1 10484*01*30*42*53*04*17*11*16*01*05*03*05-- 12862*24*24*18*35*04*12*11-*05:05-*03-DRB3- *04:02 12866*02*24*08*35*04*07*07*11--*02*03DRB3DRB4 *03:01*63:01 13363*03*24*35*51*04*15*04*11*03:03*05:05*03-DRB3DRB4 *03:01*04:01 14177*02*26*49*51*01*07*01---*05--- 14184*03*30*35*44*02*16*03*13:01/02/15--*04*06DRB3- 14195*03*26*38*51*12*15*13*16--*05*06DRB3DRB5 14226*01*02*35*44*04*16*04*11--*03*03DRB3DRB4 14227*30*30*13*51*06*15*03*07--*02-DRB3DRB4 14256*03*26*35*55*03*04*10*11--*03*05DRB3- La tipizzazione dei potenziali donatori mediante tale tecnica avrebbe inoltre, in un caso (10%), evitato di eseguire una tipizzazione aggiuntiva del locus DRB1 a causa di una ambiguità ottenuta con la tecnica di PCR-SSP a bassa risoluzione, dovuta ad alleli "common" e "well documented" e, in 4 casi (40%), avrebbe evitato da una parte l'esecuzione di tipizzazioni aggiuntive dei loci DQA1 e DPB1 e, dall'altra, avrebbe permesso l'esclusione di due pazienti dall'esecuzione dei crossmatch pre-trapianto, poiché tali pazienti avrebbero dato virtual crossmatch positivo a causa di anticorpi donatore-specifici con intensità di fluorescenza elevata (MFI >5000). Conclusioni. La nuova tecnica RT PCR-SSP ha mostrato di essere uno strumento idoneo ad ottimizzare il matching donatore/ricevente, poiché, fornendo una tipizzazione HLA estesa a tutti i loci, con una sensibile riduzione del numero di ambiguità, consente l'esecuzione di un più accurato virtual crossmatch, utilizzato al fine di selezionare i riceventi più idonei per un donatore. Permette, quindi, una più attenta valutazione del "rischio immunologico" del potenziale ricevente di trapianto d'organo e, inoltre, risulta idonea in termini di sostenibilità economica nonché di compatibilità con l'urgenza di esecuzione della tipizzazione HLA del potenziale donatore, fornendo i risultati in tempi più rapidi rispetto a quelli che sono necessari utilizzando metodiche di PCR-SSP a bassa risoluzione.
- Published
- 2016
3. A quantitative investigation in a territory of Italy on citizens' attitudes towards medicines through the COVID-19 pandemic: the importance of possible indirect effects caused by the pandemic.
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Mastino A, Pettinau F, Palla P, Ozzella G, Grosso M, and Pittau B
- Abstract
Background: The COVID-19 pandemic has greatly influenced many aspects of everyday life, particularly that of the general population health. In order to better understand the potential impacts of the COVID-19 pandemic on people's attitudes toward medicines use, a quantitative investigation was conducted in a territory of Sardinia region, Italy., Methods: Stratification of the random multilevel population sample was based on gender, age range, and territory. The methodological strategy to verify the potential approach changes towards medicines due to the COVID-19 pandemic consisted of oral interviews with adult citizens and unrecognizability preservation. Investigation, also supported by a study completed before the insurgence of the pandemic about taking medicines, interrupting treatments without consulting, and reading the information leaflet, allowed to explore citizens' attitudes before and during pandemic, and changing., Results: The most relevant findings are the tendency towards a higher occurrence of self-interruption of treatments and an increased interest in the information leaflet (package leaflet), but not an increased self-administration of medicines., Conclusions: These results indicate new indirect effects of the COVID-19 pandemic that could exert an additional impact on the state of citizens' health and health systems. The study, with reference to prophylactic medical treatments and based on some considerations concerning the pandemic from its insurgence to today, also provides solutions for related problems for the present or future periods of health emergencies., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2024 The Institute of Translational Pharmacology, C.N.R. Published by Informa UK Limited, trading as Taylor & Francis Group.)
- Published
- 2024
- Full Text
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4. Antiviral and immunomodulatory interferon-beta in high-risk COVID-19 patients: a structured summary of a study protocol for a randomised controlled trial.
- Author
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Aricò E, Castiello L, Bracci L, Urbani F, Lombardo F, Bacigalupo I, Ancidoni A, Vanacore N, Falcione A, Reggiani C, Dutti GM, Maglie MG, Papa O, Bartoletti PL, Ozzella G, Bevilacqua N, Nicastri E, Belardelli F, and Sconocchia G
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- Aged, Clinical Trials, Phase II as Topic, Female, Humans, Male, Randomized Controlled Trials as Topic, Treatment Outcome, Antiviral Agents therapeutic use, COVID-19, HIV Infections, Interferon-beta therapeutic use
- Abstract
Objectives: The primary objective of the study is to demonstrate the efficacy of low-dose IFN-β in reducing the risk of SARS-CoV-2 recently infected elderly patients to progress towards severe COVID-19 versus control group within 28 days. Secondary objectives are: 1) To assess the reduction in Intensive Care Unit (ICU) admission in patients treated with IFN-β versus control group within 28 days of randomization 2) To assess the reduction in number of deaths in IFN- β compared to control group (day 28) 3) To evaluate the increase in proportion of participants returning to negative SARS-CoV-2 RT-PCR in IFN-β -treated versus control group at Day 14 and Day 28 4) To assess the increase in SARS-CoV-2-specific binding antibody titers in IFN-β compared to control group (day 28) 5) To assess the safety of IFN-β -treated patients versus control group TRIAL DESIGN: Randomized, Open-Label, Controlled, Superiority Phase II Study. Patients, who satisfy all inclusion criteria and no exclusion criteria, will be randomly assigned to one of the two treatment groups in a ratio 2:1 (IFN-treated versus control patients). Randomization will be stratified by gender. Stratified randomization will balance the presence of male and female in both study arms., Participants: Male and female adults aged 65 years or older with newly diagnosed SARS-CoV-2 infection and mild COVID-19 symptoms are eligible for the study. The trial is being conducted in Rome. Participants will be either hospitalized or home isolated. A group of physicians belonging to the Special Unit for Regional Continued Care (USCAR), specifically trained for the study and under the supervision of the National Institute for Infectious Diseases "Lazzaro Spallanzani", will be responsible for the screening, enrolment, treatment and clinical monitoring of patients, thus acting as a bridge between clinical centers and territorial health management. Inclusion criteria are as follows: ≥ 65 years of age at time of enrolment; Laboratory-confirmed SARS-CoV-2 infection as determined by PCR, in any specimen < 72 hours prior to randomization; Subject (or legally authorized representative) provides written informed consent prior to initiation of any study procedures; Understands and agrees to comply with planned study procedures; Agrees to the collection of nasopharyngeal swabs and venous blood samples per protocol; Being symptomatic for less than 7 days before starting therapy; NEWS2 score ≤2. Exclusion criteria are as follows: Hospitalized patients with illness of any duration, and at least one of the following: Clinical assessment (evidence of rales/crackles on exam) and SpO2 ≤ 94% on room air at rest or after walking test, OR Acute respiratory failure requiring mechanical ventilation and/or supplemental oxygen; Patients currently using IFN-β (e.g., multiple sclerosis patients); Patients undergoing chemotherapy or other immunosuppressive treatments; Patients with chronic kidney diseases; Known allergy or hypersensitivity to IFN (including asthma); Any autoimmune disease (resulting from patient anamnesis); Patients with signs of dementia or neurocognitive disorders; Patients with current severe depression and/or suicidal ideations; Being concurrently involved in another clinical trial; HIV infection (based on the anamnesis); Use of any antiretroviral medication; Impaired renal function (eGFR calculated by CKD-EPI Creatinine equation < 30 ml/min); Presence of other severe diseases impairing life expectancy (e.g. patients are not expected to survive 28 days given their pre-existing medical condition); Any physical or psychological impediment in a patient that could let the investigator to suspect his/her poor compliance; Lack or withdrawal of informed consent INTERVENTION AND COMPARATOR: Control arm: No specific antiviral treatment besides standard of care. Treatment arm: 11μg (3MIU) of IFN-β1a will be injected subcutaneously at day 1, 3, 7, and 10 in addition to standard of care. The drug solution, contained in a pre-filled cartridge, will be injected by means of the RebiSmart® electronic injection device. Interferon β1a (Rebif®, Merck KGaA, Darmstadt, Germany) is a disease-modifying drug used to treat relapsing forms of multiple sclerosis (MS). The dose selected for this study is expected to exploit the antiviral and immunomodulatory properties of the cytokine without causing relevant toxicity or inducing refractoriness phenomena sometimes observed after high-dose and/or chronic IFNβ treatments., Main Outcomes: Primary endpoint of the study is the proportion of patients experiencing a disease progression, during at least 5 days, according to the National Early Warning Score (NEWS2). The NEWS2 score is a standardized approach aimed at promptly detecting signs of clinical deterioration in acutely ill patients and establishing the potential need for higher level of care. It is based on the evaluation of vital signs, including respiratory rate, oxygen saturation, temperature, blood pressure, pulse/heart rate, AVPU response. The resulting observations, compared to a normal range, are combined in a single composite "alarm" score. Any other clinical sign clearly indicating a disease worsening will be considered as disease progression., Randomization: Sixty patients will be randomized 2:1 to receive IFN-β1a plus the standard of care or the standard of care only. Eligible patients will be randomized (no later than 36 h after enrolment) by means of a computerized central randomization system. All patients will receive a unique patient identification number at enrolling visit when signing the informed consent and before any study procedure is performed. This number remains constant throughout the entire study. The randomization of patients will be closed when 60 patients have been enrolled. The randomization will be stratified by sex; for each stratum a sequence of treatments randomly permuted in blocks of variable length (3 or 6) will be generated., Blinding (masking): This is an open-label study. After the randomization, patients will be notified whether they will be in the experimental arm or in the control arm., Numbers to Be Randomised (sample Size): The study plans to enrol 60 patients: 40 in the IFN-β1a arm, 20 in the control arm, according to a 2:1 - treated: untreated ratio., Trial Status: Protocol Version: 3.0 Version Date: 18/03/2021 The study is open for recruitment since 16/04/2021.Recruitment is expected to l be completed before 15/08/2021., Trial Registration: EudraCT N°: 2020-003872-42, registration date: 19/10/2020., Full Protocol: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.", (© 2021. The Author(s).)
- Published
- 2021
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