A proof-of-concept study on the genomic evolution of Sars-Cov-2 in molnupiravir-treated, paxlovid-treated and drug-naïve patients

Little is known about SARS-CoV-2 evolution under Molnupiravir and Paxlovid, the only antivirals approved for COVID-19 treatment. By investigating SARS-CoV-2 variability in 8 Molnupiravir-treated, 7 Paxlovid-treated and 5 drug-naïve individuals at 4 time-points (Days 0-2-5-7), a higher genetic distance is found under Molnupiravir pressure compared to Paxlovid and no-drug pressure (nucleotide-substitutions/site mean±Standard error: 18.7 × 10−4 ± 2.1 × 10−4 vs. 3.3 × 10−4 ± 0.8 × 10−4 vs. 3.1 × 10−4 ± 0.8 × 10−4, P = 0.0003), peaking between Day 2 and 5. Molnupiravir drives the emergence of more G-A and C-T transitions than other mutations (P = 0.031). SARS-CoV-2 selective evolution under Molnupiravir pressure does not differ from that under Paxlovid or no-drug pressure, except for orf8 (dN > dS, P = 0.001); few amino acid mutations are enriched at specific sites. No RNA-dependent RNA polymerase (RdRp) or main proteases (Mpro) mutations conferring resistance to Molnupiravir or Paxlovid are found. This proof-of-concept study defines the SARS-CoV-2 within-host evolution during antiviral treatment, confirming higher in vivo variability induced by Molnupiravir compared to Paxlovid and drug-naive, albeit not resulting in apparent mutation selection

Effect of High-Titer Convalescent Plasma on Progression to Severe Respiratory Failure or Death in Hospitalized Patients with COVID-19 Pneumonia: A Randomized Clinical Trial

Importance: Convalescent plasma (CP) has been generally unsuccessful in preventing worsening of respiratory failure or death in hospitalized patients with COVID-19 pneumonia. Objective: To evaluate the efficacy of CP plus standard therapy (ST) vs ST alone in preventing worsening respiratory failure or death in patients with COVID-19 pneumonia. Design, Setting, and Participants: This prospective, open-label, randomized clinical trial enrolled (1:1 ratio) hospitalized patients with COVID-19 pneumonia to receive CP plus ST or ST alone between July 15 and December 8, 2020, at 27 clinical sites in Italy. Hospitalized adults with COVID-19 pneumonia and a partial pressure of oxygen-to-fraction of inspired oxygen (Pao2/Fio2) ratio between 350 and 200 mm Hg were eligible. Interventions: Patients in the experimental group received intravenous high-titer CP (≥1:160, by microneutralization test) plus ST. The volume of infused CP was 200 mL given from 1 to a maximum of 3 infusions. Patients in the control group received ST, represented by remdesivir, glucocorticoids, and low-molecular weight heparin, according to the Agenzia Italiana del Farmaco recommendations. Main Outcomes and Measures: The primary outcome was a composite of worsening respiratory failure (Pao2/Fio2ratio <150 mm Hg) or death within 30 days from randomization. Results: Of the 487 randomized patients (241 to CP plus ST; 246 to ST alone), 312 (64.1%) were men; the median (IQR) age was 64 (54.0-74.0) years. The modified intention-to-treat population included 473 patients. The primary end point occurred in 59 of 231 patients (25.5%) treated with CP and ST and in 67 of 239 patients (28.0%) who received ST (odds ratio, 0.88; 95% CI, 0.59-1.33; P =.54). Adverse events occurred more frequently in the CP group (12 of 241 [5.0%]) compared with the control group (4 of 246 [1.6%]; P =.04). Conclusions and Relevance: In patients with moderate to severe COVID-19 pneumonia, high-titer anti-SARS-CoV-2 CP did not reduce the progression to severe respiratory failure or death within 30 days. Trial Registration: ClinicalTrials.gov Identifier: NCT04716556

General practitioners as partners for a shared management of chronic HIV infection: An insight into the perspectives of Italian people living with HIV

Is it possible to achieve a collaboration between Infectious Diseases (ID) Specialists and General Practitioners (GPs) in the management of chronic HIV infection? A cross sectional survey was conducted among People Living with HIV (PLWHIV) attending the outpatient services of four Italian Infectious Diseases Centers to understand to which extent patients trust their GPs and involve them in the management of their chronic condition. Information about level of communication with GPs, subjective perception of the disease, and presence of co-medications were collected and matched with socio-demographic data using χ2statistics. A p<0.05 was considered statistically significant. From December 2019 to February 2020, 672 patients completed the survey, 59% males and 56% >50 years. Overall, 508 patients (76%) had informed GPs about HIV-positivity. Communication of diagnosis was significantly associated with age >50years, lower education level, history of disease >10 years and residency in Northern Italy. The “Undetectable = Untrasmittable” (U = U) concept was investigated as an indirect measure of perceived stigma. 23% of subjects was unaware of its meaning. Despite undetectable status, 50% of PLWHIV found difficult to communicate their condition to GPs, especially married (52% vs 48% of unmarried, p = 0.003), well-educated patients (51% vs 48, p = 0.007), living in Southern vs Northern Italy (52% vs 46%, p< 0.001). More than 75% of the participants consulted the ID specialist for co-medications and DDIs management, often complaining a lack of communication of the former with GPs. Overall, a good level of communication between PLWHIV and GPs was outlined, even if a wider involvement of the latter in HIV care is desirable

General practitioners as partners for a shared management of chronic HIV infection: An insight into the perspectives of Italian people living with HIV

Is it possible to achieve a collaboration between Infectious Diseases (ID) Specialists and General Practitioners (GPs) in the management of chronic HIV infection? A cross sectional survey was conducted among People Living with HIV (PLWHIV) attending the outpatient services of four Italian Infectious Diseases Centers to understand to which extent patients trust their GPs and involve them in the management of their chronic condition. Information about level of communication with GPs, subjective perception of the disease, and presence of co-medications were collected and matched with socio-demographic data using χ2statistics. A p<0.05 was considered statistically significant. From December 2019 to February 2020, 672 patients completed the survey, 59% males and 56% >50 years. Overall, 508 patients (76%) had informed GPs about HIV-positivity. Communication of diagnosis was significantly associated with age >50years, lower education level, history of disease >10 years and residency in Northern Italy. The “Undetectable = Untrasmittable” (U = U) concept was investigated as an indirect measure of perceived stigma. 23% of subjects was unaware of its meaning. Despite undetectable status, 50% of PLWHIV found difficult to communicate their condition to GPs, especially married (52% vs 48% of unmarried, p = 0.003), well-educated patients (51% vs 48, p = 0.007), living in Southern vs Northern Italy (52% vs 46%, p< 0.001). More than 75% of the participants consulted the ID specialist for co-medications and DDIs management, often complaining a lack of communication of the former with GPs. Overall, a good level of communication between PLWHIV and GPs was outlined, even if a wider involvement of the latter in HIV care is desirable

Generation and testing of engineered multimeric Fabs of trastuzumab

Recombinant antibodies fragments in several new formats are routinely investigated and used in diagnostic and therapeutic applications as anti-cancers molecules. New antibody formats are generated to compensate the need for multispecificity and site-specific introduction of fluorescent dyes, cytotoxic payloads or for generating semisynthetic multimeric molecules. Fabs of trastuzumab bearing transglutaminase (MTG) reactive sites were generated by periplasmic expression in E. coli and purified. Multimeric Fabs were generated by either disulfide bridge formation or by using MTG-sensitive peptide linkers. Binding to receptor was assessed by ELISA and SPR methods. Internalization and growth inhibition assays were performed on BT-474 and SKBR3 Her2+ cells. Fabs were successfully produced and dimerized or trimerized using MTG and suitably designed peptide linkers. Site-specific derivatizations with fluorophores were similarly achieved. The monomeric, dimeric and trimeric variants bind the receptor with affinities similar or superior to the full antibody. Fab and Fab2 are rapidly internalized in Her2+ cells and exhibit growth inhibition abilities similar to the full antibody. Altogether, the data show that the recombinant Fabs can be produced in E. coli and converted into multimeric variants by MTG-based bioconjugation. Similar approaches are extendable to the introduction of cytotoxic payloads for the generation of novel Antibody Drug Conjugates

Pegylated interferon α plus ribavirin for the treatment of chronic hepatitis C: A multicentre independent study supported by the Italian Drug Agency

168nononeBackground: Data on the efficacy of Peg-interferon/ribavirin therapy for chronic hepatitis C are mostly derived from treatment of selected patients enrolled in clinical trials. This study aimed to assess the effectiveness of Peg-interferon/ribavirin therapy in "real world" chronic hepatitis C patients in Italy. Methods: Independent observational multicentre study including consecutive patients receiving Peg-interferon/ribavirin in the 18 months before (retrospective phase) and after (prospective phase) the start of the study. Results: 4176 patients were eligible. The final study population consisted of 2051 patients in the retrospective and 2073 in the prospective phase.Sustained virological response was achieved by 1036 patients (50.5%) during the retrospective phase: 325 were genotypes 1/4 (34.1%) and 684 were genotypes 2/3 (67.2%) and by 800 patients (38.6%) during the prospective phase: 300 were genotypes 1/4 (28.4%) and 473 were genotypes 2/3 (51.5%).During multivariate analysis genotypes 2/3 were significantly associated with higher sustained virological response rates; cirrhosis and γ-glutamil-transpeptidase >2 times the normal limit were associated with poorer response. Conclusions: The response to Peg-interferon/ribavirin therapy in "real world" clinical practice is distinctly lower than in registration trials. The difference in response rates was more pronounced among easy-to-treat than among difficult-to-treat hepatitis C virus genotypes. © 2014 Editrice Gastroenterologica Italiana S.r.l.Rosina, Floriano; Tosti, Maria Elena; Borghesio, Elisabetta; Masocco, Maria; Mele, Alfonso; Coppola, Carmine; Milella, Michele; Borgia, Guglielmo; Andreone, Pietro; Koch, Maurizio; Zignego, Anna Linda; Romano, Mario; Carrara, Maurizio; Almasio, Piero Luigi; Azzola, Emilio; Nardone, Gerardo; Benedetti, Antonio; Carosi, Giampiero; Mazzotta, Francesco; Sagnelli, Evangelista; Rizzetto, Mario; Mascolo, M.C.; Cursaro, C.; Scuteri, A.; Crespi, C.; Gianstefani, A.; Ranieri, J.; Monti, M.; Corti, G.; Blanc, P.L.; Baragli, F.; Bellentani, S.; Gasbarrini, A.; Pompili, M.; Mecenate, F.; Picardi, A.; Vespasiani, U.; Nosotti, Null; Gasbarrini, A.; Pompili, M.; Mecenate, F.; Null, A.Picardi; Nosotti, Null; Ricci, G.L.; Paffetti, A.; Mastropietro, C.; Moretti, A.; Spagnolo, A.L.; Puoti, C.; Bellis, L.; Regazzetti, A.; Maffezzini, E.; Pietrangelo, A.; Abbati, G.; Borghi, A.; Sardini, C.; Raimondo, G.; Scribano, L.; Martines, D.; Svegliati Baroni, G.; Faraci, G.; Schi-anchi, S.; Fornaciari, G.; Massari, M.; Fabris, P.; Bertin, T.; Salvagnini, M.; Madonia, S.; Calì, A.; Civitavecchia, G.; Pirisi, M.; Smirne, C.; Andreoletti, M.; Morisco, F.; Caporaso, N.; Gentile, I.; Brancaccio, G.; Gaeta, G.B.; Liberti, A.; Iannece, M.D.; Rocco, A.; Federico, A.; Loguercio, C.; Riegler, G.; Esposito, P.; Fargion, S.; Fatta, E.; Masutti, F.; Bonaventura, M.E.; Autolitano, A.; Russello, M.; Bellia, A.; Toniutto, P.; Bitetto, D.; Pasulo, L.; Lucà, M.G.; Grattagliano, I.; Palasciano, G.; Romagno, D.; Giannelli, G.; Napoli, N.; Plattella, M.S.; Cassano, P.; Gobbo, G.; Monti, V.; Raspanti, A.; Cuccorese, Null; Colombo, A.E.; Mandelli, G.; Spinzi, G.C.; Floridia, Null; Messina, V.; Bonfante, S.; Bellissima, P.; Toti, M.; Vecchiet, J.; Falasca, K.; Portelli, V.; Stefano, G. De; Pietromatera, G.; Viganò, P.; Re, T.; Andreoni, M.; Null, G.Raineri; Grossi, P.A.; Caputo, S.; Cassola, G.; Feasi, M.; Biagio, A. Di; Nicolini, L.; Giannini, E.G.; Corbo, M.; Foti, G.; Kunkar, A.; Caterini, L.; Migliorini, D.; Chiodera, A.; Calleri, G.; Spezia, C.; Framarin, L.; Null, M.Berrutti; Ciancio, A.; Baiguera, C.; Puoti, M.; Vento, S.; Contini, C.; Boccia, S.; Casiraghi, M.A.; Simone, L.; Tacconi, D.; Caremani, M.; Almi, P.; Chimenti, M.; Cosco, Null; Messeri, D.; Esperti, F.C.; Lomonaco, L.; Pazzi, P.; Fornari, F.; Comparato, G.; Casetti, T.; Foschi, F.G.; Samori, A.; Ferretti, E.; Marin, R.; Campo, N.; Testa, R.; Rizzo, S.Rosina, Floriano; Tosti, Maria Elena; Borghesio, Elisabetta; Masocco, Maria; Mele, Alfonso; Coppola, Carmine; Milella, Michele; Borgia, Guglielmo; Andreone, Pietro; Koch, Maurizio; Zignego, Anna Linda; Romano, Mario; Carrara, Maurizio; Almasio, Piero Luigi; Azzola, Emilio; Nardone, Gerardo; Benedetti, Antonio; Carosi, Giampiero; Mazzotta, Francesco; Sagnelli, Evangelista; Rizzetto, Mario; Mascolo, M. C.; Cursaro, C.; Scuteri, A.; Crespi, C.; Gianstefani, A.; Ranieri, J.; Monti, M.; Corti, G.; Blanc, P. L.; Baragli, F.; Bellentani, S.; Gasbarrini, A.; Pompili, M.; Mecenate, F.; Picardi, A.; Vespasiani, U.; Nosotti, Null; Gasbarrini, A.; Pompili, M.; Mecenate, F.; Null, A. Picardi; Nosotti, Null; Ricci, G. L.; Paffetti, A.; Mastropietro, C.; Moretti, A.; Spagnolo, A. L.; Puoti, C.; Bellis, L.; Regazzetti, A.; Maffezzini, E.; Pietrangelo, A.; Abbati, G.; Borghi, A.; Sardini, C.; Raimondo, G.; Scribano, L.; Martines, D.; Svegliati Baroni, G.; Faraci, G.; Schi anchi, S.; Fornaciari, G.; Massari, M.; Fabris, P.; Bertin, T.; Salvagnini, M.; Madonia, S.; Calì, A.; Civitavecchia, G.; Pirisi, M.; Smirne, C.; Andreoletti, M.; Morisco, F.; Caporaso, N.; Gentile, I.; Brancaccio, G.; Gaeta, G. B.; Liberti, A.; Iannece, M. D.; Rocco, A.; Federico, A.; Loguercio, C.; Riegler, G.; Esposito, P.; Fargion, S.; Fatta, E.; Masutti, F.; Bonaventura, M. E.; Autolitano, A.; Russello, M.; Bellia, A.; Toniutto, P.; Bitetto, D.; Pasulo, L.; Lucà, M. G.; Grattagliano, I.; Palasciano, G.; Romagno, D.; Giannelli, G.; Napoli, N.; Plattella, M. S.; Cassano, P.; Gobbo, G.; Monti, V.; Raspanti, A.; Cuccorese, Null; Colombo, A. E.; Mandelli, G.; Spinzi, G. C.; Floridia, Null; Messina, V.; Bonfante, S.; Bellissima, P.; Toti, M.; Vecchiet, J.; Falasca, K.; Portelli, V.; Stefano, G. De; Pietromatera, G.; Viganò, P.; Re, T.; Andreoni, M.; Null, G. Raineri; Grossi, PAOLO ANTONIO; Caputo, S.; Cassola, G.; Feasi, M.; Biagio, A. Di; Nicolini, L.; Giannini, E. G.; Corbo, M.; Foti, G.; Kunkar, A.; Caterini, L.; Migliorini, D.; Chiodera, A.; Calleri, G.; Spezia, C.; Framarin, L.; Null, M. Berrutti; Ciancio, A.; Baiguera, C.; Puoti, M.; Vento, S.; Contini, C.; Boccia, S.; Casiraghi, M. A.; Simone, L.; Tacconi, D.; Caremani, M.; Almi, P.; Chimenti, M.; Cosco, Null; Messeri, D.; Esperti, F. C.; Lomonaco, L.; Pazzi, P.; Fornari, F.; Comparato, G.; Casetti, T.; Foschi, F. G.; Samori, A.; Ferretti, E.; Marin, R.; Campo, N.; Testa, R.; Rizzo, S

Pegylated interferon α plus ribavirin for the treatment of chronic hepatitis C: a multicentre independent study supported by the Italian Drug Agency

BACKGROUND: Data on the efficacy of Peg-interferon/ribavirin therapy for chronic hepatitis C are mostly derived from treatment of selected patients enrolled in clinical trials. This study aimed to assess the effectiveness of Peg-interferon/ribavirin therapy in "real world" chronic hepatitis C patients in Italy. METHODS: Independent observational multicentre study including consecutive patients receiving Peg-interferon/ribavirin in the 18 months before (retrospective phase) and after (prospective phase) the start of the study. RESULTS: 4176 patients were eligible. The final study population consisted of 2051 patients in the retrospective and 2073 in the prospective phase. Sustained virological response was achieved by 1036 patients (50.5%) during the retrospective phase: 325 were genotypes 1/4 (34.1%) and 684 were genotypes 2/3 (67.2%) and by 800 patients (38.6%) during the prospective phase: 300 were genotypes 1/4 (28.4%) and 473 were genotypes 2/3 (51.5%). During multivariate analysis genotypes 2/3 were significantly associated with higher sustained virological response rates; cirrhosis and γ-glutamil-transpeptidase >2 times the normal limit were associated with poorer response. CONCLUSIONS: The response to Peg-interferon/ribavirin therapy in "real world" clinical practice is distinctly lower than in registration trials. The difference in response rates was more pronounced among easy-to-treat than among difficult-to-treat hepatitis C virus genotype