Spike mutation resilient scFv76 antibody counteracts SARS-CoV-2 lung damage upon aerosol delivery.

The uneven worldwide vaccination coverage against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and emergence of variants escaping immunity call for broadly effective and easily deployable therapeutic agents. We have previously described the human single-chain scFv76 antibody, which recognizes SARS-CoV-2 Alpha, Beta, Gamma and Delta variants. We now show that scFv76 also neutralizes the infectivity and fusogenic activity of the Omicron BA.1 and BA.2 variants. Cryoelectron microscopy (cryo-EM) analysis reveals that scFv76 binds to a well-conserved SARS-CoV-2 spike epitope, providing the structural basis for its broad-spectrum activity. We demonstrate that nebulized scFv76 has therapeutic efficacy in a severe hACE2 transgenic mouse model of coronavirus disease 2019 (COVID-19) pneumonia, as shown by body weight and pulmonary viral load data. Counteraction of infection correlates with inhibition of lung inflammation, as observed by histopathology and expression of inflammatory cytokines and chemokines. Biomarkers of pulmonary endothelial damage were also significantly reduced in scFv76-treated mice. The results support use of nebulized scFv76 for COVID-19 induced by any SARS-CoV-2 variants that have emerged so far.

Human inhalable antibody fragments neutralizing SARS-CoV-2 variants for COVID-19 therapy.

As of December 2021, coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a global emergency, and novel therapeutics are urgently needed. Here we describe human single-chain variable fragment (scFv) antibodies (76clAbs) that block an epitope of the SARS-CoV-2 spike protein essential for ACE2-mediated entry into cells. 76clAbs neutralize the Delta variant and other variants being monitored (VBMs) and inhibit spike-mediated pulmonary cell-cell fusion, a critical feature of COVID-19 pathology. In two independent animal models, intranasal administration counteracted the infection. Because of their high efficiency, remarkable stability, resilience to nebulization, and low cost of production, 76clAbs may become a relevant tool for rapid, self-administrable early intervention in SARS-CoV-2-infected subjects independently of their immune status.

Delayed presentation of children to the emergency department during the first wave of COVID-19 pandemic in Italy: Area-based cohort study.

AIM: To ascertain a delay in attendances to the emergency department (ED) during 2020 lockdown. METHODS: Area-based cohort study on paediatric (0-15 years) attendances resulting in hospital admission in Tuscany, Italy, in February-May 2020, and the corresponding periods in 2018-19. We analysed times from symptom onset to arrival, the odds of arriving late (>90th centile of time) and paediatricians' judgements of a late presentation by logistic models. RESULTS: As expected, ED attendance fell sharply (-62%) in 2020 vs 2018-19. As for cases studied (455 in 2020 and 1161 in 2018-19), we documented a delay in arrival to the ED in 2020 versus 2018-19 for several groups of diagnoses, namely gastroenteritis, sepsis, wounds, burns and infections overall. Time to presentation over 90th centile was also higher in 2020 (odds ratio, OR: 1.44; 95% confidence interval: 1.00, 2.06), as were paediatricians' judgements of a late arrival (18.9% of cases in 2020 vs. 13.4% in 2018-19; OR: 1.58; 1.14, 2.19) CONCLUSION: In a population-based cohort, delayed attendances to ED ascertained both subjectively and objectively convey the message to families and to paediatricians to seek hospital care in case of severe or unremitting symptoms and not to wait longer than they normally would.

Evaluating best evidence in occupational therapy for patients with hip replacement: guidelines.

SUMMARY: Object. The following study was carried out in order to evaluate through guidelines the best evidence in occupational therapy for daily activities and quality of life of patients with hip prostheses. Methods. Recommendations were generated following the grading method of the National Program for Guidelines/National System Guidelines (PNLG-SNLG), a system for developing guidelines for recommendations in clinical practice. The Appraisal of Guidelines Research and Evaluation in Europe (AGREE) tool was also applied. Results. A total of seven studies were included in this research: one randomized controlled trial, two systematic reviews, two outcomes research studies, and two observational studies. We found that, for the three clinical questions we proposed, more research on the effectiveness of treatments is required. Conclusions. The evidence resulting from this study is not sufficient to determine whether the rehabilitation techniques under consideration are effective.

Validation and Cross-Cultural Adaptation of the Volitional Questionnaire in an Italian Population with Psychiatric Disorders: A Cross-Sectional Study.

The aims of this study were to translate and culturally adapt the Volitional Questionnaire (VQ) into Italian and to evaluate its psychometric properties in an Italian population of patients with psychiatric disorders. The translation process from English into Italian was carried out in agreement with the authors of the original scale. The study was carried out in three psychiatric facilities, with 33 patients with psychiatric diagnoses, who were older than 18 years, and two raters (6 total) for each facility in Rome, Italy. The psychometric properties were assessed via its internal consistency, test-retest and inter-rater reliability and construct validity through comparisons with the Barthel Index, Short Form-12, Beck Depression Inventory-II and Camberwell Assessment of Need (CAN). Results showed internal consistency, using Cronbach's alpha, was significant at 0.92. In the test-retest reliability evaluation, the intraclass correlation coefficient was 0.97 and the interclass correlation coefficient was 0.99. The Spearman correlation showed significant results in the comparison with the CAN (r = -0.45, p < 0.05). These results suggest that the IT-VQ may be a valid, standardized and reliable evaluation tool that can be used to detect volitional performance improvements in a population of Italian psychiatric patients.

Old treatments for new genetic conditions: Sirolimus therapy in a child affected by mosaic overgrowth with fibroadipose hyperplasia.

PIK3CA-related overgrowth spectrum (PROS) are overgrowth diseases involving mesenchymal tissues caused by postzygotic variants in the PIK3CA gene. Fibro-Adipose hyperplasia or Overgrowth (FAO) belongs to PROS. We reported the beneficial effect of oral low-dose sirolimus therapy in a child affected by progressive FAO in term of stabilization of the disease, good tolerability, and easy management.

Synthesis and preliminary in vitro evaluation of DOTA-Tenatumomab conjugates for theranostic applications in tenascin expressing tumors.

Tenatumomab is an anti-tenascin murine monoclonal antibody previously used in clinical trials for delivering radionuclides to tumors by both pre-targeting (biotinylated Tenatumomab within PAGRIT) and direct 131Iodine labeling approaches. Here we present the synthesis and in vitro characterization of three Tenatumomab conjugates to bifunctional chelating agents (NHS-DOTA, NCS-DOTA and NCS-DTPA). Results indicate ST8198AA1 (Tenatumomab-DOTAMA, derived by conjugation of NHS-DOTA), as the most promising candidate in terms of conjugation rate and yield, stability, antigen immunoreactivity and affinity. Labeling efficiency of the different chelators was investigated with a panel of cold metals indicating DOTAMA as the best chelator. Labeling of Tenatumomab-DOTAMA was then optimized with several metals and stability performed confirms suitability of this conjugate for further development. ST8198AA1 represents an improvement of the previous antibody forms because the labeling with radionuclides like 177Lu or 64Cu would allow theranostic applications in patients bearing tenascin expressing tumors.

Validation of the short version of the Van Lieshout Test in an Italian population with cervical spinal cord injuries: a cross-sectional study.

STUDY DESIGN: Psychometric study. OBJECTIVE: To validate the Italian version of the Van Lieshout Test Short Version (VLT-SV) with a spinal cord injury population. SETTING: Three Italian spinal units. METHODS: The Italian version of the VLT-SV (VLT-SV-IT) was administered to a sample of people with cervical spinal cord injuries (C-SCI) and the test-retest was performed. Reliability was assessed using Cronbach's alpha for internal consistency and the intraclass correlation coefficient for repeatability assessment (test-retest). Pearson's correlation coefficient was calculated for concurrent validity with the Italian version of the Jebsen-Taylor Hand Function Test (JTHFT) and for construct validity with the Italian version of the Spinal Cord Injury Independence Measure (SCIM III). RESULTS: The VLT-SV-IT was administered to 61 individuals and all psychometric properties were significant: Cronbach's alpha was 0.95 (left hand and right hand) and the intraclass correlation coefficient for test-retest reliability was 0.90 for the right hand, the left hand, and the total score. Pearson's correlation coefficient of the VLT-SV-IT with the JTHFT was significant, while the correlation with SCIM III was not. The obtained values are considered acceptable and consistent with international guidelines. CONCLUSIONS: The VLT-SV-IT was shown to be a reliable and valid assessment tool for measuring hand function in the Italian population with C-SCI. This result suggests that it could be used as a starting point for hand therapy and to assist in clinical decision-making regarding treatment policy.

Italian Version of the Jebsen-Taylor Hand Function Test for the Assessment of Hand Disorders: A Cross-Sectional Study.

IMPORTANCE: Having a test to evaluate hand function is fundamental to occupational therapy practice. OBJECTIVE: To assess the psychometric properties of the Italian version of the Jebsen-Taylor Hand Function Test (JTHFT). DESIGN: Cross-sectional study. SETTING: Three health care institutions in Rome, Italy. PARTICIPANTS: 136 people with injuries, burns, or neurological diseases of the hand. INTERVENTION: No intervention was provided. OUTCOMES AND MEASURES: We administered the JTHFT, an assessment of fine motor skills during performance of activities of daily living, and compared results with dynamometer readings. RESULTS: The mean ± standard deviation total time required to perform all subtests was 89.47 ± 67.98 s for the dominant hand (DH) and 167.11 ± 257.58 s for the nondominant hand (NDH). Reliability procedures were applied to data from 51 participants; mean intrarater intraclass correlation coefficient (ICC) was .814 for the DH and .981 for the NDH, and mean interrater ICC was .818 for the DH and .821 for the NDH. Pearson's correlation coefficients were significant. CONCLUSION AND RELEVANCE: Results support the use of the Italian version of the JTHFT as a measure of functional dexterity in people with upper limb disorders. WHAT THIS ARTICLE ADDS: The JTHFT is a valid and reliable assessment tool for nonspecific hand diseases. Italian health professionals can now use the JTHFT with more confidence.

Growth inhibition of human ovarian carcinoma by a novel AvidinOX-anchored biotinylated camptothecin derivative.

Oxidized form of avidin, named AvidinOX, provides stable fixation of biotinylated molecules in tissues thus representing a breakthrough in topical treatment of cancer. AvidinOX proved to be a stable receptor for radiolabeled biotin, biotinylated antibodies and cells. In order to expand applicability of the AvidinOX-based delivery platform, in the present study we investigated the possibility to hold biotinylated chemotherapeutics in AvidinOX-treated sites. A novel biotinylated gimatecan-derived camptothecin, coded ST8161AA1, was injected at suboptimal doses into human tumors xenografted in mice alone or pre-complexed to AvidinOX. Significantly higher growth inhibition was observed when the drug was anchored to AvidinOX suggesting the potential utility of this delivery modality for the local treatment of inoperable tumors.

Characterization of therapeutic protein AvidinOX by an integrated analytical approach.

AvidinOX, the oxidized derivative of Avidin, is a chemically modified glycoprotein, being currently under clinical investigation for targeted delivery of radioactive biotin to inoperable tumors. AvidinOX is produced by 4-hydroxyazobenzene-2-carboxylic acid (HABA)-assisted sodium periodate oxidation of Avidin. The peculiar property of the periodate-generated glycol-split carbohydrate moieties to form Schiff's bases with amino groups of the tissue proteins allows to achieve a tissue half-life of 2 weeks compared to 2 h of native Avidin. Carbohydrate oxidation, along with possible minor amino acid modifications, introduces additional microheterogeneity in the glycoprotein structure, making its characterization even more demanding than for native glycoproteins. Aiming at the elucidation of the effects of oxidation conditions on the AvidinOX protein backbone and sugars, this microheterogeneous glycoprotein derivative was characterized for the first time using a combination of different analytical methods, including colorimetric methods, mass spectrometry, hollow-fiber flow field-flow fractionation with UV and multi-angle laser scattering detection (HF5-UV-MALS), and NMR. The proposed integrated approach reveals structural features of AvidinOX relevant for its biological activity, e.g., oxidized sites within both carbohydrate moieties and protein backbone and conformational stability, and will be considered as an analytical tool for AvidinOX industrial preparations. It is worth noting that this study enriches also the structural data of native Avidin published up-to-date (e.g., glycan structure and distribution, peptide fingerprint, etc.). Graphical abstract Scheme of phenylacetic hydrazide/MALDI-TOF approach for quantification of aldehydes in AvidinOX based on the determination of the number of hydrazone adducts between hydrazide reagent and aldehyde groups of protein.

Cross-cultural adaptation and validation of the "Spinal Cord Injury-Falls Concern Scale" in the Italian population.

STUDY DESIGN: Psychometrics study. OBJECTIVE: The objective of this study was to develop an Italian version of the Spinal Cord Injury-Falls Concern Scale (SCI-FCS) and examine its reliability and validity. SETTING: Multicenter study in spinal units in Northern and Southern Italy. The scale also was administered to non-hospitalized outpatient clinic patients. METHODS: The original scale was translated from English to Italian using the "Translation and Cultural Adaptation of Patient-Reported Outcomes Measures" guidelines. The reliability and validity of the culturally adapted scale were assessed following the "Consensus-Based Standards for the Selection of Health Status Measurement Instruments" checklist. The SCI-FCS-I internal consistency, inter-rater, and intra-rater reliability were examined using Cronbach's alpha coefficient and the intraclass correlation coefficient, respectively. Concurrent validity was evaluated using Pearson's correlation coefficient with the Italian version of the short form of the Wheelchair Use Confidence Scale for Manual Wheelchair Users (WheelCon-M-I-short form). RESULTS: The Italian version of the SCI-FCS-I was administered to 124 participants from 1 June to 30 September 2017. The mean ± SD of the SCI-FCS-I score was 16.73 ± 5.88. All SCI-FCS items were either identical or similar in meaning to the original version's items. Cronbach's α was 0.827 (p < 0.01), the inter-rater reliability was 0.972 (p < 0.01), and the intra-rater reliability was 0.973 (p < 0.01). Pearson's correlation coefficient of the SCI-FCS-I scores with the WheelCon-M-I-short form was 0.56 (p < 0.01). CONCLUSIONS: The SCI-FCS-I was found to be reliable and a valid outcome measure for assessing manual wheelchair concerns about falling in the Italian population.

Exploring Validity, Reliability, and Factor Analysis of the Quebec User Evaluation of Satisfaction with Assistive Technology in an Italian Population: A Cross-Sectional Study.

This study explores validity, reliability, and factor analysis of the Quebec User Evaluation of Satisfaction with Assistive Technology (IT-QUEST) for users of assistive technology devices in an Italian population. It was administered to 75 people to determine which of the 24 original items were relevant to the Italian context using exploratory and confirmatory factor analyses (EFAs and CFAs) were conducted with internal consistency and test-retest reproducibility also evaluated. The EFA and CFA showed that the 24-item survey may have limited relevance to the Italian context. We individuated 12 significative items, divided into two subscales, as well as the original validation. The test-retest showed high values of the Intraclass Correlation Coefficient (ICC) (0.7-0.9), with a 0.812 ICC for the total scale providing support and evidence for Italian professionals to evaluate user satisfaction with assistive technology devices. This research also lays the foundation for developing a validation study of the IT-QUEST 2.0 that is relevant to the Italian culture. Further research may investigate psychometric properties involving more participants and different users of assistive technology devices.

PTX3 binds MD-2 and promotes TRIF-dependent immune protection in aspergillosis.

The long pentraxin 3 (PTX3) modulates different effector pathways involved in innate resistance to Aspergillus fumigatus, including complement activation or promotion of phagocytosis by interacting with FcγRs. However, whether and how TLRs modulate PTX3 mediates antifungal resistance is not known. In this study, we demonstrate that PTX3 binds myeloid differentiation protein 2 (MD-2) in vitro and exerts its protective antifungal activity in vivo through TLR4/MD-2-mediated signaling. Similar to Tlr4(-/-) mice, Md2(-/-) mice displayed high susceptibility to pulmonary aspergillosis, a phenotype associated with a proinflammatory cytokine profile and impaired antifungal activity of polymorphonuclear neutrophils. Treating Md2(-/-) mice with PTX3 failed to confer immune protection against the fungus, whereas adoptive transfer of MD-2-competent polymorphonuclear neutrophils restored it. Mechanistically, engagement of MD-2 by PTX3-opsonized Aspergillus conidia activated the TLR4/Toll/IL-1R domain-containing adapter inducing IFN-ß-dependent signaling pathway converging on IL-10. Thus, we have identified a novel receptor mechanism, involving the TLR4/MD-2/Toll/IL-1R domain-containing adapter inducing IFN-ß-mediated signaling, whereby PTX3 elicits antifungal resistance with limited immunopathology in A. fumigatus infection.

[Not Available]. / Il consenso informato in Terapia Occupazionale: proposta di una modulistica.

UNLABELLED: The legislative developments that led to the Three-year Degree of the Health Professions poses any health professional in the position of having to comply with the ethical and legal duty to obtain valid informed consent from the patient prior to treatment. In the present work, attention was focused on the figure of the occupational therapist. MATERIALS AND METHODS: Informed consent forms have been developed according to the specific disease from which the patient undergoing occupational therapy is affected. The following categories of sick were identified: amputation, developmental age, orthopedy, spinal cord injury, neurology, psychiatry, geriatry and oncology. RESULTS AND CONCLUSION: The consent forms are particularly well suited to obtaining valid consent from the patient and, at the same time, allow the occupational therapist to obtain all the information he/she needs to carry out the treatment in safety. This results improved patient compliance to therapy by facilitating a better empathic relationship with the therapist.

Efficacy of PTX3 and posaconazole combination in a rat model of invasive pulmonary aspergillosis.

Posaconazole is currently used for the prophylaxis of invasive pulmonary aspergillosis (IPA). Limitations to posaconazole usage are drug-drug interactions and side effects. PTX3 is an innate immunity glycoprotein with opsonic activity, proven to be protective in IPA animal models. This study investigated the combination of posaconazole with PTX3. The results indicate synergy between PTX3 and posaconazole against aspergillosis, suggesting that a combination of reduced doses of posaconazole with the immune response enhancer PTX3 might represent a treatment option with a higher therapeutic index than posaconazole.

Simultaneous display of two large proteins on the head and tail of bacteriophage lambda.

BACKGROUND: Consistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years. The lambda phage has been engineered to display efficiently multiple copies of peptides or even large protein domains providing a powerful tool for screening libraries of peptides, proteins and cDNA. RESULTS: In the present work we describe an original method for dual display of large proteins on the surface of lambda particles. An anti-CEA single-chain antibody fragment and green fluorescent protein or alkaline phosphatase were simultaneously displayed by engineering both gpD and gpV lambda proteins. CONCLUSIONS: Here we show that such modified phage particles can be used for the detection of target molecules in vitro and in vivo. Dual expression of functional moieties on the surface of the lambda phage might open the way to generation of a new class of diagnostic and therapeutic targeted nanoparticles.

Effect of PTX3 and voriconazole combination in a rat model of invasive pulmonary aspergillosis.

This study evaluated the pharmacological activity of PTX3, administered in combination with voriconazole, in a rat model of pulmonary aspergillosis. The data indicated additive therapeutic activities of these compounds, as demonstrated by the amelioration of respiratory function changes, reduction of lung fungal burden, and increased survival. Overall, we provide clear evidence that the combination of PTX3 with a suboptimal dose of voriconazole might represent a therapeutic option under those clinical conditions where the use of voriconazole alone is not warranted for efficacy and tolerability reasons.

Biochemical and biological characterization of a new oxidized avidin with enhanced tissue binding properties.

Chicken avidin and bacterial streptavidin are widely employed in vitro for their capacity to bind biotin, but their pharmacokinetics and immunological properties are not always optimal, thereby limiting their use in medical treatments. Here we investigate the biochemical and biological properties of a new modified avidin, obtained by ligand-assisted sodium periodate oxidation of avidin. This method allows protection of biotin-binding sites of avidin from inactivation caused by the oxidation step and delay of avidin clearance from injected tissue by generation of aldehyde groups from avidin carbohydrate moieties. Oxidized avidin shows spectroscopic properties similar to that of native avidin, indicating that tryptophan residues are spared from oxidation damage. In strict agreement with these results, circular dichroism and isothermal titration calorimetry analyses confirm that the ligand-assisted oxidation preserves the avidin protein structure and its biotin binding capacity. In vitro cell binding and in vivo tissue residence experiments demonstrate that aldehyde groups provide oxidized avidin the property to bind cellular and interstitial protein amino groups through Schiff's base formation, resulting in a tissue half-life of 2 weeks, compared with 2 h of native avidin. In addition, the efficient uptake of the intravenously injected (111)In-BiotinDOTA (ST2210) in the site previously treated with modified avidin underlines that tissue-bound oxidized avidin retains its biotin binding capacity in vivo. The results presented here indicate that oxidized avidin could be employed to create a stable artificial receptor in diseased tissues for the targeting of biotinylated therapeutics.