Recovery of activities of daily living in COVID-19 patients requiring intensive care unit or medical care unit: an observational study on the role of rehabilitation in the subacute phase

PurposeThis study aims to describe the functional status of a cohort of subacute COVID-19 patients treated in a dedicated rehabilitation unit and to compare functional outcomes between patients previously hospitalized in the intensive care unit (ICU group) and patients assisted in the medical care unit (MCU group).Materials and methodsClinical and functional evaluations were performed at admission and discharge. The functional status was assessed using Barthel index (BI), functional ambulation categories (FAC), trunk control test (TCT), and dysphagia outcome and severity score (DOSS). All patients received multidisciplinary tailored rehabilitation.ResultsWe evaluated 171 patients (with a mean age of 67.7 ± 11.9 years, 117 were males), 110 coming from the ICU (with a mean age of 63.24 ± 10.9 years), and 61 coming from the MCU (with a mean age of 75.75 ± 9.09 years). The ICU group showed a worse functional status at admission compared with the MCU group [BI 2.5 (0–20) vs. 20 (10–60), FAC 0 (0–0) vs. 0 (0–2), TCT 61 (42–100) vs. 100 (61–100), DOSS 5 (1–7) vs. 7 (7–7)] and had significantly longer hospital stay. At discharge, all functional scales were improved with no statistically significant differences between the two groups.ConclusionEarly rehabilitation of COVID-19 survivors improves functional recovery closing the initial gap between the ICU and MCU groups. In addition, it is effective to improve the functional outcome reducing the costs for longer-term assistance of COVID-19 patients

Cytotoxic Effects of 5-Azacytidine on Primary Tumour Cells and Cancer Stem Cells from Oral Squamous Cell Carcinoma: An In Vitro FTIRM Analysis

In the present study, the cytotoxic effects of 5-azacytidine on primary Oral Squamous Cell Carcinoma cells (OSCCs) from human biopsies, and on Cancer Stem Cells (CSCs) from the same samples, were investigated by an in vitro Fourier Transform InfraRed Microscospectroscopy (FTIRM) approach coupled with multivariate analysis. OSCC is an aggressive tumoral lesion of the epithelium, accounting for ~90% of all oral cancers. It is usually diagnosed in advanced stages, and this causes a poor prognosis with low success rates of surgical, as well as radiation and chemotherapy treatments. OSCC is frequently characterised by recurrence after chemotherapy and by the development of a refractoriness to some employed drugs, which is probably ascribable to the presence of CSCs niches, responsible for cancer growth, chemoresistance and metastasis. The spectral information from FTIRM was correlated with the outcomes of cytotoxicity tests and image-based cytometry, and specific spectral signatures attributable to 5-azacytidine treatment were identified, allowing us to hypothesise the demethylation of DNA and, hence, an increase in the transcriptional activity, together with a conformational transition of DNA, and a triggering of cell death by an apoptosis mechanism. Moreover, a different mechanism of action between OSSC and CSC cells was highlighted, probably due to possible differences between OSCCs and CSCs response

SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

Reliable point-of-care (POC) rapid tests are crucial to detect infection and contain the spread of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The emergence of several variants of concern (VOC) can reduce binding affinity to diagnostic antibodies, limiting the efficacy of the currently adopted tests, while showing unaltered or increased affinity for the host receptor, angiotensin converting enzyme 2 (ACE2). We present a graphene field-effect transistor (gFET) biosensor design, which exploits the Spike-ACE2 interaction, the crucial step for SARS-CoV-2 infection. Extensive computational analyses show that a chimeric ACE2-Fragment crystallizable (ACE2-Fc) construct mimics the native receptor dimeric conformation. ACE2-Fc functionalized gFET allows in vitro detection of the trimeric Spike protein, outperforming functionalization with a diagnostic antibody or with the soluble ACE2 portion, resulting in a sensitivity of 20 pg/mL. Our miniaturized POC biosensor successfully detects B.1.610 (pre-VOC), Alpha, Beta, Gamma, Delta, Omicron (i.e., BA.1, BA.2, BA.4, BA.5, BA.2.75 and BQ.1) variants in isolated viruses and patient's clinical nasopharyngeal swabs. The biosensor reached a Limit Of Detection (LOD) of 65 cps/mL in swab specimens of Omicron BA.5. Our approach paves the way for a new and reusable class of highly sensitive, rapid and variant-robust SARS-CoV-2 detection systems

Le alterazioni del cammino nelle donne trattate per carcinoma della mammella

Il Carcinoma Mammario (CM) è il tumore più comune nel sesso femminile a livello mondiale. Nonostante la disponibilità di strategie di trattamento sempre più efficaci e il progressivo aumento della sopravvivenza, le donne sopravvissute al CM (Brest Cancer Survivors-BCs) convivono con le sequele funzionali dei trattamenti oncologici. Molti di questi effetti collaterali come fatigue, disturbi posturali, limitazione funzionale dell’arto superiore o neuropatia periferica da chemioterapici, possono influenzare negativamente la capacità di deambulazione. Studi osservazionali hanno dimostrato che le donne sottoposte a intervento chirurgico per CM presentano una ridotta velocità ai test del cammino rispetto ai valori normativi. Tuttavia, fino ad oggi, la letteratura scientifica non ha fornito dati di valutazione oggettiva della deambulazione nelle BCs. Lo scopo di questo studio osservazionale è quello di valutare quantitativamente il pattern di deambulazione di una popolazione di BCs, confrontando i parametri Spazio-Temporali (ST) con quelli di donne sane con anamnesi negativa per neoplasia mammaria (Healthy Controls, HC). Sono state incluse 23 BCs reclutate presso il Centro Senologico dell’Azienda Ospedaliera Universitaria Pisana e 16 donne sane appaiate per età (HC). I criteri di inclusione per le BCs sono stati: donne di età >18 anni sottoposte a trattamento chirurgico del CM e che avevano completato i trattamenti chemioterapici e radioterapici almeno 3 mesi prima dell’arruolamento nello studio. Sono state escluse le pazienti con malattia metastatica o non controllata e pazienti con compromissione della deambulazione da patologie ortopediche e neurologiche pregresse. Tutti i soggetti hanno eseguito l’analisi del cammino con la valutazione dei seguenti parametri ST: velocità, lunghezza del passo e del semipasso, larghezza del passo, cadenza, tempo di appoggio e di doppio supporto. Tramite l’analisi statistica sono stati posti a confronto i parametri ST delle pazienti BCs con quelli dei soggetti sani. Per valutare il tempo di insorgenza delle modifiche del pattern deambulatorio, Il gruppo BCs è stato stratificato in due sottogruppi in base al tempo trascorso dall’intervento chirurgico (12 mesi) e confrontato con i soggetti sani. Infine, nell’ipotesi che le alterazioni dei parametri ST fossero più pronunciate nell’emisoma sottoposto a intervento chirurgico è stata eseguita un’analisi intrapaziente per confrontare i parametri ST del lato operato e con il controlaterale. I dati ottenuti dimostrano che le BCs presentano un alterato schema del passo, con ridotta velocità dell’andatura (0,66±0,21 vs 0,89±0,22, p<0,01), ridotta lunghezza del semipasso (0,48±0,10 vs 0,54±0,07; p<0,05) e del passo (0,90±0,16vs1,07±0,14; p<0,01) e aumento della larghezza del passo (0,12±0,03 vs 0,10±0,03; p<0,01). La cadenza è ridotta (81,12±13,68 vs 98,78±12,48; p<0,01), mentre il tempo di appoggio (63,1±3,7 vs 57,69±3,69; p<0,01) e di doppio supporto (14,53±4,85 vs 8,39±2,87; p<0,01) sono prolungati. Le modificazioni dei parametri ST sono già riscontrabili entro 12 mesi dall’intervento chirurgico, suggerendo che le alterazioni del cammino, caratterizzate da un pattern di deambulazione lento e instabile, si sviluppino precocemente, in seguito ai trattamenti oncologici. L'analisi intrapaziente dei parametri ST tra il lato operato e il controlaterale non ha rivelato differenze statisticamente significative, anche se la riduzione della lunghezza del passo e l’aumento della larghezza del passo rispetto ai valori dei soggetti normali sono state riscontrate solo sul lato operato. In conclusione, il nostro studio fornisce importanti informazioni riguardo le alterazioni del pattern deambulatorio delle pazienti operate per CM. L’analisi quantitativa del cammino evidenzia un’andatura più lenta e meno stabile riscontrabile già entro i 12 mesi dall’intervento chirurgico. Ulteriori studi che includano l’analisi cinematica e l’elettromiografia di superficie sono necessari per caratterizzare l’alterata biomeccanica deambulatoria delle BCs. Interventi riabilitativi personalizzati e precoci, entro il primo anno dalla diagnosi, potrebbero rivelarsi utili a prevenire il declino funzionale delle pazienti sopravvissute al CM

Identificazione di subsets clinico-sierologici nella sindrome di Sjögren primitiva: analisi retrospettiva di una casistica monocentrica

Introduzione: La Sindrome di Sjögren (pSS) è una malattia autoimmune, relativamente frequente e a lenta evoluzione, caratterizzata da un ampio spettro di manifestazioni cliniche organo-specifiche e sistemiche. L’espressione clinica più peculiare è l’esocrinopatia autoimmune riconducibile all’infiltrazione linfocitaria focale che compromette da un punto di vista anatomico e funzionale le ghiandole lacrimali e salivari. Mentre la sindrome sicca è un aspetto ben caratterizzato e di riscontro praticamente costante nella quasi totalità dei pazienti, i dati della letteratura relativi a prevalenza, distribuzione temporale e severità delle manifestazioni extraghiandolari della malattia sono ad oggi incompleti e derivano da studi condotti su casistiche in genere poco numerose ed eterogenee. Vi è pertanto un crescente interesse nel caratterizzare i diversi volti dell’espressione fenotipica della pSS e nell’identificare fattori prognostici sfavorevoli che possano permettere di individuare precocemente i pazienti più a rischio, candidati a ricevere trattamenti terapeutici più aggressivi. Scopo del lavoro: Lo scopo di questo studio osservazionale, condotto su una casistica monocentrica di 495 pazienti con pSS, è stato in primis la definizione della prevalenza e della distribuzione temporale delle manifestazioni extraghiandolari al fine di identificare fattori di ordine epidemiologico, clinico e sierologico che possano influenzare la storia naturale della malattia definendo, in ultima analisi, le caratteristiche del subset di pazienti ad alto rischio di complicanze potenzialmente gravi. In secondo luogo, questo studio si è proposto di descrivere le strategie terapeutiche adottate e l’adesione al trattamento focalizzando, in particolare, l’attenzione sull’idrossiclorochina in quanto farmaco più frequentemente somministrato per il controllo della malattia. Risultati: La coorte ha incluso 480 donne e 15 uomini (ratio 32:1) con una età media alla diagnosi di 51,5±14 anni seguiti per un follow up medio 6±7 anni. Nel 55% dei pazienti la malattia si è limitata all'impegno ghiandolare mentre nel corso del follow up il 45% ha sviluppato almeno una complicanza viscerale. Nell’ambito delle manifestazioni extraghiandolari, quelle più severe (impegno renale a livello glomerulare, cutaneo, neurologico e malattie linfoproliferative maligne) sono risultate presenti in meno del 15% dei pazienti inclusi. Tali manifestazioni si sono presentate più tardivamente nel decorso di malattia, dopo circa 5 anni di follow up. L'analisi multivariata ha evidenziato che la presenza di ipergammaglobulinemia, di ipocomplementemia e di autoanticorpi non organo specifici (anti-Ro/SS-A anti-La/SS-B, del Fattore Reumatoide e delle crioglobuline) si associa più frequentemente alle manifestazioni di tipo sistemico. Le manifestazioni cliniche più severe sono risultate correlate a fattori epidemiologici come l'esordio di malattia prima dei 40 anni e a fattori sierologici suggestivi dell'attivazione B linfocitaria quali l'ipocomplemantemia, la positività del Fattore Reumatoide, delle crioglobuline e l'ipergammaglobulinemia. Per quanto concerne la terapia, il 42% dei pazienti della nostra casistica è stato trattato solo con terapia sintomatica e il 41% con farmaci immunomodulanti come l'idrossiclorochina; solo l’11% dei pazienti ha richiesto una terapia con immunosoppressori. In conclusione, questo studio ha permesso di definire la prevalenza delle manifestazioni ghiandolari e sistemiche della pSS nella storia naturale della malattia, dimostrando come fattori di ordine epidemiologico e sierologico possano influenzarne l’espressione fenotipica e la severità. Lo studio conferma inoltre che la pSS nel tempo rimane una malattia relativamente benigna seppure “sistemica” e che solo una minoranza dei pazienti sono candidati ad assumere una terapia più aggressiva

Adherence to a home physical exercise program in patients with osteoporotic vertebral fractures: A retrospective observational study

Background: Adherence to treatment is one of the most common problems in patients suffering from chronic disease such as osteoporosis, and special commitment is required to patients, especially regarding rehabilitation. There is increasing evidence that physical interventions aimed at relieving pain and reducing physical impairments could play a crucial role in improving the quality of life and reducing the risk of fractures in patients with severe osteoporosis. Objective: The aim of this study was to assess the compliance and determine the acceptability of a home-self-managed exercises program in patients with vertebral fractures, one of the most frequent and serious consequences of osteoporosis. Methods: We conducted a retrospective observational study of patients undergoing a home exercise program, monitoring them with clinical scales, questionnaires, and routine visits. Results: 62.86% of the patients were compliant with the treatment; the absence of supervision by health personnel was the primary cause of non-compliance, followed by the lack of time and the lack of motivation. Compliant patients showed a significant reduction in lumbar pain (p 0.011), an improvement in posture with a reduction of dorsal kyphosis (occipital-wall distance T0-T1, p-value = 0.02) and an improvement in QoL (p-value = 0.001) and physical performance at the 20 m walking test (p-value = 0.003). Conclusions: A home exercise program is feasible and could improve signs and symptoms in patients with vertebral fractures due to OP

Overlap of ACA-positive systemic sclerosis and Sjögren's syndrome: a distinct clinical entity with mild organ involvement but at high risk of lymphoma

OBJECTIVES: We aimed to assess the prevalence of patients with either primary Sjögren's syndrome (pSS) and positive anticentromere antibodies (ACA) and secondary Sjögren's syndrome (sSS) and limited cutaneous ACA positive-systemic sclerosis (SSc) in two large cohorts of patients with pSS and SSc¸ and also to compare the clinical features of these two subsets with those of patients affected by 'ACA-positive SSc without sicca symptoms' and 'pSS'. METHODS: In this retrospective monocentric study, the case records of 'overlap' patients fulfilling both the classification criteria for SS and the LeRoy criteria for early SSc were identified from two datasets of patients with limited cutaneous ACA positive SSc (209 subjects) and with pSS (402 subjects) who attended our Rheumatology Unit in the years between 1989 and 2011. Control groups were represented by SSc subjects without sicca symptoms ('SSc group') and ACA negative Pss patients ('pSS group'). SSc patients with sicca symptoms ('Sicca-SSc group') who did not complete the diagnostic algorithm for SS were excluded from the analysis. Demographic, clinical and immunological data of the patients enrolled were collected cumulatively over the entire follow up period. Statistical analysis was performed using SPSS 13 (SPSS Inc., Chicago IL, USA). RESULTS: Out of the two datasets 41 'overlap' patients were selected. The control groups were represented by 102/209 SSc subjects without sicca symptoms ('SSc group') and 387/402 pSS patients ('pSS group'). Eighty-one 'sicca-SSc' with an incomplete work-up for SS were excluded from the analysis. The prevalence of ACA positive pSS patients among pSS was 3.7% (15/402), while the frequency of patients with definite sSS in the SSc cohort was 20% (26/128). No differences were detected between 'overlap' patients and control groups, relatively to demographic characteristics. 'Overlap patients' were characterised by a milder SSc disease (i.e. lower frequency of sclerodactily, negative evolution of the capillaroscopy pattern or absence of severe systemic involvement) whereas, as far as the SS-related manifestations were concerned, although often lacking in specific autoantibodies (i.e. rheumatoid factor, anti-Ro/SSA, anti-La/SSB), the 'overlap patients' displayed a full blown SS phenotype with recurrent salivary gland enlargement, purpura, fatigue, arthralgias, and leukocytopenia. It is noteworthy that the prevalence of non-Hodgkin's lymphoma in the 'overlap patients' was higher than in pSS. CONCLUSIONS: Taken together, the results of our work emphasise the existence of a novel distinct clinical entity which might tentatively be called 'ACA-positive limited scleroderma/SS overlap syndrome' characterised by a benign SSc clinical course but at a high risk of non-Hodgkin's lymphom

Altered type I collagen networking in osteoporotic human femoral head revealed by histomorphometric and Fourier transform infrared imaging correlated analyses

Bone homeostasis is the equilibrium between organic and inorganic components of the extracellular matrix (ECM) and cells. Alteration of this balance has consequences on bone mass and architecture, resulting in conditions such as osteoporosis (OP). Given ECM protein mutual regulation and their effects on bone structure and mineralization, further insight into their expression is crucial to understanding bone biology under normal and pathological conditions. This study focused on Type I Collagen, which is mainly responsible for structural properties and mineralization of bone, and selected proteins implicated in matrix composition, mineral deposition, and cell-matrix interaction such as Decorin, Osteocalcin, Osteopontin, Bone Sialoprotein 2, Osteonectin and Transforming Growth Factor beta. We developed a novel multidisciplinary approach in order to assess bone matrix in healthy and OP conditions more comprehensively by exploiting the Fourier Transform Infrared Imaging (FTIRI) technique combined with histomorphometry, Sirius Red staining, immunohistochemistry, and Western Blotting. This innovatory procedure allowed for the analysis of superimposed tissue sections and revealed that the alterations in OP bone tissue architecture were associated with warped Type I Collagen structure and deposition but not with changes in the total protein amount. The detected changes in the expression and/or cooperative or antagonist role of Decorin, Osteocalcin, Osteopontin, and Bone Sialoprotein-2 indicate the deep impact of these NCPs on collagen features of OP bone. Overall, our strategy may represent a starting point for designing targeted clinical strategies aimed at bone mass preservation and sustain the FTIRI translational capability as upcoming support for traditional diagnostic methods

Hyperspectral characterization of the MSTO-211H cell spheroid model: A FPA–FTIR imaging approach

none9noneValentina Notarstefano, Simona Sabbatini, Maurizio Sabbatini, Aldo Arrais, Alessia Belloni, Chiara Pro, Lisa Vaccari, Domenico Osella, Elisabetta GiorginiNotarstefano, Valentina; Sabbatini, Simona; Sabbatini, Maurizio; Arrais, Aldo; Belloni, Alessia; Pro, Chiara; Vaccari, Lisa; Osella, Domenico; Giorgini, Elisabett

Exploiting fourier transform infrared and Raman microspectroscopies on cancer stem cells from oral squamous cells carcinoma: new evidence of acquired cisplatin chemoresistance

Oral Squamous Cells Carcinoma (OSCC) is characterised by the risk of recurrence and the onset of a refractoriness response to chemotherapy drugs. These phenomena have been recently related to a subpopulation of Cancer Stem Cells (CSCs), which have either an innate or acquired drug resistance, triggered by chemotherapy treatments. In this light, to precisely target chemotherapy regimens, it is essential to improve knowledge on CSCs, with a particular focus on their molecular features. In this work, a subpopulation of CSCs, isolated by tumour sphere formation from primary OSCC cells, were treated with cisplatin for 16, 24 and 48 hours and analysed by infrared absorption and Raman microspectroscopies. CSC spectral data were compared with those obtained in previous work, for primary OSCC cells treated under the same conditions. Routine viability/apoptosis cell-based assays evidenced in CSCs and primary OSCCs, a similar degree of sensitivity to the drug at 24 hours, while a reversion of the conventional monotonic time response exhibited by OSCCs was shown by CSCs at 48 hours. This peculiar time response was also supported by the analysis of IR and Raman data, which pinpointed alterations in the lipid composition and DNA conformation in CSCs. The results obtained suggest that CSCs, although sharing with OSCC cells a similar sensitivity to cisplatin, display the onset of a mechanism of chemoresistance and enrichment of resistant CSCs as a result of drug treatment, shedding new light on the severe issue of refractoriness of some patients to chemotherapy conventionally used for OSCC