Immunophenotyping of peripheral blood cells allows to discriminate MIS-C and Kawasaki disease

Background: The pathogenesis of the novel described multisystem inflammatory syndrome in children (MIS-C) and Kawasaki disease (KD) is still debated as it is not clear if they are the same or different nosological entities. However, for both the diseases a rapid and unequivocal diagnosis is mandatory to start the therapy before the onset of severe complications. In this study, we aimed to evaluate the white cell populations in MIS-C and KD as potential markers to discriminate between the two diseases. Methods: We studied white cell populations by flow cytometry in 46 MIS-C and 28 KD patients in comparison to 70 age-matched healthy children. Results: MIS-C patients had a significant lymphopenia that involved both B and T populations while KD patients showed a significant neutrophilia and thrombocythemia. Granulocyte/lymphocyte ratio helped to diagnose both MIS-C and KD with a high diagnostic sensitivity, while a multivariate analysis of granulocyte and T lymphocyte number contributed to discriminate between the two diseases. Conclusions: The relevant lymphopenia observed in MIS-C patients suggests that the disease would be a post-infectious sequel of COVID-19 immunologically amplified by a massive cytokine release, while the significant neutrophilia and thrombocythemia observed in KD confirmed that the disorder has the genesis of a systemic vasculitis. The analysis of a panel of circulating cells may help to early diagnose and to discriminate between the two diseases. Supplementary information: The online version contains supplementary material available at 10.1186/s41231-022-00128-2

Antifungal and Antibiofilm Activity of Cyclic Temporin L Peptide Analogues against Albicans and Non-Albicans Candida Species

Temporins are one of the largest families of antimicrobial peptides with both anti-inflammatory and antimicrobial activity. Herein, for a panel of cyclic temporin L isoform analogues, the antifungal and antibiofilm activities were determined against representative Candida strains, including C. albicans, C. glabrata, C. auris, C. parapsilosis and C. tropicalis. The outcomes indicated a significant anti-candida activity against planktonic and biofilm growth for four peptides (3, 7, 15 and 16). The absence of toxicity up to high concentrations and survival after infection were assessed in vivo by using Galleria mellonella larvae, and the correlation between conformation and cytotoxicity was investigated by fluorescence assays and circular dichroism (CD). By combining fluorescence spectroscopy, CD, dynamic light scattering, confocal and atomic force microscopy, the mode of action of four analogues was hypothesized. The results pinpointed that peptide 3 emerged as a non-toxic compound showing a potent antibiofilm activity and represents a promising compound for biomedical applications

MIS-C: A COVID-19-associated condition between hypoimmunity and hyperimmunity

: Multisystem inflammatory syndrome in children (MIS-C) is a rare, severe complication of COVID-19. A better knowledge of immunological, cellular, and genetic characteristics of MIS-C could help better understand the pathogenesis of the disease and contribute to identifying specific diagnostic biomarkers and develop targeted therapies. We studied 37 MIS-C children at hospital admission and 24 healthy controls analyzing serum cytokines (IFN-α, IFN-β, IFN-γ, IL-6, IL-10, IL-17A, IL-12p70 and TNF), lymphocyte populations by flow cytometry and 386 genes related to autoimmune diseases, autoinflammation and primary immunodeficiencies by NGS. MIS-C patients showed a significant increase of serum IFNγ (despite a significant reduction of activated Th1) and ILs, even if with a great heterogeneity among patients, revealing different pathways involved in MIS-C pathogenesis and suggesting that serum cytokines at admission may help to select the inflammatory pathways to target in each patient. Flow cytometry demonstrated a relevant reduction of T populations while the percentage of B cell was increased in agreement with an autoimmune pathogenesis of MIS-C. Genetic analysis identified variants in 34 genes and 83.3% of patients had at least one gene variant. Among these, 9 were mutated in more patients. Most genes are related to autoimmune diseases like ATM, NCF1, MCM4, FCN3, and DOCK8 or to autoinflammatory diseases associated to the release of IFNγ like PRF1, NOD2, and MEF. Thus, an incomplete clearance of the Sars-CoV2 during the acute phase may induce tissue damage and self-antigen exposure and genetic variants can predispose to hyper-reactive immune dysregulation events of MIS-C-syndrome. Type II IFN activation and cytokine responses (mainly IL-6 and IL-10) may cause a cytokine storm in some patients with a more severe acute phase of the disease, lymphopenia and multisystemic organ involvement. The timely identification of such patients with an immunocytometric panel might be critical for targeted therapeutic management

Single and Interactive Effects of Unmalted Cereals, Hops, and Yeasts on Quality of White-Inspired Craft Beers

White beers owe their name to their straw yellow colour deriving from the use of unmalted wheat, which also supplies a relatively high protein content causing haze formation. This study aimed to develop white-inspired craft beers made with combinations of three mixtures of barley malt/unmalted wheat (alternatively durum-var. Dauno III, soft-var. Risciola, or emmer-var. Padre Pio), two hop varieties (Cascade or Columbus), and two Saccharomyces cerevisiae strains (Belgian yeast and a high-ester producing yeast); and assess the single and interactive effects of these ingredients on physical, chemical, and sensory characteristics of the beers. According to the graphical representation of the results for the Principal Component Analysis, most of the samples appear overlapped since they had similar characteristics, but it was possible to highlight two clusters of beers different from the others: those produced with (a) Risciola wheat and Columbus hop and (b) Dauno III wheat, Cascade hop, and the Belgian yeast. The beers of these clusters obtained the highest scores for their overall quality that, in turn, was positively correlated with concentrations of citric acid, 4-hydroxybenzoic acid, syringic acid, and epicatechin; alcohol %, colour, amount and persistence of foam, intensity of fruity flavour, and body

An innovative oligonucleotide microarray to detect spoilage microorganisms in wine

The aim of this investigation has been the design and validation of an oligonucleotide microarray in order to detect 17 different wine-spoilage microorganisms, i.e. 9 yeasts, 5 lactic bacteria and 3 acetic acid bacteria species. Furthermore, several strains belonging to these species has been found to produce undesirable compounds for wine consumers. Oligonucleotide probes specific for each microorganism were designed to target the intergenic spacer regions (ISR) between 18S-5.8S region for yeasts and 16S-ITS1 region for bacteria. Prior to hybridization the ISR were amplified by polymerase chain reactions using designed consensus primers. Each oligonucleotide-probes exclusively recognized its target without undesired aspecific cross-hybridizations. Under our experimental condition, the microarray assay analysis was able to detect the amount of DNA equivalent to 24 (Saccharomyces cerevisiae), 160 (Lactobacillus brevis) and 124 (Gluconobacter oxydans) cells, three species chosen as experimental models for the three studied microbial classes. Moreover, a novel procedure that allowed the extraction of genomic DNA from a mixture of eukaryotic and prokaryotic cells from contaminated wine was developed. The obtained results confirm that the microarray assay is able to detect specifically different spoilage microorganisms present in mixture in contaminated wines. For the first time the microarray methodology has been applied for the simultaneous identification of different mixed population of spoilage yeast and bacteria directly isolated from wine, thus indicating the practicability of oligonucleotide microarrays as a contamination control in wine industry

Biomarkers of Endothelial Damage in Distinct Phases of Multisystem Inflammatory Syndrome in Children

Endothelial hyperinflammation and vasculitis are known hallmarks of acute COVID-19 and multisystem inflammatory syndrome in children (MIS-C). They are due to the direct effect of the virus on endothelial cells enhanced by pro-inflammatory modulators and may cause venous/arterial thrombosis. Therefore, it is essential to identify patients with endothelial damage early in order to establish specific therapies. We studied the monocyte chemoattractant protein 1 (MCP-1), the perinuclear anti-neutrophil cytoplasmic antibodies (pANCA), and the vascular endothelial growth factor A (VEGF-A) in serum from 45 MIS-C patients at hospital admission and 24 healthy controls (HC). For 13/45 MIS-C patients, we measured the three serum biomarkers also after one week from hospitalization. At admission, MIS-C patients had significantly higher levels of MCP-1 and VEGF-A than the HC, but no significant differences were observed for pANCA. While after one week, MCP-1 was significantly lower, pANCA was higher and VEGF-A levels were not significantly different from the admission values. These findings suggest an involvement of epithelium in MIS-C with an acute phase, showing high MCP-1 and VEGF-A, followed by an increase in pANCA that suggests a vasculitis development. The serum biomarker levels may help to drive personalized therapies in these phases with anticoagulant prophylaxis, immunomodulators, and/or anti-angiogenic drugs

Urotensin II receptor expression in patients with ulcerative colitis: a pilot study

Urotensin II (U-II) is a vasoactive peptide that interacts with a specific receptor named UTR. Recently, our group has demonstrated increased UTR expression in both human colon adenocarcinoma cell lines and adenomatous polyps, as well as in colon carcinoma samples if compared to healthy colon samples of the same patients. We also showed that an UTR agonist induced an increase in colon adenocarcinoma cell growth in vitro, whereas the UTR block with a specific antagonist caused an inhibition of their growth and an inhibition of about 50% of both motility and cell invasion. Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) associated with an increased baseline risk for colon cancer compared with the general population, and this risk is mostly attributed to chronic inflammation and immune dysregulation. This risk increases along with the duration of the disease, as demonstrated by many studies. There are no UTR expression data related to UC, and we therefore evaluated UTR expression in ill colon biopsies and in healthy colon ones of patients with UC and colon biopsies of healthy patients

The urotensin-II receptor: A marker for staging and steroid outcome prediction in ulcerative colitis

Background: Urotensin-II receptor- (UTR) related pathway exerts a key-role in promoting inflammation. The aim was to assess the relationship between UTR expression and clinical, endoscopic and biochemical severity of ulcerative colitis (UC), exploring its predictivity of intravenous (iv) steroid administration therapeutic outcome. Methods: One-hundred patients with first diagnosis of UC and 44 healthy subjects were enrolled. UTR expression was assessed by qPCR, Western Blot (WB) and immunohistochemistry (IHC). Clinical, endoscopic and histological activity of UC were evaluated by using Truelove and Witts (T&W) severity index, Mayo Endoscopic Score (MES), and Truelove and Richards Index (TRI). The partial and full Mayo scores (PMS and FMS) were assessed to stage the disease. Results: The UTR expression, resulted higher in the lesioned mucosa of UC patients in comparison to healthy subjects (p < .0001 all). Direct relationship between UTR (mRNA and protein) expression and disease severity assessment (T&W, PMS, MES and TRI) was highlighted (p < .0001 all). UTR expression resulted also higher in the 72 patients requiring iv steroids administration compared to those who underwent alternative medications, (p < .0001). The 32 steroid-non-responders showed an increased UTR expression (WB, IHC and qPCR from lesioned mucosa), compared to 40 steroid-responders (p: .0002, .0001, p < .0001 respectively). The predictive role of UTR expression (p < .05) on the negative iv steroids administration therapeutic outcome was highlighted and ROC curves identified the thresholds expressing the better predictive performance. Conclusions: UTR represents a promising inflammatory marker related to clinical, endoscopic, and histological disease activity as well as a predictive marker of steroid administration therapeutic outcome in the UC context

Semeiotica Medica e Metodologia Clinica, 3ª ed

L’ esperienza maturata nel corso della nostra lunghissima vita professionale di medici in prima-linea, ci consente di riaffermare l’ assoluta necessità da parte dei docenti della nostra Facoltà, di formare i futuri medici, non solo attraverso le necessarie conoscenze cliniche, sempre più in espansione, sia in ambito diagnostico che terapeutico, ma anche attraverso la rivalorizzazione di quelle finalità etiche e morali che stanno alla base del rapporto medico-paziente, molto spesso disattese nel corso degli ultimi decenni. E’ nostra convinzione che uno dei compiti insostituibili di un professore di Medicina, sia quello di saper infondere nella coscienza dei nostri studenti tutti i fondamentali princìpi e valori dell’Humanitas. E non vi è alcun dubbio che il rapporto medico-paziente venga garantito proprio dalla Semeiotica, attraverso la fondamentale tappa dell’Anamnesi, che rappresenta la principale fase dell’approccio clinico, costituendo il maggiore “input” informativo, alla diagnostica e contribuendo in modo decisivo, all’istituzione di un rapporto umano ed efficace, tra il malato ed il garante del suo stato di salute. L’ incontro con il paziente rappresenta la parte cruciale del percorso del medico; infatti, il malato gli pone 3 precise domande: a) che tipo di male lo affligge; b) quale evoluzione esso potrà avere; c) cosa si può fare per eliminarlo. Non v’è dubbio come la risposta ai quesiti b) e c) sia direttamente connessa con l’ individuazione da parte del medico ei problemi impliciti nel quesito a). Cioè, solo l’esatta identificazione dello stato morboso renderà possibile la più idonea terapia. Per raggiungere il risultato finale, cioè la sintesi diagnostica, la Semeiotica si avvale di diverse procedure rappresentate da anamnesi, esame fisico del paziente (o esame obiettivo), esami di laboratorio, radiologici e strumentali: risulta quindi evidente l’assoluta importanza di questa disciplina per il raggiungimento di una corretta diagnosi. Ci auguriamo che la nuova stesura di quest’opera possa servire allo studente per appassionarlo ancora di più allo studio del paziente, per stimolarne il ragionamento clinico, e per convincerlo che l’anamnesi e l’esame obiettivo continuano a conservare, a dispetto delle più sofisticate indagini tecnologiche, un’importanza cruciale nell’interpretazione della patologia e nella visione “globale” del malato. E ‘non da ultimo’, vogliamo ringraziare di cuore tutti i validi Autori di questo libro che, con grande competenza e professionalità, ci hanno affiancato nel compimento di questa “fatica” fisica, organizzativa e mentale, allo scopo di raggiungere in concert l’ obiettivo prefissato: e cioè quello di essere stati in grado di realizzare un’opera idonea a rispondere alle più attuali necessità diagnostico-cliniche e, soprattutto…, “di aver saputo raccontare agli Studenti della nostra Facoltà la Semeiotica Medica”