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Background: Atrial fibrillation is the most common sustained cardiac arrhythmia in adults and it is associated with a high burden of mortality and morbidity worldwide. Catheter ablation is increasingly used to improve symptoms and prognosis in selected patients. Lower limb venous access with subsequent transseptal approach to the left atrium is the standard procedure for atrial fibrillation catheter ablation., Case Presentation: We report an unusual case of complex venous anomaly with a left-sided inferior vena cava with hemiazygos continuation to a persistent left superior vena cava draining in an enlarged coronary sinus in a patient with persistent atrial fibrillation scheduled for transcatheter ablation., Discussion: Lower limb venous anomalies may limit a standard transseptal approach to the left atrium thus precluding an effective catheter ablation procedure for atrial fibrillation. Alternative interventions, such as unconventional percutaneous access, thoracoscopic approach and "ablate and pace" procedures, may be necessary in patients with symptomatic atrial fibrillation and complex venous anomalies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Purpose: We aimed to investigate the clinical performance of edoxaban for the treatment of pulmonary embolism (PE) in hospitalized COVID-19 patients., Methods: We conducted a retrospective analysis selecting hospitalized patients with COVID-19 admitted to our Institution from 20 May 2020 to 20 November 2020 with computer tomography (CT) detected PE at admission, treated with edoxaban after initial parenteral therapy. Clinical outcomes were compared between patients with and without ARDS at admission and between those with and without CT confirmed PE resolution., Results: 50 patients were included. Mean follow-up was 42.5 ± 10 days. No baseline differences were found between patients with ARDS (30%) and those without ARDS at admission. Patients with PE resolution (84%) were younger ( P = 0.03 ), had a shorter duration of fondaparinux therapy (9.9 ± 3.8 vs 15.8 ± 7.5 days; P = 0.0015) and length of hospitalization (36 ± 8 vs 46 ± 9 days: P =  0.0023) compared with those without PE resolution. 2 patients experienced major bleedings. At multivariate analysis the time to edoxaban switch was the only predictor of the PE resolution (HR: 0.92; 95% C.I. 0.86 to 0.99 ) ., Conclusion: Edoxaban was an effective and safe treatment for acute PE in COVID-19 setting.

The efficacy of hydroxychloroquine (HCQ) in treating SARS-CoV-2 infection is harshly debated, with observational and experimental studies reporting contrasting results. To clarify the role of HCQ in Covid-19 patients, we carried out a retrospective observational study of 4,396 unselected patients hospitalized for Covid-19 in Italy (February-May 2020). Patients' characteristics were collected at entry, including age, sex, obesity, smoking status, blood parameters, history of diabetes, cancer, cardiovascular and chronic pulmonary diseases, and medications in use. These were used to identify subtypes of patients with similar characteristics through hierarchical clustering based on Gower distance. Using multivariable Cox regressions, these clusters were then tested for association with mortality and modification of effect by treatment with HCQ. We identified two clusters, one of 3,913 younger patients with lower circulating inflammation levels and better renal function, and one of 483 generally older and more comorbid subjects, more prevalently men and smokers. The latter group was at increased death risk adjusted by HCQ (HR[CI95%] = 3.80[3.08-4.67]), while HCQ showed an independent inverse association (0.51[0.43-0.61]), as well as a significant influence of cluster∗HCQ interaction ( p < 0.001). This was driven by a differential association of HCQ with mortality between the high (0.89[0.65-1.22]) and the low risk cluster (0.46[0.39-0.54]). These effects survived adjustments for additional medications in use and were concordant with associations with disease severity and outcome. These findings suggest a particularly beneficial effect of HCQ within low risk Covid-19 patients and may contribute to clarifying the current controversy on HCQ efficacy in Covid-19 treatment., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Augusto Di Castelnuovo et al.)

Chemical modifications of galactomannans are applied to improve and/or modify their solubility, rheological and functional properties, but have limited specificity and are often difficult to control. Enzymatic reactions, catalyzed under mild process conditions, such as depolymerization, debranching and oxidation, represent a viable and eco-friendly alternative. In this study, we describe oxidation of guar galactomannan primary hydroxyl groups by a fungal laccase using the stable radical TEMPO as mediator. Four fungal laccases were investigated from: Trametes versicolor, Myceliophthora thermophila, Thielavia arenaria, Cerrena unicolor. The laccase from T. versicolor was found to efficiently oxidize TEMPO and to be free of mannanase side activity. Oxidation of galactomannan with this enzyme plus TEMPO brought about a ten-fold increase in viscosity of a guar galactomannan solution and altered its rheological profile, by converting a viscous polysaccharide solution into an elastic gel. This structural modification is presumably due to formation of inter-chain hemiacetalic bonds between newly generated carbonyl groups and free OH groups, yielding a cross-linked gel. These findings could be of practical importance, considering that polysaccharides with high viscosity, gelling and elastic properties can find interesting and novel applications as thickeners, viscosifiers and emulsion stabilizers in several industrial applications such as: personal care, oil operations, paper coating, paints, construction and mining., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Macrophages are pivotal cells of the immune system and play a key role in the host defence mechanism against pathogens. To date, the importance of macrophages and the role of humoral response in eliciting macrophage activity against Mycoplasma mycoides subsp. mycoides small colony (Mmm-SC), the causative agent of contagious bovine pleuropneumonia (CBPP), have only been marginally elucidated or are almost unknown. The present study was undertaken to investigate the changes in surface morphology of macrophages after in vitro infection with Mmm-SC in the presence of bovine immune serum. Morphological analysis was performed on macrophage cultures at 6 h post infection using the three-dimensional vision of scanning electron microscopy. Non-infected macrophages in the presence of negative or immune serum and macro phages infected with Mmm-SC in the absence of serum showed only minor cell surface changes. In contrast, clear surface modifications, broad veils, fine philopodia highlighting cell activation and small aggregates of mycoplasma closely attached to the macrophage membrane, were observed in infected macrophage cultures in the presence of immune serum. Our results suggest that specific humoral response to Mmm-SC may contribute and support phagocytic activity of macrophages.

American foulbrood, caused by Paenibacillus larvae subsp. larvae (White 1906) is one of the most serious diseases of honey bees, causing beekeepers and health workers to make difficult, complex decisions and leading to the development of 'organic' treatments, such as grapefruit seed extract, with minor residue problems in the end product. This study evaluates the chemical composition of grapefruit seed extracts using gas chromatography/mass spectrometry for the detection of benzethonium chloride, cetrimonium bromide and decyltrimethylammonium chloride. The results obtained suggest a close correlation between the microbial effect and the presence of chemical additives in the samples analysed.

The precise role of bovine interferon-gamma (BoIFN-gamma) in disease and therapy is still poorly defined. Clearly it is involved in defence against parasites, bacteria, viruses and possibly tumor cells. This paper reports the expression of BoIFN-gamma in a baculovirus system to generate a fully functional recombinant protein. Bovine interferon-gamma cDNA was cloned from mitogen stimulated peripheral blood mononuclear cell (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The cDNA open reading frame (ORF) encoding for a putative 166 amino acid protein (22KDa) was cloned and expressed into baculovirus transfer vector pBlueBac 4.5/V5 His. This vector was co-transfected with Autografa californica multiple nuclear polyhedrosis virus (AcMNPV) DNA into Spodoptera frugiperda cells (Sf9) and the recombinant virus, named AcBoIFN-gamma, was then recovered. Recombinant BoIFN-gamma (rBoIFN-gamma His) was accumulated in the serum-free medium of AcBoIFN-gamma-infected cells. The nickel affinity spin column purified rBoIFN-gamma His was shown to be a glycosylated 20-22 KDa protein as confirmed by SDS-PAGE glycan determination and showed antiviral activity in vitro against the bovine viral diarrhoea-mucosal disease virus (BVD/MD). The production of this bioactive rBoIFN-gamma His will allow us to explore this cytokine as a potential vaccine adjuvant or therapeutic agent for bovine diseases.

Deletion of megakaryocytic-specific regulatory sequences of GATA-1 (Gata1(tm2Sho) or GATA-1(low) mutation) results in severe thrombocytopenia, because of defective thrombocytopoiesis, and myelofibrosis. As documented here, the GATA-1(low) mutation blocks megakaryocytic maturation between stage I and II, resulting in accumulation of defective megakaryocytes (MKs) in the tissues of GATA-1(low) mice. The block in maturation includes failure to properly organize alpha granules because von Willebrand factor is barely detectable in mutant MKs, and P-selectin, although normally expressed, is found frequently associated with the demarcation membrane system (DMS) instead of within granules. Conversely, both von Willebrand factor and P-selectin are barely detectable in GATA-1(low) platelets. Mutant MKs are surrounded by numerous myeloperoxidase-positive neutrophils, some of which appear in the process to establish contact with MKs by fusing their membrane with those of the DMS. As a result, 16% (in spleen) to 34% (in marrow) of GATA-1(low) MKs contain 1 to 3 neutrophils embedded in a vacuolated cytoplasm. The neutrophil-embedded GATA-1(low) MKs have morphologic features (high electron density and negativity to TUNEL staining) compatible with those of cells dying from para-apoptosis. We suggest that such an increased and pathologic neutrophil emperipolesis may represent one of the mechanisms leading to myelofibrosis by releasing fibrogenic MK cytokines and neutrophil proteases in the microenvironment.

A method to determine residues of the insecticide rotenone in honey using high-performance liquid chromatography (HPLC) is described. The sample was extracted with an acetone/water mixture, followed by a liquid/liquid partition with diethyl ether. A solid-phase extraction (SPE) clean-up step on alumina neutral cartridge was then performed. The chromatographic analysis was conducted on a C18 column (250 x 4 mm, 5 microm) using acetonitrile-water (65:35, v/v) as mobile phase. Rotenone was detected in the ultraviolet range at a wavelength of 295 nm. The specificity of the method was demonstrated through analyses of raw and commercial honey samples. The limit of detection was equal to 40 microg kg(-1). The precision and accuracy of the method were evaluated trueness honey samples spiked at three concentration levels (100-250-500 microg kg(-1)). The intra-laboratory coefficient of variation (from 9.2 to 10.6%) and mean recovery values (from 81.4 to 86.6%) were satisfactory. The calibration curve was linear in the range 0.125-2 microg ml(-1), with a determination coefficient R2 of 0.9999. Rotenone levels in honey samples from bees treated with this miticide were in the range 120-430 microg kg(-1).

Twenty-one samples of grapefruit seed extract (GSE) either from marketed products or provided by an apiculturist were analysed to verify their inhibition activity, in particular against Paenibacillus larvae subsp. larvae, responsible for American foulbrood. The bactericide capacity of GSE has been measured in Bacillus subtilis BGA, Bacillus cereus 11778, Bacillus cereus K250 and Micrococcus luteus 9341a; these bacteria are normally used in the laboratory to study inhibitors. The results showed that not all GSE have the same inhibitory activity and two of those analysed do not inhibit the five bacteria used. Considering that 19 samples inhibited American foulbrood bacillus, the authors conclude that the use of a natural product (such as GSE) to control this important disease of bees, can be used as a substitute for chemotherapeutic products, after appropriate expedients.