404 research outputs found

    Humoral and cellular response to BoHV-1 in buffalo and cattle treated with an inactivated marker vaccine

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    The study is aimed at assessing and comparing the immune response to BoHV-1 elicited by an inactivated marker vaccine in buffaloes and cattle. Vaccination did not produced any local or general reactions in buffaloes. Seroneutralizing antibodies and cellular response by IFN-Îł- test have been detected in buffaloes and cattle after a prime/ booster vaccination strategy. Humoral and cellular responses were significantly higher in cattle than in buffaloes. Data pointed out the possibility to use the marker vaccine in buffaloes. However, further studies must be planned to assess the immune pressure of marker vaccines in terms of IBR eradicative attitude in infected buffalo herds

    Efficient Delivery of MicroRNA and AntimiRNA Molecules Using an Argininocalix[4]arene Macrocycle

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    MicroRNAs (miRNAs) are short non-coding RNA molecules acting as gene regulators by repressing translation or by inducing degradation of the target RNA transcripts. Altered expression of miRNAs may be involved in the pathogenesis of many severe human diseases, opening new avenues in the field of therapeutic strategies, i.e., miRNA targeting or miRNA mimicking. In this context, the efficient and non-toxic delivery of premiRNA and antimiRNA molecules might be of great interest. The aim of the present paper is to determine whether an argininocalix[4]arene is able to efficiently deliver miRNA, premiRNA, and antimiRNA molecules to target cells, preserving their biological activity. This study points out that (1) the toxicity of argininocalix[4]arene 1 is low, and it can be proposed for long-term treatment of target cells, being that this feature is a pre-requisite for the development of therapeutic protocols; (2) the delivery of premiRNA and antimiRNA molecules is efficient, being higher when compared with reference gold standards available; and (3) the biological activity of the premiRNAs and antimiRNAs is maintained. This was demonstrated using the argininocalix[4]arene 1 in miRNA therapeutic approaches performed on three well-described experimental model systems: (1) the induction of apoptosis by antimiR-221 in glioma U251 cells; (2) the induction of apoptosis by premiR-124 in U251 cells; and (3) the inhibition of pro-inflammatory IL-8 and IL-6 genes in cystic fibrosis IB3-1 cells. Our results demonstrate that the argininocalix[4]arene 1 should be considered a very useful delivery system for efficient transfer to target cells of both premiRNA and antimiRNA molecules, preserving their biological activity

    Short communication: Characterization of a monoclonal antibody for Îș-casein B of cow's milk1

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    A monoclonal antibody (antik-B) against an oligopeptide of 23 AA corresponding to the region 131-153 of bovine kappa-casein (kappa-CN) B was generated using the Human Combinatorial Antibody Library (HuCAL) technology. Both AA substitutions distinguishing kappa-CN A and B are located in that region (positions 136 and 148). In this study, the reactivity of antik-B to milk samples collected from cows previously genotyped as CSN3*AA, CSN3*AB, and CSN3*BB was tested. According to Western blot results, antik-B recognized kappa-CN B and it showed no cross-reactivity toward kappa-CN A and other milk proteins. Furthermore, a modified Western blot method, urea-PAGE Western blot, was set up to assess the reactivity of antik-B toward all isoforms of kappa-CN B. In conclusion, antik-B was specific to kappa-CN B in milk and it seemed to be reactive toward all its isoforms

    A simplified sars-cov-2 pseudovirus neutralization assay

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    COVID-19 is an ongoing pandemic caused by the highly infectious coronavirus SARS-CoV-2 that is engaging worldwide scientific research to find a timely and effective eradication strategy. Great efforts have been put into anti-COVID-19 vaccine generation in an effort to protect the world population and block SARS-CoV-2 spread. To validate the protective efficacy of the vaccination campaign and effectively control the pandemic, it is necessary to quantify the induction of neutralizing antibodies by vaccination, as they have been established to be a correlate of protection. In this work, a SARS-CoV-2 pseudovirus neutralization assay, based on a replication-incompetent lentivirus expressing an adapted form of CoV-2 S protein and an ACE2/TMPRSS2 stably expressing cell line, has been minimized in terms of protocol steps without loss of accuracy. The goal of the present simplified neutralization system is to improve SARS-CoV-2 vaccination campaign by means of an easy and accessible approach to be performed in any medical laboratory, maintaining the sensitivity and quantitative reliability of classical serum neutralization assays. Further, this assay can be easily adapted to different coronavirus variants by simply modifying the pseudotyping vector

    Evidence for a Novel Reaction Mechanism of a Prompt Shock-Induced Fission Following the Fusion of 78Kr and 40Ca Nuclei at E/A =10 MeV

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    An analysis of experimental data from the inverse-kinematics ISODEC experiment on 78Kr+40Ca reaction at a bombarding energy of 10 AMeV has revealed signatures of a hitherto unknown reaction mechanism, intermediate between the classical damped binary collisions and fusion-fission, but also substantially different from what is being termed in the literature as fast fission or quasi fission. These signatures point to a scenario where the system fuses transiently while virtually equilibrating mass asymmetry and energy and, yet, keeping part of the energy stored in a collective shock-imparted and, possibly, angular momentum bearing form of excitation. Subsequently the system fissions dynamically along the collision or shock axis with the emerging fragments featuring a broad mass spectrum centered around symmetric fission, relative velocities somewhat higher along the fission axis than in transverse direction, and virtually no intrinsic spin. The class of massasymmetric fission events shows a distinct preference for the more massive fragments to proceed along the beam direction, a characteristic reminiscent of that reported earlier for dynamic fragmentation of projectile-like fragments alone and pointing to the memory of the initial mass and velocity distribution.Comment: 5 PAGES, 6 FIGURE

    Omics sciences and precision medicine in melanoma

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    Background: This article provides an overview of the application of omics sciences in melanoma research. The name omics sciences refers to the large-scale analysis of biological molecules like DNA, RNA, proteins, and metabolites. Methods: In the course of this review, we have adopted a focu-sed research strategy, meticulously selecting the most pertinent and emblematic articles related to the topic. Our methodology included a systematic examination of the scientific literature to guarantee a thorough and precise synthesis of the existing sources. Results: With the advent of high-throughput technologies, omics have become an essential tool for understanding the complexity of melanoma. In this article, we discuss the different omics approaches used in melanoma research, including genomics, transcriptomics, proteomics, and metabolomics. We also highlight the major findings and insights gained from these studies, including the identification of new therapeutic targets and the development of biomarkers for diagnosis and prognosis. Finally, we discuss the challenges and future directions in omics-based melanoma research, including the integration of multiple omics data and the development of personalized medicine approaches. Conclusions: Overall, this article emphasizes the importance of omics science in advancing our understanding of melanoma and its potential for improving patient outcomes

    Assessing Helicobacter pylori motility and biofilm formation in subinhibitory concentrations of antimicrobials.

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    Numerous studies have shown that subinhibitory concentrations of antimicrobials can alter bacterial virulence factors. This study evaluates motility and biofilm formation by H. pylori 43504 grown in subinhibitory concentrations of amoxicillin (AMX), clarithromycin (CLA), or tetracycline (TET). For the swimming and swarming motility assays, H. pylori 43504 suspensions were prepared with the strain alone or with the strain in AMX, CLA, or TET at Âœ MIC. Next, the media were incubated at 37 ÂșC, under microaerophilia. To assess biofilm formation in the presence of one of the antimicrobials at subinhibitory antimicrobial concentrations, bacterial suspensions (109 CFU/mL) were prepared in 2.5% FBS containing AMX, CLA, or TET at Âœ MIC. After incubation for 10 days, H. pylori 43504 grown in medium containing AMX, CLA, or TET at Âœ MIC presented greater swimming motility and lower swarming motility than the non-treated strain. H. pylori 43504 grown in medium containing AMX, CLA, or TET at Âœ MIC showed stronger biofilm production than the non-treated strain. Our results showed that AMX, CLA, or TET at subinhibitory concentrations favors H. pylori 43504 swimming motility and biofilm formation after incubation for 3 days. This may have clinical consequences and make the microorganism difficult to eradicate

    Evaluation of the finger wrinkling test: a pilot study

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    Purpose: Tilt table testing mainly evaluates the systemic cardiovascular part of the autonomic nervous system, while it is assumed that the finger wrinkling test assesses the peripheral part of the autonomic nervous system. In this study we explored whether the finger wrinkling test could be a useful test for autonomic dysfunction and whether the clinical evaluation of wrinkling can be improved by digital analysis of photographs. Methods: As much as 20 healthy subjects and 15 patients underwent tilt table testing and finger wrinkling testing. During the finger wrinkling test the right hand was immersed in water at 40°C. The degree of wrinkling was assessed with a 5-point clinical scale at baseline, 5, 15 and 30 min of immersion. Photographs were taken at the same intervals. Several features were evaluated using digital analysis: length and gradient of automatically detected wrinkle and mean, maximum, minimum, variance and derivative of grey value of pixels. Results: Clinical scoring of wrinkling allowed differentiation between healthy subjects and patients with a normal and an abnormal response to tilt table testing. Relevant features obtained with digital analysis were mean grey value and the gradient of automatically detected wrinkle. McNemar’s test showed no difference in test results between the tilt table test and the finger wrinkling test with a kappa of 0.68. Conclusion: The finger wrinkling test can be used as a screening test before tilt table testing. Visual evaluation of wrinkling is still superior to digital analysis of photographs

    Angiocentric glioma-associated seizures: The possible role of EATT2, pyruvate carboxylase and glutamine synthetase

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    Purpose: Our purpose was to better understand the pathogenesis of seizures associated with angiocentric glioma. Angiocentric glioma is an indolent and rare low-grade glioma. Its typical clinical presentation is with epileptic seizures. The pathogenesis of tumor-associated seizures is poorly understood. Among the possible pathomechanisms, the increased neurotoxic concentrations of the glutamate has been proposed. Glutamate transporters, pyruvate carboxylase and glutamine synthetase are involved in maintaining the physiological concentration of glutamate in the inter synaptic spaces. Methods: We evaluated the immunohistochemical expression of EAAT2 (the most important glutamate transporter), pyruvate carboxylase and glutamine synthetase in 17 angiocentric gliomas. Results: EAAT2 was never expressed (0%) in the neoplastic cells in none of the cases studied. Pyruvate carboxylase was expressed in the cytoplasm of the neoplastic cells in 16/17 cases (94 %). Glutamine synthetase was expressed in the cytoplasm of the neoplastic cells in 15/17 cases (88 %). Conclusion: The net result of this enzymatic expression, in particular considering the loss of EAAT2, could be an increased glutamate concentration in the synaptic clef, which might increase local network excitability initially involving intratumoral neurons. The observation that the angiocentric glioma-associated epilepsy might be at least in part related to EAAT2 deficiency opens up interesting therapeutic perspectives

    Azithromycin inhibits nuclear factor-κB activation during lung inflammation : an in vivo imaging study

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    We studied in vivo the potential involvement of nuclear factor-\u3baB (NF-\u3baB) pathway in the molecular mechanism of the anti-inflammatory and immunomodulatory activity of azithromycin in the lung. Mice transiently transfected with the luciferase gene under the control of a NF-\u3baB responsive element were used to assess in vivo NF-\u3baB activation by bioluminescence imaging. Bioluminescence as well as inflammatory cells and concentrations of proinflammatory cytokines in bronchoalveolar lavage fluids, were monitored in an acute model of pulmonary inflammation resulting from intratracheal instillation of lipopolysaccharide. Lipopolysaccharide (LPS) instillation induced a marked increase in lung bioluminescence in mice transiently transfected with the luciferase gene under the control of an NF-\u3baB responsive element, with significant luciferase expression in resident cells such as endothelial and epithelial cells, as assessed by duoplex immunofluorescence staining. Activation of NF-\u3baB and inflammatory cell lung infiltration linearly correlated when different doses of bortezomib were used to inhibit NF-\u3baB activation. Pretreatment with azithromycin significantly decreased lung bioluminescence and airways cell infiltration induced by LPS, also reducing proinflammatory cytokines concentrations in bronchoalveolar lavages and inhibiting NF-\u3baB nuclear translocation. The results obtained using a novel approach to monitor NF-\u3baB activation, provided, for the first time, in vivo evidence that azithromycin treatment results in pulmonary anti-inflammatory activity associated with the inhibition of NF-\u3baB activation in the lung
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