A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20 degrees C after spotting on filter paper, for subsequent ...serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA.
Monoclonal antibodies (Mabs) were spotted within a 0.5-1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20 degrees C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20 degrees C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin.
Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20 degrees C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper.
The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked.
Biflorin is an o-naphthoquinone isolated from the roots of the plant Capraria biflora L. (Scrophulariaceae). In this study, the cytotoxic effects of biflorin were verified, and late apoptosis was ...detected in various cancer cell lines by in situ analysis. The cytotoxicity was further evaluated exclusively for 48 h of treatment in different tumor and non-tumor cell lines (Hep-2, HeLa, HT-29, A-375, and A-549, and HEK-293, respectively). The results indicated that biflorin induced selective cytotoxicity in tumor cells. HeLa cells were more susceptible to biflorin, followed by HT-29, A-549, A-375, and Hep-2 at all concentrations (range 5-50 μg/mL), and the highest half-maximal inhibitory concentration IC50 (56.01 ± 1.17 μg/mL) was observed in HEK-293 cells. Late apoptotic/necrotic events, observed by in situ immunostaining with Annexin V, varied with each cell line; an increase in late apoptotic events was observed corresponding to the increase in biflorin dosage. Hep-2 cells showed a greater percentage of late apoptotic events among the tumor cell lines when treated with higher concentrations of biflorin (69.63 ± 2.28%). The non-tumor HEK-293 line showed greater resistance to late apoptotic events, as well as a lower level of cytotoxicity (77.69 ± 6.68%) than the tested tumor lines. The data presented indicate that biflorin showed an important, possibly selective, cytotoxicity against tumor cell lines, thereby revealing a promising novel substance with potential anticancer activity for tumor therapy.
Abstract
Background
Triage is the crucial point for overall emergency department (ED) management. Its purpose is to quickly and safely identify among the many patients arriving those who need rapid ...medical evaluation 1. Present triage systems are based on symptomatic assessment and clinical impression to assess patient severity on admission to the ED. Some clinical tools that are widely validated in practice, such as ECG, could be implemented in triage to improve the stratification activity of the system 2.
Purpose
To evaluate whether the use of ECG can improve nursing triage prediction for patients admitted to the ED for symptoms related to suspected cardiac pathology.
Methods
A prospective observational single-centered study was conducted. All patients who visited the ED for chest pain, syncope, dyspnoea or palpitation and underwent ECG at the time of triage were considered. The study was conducted by a pool of nurses who had previously received special training in ECG reading. Background characteristics, comorbidities and clinical conditions were recorded for each enrolled patient. In addition, the priority level assigned according to the application of the 5-level Manchester Triage System (1 to 5) and the level of priority after ECG performance (1 to 5) were recorded. The study outcome was composite: the presence of a major cardiac event within 6 weeks after triage, a cardiogenic syncope, the presence of a pulmonary embolism or aortic dissection, or an arrhythmia requiring specific emergency therapy 3. The priority level assigned in triage and the priority level given after performing the ECG were compared by analysis of the receiver operating characteristic.
Results
A total of 1,211 patients were included in the study. Of these, 62.4% (756/1,211) presented chest pain, 13.8% (167/1,211) syncope, 18.2% (221/1,211) palpitation and 5.5% (67/1,211) dyspnoea. Overall, 10.5% (127/1,211) had a positive outcome. The concordance between triage nurses and cardiologists in detecting pathological and non-pathological ECGs presented a Cohen's kappa of 0.821 (p<0.001). ECG performed by nurses increased the patient's priority in 7.5% (91/1,211) of cases and reduced it in 39.6% (480/1,211) of cases. The discriminatory ability of the triage system before ECG was performed showed an area under the receiver operating characteristic curve of 0.708 (95% CI 0.659–0.757), whereas the discriminatory ability of triage after ECG was 0.845 (95% CI 0.808–0.883).
Conclusions
The use of ECG in triage can be a simple and safe tool that improves the assessment of the patient's priority when presenting to the ED for a cardiac symptom. The presence of a good diagnostic concordance and a good improvement in discriminatory ability may support the implementation of ECG in nursing triage.
Funding Acknowledgement
Type of funding sources: None.
We investigated the incidence of multiple sclerosis (MS) in a Sicilian community located at sea level. The study was a retrospective search for MS patients. Incidence was studied in the period from 1 ...January 1985 to 31 December 1994. We searched for definite MS patients, according to Poser's criteria, among the population resident in Bagheria (Palermo province). There were 25 subjects affected by MS, of which 20 were incident MS patients. The average annual incidence was 4.4 per 100,000 persons (n = 453,385 person-years). The incidence increased over time (1985-1989 = 3.5, 1990-1994 = 5.3). A parallel decrease of the interval between onset and diagnosis of MS was observed (1985-1989, 3.7 years, 95% CI = 1.6-7.3; 1990-1994, 1.9 years, 95% CI = 1.0-3.3). These results confirm that MS is frequent in Sicily.
The phenolic content, antioxidant potential, and antimicrobial activity of extracts of different parts of the fruit from Capsicum baccatum L. var. pendulum were investigated. The analysis of phenolic ...content was performed by the Folin-Ciocalteu method and reversed-phase high-performance liquid chromatography. The in vitro antioxidant activity was assessed by the total reactive antioxidant potential and total antioxidant reactivity index. The antioxidant activity was positively correlated with the amount of phenolics found in each sample. The ex vivo antioxidant potential was assessed using the rat liver slice model. The antimicrobial activity was screened using Gram-positive and Gram-negative bacteria and fungi. All the extracts revealed antioxidant activity and a weak antimicrobial activity.
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