Accelerated and Improved Quantification of Lymphocytic Choriomeningitis Virus (LCMV) Titers by Flow Cytometry

Lymphocytic choriomeningitis virus (LCMV), a natural murine pathogen, is a member of the Arenavirus family, may cause atypical meningitis in humans, and has been utilized extensively as a model pathogen for the study of virus-induced disease and immune responses. Historically, viral titers have been quantified by a standard plaque assay, but for non-cytopathic viruses including LCMV this requires lengthy incubation, so results cannot be obtained rapidly. Additionally, due to specific technical constraints of the plaque assay including the visual detection format, it has an element of subjectivity along with limited sensitivity. In this study, we describe the development of a FACS-based assay that utilizes detection of LCMV nucleoprotein (NP) expression in infected cells to determine viral titers, and that exhibits several advantages over the standard plaque assay. We show that the LCMV-NP FACS assay is an objective and reproducible detection method that requires smaller sample volumes, exhibits a ∼20-fold increase in sensitivity to and produces results three times faster than the plaque assay. Importantly, when applied to models of acute and chronic LCMV infection, the LCMV-NP FACS assay revealed the presence of infectious virus in samples that were determined to be negative by plaque assay. Therefore, this technique represents an accelerated, enhanced and objective alternative method for detection of infectious LCMV that is amenable to adaptation for other viral infections as well as high throughput diagnostic platforms

Perinatal asphyxia: current status and approaches towards neuroprotective strategies, with focus on sentinel proteins

Delivery is a stressful and risky event menacing the newborn. The mother-dependent respiration has to be replaced by autonomous pulmonary breathing immediately after delivery. If delayed, it may lead to deficient oxygen supply compromising survival and development of the central nervous system. Lack of oxygen availability gives rise to depletion of NAD+ tissue stores, decrease of ATP formation, weakening of the electron transport pump and anaerobic metabolism and acidosis, leading necessarily to death if oxygenation is not promptly re-established. Re-oxygenation triggers a cascade of compensatory biochemical events to restore function, which may be accompanied by improper homeostasis and oxidative stress. Consequences may be incomplete recovery, or excess reactions that worsen the biological outcome by disturbed metabolism and/or imbalance produced by over-expression of alternative metabolic pathways. Perinatal asphyxia has been associated with severe neurological and psychiatric sequelae with delayed clinical onset. No specific treatments have yet been established. In the clinical setting, after resuscitation of an infant with birth asphyxia, the emphasis is on supportive therapy. Several interventions have been proposed to attenuate secondary neuronal injuries elicited by asphyxia, including hypothermia. Although promising, the clinical efficacy of hypothermia has not been fully demonstrated. It is evident that new approaches are warranted. The purpose of this review is to discuss the concept of sentinel proteins as targets for neuroprotection. Several sentinel proteins have been described to protect the integrity of the genome (e.g. PARP-1; XRCC1; DNA ligase IIIα; DNA polymerase β, ERCC2, DNA-dependent protein kinases). They act by eliciting metabolic cascades leading to (i) activation of cell survival and neurotrophic pathways; (ii) early and delayed programmed cell death, and (iii) promotion of cell proliferation, differentiation, neuritogenesis and synaptogenesis. It is proposed that sentinel proteins can be used as markers for characterising long-term effects of perinatal asphyxia, and as targets for novel therapeutic development and innovative strategies for neonatal care

Advancing biomedical substrate engineering: An eco-friendly route for synthesizing micro- and nanotextures on 3D printed Ti–6Al–4V

This study investigates the impact of a combined approach involving sandblasting and electrochemical surface treatment using a deep eutectic solvent Ethaline, a eutectic mixture of choline chloride and ethylene glycol, on the surface characteristics of Ti–6Al–4V biomedical substrates fabricated through direct selective laser melting (DSLM). Research has focused on surface morphology, topography, chemical composition, and cell adhesion. The novel approach demonstrated the ability to create a hierarchical surface structure with both micro and nanopatterns. The rough edges resulting from the sandblasting process were effectively smoothed through subsequent electrochemical processing. Additionally, the issue of residual sand particles, which commonly arise in sandblasting procedures, was successfully addressed with the new method. The results indicated that Ti alloy samples subjected to sandblasting and electrochemical treatment in Ethaline exhibited improved surface hydrophilicity. In-vitro cell adhesion tests confirmed the potential for bio-inspired properties of DSLM-printed Ti–6Al–4V biomedical substrates achieved through the combination of sandblasting and electrochemical processing in Ethaline

The effect of cardiorehabilitation on the quality of life by congestive heart failure patients

Abstract Funding Acknowledgements Type of funding sources: None. Introduction To examine selected parameters of quality of life in patients with cardiac failure after cardiorehabilitation in outpatient and inpatient form and to compare the effect with each other. Methods and methodology: The survey included 64 respondents without age restrictions with chronic heart failure NYHA I. - III. after undergoing inpatient or outpatient cardiorehabilitation, polymorbid, oncology and psychiatric patients were excluded. The questionnaire with 20 questions focused on 12 quality of life domains. Statistical processing: t-test, ANOVA, Chi-square test, Spearman correlation coefficient (p &amp;lt;0.05). Results 35 men (54.7%) and 29 women (45.3%) were included, with a mean age of 58.07 years, with a mean duration of CHF treatment of 3.99 years, subgroups (internal 25, cardiology 21 and institute 18 probands) did not differ in age. We found that cardiorehabilitation has a positive effect on the quality of life of patients with CHF, if it is associated with social support, appropriate education and meets the conditions of a comprehensive approach. Patients rated their quality of life better compared to their quality of life a year ago. Outpatients are more satisfied with their health than patients in inpatient treatment, patients with a more severe manifestation of CHF have a greater anxiety from physical activity and therefore less motivation to move. All patients expressed dissatisfaction with their sex lives. Patients in outpatient treatment experience more depression. There were no differences between the subgroups in the evaluation of the degree of education by the nurse and the effect of the exercise on the subjective condition. All subgroups perceived the other person"s help more valuable than medical help. Patients from inpatient treatment have been more motivated to change their lifestyle on their own initiative and through the influence of information from healthcare professionals. Conclusion Comprehensive cardiovascular rehabilitation of patients with heart failure improves many of the prognostic indicators of heart failure, reduces mortality and the frequency of rehospitalizations, reduces treatment costs, improves quality of life. nevertheless, rehabilitation programs for chronic heart failure are being promoted only very slowly. The reason may be the absence of standard procedures, as well as organizational, economic and personnel demands. </jats:sec