Development of a method for molecular subtyping <i>Bacillus anthracis</i> using HRM PCR

Introduction. Bacillus anthracis is the causative agent of anthrax, a pathogen characterized by high genetic monomorphism that complicates differentiation of strains. Thus, molecular methods for pathogen typing require the improvement. The aim of the study. To select marker SNPs for new genetic groups of B. anthracis and to develop a method for their laboratory identification using HRM PCR. Materials and methods. The core genome of 222 strains of B. anthracis from the GenBank database and 66 strains from the collection of pathogenic microorganisms of the Stavropol Anti-Plague Institute was aligned using the parsnp software. A dendrogram based on 7242 core genome SNPs was built in MEGA X software. The strains for validation of the HRM method included representatives of various genetic groups. The HRM PCR reaction was performed using the "Type-it HRM PCR Kit" and "KAPA HRM FAST qPCR Kit" and a Rotor Gene DNA thermocycler with HRM function. Data analysis and visualization were performed using custom scripts in the Python and R development environments. Results and discussion. Marker SNPs for 6 genetic groups have been identified, which make it possible to determine whether strains belong to one of 7 new subclusters. Pairs of primers were selected for the loci containing them, HRM PCR parameters were optimized for discrimination of different alleles of SNP loci, and an analysis scheme was developed. Conclusion. Thus, marker SNPs were selected to determine the genetic subclusters A.Br.CEA, A.Br.STI, A.Br.Tsiankovskii, B.Br.Europe, B.Br.Siberia, B.Br.Asia, B.Br.018, and a new laboratory method was developed for molecular subtyping of B. anthracis using HRM PCR

Short-Term Therapies for Treatment of Acute and Advanced Heart Failure—Why so Few Drugs Available in Clinical Use, Why Even Fewer in the Pipeline?

Both acute and advanced heart failure are an increasing threat in term of survival, quality of life and socio-economical burdens. Paradoxically, the use of successful treatments for chronic heart failure can prolong life but—per definition—causes the rise in age of patients experiencing acute decompensations, since nothing at the moment helps avoiding an acute or final stage in the elderly population. To complicate the picture, acute heart failure syndromes are a collection of symptoms, signs and markers, with different aetiologies and different courses, also due to overlapping morbidities and to the plethora of chronic medications. The palette of cardio- and vasoactive drugs used in the hospitalization phase to stabilize the patient’s hemodynamic is scarce and even scarcer is the evidence for the agents commonly used in the practice (e.g., catecholamines). The pipeline in this field is poor and the clinical development chronically unsuccessful. Recent set backs in expected clinical trials for new agents in acute heart failure (AHF) (omecamtiv, serelaxine, ularitide) left a field desolately empty, where only few drugs have been approved for clinical use, for example, levosimendan and nesiritide. In this consensus opinion paper, experts from 26 European countries (Austria, Belgium, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Israel, Italy, The Netherlands, Norway, Poland, Portugal, Russia, Slovenia, Spain, Sweden, Switzerland, Turkey, U.K. and Ukraine) analyse the situation in details also by help of artificial intelligence applied to bibliographic searches, try to distil some lesson-learned to avoid that future projects would make the same mistakes as in the past and recommend how to lead a successful development project in this field in dire need of new agents

Short-Term Therapies for Treatment of Acute and Advanced Heart Failure—Why so Few Drugs Available in Clinical Use, Why Even Fewer in the Pipeline?

Both acute and advanced heart failure are an increasing threat in term of survival, quality of life and socio-economical burdens. Paradoxically, the use of successful treatments for chronic heart failure can prolong life but—per definition—causes the rise in age of patients experiencing acute decompensations, since nothing at the moment helps avoiding an acute or final stage in the elderly population. To complicate the picture, acute heart failure syndromes are a collection of symptoms, signs and markers, with different aetiologies and different courses, also due to overlapping morbidities and to the plethora of chronic medications. The palette of cardio- and vasoactive drugs used in the hospitalization phase to stabilize the patient’s hemodynamic is scarce and even scarcer is the evidence for the agents commonly used in the practice (e.g., catecholamines). The pipeline in this field is poor and the clinical development chronically unsuccessful. Recent set backs in expected clinical trials for new agents in acute heart failure (AHF) (omecamtiv, serelaxine, ularitide) left a field desolately empty, where only few drugs have been approved for clinical use, for example, levosimendan and nesiritide. In this consensus opinion paper, experts from 26 European countries (Austria, Belgium, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Israel, Italy, The Netherlands, Norway, Poland, Portugal, Russia, Slovenia, Spain, Sweden, Switzerland, Turkey, U.K. and Ukraine) analyse the situation in details also by help of artificial intelligence applied to bibliographic searches, try to distil some lesson-learned to avoid that future projects would make the same mistakes as in the past and recommend how to lead a successful development project in this field in dire need of new agents

Short-Term Therapies for Treatment of Acute and Advanced Heart Failure—Why so Few Drugs Available in Clinical Use, Why Even Fewer in the Pipeline?

Both acute and advanced heart failure are an increasing threat in term of survival, quality of life and socio-economical burdens. Paradoxically, the use of successful treatments for chronic heart failure can prolong life but—per definition—causes the rise in age of patients experiencing acute decompensations, since nothing at the moment helps avoiding an acute or final stage in the elderly population. To complicate the picture, acute heart failure syndromes are a collection of symptoms, signs and markers, with different aetiologies and different courses, also due to overlapping morbidities and to the plethora of chronic medications. The palette of cardio- and vasoactive drugs used in the hospitalization phase to stabilize the patient’s hemodynamic is scarce and even scarcer is the evidence for the agents commonly used in the practice (e.g. catecholamines). The pipeline in this field is poor and the clinical development chronically unsuccessful. Recent set backs in expected clinical trials for new agents in acute heart failure (AHF) (omecamtiv, serelaxine, ularitide) left a field desolately empty, where only few drugs have been approved for clinical use, for example, levosimendan and nesiritide. In this consensus opinion paper, experts from 26 European countries (Austria, Belgium, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Israel, Italy, The Netherlands, Norway, Poland, Portugal, Russia, Slovenia, Spain, Sweden, Switzerland, Turkey, U.K. and Ukraine) analyse the situation in details also by help of artificial intelligence applied to bibliographic searches, try to distil some lesson-learned to avoid that future projects would make the same mistakes as in the past and recommend how to lead a successful development project in this field in dire need of new agents

Measurement of beauty production via non-prompt D0{\rm D}^{0} mesons in Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 5.02 TeV

The production of non-prompt ${\rm D}^{0}$ mesons from beauty-hadron decays was measured at midrapidity ($\left| y \right| 5~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions. The data are described by models that include both collisional and radiative processes in the calculation of beauty-quark energy loss in the quark-gluon plasma, and quark recombination in addition to fragmentation as a hadronization mechanism. The ratio of the non-prompt to prompt ${\rm D}^{0}$-meson $R_{\rm AA}$ is larger than unity for $p_{\rm T} > 4~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions, as predicted by models in which beauty quarks lose less energy than charm quarks in the quark-gluon plasma because of their larger mass

Measurement of beauty production via non-prompt D0{\rm D}^{0} mesons in Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 5.02 TeV

The production of non-prompt ${\rm D}^{0}$ mesons from beauty-hadron decays was measured at midrapidity ($\left| y \right| 5~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions. The data are described by models that include both collisional and radiative processes in the calculation of beauty-quark energy loss in the quark-gluon plasma, and quark recombination in addition to fragmentation as a hadronization mechanism. The ratio of the non-prompt to prompt ${\rm D}^{0}$-meson $R_{\rm AA}$ is larger than unity for $p_{\rm T} > 4~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions, as predicted by models in which beauty quarks lose less energy than charm quarks in the quark-gluon plasma because of their larger mass

Measurement of beauty production via non-prompt D0{\rm D}^{0} mesons in Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 5.02 TeV

The production of non-prompt ${\rm D}^{0}$ mesons from beauty-hadron decays was measured at midrapidity ($\left| y \right| 5~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions. The data are described by models that include both collisional and radiative processes in the calculation of beauty-quark energy loss in the quark-gluon plasma, and quark recombination in addition to fragmentation as a hadronization mechanism. The ratio of the non-prompt to prompt ${\rm D}^{0}$-meson $R_{\rm AA}$ is larger than unity for $p_{\rm T} > 4~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions, as predicted by models in which beauty quarks lose less energy than charm quarks in the quark-gluon plasma because of their larger mass

Measurement of beauty production via non-prompt D0{\rm D}^{0} mesons in Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 5.02 TeV

The production of non-prompt ${\rm D}^{0}$ mesons from beauty-hadron decays was measured at midrapidity ($\left| y \right| 5~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions. The data are described by models that include both collisional and radiative processes in the calculation of beauty-quark energy loss in the quark-gluon plasma, and quark recombination in addition to fragmentation as a hadronization mechanism. The ratio of the non-prompt to prompt ${\rm D}^{0}$-meson $R_{\rm AA}$ is larger than unity for $p_{\rm T} > 4~\mathrm{GeV}/c$ in the $0-10$% central Pb-Pb collisions, as predicted by models in which beauty quarks lose less energy than charm quarks in the quark-gluon plasma because of their larger mass