Assessment of physicians’ and senior medical students’ knowledge in treatment of patients with community acquired pneumonia: Current results of the KNOCAP project

Introduction. Community-acquired pneumonia (CAP) remains one of the most acute problems of bronchopulmonary pathology being the 4th in the mortality structure (after cardiovascular, cerebrovascular diseases and malignant neoplasms) and the 1st among all fatalities from infectious diseases. Thanks to the scientific progress achieved in the antibiotic therapy and vaccine prophylaxis, the death toll has decreased four times compared to its rate during the “pre-antibiotic era“. However, nowadays there is a steadily increasing trend in the pneumonia mortality rate in Russia. The only possible way to increase efficacy of CAP treatment is timely initiated rational antibiotic therapy, considering the possible etiologies, risk factors and the severity of the patient’s condition. Materials and methods. The article represents the results of anonymous prospective surveys within the framework of the KNOCAP multi-centered research project aimed at accessing the knowledge on the fundamental issues in diagnosis and treatment of community-acquired pneumonia. The survey involved 222 students in their fifth- and sixth years in medical institute from Belgorod, Dnepr (Dnipro), Voronezh, Kiev (Kyiv) and Saratov and 110 physicians from Krasnodar, Saratov, Belgorod and Dnepr.Results and discussion. According to the results of the survey, such levels of correct answers were given by doctors and students, respectively: the inadmissibility of antimicrobial therapy (AMT) delay in CAP - 82% and 59%; the main criterion for withholding AMT - 56% and 37%; “sequential therapy” - 61% and 59%. At the same time, only 24% of the students and 23% of the physicians surveyed correctly reported typical mistakes in the treatment of a non-severe CAP with 50% or more accuracy; and in case of initial treatment, the number of correct responses was less than 28% for students and 45% for doctors. Conclusion. The survey showed that both senior medical students majoring in Medical Care and general practitioners had a low level of knowledge in CAP treatment. Hence, curricula need to be adjusted both in medical universities and in health institutions for practitioners in order to inform them and, thus, improve the quality of their knowledge in this field

Assessment of physicians’ and senior medical students’ knowledge in treatment of patients with community-acquired pneumonia: Current results of the KNOCAP project

Community-acquired pneumonia (CAP) remains one of the most acute problems of bronchopulmonary pathology being the 4th in the mortality structure (after cardiovascular, cerebrovascular diseases and malignant neoplasms) and the 1st among all fatalities from infectious diseases. Thanks to the scientific progress achieved in the antibiotic therapy and vaccine prophylaxis, the death toll has decreased four times compared to its rate during the “pre-antibiotic era“. However, nowadays there is a steadily increasing trend in the pneumonia mortality rate in Russia. The only possible way to increase efficacy of CAP treatment is timely initiated rational antibiotic therapy, considering the possible etiologies, risk factors and the severity of the patient’s condition

Study of Fatty-acid Composition of Goat and Sheep Milk and Its Transformation in the Production of Yogurt

The information on the structure of fatty acids is crucial for the production and promotion of goat and sheep milk in the manufacture of dairy products. The fatty-acid profile of milk fat can affect the nutritional value and market value of dairy products. The purpose of the research is to study the properties of fatty-acid composition of goat and sheep milk and its transformation in the process of yogurt production. The study of fatty-acid composition was performed using the method of gas chromatography. The milk of goats of Zaanen breed and sheep of North Caucasian breed, as well as yogurt produced from a mixture of goat and sheep milk in a ratio of 1:1 have become the objects of the study. It was found that goat's milk contains 12% less saturated fatty acids than sheep's milk. Oleic, stearic and palmitic acids are the main fatty acids found in the fat phase of milk and yogurt production. Changes in the concentration of individual fatty acids during milk processing and in the period of yogurt storage were noted. As a result of maturation and seven-day storage, the amount of saturated fatty acids in yogurt increased by 5% compared to the original milk mixture. The content of polyunsaturated fatty acids in yogurt decreased by 19.27 %. The highest ratio value of hypocholesterolemic and hypercholesterolemic fatty acids was noted in goat milk. It is proved that goat milk is characterized by the most acceptable fatty acid composition in terms of healthy nutrition and prevention of atherosclerosis and thrombosis. A tendency to reduce the amount of monounsaturated and polyunsaturated fatty acids, with a simultaneous increase in the content of saturated fatty acids is noted in the process of yogurt production and storage. It was found that the production of yogurt with the use of goat's milk in the mixture allows reducing the values of atherogenicity and thrombogenicity indices. The results of the study provide an information basis for the production of qualitatively new fermented milk drinks with a fatty-acid profile favorable for human health

CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents

<div><p>Binding of the Ca²⁺/calmodulin(CaM)-dependent protein kinase II (CaMKII) to the NMDA-type glutamate receptor (NMDAR) subunit GluN2B controls long-term potentiation (LTP), a form of synaptic plasticity thought to underlie learning and memory. Regulation of this interaction is well-studied biochemically, but not under conditions that mimic the macromolecular crowding found within cells. Notably, previous molecular crowding experiments with lysozyme indicated an effect on the CaMKII holoenzyme conformation. Here, we found that the effect of molecular crowding on Ca²⁺/CaM-induced CaMKII binding to immobilized GluN2B <i>in vitro</i> depended on the specific crowding reagent. While binding was reduced by lysozyme, it was enhanced by BSA. The ATP content in the BSA preparation caused CaMKII autophosphorylation at T286 during the binding reaction; however, enhanced binding was also observed when autophosphorylation was blocked. Importantly, the positive regulation by nucleotide and BSA (as well as other macromolecular crowding reagents) did not alleviate the requirement for CaMKII stimulation to induce GluN2B binding. The differential effect of lysozyme (14 kDa) and BSA (66 kDa) was not due to size difference, as both dextran-10 and dextran-70 enhanced binding. By contrast, crowding with immunoglobulin G (IgG) reduced binding. Notably, lysozyme and IgG but not BSA directly bound to Ca²⁺/CaM in an overlay assay, suggesting a competition of lysozyme and IgG with the Ca²⁺/CaM-stimulus that induces CaMKII/GluN2B binding. However, lysozyme negatively regulated binding even when it was instead induced by CaMKII T286 phosphorylation. Alternative modes of competition would be with CaMKII or GluN2B, and the negative effects of lysozyme and IgG indeed also correlated with specific or non-specific binding to the immobilized GluN2B. Thus, the effect of any specific crowding reagent can differ, depending on its additional direct effects on CaMKII/GluN2B binding. However, the results of this study also indicate that, in principle, macromolecular crowding enhances CaMKII binding to GluN2B.</p></div

The Reduced Level of Inorganic Polyphosphate Mobilizes Antioxidant and Manganese-Resistance Systems in <i>Saccharomyces cerevisiae</i>

Inorganic polyphosphate (polyP) is crucial for adaptive reactions and stress response in microorganisms. A convenient model to study the role of polyP in yeast is the Saccharomyces cerevisiae strain CRN/PPN1 that overexpresses polyphosphatase Ppn1 with stably decreased polyphosphate level. In this study, we combined the whole-transcriptome sequencing, fluorescence microscopy, and polyP quantification to characterize the CRN/PPN1 response to manganese and oxidative stresses. CRN/PPN1 exhibits enhanced resistance to manganese and peroxide due to its pre-adaptive state observed in normal conditions. The pre-adaptive state is characterized by up-regulated genes involved in response to an external stimulus, plasma membrane organization, and oxidation/reduction. The transcriptome-wide data allowed the identification of particular genes crucial for overcoming the manganese excess. The key gene responsible for manganese resistance is PHO84 encoding a low-affinity manganese transporter: Strong PHO84 down-regulation in CRN/PPN1 increases manganese resistance by reduced manganese uptake. On the contrary, PHM7, the top up-regulated gene in CRN/PPN1, is also strongly up-regulated in the manganese-adapted parent strain. Phm7 is an unannotated protein, but manganese adaptation is significantly impaired in &#916;phm7, thus suggesting its essential function in manganese or phosphate transport

Ca²⁺/CaM stimulation is still required for CaMKII to GluN2B binding.

<p><i>A</i>, CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096522#pone-0096522-g001" target="_blank">Figure 1A</a>, but in the presence or absence of Ca²⁺/CaM (1 mM/1 µM), ADP (100 µM), or BSA (100 mg/ml). Bound CaMKII was eluted and detected by Western-analysis. BSA and ADP alone were insufficient to induce CaMKII to GluN2B binding. GST-GluN2B detection is shown as a loading control. Representative images are from the same experiment and Western-blots. <i>B</i>, CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in panel A. Different crowding agents, BSA, dextran-10 (DEX10) and dextran-70 (DEX70) (all at 100 mg/ml) in the presence of ADP (100 µM) were not sufficient to induce detectable CaMKII binding to GluN2B without CaMKII stimulation.</p

Induction of binding by T286 phosphorylation, instead of Ca²⁺/CaM.

<p>CaMKIIα was pre-incubated with ATP and Ca²⁺/CaM to induce autophosphorylation at T286; then, Ca²⁺ was chelated with EGTA and further phosphorylation was inhibited by staurosporine. Phospho-T286 CaMKIIα (40 nM subunits) was bound to GST-GluN2B as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096522#pone-0096522-g001" target="_blank">Figure 1A</a>, but in the presence of 1.5 mM EGTA, 2 µM staurosporine, and without addition of Ca²⁺/CaM or nucleotide. Lysozyme (100 mg/ml) decreased CaMKII binding to GluN2B. T286-phosphorylation was verified using a phospho-T286-specific antibody. Phospho-T286 immunoblots are from the same experiment run on a separate gel to avoid reprobing blots at the same molecular weight. Dotted lines represent IDVs from CaMKII/GluN2B binding when instead stimulated with Ca²⁺/CaM and nucleotide (from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096522#pone-0096522-g001" target="_blank">Figure 1A</a>). n = 6 or 7 per group; **: p<0.01, difference from control, one-way ANOVA followed by Tukey's HSD. #: no difference compared to the same binding condition, but with binding induced by Ca²⁺/CaM (compare Fig. 1A). Bar graphs indicate mean ± s.e.m, and GST-GluN2B detection is shown as a loading control.</p

Two differently sized dextrans both enhance CaMKII binding to GluN2B, indicating that the differential effects of BSA and lysozyme were not due to size differences.

<p>Ca²⁺/CaM-stimulated CaMKII binding to GluN2B was tested as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096522#pone-0096522-g001" target="_blank">Figure 1A</a>. Molecular crowding with dextran-10 (DEX10) or dextran-70 (DEX 70), both at 100 mg/ml, both increased CaMKII binding to GluN2B to an equal extent. n = 6–7; *: p<0.05, **: p<0.01 in one-way ANOVA followed by Tukey's HSD; n.s.: no significant difference between the two dextrans. Bar graphs indicate mean ± s.e.m, and GST-GluN2B detection is shown as a loading control.</p