[Problems associated with effective pharmacotherapy of the elderly patients (geriatrics): A review].

The worlds older population is growing dramatically. At the same time, ensuring an appropriate high standard of living for the elderly by reducing of morbidity and disability of geriatric patients is one of the main objectives of the modern healthcare system. However, changes associated with body aging necessitate application of novel approaches to the correction of pharmacotherapy and usage of specialized dosage forms. Such medicinal products provide both an appropriate therapeutic effect and facilitate their use. Presented review considers several features of pharmacotherapy of geriatric patients.

[Modern extemporaneous formulations in the geriatric care management: current opportunities and future challenges. A review].

Age-associated disorders, including cognitive functions, that often occur in geriatric patients, necessitate the use of novel approaches to provide appropriate medical care, pharmacoprophylaxis and pharmacotherapy among them. At the same time, an important objective of the national healthcare system is not only stimulating of pharmaceutical companies and pharmacies to expand the range of medicines intended for elderly patients, but also increasing availability of medicinal products, including the integration of extemporaneous formulations into clinical practice. Presented review considers several features of the regulation of the use of extemporaneous formulations in the treatment of geriatric patients. Examples of prescriptions that are used in Russian medical practice and are of the greatest interest in the treatment of elderly patients are also presented.

[Familial case of hypogonadotropic hypogonadism as the CHARGE syndrome manifestation].

CHARGE syndrome is a rare autosomal dominant disease caused by CHD7 gene mutations. Individuals with CHARGE display a wide spectrum of clinical features. It might be presented only as a delay puberty, which does not require any hormone replacement therapy to severe CHARGE phenotype, requiring a multidisciplinary therapeutic approach. Wild spectrum of clinical presentation can be seen even among the patients with identical mutation. Diagnosis might be suspected by a combination of major and minor clinical criteria of this disorder, but molecular genetic analysis is mandatory for final verification. Accurate diagnosis is essential to informing patients about all possible clinical features, reproductive status and choosing the correct treatment approach. The most common endocrine abnormality in patients with CHARGE syndrome is the disturbance in gonadotropins function ranged from delay puberty to persistent hypogonadotropic hypogonadism with different olfactory phenotypes, resulted by specific role of CHD7 in GnRH neuronal embryogenesis.We describe a familial case of CHARGE syndrome with significant intrafamilial clinical heterogeneity due to CHD7 gene mutation.

Malyl-CoA lyase provides glycine/glyoxylate synthesis in type I methanotrophs.

The biochemical routes for assimilation of one-carbon compounds in bacteria require many clarifications. In this study, the role of malyl-CoA lyase in the metabolism of the aerobic type I methanotroph Methylotuvimicrobium alcaliphilum 20Z has been investigated by gene inactivation and biochemical studies. The functionality of the enzyme has been confirmed by heterologous expression in Escherichia coli. The mutant strain lacking Mcl activity demonstrated the phenotype of glycine auxotrophy. The genes encoding malyl-CoA lyase are present in the genomes of all methanotrophs, except for representatives of the phylum Verrucomicrobium. We suppose that malyl-CoA lyase is the enzyme that provides glyoxylate and glycine synthesis in the type I methanotrophs supporting carbon assimilation via the serine cycle in addition to the major ribulose monophosphate cycle.

Approbation of russian test strips for antimicrobial susceptibility testing of microorganisms by gradient diffusion method (E-test).

The most reliable criterion for the microorganisms antibiotic susceptibility is the value of the minimum inhibitory concentration (MIC). The gradient diffusion method (epsilometric test, e-test), carried out using test strips impregnated with an antimicrobial agent, allows to obtain the quantitative value of MIC, bypassing the time-consuming steps of the traditional method of serial dilutions. We tested strips for the epsilometric test, developed at Saint-Petersburg Pasteur Institute. The quality control, carried by testing the reference strains of E. coli ATCC 25922 and S. aureus 29213, confirmed compliance of the manufacturer's declared antibiotic concentration ranges. The MIC values obtained in the study of the antibiotic susceptibility of 101 clinical isolates of microorganisms of the ESKAPE group, isolated from patients of various hospitals in St. Petersburg, were compared to 8 antimicrobial agents using tested test strips and the corresponding M.I.C. Evaluator of the Oxoid (UK). A high percentage of compliance of MIC values was demonstrated, as well as full compliance of susceptibility categories for all microorganism/antibiotic combinations. The resulting divergences in the MIC values did not exceed one step of a double serial dilution, which is permissible according to GOST R ISO 20776-1-2010. The study showed that the test strips developed at the Saint-Petersburg Pasteur Institute DNT are an acceptable alternative to import tests and, after the registration certificate will be completed, can be offered to determine the susceptibility of microorganisms to antimicrobial agents in bacteriological laboratories.

Antimicrobial resistance of Escherichia coli, isolated from children's intestinal microbiota.

Recent studies have shown that bacterial resistance existed long before antimicrobials were used in medicine, and not only pathogens are resistant to antibiotics. 511 strains of E. coli isolated from the intestinal microbiota of children aged 1 month to 17 years living in St. Petersburg were studied: the susceptibility to 15 antibiotics was determined by the disk diffusion method, as well as the susceptibility to 6 commercial bacteriophages produced by «Microgen¼ (Russia). The b-lactamase genes of molecular families TEM, SHV, OXA, and CTX-M were detected by multiplex PCR. 39,3% E. coli isolates were resistant to one or more antimicrobial classes. The proportion of multidrug resistant isolates (resistant to 3 or more classes) was 16,6%. Multidrug resistance to clinically significant antimicrobial classes (extended-spectrum cephalosporins (ESC) + fluoroquinolones + aminoglycosides) was detected in 0,8% isolates. Resistance to aminopenicillins was detected in 29,5%, ESC - 11,2%, fluoroquinolones - 13,3%, tetracycline - 20,0%, chloramphenicol - 9,8%, aminoglycosides - 2,5% isolates. b-lactam resistance was due to the beta-lactamase production: to ampicillin - the molecular family TEM (81,9%), ESC - the CTX-M molecular family (87,7%) CTX-M1 - (66%) and CTX-M9 groups (34%). 43,5% multidrug resistant E. coli isolates were susceptible to at least one of the six commercial bacteriophages produced by «Microgen¼. The study showed that the intestinal microbiota of children is an important reservoir of E. coli resistant (including multidrug resistance) to various classes of antibiotics, and bacteriophage therapy is an alternative method for eradication of antibiotic-resistant E. coli.

[Current trends in the development of resistance to clinically significant antibiotics in Salmonella (review of literature).]

Salmonella is one of the leading bacterial pathogens of acute diarrhea as well as foodborne outbreaks. Salmonellosis can occur as gastroenteritis with the development of complications and generalization of infection, also the extra intestinal diseases that require antibiotic therapy are often registered. Currently, the effectiveness of many antibiotics is reduced due to the development of resistance in Salmonella. National Salmonella surveillance systems monitor Salmonella resistance to «critically important for medicine¼ antibiotics (extended-spectrum cephalosporins and fluoroquinolones), as well as multidrug resistance. Quinoloneresistant Salmonella is considered as a high-priority resitant pathogen by the World Health Organization. The article describes the current situation on salmonellosis in the world. Foreign and Russian current data about the leading Salmonella serotypes in different regions of the world are presented. The prevalence of clinically significant resistance depending of the Salmonella serotypes in countries with state monitoring systems is shown. The authors described the leading molecular resistance mechanisms (chromosomal and plasmid mediated) and showed their prevalence in different Salmonella serotypes. The article gives the information about Salmonella successful international multidrug resistant clones with specific resistance phenotypes and genotypes. The authors describe the molecular methods for detection of resistance mechanisms, and show the necessity and significance of antimicrobial susceptibility monitoring in the epidemiological Salmonella surveillance.

[Antimicrobial resistance and clinical significant resistance mechanisms of Salmonella isolated in 2014-2018 in St.Petersburg, Russia.]

The article presents the results of antimicrobial resistance monitoring of Salmonella isolated from children and adults with diarrhea in St. Petersburg in 2014-2018. In 746 isolates of 42 serovars more than 90,0% belonged to three: S. enteritidis (79,6%), S. typhimurium (6,8%) and S. infantis (3,8%). The antimicrobial susceptibility testing (according the EUCAST) to 7 classes of antimicrobials revealed the resistance in 78,6% of Salmonella. Low-level quinolone resistance (MIC of ciprofloxacin 0,12-0,5 mg/l) was detected in 63,3% isolates (S. enteritidis -71,0%, S. typhimurium - 15,7%, S. infantis - 89,3%) and was due to five kinds of single nucleotide substitutions in gyrA: Asp87Tyr - 36,1% of studied isolates (only S. infantis); Ser83Phe - 22,2% (only S. enteritidis); Asp87Asn - 19,4% (S. enteritidis, S. typhimurium, S. hadar, S. newport); Ser83Tyr -11,1% (S. enteritidis and S. infantis) and Asp87Gly - 8,3% (only S. enteritidis). Only in one S. kentucky isolate with high-level fluoroquinolone resistance (MIC of ciprofloxacin > 8,0 mg/l) two substitutions (Ser83Phe and Asp87Asn) were detected. Two Salmonella isolates (S. typhimurium and S. corvallis) had plasmid-mediated quinolone resistance (qnrS). Extended-spectrum cephalosporin resistance was found in 6 Salmonella serovars (1,6%). The bla-genes were detected: of genetic group CTX-M1 - in 10 isolates (serovars S. typhimurium, S. enteritidis, S. abony, S. coeln and S. virchow), CTX-M2 - in 2 S. typhimurium isolates, CTX-M9 - in three S. enteritidis isolates. In one S. typhimurium CTX-M1 and CTX-M2 were detected. The gene of CMY-2 (molecular class C cephalosporinase) was revealed in two isolates (S. newport and S. enteritidis). Our study showed that Salmonella (the main bacterial pathogen of acute diarrhea in children and adults) isolated in Saint-Petersburg had antimicrobial resistance to drugs of choice for salmonellosis treatment.

[Methodical aspects of antimicrobial susceptibility testing of Salmonella (review of literature).]

Salmonella is one of the leading food-borne infection pathogen: annually in the Russian Federation about 50 thousand cases of salmonellosis are registered. Antimicrobial therapy is necessary in the case of severe infection in children under 6 years and persons over 50 years, in patients with severe accompanying disease, as well as in the case of generalization of the infection. Beta-lactam antibiotics, quinolones and azithromycin are included in the list of drugs recommended for antimicrobial therapy of salmonellosis, including typhoid fever. The effectiveness of therapy largely depends on the appropriate antimicrobial susceptibility testing: the choice of testing method, indicator antibiotics and result interpretation. Salmonella belong to the Enterobacteriacae family and are characterized by common mechanisms of resistance to quinolones and beta-lactams, but antimicrobial susceptibility testing of Salmonella to these groups of antibiotics has a number of features. The article presents current data on the susceptibility of Salmonella, including S. Typhi, to antibiotics and leading clinically significant resistance mechanisms. The methodical aspects of Salmonella antimicrobial susceptibility testing of the drugs used for the treatment of salmonellosis (quinolones, beta-lactams and azithromycin) are described in detail. Interpretation of Salmonella testing results according the modern international and Russian recommendations are presented. The authors propose the algorithms for Salmonella antimicrobial susceptibility testing of quinolones, cephalosporins and carbapenems, as well as criteria for result interpretation, allowing the detection of clinically significant mechanisms of resistance to beta-lactams (production of beta-lactamases of different molecular classes) and quinolones (chromosomal mutations and acquired resistance genes).

Serine-glyoxylate aminotranferases from methanotrophs using different C1-assimilation pathways.

The indicator enzyme of the serine pathway of assimilation of reduced C1 compounds, serine-glyoxylate aminotransferase (Sga), has been purified from three methane-oxidizing bacteria, Methylomicrobium alcaliphilum 20Z, Methylosinus trichosporium OB3b and Methylococcus capsulatus Bath. The native enzymes were shown to be dimeric (80 kDa, strain 20Z), tetrameric (~ 170 kDa, strain OB3b) or trimeric (~ 120 kDa, strain Bath). Sga from the three methanotrophs catalyse the pyridoxal phosphate-dependent transfer of an amino group from serine to glyoxylate and pyruvate; the enzymes from strains 20Z and Bath also transfer an amino group from serine to α-ketoglutarate and from alanine to glyoxylate. No other significant differences between the Sga from the three methanotrophs were found. The three methanotrophic Sga have their highest catalytic efficiencies in the reaction between glyoxylate and serine, which is in agreement with their function to provide circulation of the serine assimilation pathway.The disruption of the sga gene in Mm. alcaliphilum resulted in retardation of growth rate of the mutant cells and in a prolonged lag-phase after passaging from methane to methanol. In addition, the growth of the mutant strain is accompanied by formaldehyde accumulation in the culture liquid. Hence, Sga is important in the serine cycle of type I methanotrophs and this pathway could be related to the removal of excess formaldehyde and/or energy regulation.

Biochemical Properties and Phylogeny of Hydroxypyruvate Reductases from Methanotrophic Bacteria with Different C1-Assimilation Pathways.

In the aerobic methanotrophic bacteria Methylomicrobium alcaliphilum 20Z, Methylococcus capsulatus Bath, and Methylosinus trichosporium OB3b, the biochemical properties of hydroxypyruvate reductase (Hpr), an indicator enzyme of the serine pathway for assimilation of reduced C1-compounds, were comparatively analyzed. The recombinant Hpr obtained by cloning and heterologous expression of the hpr gene in Escherichia coli catalyzed NAD(P)H-dependent reduction of hydroxypyruvate or glyoxylate, but did not catalyze the reverse reactions of D-glycerate or glycolate oxidation. The absence of the glycerate dehydrogenase activity in the methanotrophic Hpr confirmed a key role of the enzyme in utilization of C1-compounds via the serine cycle. The enzyme from Ms. trichosporium OB3b realizing the serine cycle as a sole assimilation pathway had much higher special activity and affinity in comparison to Hpr from Mm. alcaliphilum 20Z and Mc. capsulatus Bath assimilating carbon predominantly via the ribulose monophosphate (RuMP) cycle. The hpr gene was found as part of gene clusters coding the serine cycle enzymes in all sequenced methanotrophic genomes except the representatives of the Verrucomicrobia phylum. Phylogenetic analyses revealed two types of Hpr: (i) Hpr of methanotrophs belonging to the Gammaproteobacteria class, which use the serine cycle along with the RuMP cycle, as well as of non-methylotrophic bacteria belonging to the Alphaproteobacteria class; (ii) Hpr of methylotrophs from Alpha- and Betaproteobacteria classes that use only the serine cycle and of non-methylotrophic representatives of Betaproteobacteria. The putative role and origin of hydroxypyruvate reductase in methanotrophs are discussed.

Pharmaceutical counseling: Between evidence-based medicine and profits.

BACKGROUND: The number of pharmacies, which produce drug formulations locally, has recently considerably reduced in Russia. Pharmacies mainly operate as retailers of industrially manufactured drugs.Pharmaceutical consultation of customers at pharmacies aimed at responsible self-medication is the most popular and accessible feature of pharmaceutical care. In Russia there is a significant list of medicines approved for sale in pharmacies on a non-prescription basis that is specified in the product label. In this regard, the role of pharmacists in public health in Russia increases. Pharmacist, working directly with population, is an important figure for the rational use of medicines. This type of work requires high level of professional training and appropriate ethics. OBJECTIVE: To explore the current status of pharmaceutical counseling in Russia. METHODS: Situation analysis, surveys of pharmacists. RESULTS: Our experience in the system of postgraduate professional education, the results of the survey of pharmacists, and the long-term dialogue with pharmacists allowed us to identify several unresolved issues in the work of a pharmacist selling non-prescription drugs.Lack of differentiation in the functions of a pharmacist with a higher education and pharmaceutical technologist: In production/industrial pharmacy technicians are engaged in manufacturing of pharmaceutical formulations. However, due to the loss of production functions technologists had to move away from production laboratories of apothecaries to the sales area. Currently, the apothecary's assignment to receive prescriptions and dispense medications can be fulfilled by either a pharmacist or a pharmaceutical technician. It significantly discerns the pharmacy from the medical organization with clearly delineated functions of doctors and nurses. Russian regulations should consider the level of education required for high-quality pharmaceutical counseling.Contradiction between the pharmacist's special functions and trade procedure with the lack of pharmaceutical counseling standards: Article 1.1 "Code of Ethics of the pharmaceutical worker of Russia" states: "The main task of the professional activity of the pharmaceutical worker - protection of human health", Article 1.3 states that a pharmaceutical worker must take professional decisions solely in the interests of a patient [1]. However, the pharmacy is a trade organization, thus as a retailer the pharmacy is directly interested in making profits and increasing sales of pharmaceutical products, including non-prescription medicines. Moreover, while the clinical medicine is monitored for unjustified prescribing and measures are being taken to prevent polypharmacy, for a pharmacist the growing sales of over-the-counter drugs, active promotion of dietary supplements, homeopathic medicines, medical devices, and, consequently, an increase of financial indicators (particularly "average purchase size") - all are characteristics of success [2].Rational use of over-the-counter medicines requires introduction of pharmaceutical counseling standards (pharmaceutical care) according to symptoms - major reasons to visit a pharmacy as part of responsible self-medication (cold, sore throat, headache, diarrhea, etc.). Standards of pharmaceutical counseling should be objective, reliable and up-to-date and contain recommendations for the rational use of over-the-counter drugs as well as indications requiring treatment to the doctor. Standardization of pharmaceutical counseling in terms of Evidence-based Pharmacy would enhance the efficiency, safety and cost-effectiveness of over-the-counter medicines.Currently, the lack of clinical component in the higher pharmaceutical education and the lack of approved standards of pharmaceutical counseling lead to the introduction of cross-selling technologies (which are broadly applied in other areas of trade, for example, the offer of a boot-polish during the sale of shoes) to the pharmaceutical practice [2, 3]. However, drugs belong to a special group of products, proper selection of which requires special education, and the consumer is not always able to evaluate the quality of the recommendations. Marketing cross-selling recommendations are aimed at promotion of the over-the-counter medicines for customers buying prescription drugs. For example, business coaches recommend the pharmacists to make additional offers: with the purchase of physician-prescribed antibiotics - offer of vitamins, with prescribed nonsteroidal anti-inflammatory drugs - commercially available ointment with non-steroidal topical formulation ("to enhance the effect") and others. These recommendations do not agree with evidence-based medicine and lead to inefficient use of over-the-counter drugs and unjustified financial expenses. CONCLUSIONS: Thus, to ensure the rational use of medicines permitted for free (non-prescription) dispensing at the pharmacies, pharmaceutical information needs standardization on the basis of evidence-based medicine as well as standardization of the pharmaceutical counseling service. The development of practical recommendations on the rational use of over-the counter medicines by doctors and pharmacists with further adoption at the state level, the recommendation of most secure, efficient and cost-effective over-the-counter medications during pharmaceutical counseling in pharmacies will contribute to the restoration and preservation of public health.

Detection of highly conductive surface electron states in topological crystalline insulators Pb(1-x)SnxSe using laser terahertz radiation.

We suggest a method for detection of highly conductive surface electron states including topological ones. The method is based on measurements of the photoelectromagnetic effect using terahertz laser pulses. In contrast to conventional transport measurements, the method is not sensitive to the bulk conductivity. The method is demonstrated on an example of topological crystalline insulators Pb(1-x)SnxSe. It is shown that highly conductive surface electron states are present in Pb(1-x)SnxSe both in the inverse and direct electron energy spectrum.

Hypotensive effect of retabolil correcting the concentration of aldosterone during stress exposures.

Male rats were exposed to single or repeated (19 days) cold treatment (4°C) and non-cold stress (60-min shaking on a laboratory shuttle device). Retabolil had a hypotensive effect, which was accompanied by the prevention of a stress-induced increase in the concentration of a hypertensive hormone aldosterone. Under conditions of repeated stress, these effects were realized via µ-opioid receptors. Our results suggest that retabolil can be used as a hypotensive and aldosterone-blocking agent, at least during stress exposure in animals (and probably in humans).

[Farmacological effect of retabolil on aldosterone level and arterial pressure in rats under the action of vibrations].

The experiments were performed on male rats, which were subjected to single and multiply repeated vibrations (low-frequency, horizontal, high-amplitude) analogous to the action of motor transport vibrations. It is established that the administration of retabolil produces a hypotensive effect and blocks the vibration-induced increase in the level of hypertensive hormone aldosterone. Under conditions of the multiply repeated action of vibrations, both effects were realized via micro-opioid receptors. In the case of a single action, these receptors were only involved in a hypotensive effect but not mediated in aldosterone suppression. Both these effects were absent in the control group of animals (not subjected to vibrations). Therefore, retabolil can be used as a hypotensive and aldosterone-blocking drug for vibration-induced hypertension in animals and, probably, in humans.

Effects of dehydroepiandrosterone sulfate on blood pressure and aldosterone level via µ-opioid receptors.

Dehydroepiandrosterone sulfate (30 mg/kg) produced a hypotensive effect by preventing stress-induced surge of hypertensive hormone aldosterone in rats after manifold repeated, but not single stress exposure. Both effects were realized via µ-opioid receptors. Thus, µ-opioid mechanism of blockage of aldosterone surge can underlay the hypotensive effects of dehydroepiandrosterone sulfate at least under conditions of manifold repeated exposures.

Cytotoxic and Immunochemical Properties of Viscumin Encapsulated in Polylactide Microparticles.

Biodegradable polylactide microparticles with encapsulated cytotoxic protein viscumin were obtained via the ultrasound-assisted supercritical fluid technique. The size of the microparticles was 10-50 µM, as shown by electron microscopy. The time course of viscumin release from microparticles was studied using an immunoenzyme test system with anti-viscumin monoclonal antibodies. It was found that 99.91% of the cytotoxic protein was incorporated into polymer microparticles. Only 0.08% of the initially encapsulated viscumin was released from the microparticles following incubation for 120 h in a phosphate-buffered saline at neutral pH. Importantly, the method of ultrasonic dry supercritical fluid encapsulation failed to alter both the cytotoxic potency and the immunochemical properties of the encapsulated viscumin. Thus, this procedure can be used to generate biodegradable polylactide microparticles with encapsulated bioactive substances.

[The characteristics of biological properties of E. coli O104:H4--the causative agent of large-scale alimentary ictus in Germany may 2001].

The review presents the characteristics of E. coli O104:H4, the causative agent of large-scale alimentary ictus in Germany in spring time 2011. The antigenic characteristics and factors of E. coli pathogenicity are taken into account. The causative agent has a combination of pathogenic factors of two groups of diarrheigenic Escherichia: shigella similar toxin, specific for entero-hemorrhagic E. coli and adhesins of enteroaggregative E. coli.

[Study of photodynamic treatment for P-388 lympho-leukemia in mice].

Data are presented on the results of photodynamic treatment (PDT) of mice DBA2 with transplantable lympho-leukemia P-388. Different regimens of photosensitizer Dimegin and emission were used. Both intravenous PDT and in combination with local PDT should be recommended.

CTLA4Ig treatment in patients with multiple sclerosis: an open-label, phase 1 clinical trial.

BACKGROUND: The modulation of costimulatory pathways represents an original therapeutic approach to regulate T cell-mediated autoimmune diseases by preventing or reducing autoantigen-driven T-cell activation in humans. Autoreactive CD4(+) T cells play a critical role in initiating the immune response leading to the chronic inflammation and demyelination characteristic of multiple sclerosis (MS). METHODS: We used IV infusions of CTLA4Ig to block the CD28/B7 T-cell costimulatory pathway in a phase 1 dose-escalation study in MS. Sixteen patients with relapsing-remitting MS received a single CTLA4Ig infusion and were monitored for up to 3 months after treatment. In an extension study, four additional subjects received four doses of CTLA4Ig. RESULTS: CTLA4Ig was well tolerated in patients with MS, and most adverse events were rated as mild. Immunologic assessment of the patients showed a reduction in myelin basic protein (MBP) proliferation within 2 months of infusion and decreased interferon-gamma production by MBP-specific lines. CONCLUSIONS: Inhibiting costimulatory molecule interactions by using CTLA4Ig seems safe in multiple sclerosis (MS), and the immunologic effects suggest that it may be a promising approach to regulate the inflammatory process associated with MS.