Plasma pharmacokinetics and tissue distribution of L-lysine α-oxidase from Trichoderma cf. aureoviride RIFAI VKM F- 4268D in mice.

L-lysine α-oxidase (LO) is an L-amino acid oxidase with antitumor, antimicrobial and antiviral properties. Pharmacokinetic (PK) studies were carried out by measuring LO concentration in plasma and tissue samples by enzyme immunoassay. L-lysine concentration in samples was measured spectrophotometrically using LO. After single i.v. injection of 1.0, 1.5, 3.0 mg/kg the circulating T1/2 of enzyme in mice varied from 51 to 74 min and the AUC0-inf values were 6.54 ± 0.46, 8.66 ± 0.59, 9.47 ± 1.45 µg/ml × h, respectively. LO was distributed in tissues and determined within 48 h after administration with maximal accumulation in liver and heart tissues. Mean time to reach the maximum concentration was highest for the liver-9 h, kidney-1 h and 15 min for the tissues of heart, spleen and brain. T1/2 of LO in tissues ranged from 7.75 ± 0.73 to 26.10 ± 2.60 h. In mice, plasma L-lysine decreased by 79% 15 min after LO administration in dose 1.6 mg/kg. The serum L-lysine levels remained very low from 1 to 9 h (< 25 µM, 17%), indicating an acute lack of L-lysine in animals for at least 9 h. Concentration of L-lysine in serum restored only 24 h after LO administration. The results of LO PK study show that it might be considered as a promising enzyme for further investigation as a potential anticancer agent.

Human seroprevalence data indicate other factors than climatic conditions influencing dirofilariosis transmission in the Russian Federation.

The species of the genus Dirofilaria are filarial parasites causing zoonotic infections in humans with an increasing incidence in temperate and tropical areas of the world. Due to its classification as a vector-borne disease, the most important factors influencing dirofilariosis transmission are those related to climate, such as temperature and humidity. However, other factors linked with human behaviour can influence the distribution of the parasite. Although the Russian Federation could be considered as a non-suitable area for Dirofilaria spp. transmission due to its climatic conditions, one third of the human cases of dirofilariosis have been declared in this country. Here, seroepidemiological data on human dirofilariosis for five different regions distributed throughout the Russian Federation (Rostov, Moscow, Ekaterinburg, Yakutia and Khabarovsk) were obtained. A total of 940 serum samples from totally random donors living in these areas were analysed by enzyme-linked immunosorbent assay for the detection of anti-Dirofilaria immitis immunoglobulin G antibodies. Similar seroprevalence data ranging from 3.41% in Yakutia to 6.95% in Khabarovsk, with no significant correlation with climatic data of yearly average temperature and rainfall from these regions were found. These results suggest that other factors probably related to human behaviour, and not only climatic conditions, might be facilitating the spread of human dirofilariosis in these areas.

Nine cases of human dipylidiasis in Moscow region during 1987 to 2017.

Dipylidium caninum is a parasite that commonly infects dogs and cats worldwide. The large population of wild and stray dogs and cats may potentially transmit D. caninum to humans via their flea and lice. Humans are an accidental host, and dipylidiasis is more commonly seen in infants and children. There is scant information about human dipylidiasis in Russia. We report nine cases of dipylidiasis - eight in children and one in an adult. The patients were asymptomatic, except for excreting active proglottids in their faeces, which was the most common complaint. The clinical significance of asymptomatic dipylidiasis is not understood, except mothers were anxious because of the continuous appearance of active worms in the faeces of their children. The patients were successfully treated with praziquantel (15 mg/kg). Preventing dipylidiasis in pets and humans requires the control of fleas and lice, avoiding the outdoor defecation of definitive hosts, deworming pets, preventing children from playing with stray animals and spread of information about dipylidiasis among pet owners. Dogs and cats in many places in Russia breed freely, defecate outdoors in any area, and are not subjected to deworming and insect control. These circumstances favour the fact that, although this zoonosis is rare, it is a re-emerging disease and might reach important levels in Russia.

Possible case of trichinellosis associated with beaver (Castor fiber) meat.

Although there have been occasional reports of rare and low-level trichinellae infestation in beavers, no human cases of beaver-associated trichinellosis have been described. This report presents a possible case of human trichinellosis linked to beaver meat. Increasing consumption of beaver meat necessitates raising awareness of this potential source of trichinellosis.

Genetic characterization of Plasmodium vivax in the Kyrgyz Republic.

At the end of 2016, Kyrgyz Republic was certified by the World Health Organization as a malaria-free country, while only a decade ago this disease posed a serious health threat. The progress achieved by Kyrgyz Republic provides a unique example of tertian (Plasmodium vivax) malaria elimination. This success was based on an integrated approach, including measures for the treatment of infected people and disease prevention, vector control and the development of an effective national epidemiological surveillance system. Lower P. vivax msp-1, msp-3α, csp and dbpII genes polymorphism was revealed in Kyrgyz Republic in compare with that in Tajikistan. Molecular characterization of the causative agent found that P. vivax populations in Kyrgyz Republic was comprised by several lineages, highly divergent in the south-western and genetically homogeneous in the northern regions of Kyrgyz Republic, d. Such profile in the northern regions was compatible with several recent introductions rather than a long-term endemic circulation of the parasite. A low level of genetic variability suggested that the parasitic systems of tertian malaria, were not adapted, which, along with other factors, largely determined the possibility of malaria elimination in northern Kyrgyz Republic. Other determinants included environmental, social, and epidemiological factors that limited the spread of malaria. South-western Kyrgyz Republic, a region with a high level of interstate migration, requires considerable attention to prevent the spread of malaria.

Phylodynamics of Crimean Congo hemorrhagic fever virus in South Russia.

Phylodynamics of Crimean Congo Hemorrhagic fever virus (CCHFV) genotype V in South Russia was analyzed using 244 partial (452-571 nt) sequences in all three genomic segments and 38 complete genomic sequences. Despite increased number of sequences, the Russian lineage of the European genotype V (commonly termed GtVa) was distinct from GtV isolates from Turkey and the Balkan countries. No geographic pattern was observed in phylogenetic subgrouping of CCHFV within South Russia. Identical isolates could be found at distant locations spaced by hundreds of kilometers, while relatively divergent viruses circulated in the same region. Full genome analysis indicated that reassortment events within GtVa occurred every few decades (median half-life of a non-reassortant node 30-40 years) and involved M and S segments. Therefore, in South Russia CCHFV represents a highly dynamic population of frequently reassorting viruses.

Viral Vectors for Gene Therapy: Current State and Clinical Perspectives.

Gene therapy is the straightforward approach for the application of recent advances in molecular biology into clinical practice. One of the major obstacles in the development of gene therapy is the delivery of the effector to and into the target cell. Unfortunately, most methods commonly used in laboratory practice are poorly suited for clinical use. Viral vectors are one of the most promising methods for gene therapy delivery. Millions of years of evolution of viruses have resulted in the development of various molecular mechanisms for entry into cells, long-term survival within cells, and activation, inhibition, or modification of the host defense mechanisms at all levels. The relatively simple organization of viruses, small genome size, and evolutionary plasticity allow modifying them to create effective instruments for gene therapy approaches. This review summarizes the latest trends in the development of gene therapy, in particular, various aspects and prospects of the development of clinical products based on viral delivery systems.

Retrospective diagnosis of two rabies cases in humans by high throughput sequencing.

BACKGROUND: Rabies is prevalent in 150 countries and is definitely found in Russian Federation. The average registered incidence of rabies infection among animals in Russia is 3000 cases per year. At least 500,000 cases of animal bites and scratches are registered in the Russia every year, but only 2-4 cases of rabies infection in humans are reported per year. This relatively low incidence of rabies infection among humans is the result of a well-organized program of rabies surveillance and control as well as the readily available vaccination and immunoglobulin therapies. However, physician awareness of rabies infection in patients with acute encephalopathy is low, and some cases of rabies remain undiagnosed. OBJECTIVES: A retrospective study of autopsy materials from patients who died of encephalitis of unknown etiology in the Astrakhan region of Russia in 2003. STUDY DESIGN: A broad-range polymerase chain reaction (PCR) analysis followed by high throughput sequencing were used for the diagnosis. RESULTS: Two cases of rabies were detected and subsequently confirmed using a fluorescent antibody test, an enzyme-linked immunosorbent assay (ELISA) and a mouse inoculation test. Two strains of rabies virus were isolated and characterized using virological methods. The entire genome of each strain was sequenced.

[Molecular-epidemiological monitoring of enterovirus circulation in the Far East and Zabaikalye].

6 year molecular-biological monitoring of enteroviruses in the Far East and Zabaikalye was carried out. Nucleotide sequence of 125 strains was determined from 2006 to 2011. Molecular analysis was carried out in VP1 virus genome region. Phylogenetic interactions for ECHO-6, ECHO-30, ECHO-11, Coxsackie B-5 (CB-5), Coxsackie B-1 (CB-1) and Coxsackie A-9 (CA-9) were analyzed. Highly dynamic epidemiology was shown to be inherent for ECHO-6 and ECHO-30 viruses and is characterized by genetic heterogeneity and consequent change of virus variants. On the contrary a relative stability of circulating genotypes is intrinsic for CB-1, CB-5 and ECHO-11 enteroviruses. The results of molecular-biological studies indicate frequent introduction of new enterovirus variants from countries of Europe and Asia.

[Seroepidemiology and molecular epidemiology of enterovirus type 71 in the world and the Russian Federation].

A review of recent publications on epidemiology and seroepidemiology of enterovirus type 71 in various regions of the world and authors' own results of study of seroepidemiology and molecular epidemiology of EV71 in Russia are presented.

[Experience of application of multiplex qPCR for differential diagnostics of intestinal viral infections].

AIM: Evaluate the effectiveness of multiplex reverse transcription (RT) and polymerase chain reaction with fluorescence detection in real time mode (qPCR) methods for differential detection of 11 groups of intestine viruses (adenoviruses, enteroviruses, polioviruses, hepatitis A and E viruses, group A and C rotaviruses, orthoreoviruses, noroviruses, sapoviruses and astroviruses) in various biological samples. MATERIALS AND METHODS: Panels of virus isolates and clinical samples characterized by reference methods were used to evaluate sensitivity of detection of various intestine viruses. Nucleic acids were isolated from study samples and multiplex RT and qPCR were carried out. RESULTS: Sensitivity of laboratory reagent kit (LRK) when compared with results obtained from reference methods was 100% for rotavirus A, adenovirus, enterovirus and norovirus, 88.9% for hepatitis E virus and 92.3% for hepatitis A virus, and diagnostic specificity - 99.4%. During analysis of 697 clinical samples from patients with acute intestine infection symptoms nucleic acids of various intestine viruses were isolated in 71.7%. CONCLUSION: Multiplex qRT-PCR was shown as an effective method of etiologic diagnostics of an intestine viral infection. Use of LRK was demonstrated to establish etiology of intestine diseases in 63 - 72% and in children with watery diarrhea - in approximately 90% of cases.

[Social and economic significance of enterovirus infection and its role in etiologic structure of infectious diseases in the world].

Human enteroviruses comprised by more than 100 serotypes, they spread everywhere and can cause wide spectrum of diseases as well as significant social and economic loss. Influenza-like illness and mild forms of enterovirus infection (herpangina, exanthema) are widespread and causes of significant number of visits in clinics. Economic cost of mild form of enterovirus infection is not high although great number of cases (10 - 15 mln cases yearly in USA) determines its important economic significance. Single cases and outbreaks of enterovirus aseptic meningitis occur less frequently but lead to significant economic burden due to hospitalization costs. Enteroviruses are also cause up to 30% of sepsis-like disease in newborns and play important role in infant morbidity and mortality. Potential of enteroviruses as a source of new diseases in humans has a special significance for practical healthcare. In XX century enteroviruses became a cause of pandemics of paralytic poliomyelitis, hemorrhagic conjunctivitis, and foot-and-mouth-like disease, which caused vast social and economic loss, and emergence of new forms of enterovirus infection is quite possible in XXI century.

Recombination among picornaviruses.

Picornaviruses are small non-enveloped positive strand RNA viruses that can cause a wide range of clinical manifestations in humans and animals. Many of these viruses are highly diversified and globally prevalent. Natural recombination has been reported in most picornavirus genera and is a key genetic feature of these infectious agents. In several socially relevant picornavirus genera, such as enteroviruses, aphthoviruses, parechoviruses and cardioviruses, recombination, combined with dynamic global epidemiology, maintains virus species as a worldwide pool of genetic information. It can be suggested that on a short time scale recombination acts to promote virus diversity, and new recombinant forms of picornaviruses emerge frequently as 'snapshots' of this global pool. On a longer time scale, recombination maintains stability of a gene pool of a species by shuffling sequences and thus limiting divergence and speciation. This review covers existing evidence of recombination in most genera of the family Picornaviridae and possible implications for diagnostics, epidemiology and classification.

[Encephalomyelitis caused by enterovirus type 71 in children].

Enterovirus type 71 (EV71) is a causative agent of large outbreaks of hand, foot, and mouth disease (HFMD) in Europe (Bulgaria, 1975; Hungary, 1978) and South-East Asia (Malaysia, 1977; Taiwan, 1998; Singapore, 2000-2007; People's Republic of China, 2007-2009). HFMD afflicted children less than 10 years of age and resulted in recovery within 3-7 days. In a small percentage of infants (aged 6 months to 3 years), HFMD was accompanied by acute neurological complications, such as serous meningitis, poliomyelitis-like syndrome (extremity pareses and muscle paralyses); brain stem encephalitis (myoclonic jerks, tremor, lethargy, swallowing and speech disorders, cardiopulmonary failure, pulmonary edema, shock, coma, death). X-ray study revealed pulmonary hemorrhages and edema. Mortality rates were as high as 82-94% in severe cases. Incapacitating motor, respiratory, and psychoemotional disorders persisted in some surviving children. Pathomorphologically, patients with central nervous system disease and cardiopulmonary failure were found to have acute inflammation of the grey matter of the brain stem (medulla oblongata, pons) and spinal cord. Inflammatory changes in the lung and myocardial tissues were negligible or absent. Fatal pulmonary edema was neurogenic in origin and resulted from damage to the respiratory and vasomotor centers of the brain stem.

[Enterovirus 71: epidemiology and diagnostics].

In 1970s enterovirus type 71 (EV71) caused several epidemics of poliomyelitis-like disease with severe neurologic sequelae. In the last 20 years EV71 was the cause of series of outbreaks and epidemics of foot-and-mouth disease-like conditions with neurologic sequelae in countries of South-East Asia. During the last epidemic of EV71 infection, which occurred in China in 2008, more than 60,000 cases was registered, 38 of which were lethal. Some aspects of epidemiology and laboratory diagnostics of disease caused by EV71 are considered in this review.

[Molecular epidemiology of ECHO 6 virus, the causative agent of the 2006 outbreak of serous meningitis in Khabarovsk].

A total of 3194 cases of enterovirus meningitis were notified in the Russian Federation in 2005, of them there were 1434 cases in the Khabarovsk Territory. Enteroviruses were isolated from 1020 out of the virologically studied 1362 patients from the Khabarovsk Territory. Viruses E6 and E30 were isolated in 80 and 14.7% of cases, respectively. E1, E3, E7, E33, Coxsackie virus B1, B4, B5, and A10 were sporadically detected. The E6 strains isolated in Komsomolsk-on-Amur were identical while E6 strains isolated in Khabarovsk belonged to two different genotypes and greatly differed from those isolated in Konsomolsk-on-Amur. The virus E30 strains isolated in Khabarovsk and Komsomolsk-on-Amur had a 1% difference in VP1 genome nucleotide sequence and belonged to E30 subtype that circulated in Russia and Kazakhstan in 2004-2005.

[Epidemiologic and etiologic characteristic of enterovirus infections in Khabarovsk region].

Results of epidemiologic, virologic, and serologic studies of enterovirus infections in Khabarovsk region from 1975 to 2006 were analyzed. Patterns of epidemic process of these infections were established: periodic change of dominating type of pathogen in the population; onset of the large epidemic peaks of incidence during emergence of circulation of new for the given area serotypes of enteroviruses; possibility of realization of several routes of virus transmission. Role of water factor in the progress of the epidemic process was revealed. Etiology of the large epidemic rise of aseptic meningitis incidence in Khabarovsk region in 2006 was established--the leading pathogens were ECHO viruses serotypes E6 and E30.

[Acute enterovirus uveitis in infants].

Enterovirus uveitis (EU) is a new infant eye disease that was first detected and identified in Russia in 1980-1981. Three subtypes of human echoviruses (EV19K, EV11A, and EV11/B) caused 5 nosocomial outbreaks of EU in different Siberian cities and towns in 1980-1989, by affecting more than 750 children mainly below one year of age. Sporadic and focal EU cases (more than 200) were also retrospectively diagnosed in other regions of Russia and in different countries of the former Soviet Union. There were following clinical manifestations: common symptoms of the infection; acute uveitis (rapid focal iridic destruction, pupillary deformities, formation of membranes in the anterior chamber of the eye); and in 15-30% of cases severe complications, cataract, glaucoma, vision impairments. Uveitis strains EV19 and EV11 caused significant uveitis in primates after inoculation into the anterior chamber of the eye, as well as sepsis-like fatal disease with liver necrosis after venous infection. The uveitis strains are phylogenetically and pathogenetically close for primates to strains EV19 and EV11 isolated from young children with sepsis-like disease. The contents of this review have been published in the Reviews in Medical Virology, 2004, vol. 14, p. 241-254.

[A role of recombination in the evolution of enteroviruses].

Enteroviruses, members of the family Picornaviridae (more than 70 serotypes), are distributed throughout the world and cause a broad spectrum of clinical manifestations in man and animals. Human enteroviruses are classified into 5 species: human enteroviruses A to D, and Poliovirus. Recombination has long been known to be an important property of poliovirus genetics; however, recombination has been recently shown to be also ubiquitous in non-polio enteroviruses. Prototype enterovirus strains have complex phylogenetic relations and all currently available enterovirus strains are recombinant to prototype strains. The extremely high rate of recombination allows the fragments of the enterovirus genome to evolve independently at a microevolutionary scale. Recombination strictly takes place between the members of the same species, usually outside the capsid-encoding genome region. Therefore, it can be concluded that the species enterovirus exists as a worldwide reservoir of the genetic information that shuffles frequently to produce new virus variants. This new model of the genetics of enterovirus accounts for failures to associate the serotype of enteroviruses and the form of the disease caused. The contents of this review have been published in the Reviews in Medical Virology, 2005.

[Molecular epidemiology of the ECHO 30 virus in Russia and CIS countries]. / Molekuliarnaia épidemiologiia virusa ECHO 30 na territorii Rossii i stran SNG.

Nucleotide sequences of the VP1 genome of 28 ECHO 30 strains isolated in 1998-2002 in Russia and several CIS countries were determined. The EV30 studies strain were divided into 4 groups according to isolation place and time. Group 1 is presented by 2 strains isolated in 1998 in Russia and Byelorussia. Group 2 comprises 17 strains isolated in 1999-2000 in Russia (its southern regions and Stavropol Territory), Ukraine, Georgia, and Azerbaijan. Five strains isolated in 2002 in Russia (Kalmykia) belong to Group 3; and Group 4 has 4 strains isolated in 2002 in Moldova and Russia (Magadan). A frequently changing EV30 subtype was simultaneously detected in extensive territories.