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Dystrophin is essential for muscle health: its sarcolemmal absence causes the fatal, X-linked condition, Duchenne muscular dystrophy (DMD). However, its normal, spatial organization remains poorly understood, which hinders the interpretation of efficacy of its therapeutic restoration. Using female reporter mice heterozygous for fluorescently tagged dystrophin ( Dmd EGFP ), we here reveal that dystrophin distribution is unexpectedly compartmentalized, being restricted to myonuclear-defined sarcolemmal territories extending ~80 µm, which we called “basal sarcolemmal dystrophin units (BSDUs).” These territories were further specialized at myotendinous junctions, where both Dmd transcripts and dystrophin protein were enriched. Genome-level correction in X-linked muscular dystrophy mice via CRISPR/Cas9 gene editing restored a mosaic of separated dystrophin domains, whereas transcript-level Dmd correction, following treatment with tricyclo-DNA antisense oligonucleotides, restored dystrophin initially at junctions before extending along the entire fiber—with levels ~2% sufficient to moderate the dystrophic process. We conclude that widespread restoration of fiber dystrophin is likely critical for therapeutic success in DMD, perhaps most importantly, at muscle–tendon junctions.

Summary Rheumatoid arthritis (RA) is a systemic inflammatory autoimmune disease that leads to joint destruction and disability. Despite a significant progress in administration of biological agents for RA patients, there is still a need for improved therapy. Intravenous immunoglobulins (IVIG), a pooled polyspecific immunoglobulin (Ig)G extracted from 5000 to 20 000 healthy subjects, showed beneficial therapeutic effect in patients with immune deficiency, sepsis and autoimmune diseases. The current study aimed to investigate the beneficial effect of treatment with IVIG in established collagen-induced arthritis in DBA/1j mice. Murine arthritis was induced in DBA/1j mice. Treatment with IVIG began when the disease was established. The clinical score was followed twice a week until day 48. The mice were bled for plasma and the paws were hematoxylin and eosin (H&E)-stained. Cytokine profile in the plasma was analyzed by Luminex technology and titers of circulating anti-collagen antibodies in the plasma was tested by enzyme-linked immunosorbent assay. Our results show that treatment with IVIG in murine significantly reduced the clinical arthritis score (P

International audience; Soluble guanylate cyclase (sGC) is the human receptor of nitric oxide (NO) in numerous kinds of cells and produces the second messenger 3',5'-cyclic guanosine monophosphate (cGMP) upon NO binding to its heme. sGC is involved in many cell signaling pathways both under healthy conditions and under pathological conditions, such as angiogenesis associated with tumor growth. Addressing the selective inhibition of the NO/cGMP pathway is a strategy worthwhile to be investigated for slowing down tumoral angiogenesis or for curing vasoplegia. However, sGC inhibitors are lacking investigation. We have explored a chemical library of various natural compounds and have discovered inhibitors of sGC. The selected compounds were evaluated for their inhibition of purified sGC in vitro and sGC in endothelial cells. Six natural compounds, from various organisms, have IC50 in the range 0.2-1.5 μM for inhibiting the NO-activated synthesis of cGMP by sGC, and selected compounds exhibit a quantified antiangiogenic activity using an endothelial cell line. These sGC inhibitors can be used directly as tools to investigate angiogenesis and cell signaling or as templates for drug design.

Measles is a disease caused by the highly infectious measles virus (MeV) that results in both viremia and lymphopenia. Lymphocyte counts recover shortly after the disappearance of measles-associated rash, but immunosuppression can persist for months to years after infection, resulting in increased incidence of secondary infections. Animal models and in vitro studies have proposed various immunological factors underlying this prolonged immune impairment, but the precise mechanisms operating in humans are unknown. Using B cell receptor (BCR) sequencing of human peripheral blood lymphocytes before and after MeV infection, we identified two immunological consequences from measles underlying immunosuppression: (i) incomplete reconstitution of the naive B cell pool leading to immunological immaturity and (ii) compromised immune memory to previously encountered pathogens due to depletion of previously expanded B memory clones. Using a surrogate model of measles in ferrets, we investigated the clinical consequences of morbillivirus infection and demonstrated a depletion of vaccine-acquired immunity to influenza virus, leading to a compromised immune recall response and increased disease severity after secondary influenza virus challenge. Our results show that MeV infection causes changes in naive and memory B lymphocyte diversity that persist after the resolution of clinical disease and thus contribute to compromised immunity to previous infections or vaccinations. This work highlights the importance of MeV vaccination not only for the control of measles but also for the maintenance of herd immunity to other pathogens, which can be compromised after MeV infection.

To study ultrastructural changes in stria's vascularis cells of inner ear and determine possible ways of correction.The work was carried out on male guinea pigs. After completion of the experiment, stria vascularis of inner ear was subjected to electron microscopic examination.In the control group (receiving gentamycin sulfate in an ototoxic dose), signs of blood flow disturbance were revealed, as well as ultrastructural changes in stria's vascularis cells (expansion of intercellular space, deformation of organelles, thinning of glycocalyx, blebbing). Also, fragmented cells were found. These changes are characteristics for apoptosis. In experimental group (receiving gentamycin sulfate and melaxen), degenerative changes were less pronounced. An increase of cell's secretory activity was observed.Changes in stria's vascularis cells by using melaxen are less pronounced. Increase of cell's secretory activity in stria vascularis is a compensatory reaction and saves auditory function.Цель исследования - изучение ультраструктурных изменений в клетках сосудистой полоски внутреннего уха и определение возможных путей коррекции. Результаты и обсуждение. В контрольной группе (животные получали гентамицина сульфат в ототоксической дозе) были выявлены признаки нарушения кровотока, а также ультраструктурные изменения клеток сосудистой полоски (расширение межклеточного пространства, деформация органелл, истончение гликокаликса, блеббинг). Также были обнаружены фрагментированные клетки. Данные изменения характерны для апоптоза. В экспериментальной группе (животные получали гентамицина сульфат и мелатонин) дегенеративные изменения были менее выраженными. Наблюдалось увеличение секреторной активности клеток. Заключение. Таким образом, ототоксические изменения в сосудистой полоске при применении мелатонина носят менее выраженный характер. Повышение секреторной активности клеток сосудистой полоски является компенсаторной реакцией и морфологическим субстратом сохранения слуховой функции.

Rift valley fever virus (RVFV) is a causative agent of a viral zoonosis that constitutes a major clinical burden in wild and domestic ruminants. The virus causes major outbreaks in livestock (sheep, goats, cattle and camels) and can be transmitted to humans by contaminated animal products or via arthropod vectors. Human-to-human transmission has not been reported to date, but spill-over events from animals have led to outbreaks in humans in Africa and the Arabian Peninsula. Currently, there is no licensed human vaccine against RVFV and the virus is listed as a priority pathogen by the World Health Organisation (WHO) due to the high epidemic potential and the lack of effective countermeasures. Multiple large RVFV outbreaks have been reported since the virus was discovered. During the last two decades, over 4000 cases and ~1000 deaths have been reported. The lack of systematic surveillance to estimate the true burden and incidence of human RVF disease is a challenge for planning future vaccine efficacy evaluation. This creates a need for robust diagnostic methodologies that can be deployed in remote regions to aid case confirmation, assessment of seroprevalence as well as pathogen surveillance required for the different stages of vaccine evaluation. Here, we perform comprehensive landscaping of the available diagnostic solutions for detection of RVFV in humans. Based on the identified gaps in the currently available in-house and commercially available methods, we highlight the specific investment needs for diagnostics that are critical for accelerating the development of effective vaccines against RVFV.

Despite decades of surveillance and pharmaceutical and non-pharmaceutical interventions, seasonal influenza viruses continue to cause epidemics around the world each year. The key process underlying these recurrent epidemics is the evolution of the viruses to escape the immunity that is induced by prior infection or vaccination. Although we are beginning to understand the processes that underlie the evolutionary dynamics of seasonal influenza viruses, the timing and nature of emergence of new virus strains remain mostly unpredictable. In this Review, we discuss recent advances in understanding the molecular determinants of influenza virus immune escape, sources of evolutionary selection pressure, population dynamics of influenza viruses and prospects for better influenza virus control.

Velichka Andonova,1,2 Petya Peneva,1,2 George S Georgiev,3 Vencislava T Toncheva,3 Elisaveta Apostolova,4 Zhivko Peychev,5 Stela Dimitrova,6 Mariana Katsarova,6 Nadia Petrova,7 Margarita Kassarova1,2 1Department of Pharmaceutical Sciences, Faculty of Pharmacy, Medical University-Plovdiv, 2Technological Center for Emergency Medicine (TCEMED), Plovdiv, 3Faculty of Chemistry and Pharmacy, Sofia University “St Kliment Ohridski”, Sofia, 4Department of Pharmacology and Drug Toxicology, Faculty of Pharmacy, Medical University-Plovdiv, 5Department of Medical Informatics, Biostatistics and e-learning, Faculty of Public Health, Medical University-Plovdiv, 6Department of Chemistry and Biochemistry, Faculty of Pharmacy, Medical University-Plovdiv, Plovdiv, 7Institute of Mineralogy and Crystallography, Bulgarian Academy of Sciences, Sofia, Bulgaria Abstract: The purpose of the study was to investigate the stability and biopharmaceutical characteristics of ketoprofen, loaded in polymeric carriers, which were included into a bigel in a semisolid dosage form. The polymer carriers with in situ-included ketoprofen were obtained by emulsifier-free emulsion polymerization of the monomers in aqueous medium or a solution of the polymers used. The morphological characteristics of the carriers, the in vitro release and the photochemical stability of ketoprofen were evaluated. The model with optimal characteristics was included in a bigel formulation. The bigel was characterized in terms of pH, rheological behavior, spreadability, and in vitro drug release. Acute skin toxicity, antinociceptive activity, anti-inflammatory activity, and antihyperalgesic effects of the prepared bigel with ketoprofen-loaded polymer carrier were evaluated. The carriers of ketoprofen were characterized by a high yield and drug loading. The particle size distribution varied widely according to the polymer used, and a sustained release was provided for up to 6 hours. The polymer mixture poly(vinyl acetate) and hydroxypropyl cellulose as a drug carrier, alone or included in the bigel composition, improved the photostability of the drug compared with unprotected ketoprofen. The bigel with ketoprofen-loaded particles provided sustained release of the drug and had optimal rheological parameters. In vivo experiments on the bigel showed no skin inflammation or irritation. Four hours after its application, a well-defined analgesic, anti-inflammatory, and antihyperalgesic effect was registered. The polymer mixture of poly(vinyl acetate) and hydroxypropyl cellulose as a carrier of ketoprofen and the bigel in which it was included provided an enhanced photostability and sustained drug release. Keywords: antihyperalgesic effect, antinociceptive activity, biphasic systems, carrageenan-induced edema, emulsifier-free radical polymerization

A novel betacoronavirus associated with lethal respiratory and renal complications was recently identified in patients from several countries in the Middle East. We report the deep genome sequencing of the virus directly from a patient’s sputum sample. Our high-throughput sequencing yielded a substantial depth of genome sequence assembly and showed the minority viral variants in the specimen. Detailed phylogenetic analysis of the virus genome (England/Qatar/2012) revealed its close relationship to European bat coronaviruses circulating among the bat species of the Vespertilionidae family. Molecular clock analysis showed that the 2 human infections of this betacoronavirus in June 2012 (EMC/2012) and September 2012 (England/Qatar/2012) share a common virus ancestor most likely considerably before early 2012, suggesting the human diversity is the result of multiple zoonotic events., link_to_OA_fulltext

Background: Oxidative stress and amyloid deposition are tightly interconnected pathological features of Alzheimer disease. In this respect, both amyloid production and aggregation may be stimulated by oxidative stress and also the increase of pathogenic β-amyloid and its aggregated form lead to oxidative stress progression. Therefore, the search for potential drugs with both antioxidant and antiaggregation properties are of great interest. Methods: In this study, we described the stereospecific synthesis of alkaloid securinine aminoderivatives. Results: We showed that the newly synthesized compounds possess antioxidant and metal-chelating properties. Indeed, we report that one compound has inhibitory effects towards μ-amyloid aggregation. Conclusion: Based on these results, aminoderivatives of securinine scaffold are promising compounds for development of new drugs for the treatment of neurodegenerative diseases.

Four males and three females ranging in age from 20 to 35 years and afflicted with complicated Type 1-diabetes for more than 8 years underwent islet cell allotransplantation (ATx, 6 cases) and xenotransplantation (XTx, 1 case). Precultured islet cells derived from human or bovine fetal pancreata were injected into the m. rectus abdominis. Immunosuppression was not applied. Plasma C-peptide and islet cell surface antibodies (ICSA) were continually measured both before and until the twentienth week following islet cell transplantation. All recipients were subdivided as "responsive" (RR, 3 males) or "non-responsive" (NRR, 1 male and 3 females), according to the dynamics of their ICSA levels. All 3 RR (1XTx and 2 ATx) showed a peak of ICSA two weeks after cell injection. Subsequent ICSA levels had the tendency to either diminish or increase. Heterogeneity of preoperative antibody level, especially in NRR, was also observed. No associations between ICSA and ATx or XTx, age at diabetes onset, or duration of the disease was found. Only one RR with XTx had a reduced daily insulin requirement and a significant C-peptide response similar to the dynamics of ICSA levels. A greater mass of available bovine islet cells might be responsible for this effect.

We studied the effect of corticotropin-like intermediate lobe peptide (CLIP) on presynaptic NMDA receptors and postsynaptic GABA, NMDA, and AMPA receptors in rat brain. CLIP inhibited presynaptic and postsynaptic NMDA receptors, but potentiated postsynaptic GABA and AMPA receptors. Our results indicate that CLIP modulates function of ionotropic receptors for glutamate and GABA.

We analyzed the effects of weak combined magnetic fields, tuned to the cyclotron resonance condition for calcium ions, obtained in different phases of planarian regeneration. We showed that the result of regeneration in 72 hours after decapitation depends on the length of exposure, and the time between decapitation and initiation of a half-hour exposure. The experimental dependence can be explained by a multiplicity of enzymatic targets activated in different phases of the regeneration process.

The main dogma in reproductive medicine claims that ovarian reserve in mammals is formed during prenatal period and the decline in the existing oocyte pool can not be compensated. The thesis that girls are born with a certain number of eggs, which gradually decrease with age, and the ovaries can not produce more reproductive cells was greatly "shaken" during the last decade. Some reports have appeared for the presence of germline stem cells in the adult ovaries and for their capacity to form oocytes in the postnatal period. In this review are provided summarized literature data on this matter.

This article reviews accumulating knowledge on the signs and pathogenesis of cognitive impairment in schizophrenia. The existing data indicate that cognitive disorders, along with positive and negative symptoms, are key symptoms of schizophrenia. Several neurotransmitter systems, including prefrontal dopaminergic transmission, are involved in the pathological process that causes cognitive disorders. GABAergic and glutamatergic pathophysiological models of schizophrenia are discussed. The main subtype of glutamate receptor NMDA receptor hypofunction is one of the causes of cognitive impairment in schizophrenia. Recent studies revealed alterations in gene expression and activity of kynurenine pathway enzymes in patients with schizophrenia. Concentration of kynurenic acid (KYNA) is increased in the prefrontal cortex of schizophrenca patients and correlated with glutamate, dopamine and acetylcholine levels that reflects the pathophysiological mechanism and degree of cognitive impairment. An overview of animal models of schizophrenia, in particular pharmacological models, is presented. Treatment approaches that promote the recovery of cognitive function are described.

We studied the effect of somatostatin on presinaptic NMDA receptors and postsinaptic GABA, NMDA, and AMPA receptors in rat brain. It was shown that somatostatin inhibits NMDA-induced (45)Ca(2+) uptake into synaptosomes isolated from rat brain cortex (IC50=2.8×10(-11) M). Somatostatin potentiates AMPA receptors and inhibits hippocampal NMDA receptors in the entire range of examined concentrations (10(-14)-10(-7) M); it also potentiates or inhibits GABA receptor currents in a concentration-dependent manner. Our results suggest that somatostatin modulates the function of ionotropic glutamate and GABA receptors and is involved in cognitive and neurodegenerative processes in the mammalian brain.

We studied the effect of muramyl dipeptides on postsynaptic GABA, NMDA, and AMPA receptors and presynaptic NMDA receptors. L,D-Dipeptide more potently than L,L-dipeptide inhibited postsynaptic NMDA receptors, potentiated GABA and AMPA receptors, and inhibited (45)Ca(2+) uptake in synaptosomes from of rat brain cortex. Our results indicate that muramyl dipeptides modulate function of glutamate and GABA receptors.

We studied the effect of prostaglandins on presynaptic NMDA receptors. Prostaglandin E2 inhibited NMDA-induced (45)Ca2+ uptake by synaptosomes in low concentrations (IC50 approximately 10 microM), but potentiated it in higher concentrations. Prostaglandin D2 increased (45)Ca2+ uptake by synaptosomes during stimulation of NMDA receptors. Our results indicate that prostaglandins D2 and E2 modulate function of presynaptic NMDA receptors.

African swine fever is viral disease of domestic and wild pigs which leads to almost total mortality and causes great economic losses due to absence of vaccine. Having been introduced into the Russian Federation in 2007 the disease has spread widely in the southern region of the country and since 2011 has demonstrated a tendency to form a secondary endemic zone in the central part of the country. In the present study spatio-temporal patterns of ASF diffusion in the populations of wild and domestic pigs are analyzed. The structure of the domestic swine population is conventionally divided into a sub-population at low biosecurity (77% of the total number of outbreaks in domestic pigs) and a population at high biosecurity (23%). The statistics of ASF cases registered in each of these sub-populations is presented. The possible causes of ASF diffusion across the country are discussed. The use of geo-information technologies (GIS) enabled confirmation of the conclusion that an epidemic center has shifted into the central part of Russia. The main conclusions of this study are that: (1) anthropogenic factors play the leading role in the spread of ASF across the territory of the RF; (2) small-scale private holdings (low biosecurity population) are more exposed to ASF virus introduction; (3) there is a high risk of diffusion of ASFV from the secondary endemic zone in the central part of the RF to neighboring regions.

Novel 1,2,4-thiadiazole derivatives as potent neuroprotectors were synthesized and identified. Their ability to inhibit the glutamate stimulated Ca uptake was measured. Permeation experiments on the phospholipid membranes were conducted, and the apparent permeability coefficients were obtained. The partition coefficients in n-octanol/buffer (pH 7.4) and n-hexane/buffer (pH 7.4) immiscible phases (as model systems for characterizing gastrointestinal tract membranes and BBB) were determined. A classification of the studied compounds from the standpoint of “permeability–activity” properties was proposed.

The neuroprotective action of hybrid structures based on fullerene C60 with attached proline amino acid has been studied. Hybrid structures contained natural antioxidant carnosine or addends with one or two nitrate groups. It has been shown that all studied compounds had antioxidant activity and decreased the concentration of malondialdehyde in homogenates of the rat brain. Compound 1, which contained the antioxidant carnosine, has been found to be the most effective antioxidant. All compounds except IV and V inhibited the activity of monoamine oxidase B, while compounds I-IV increased the activity of monoamine oxidase A. All investigated compounds inhibited glutamate-induced Ca2+ uptake into synaptosomes of the rat brain cortex. Compound III, containing two nitrate groups, has been found to be the most effective inhibitor. This compound caused a significant increase of the currents of AMPA receptors (AMPA, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid).

Various salts of 3-allyl-1,1-dibenzyl-2-ethyl-isothiourea, 1 (hydrochloride), 2 (hydrobromide), and 3 (hydroiodide), were synthesized. Ca-blocking properties of these salts were studied. Comparative analysis of the potentiating effects of 3 and cyclothiazide (CT) on transmembrane currents caused by kainic acid (KA) and glutamate in Purkinje neurons was performed. Analysis of the effects of 1 on N-methyl-D-aspartate (NMDA) receptors was performed on primary culture of heterogeneous neurons of rat cerebral cortex. Single crystals were grown and X-ray diffraction experiments solving the crystal structures of 1-3 were carried out. Analysis of conformations of the molecules in the crystal lattices was performed. The temperature dependencies of the solubility of 1-3 in water and n-octanol were obtained, and the thermodynamic parameters of solubility process were calculated. The effect of halogen atoms on the solubility was analyzed. The partitioning processes in the water-octanol system were studied at 25 degrees C. Chemical stability of tested salts in pH 7.4 phosphate buffer was determined at 25 degrees C.

It was demonstrated that uridine affects presynaptic NMDA and kainite receptors of rat brain cortex. Uridine considerably inhibited (45)Ca2+ uptake into synaptoneurosomes (IC50 = 7.1 x 10(-12) M) under conditions NMDA stimulation and increased it under conditions AMPA stimulation (157.8%).

We studied the effect of delta sleep-inducing peptide on GABA receptors of hippocampal and cerebellar neurons in rats. It was shown that delta sleep-inducing peptide considerably and dose-dependently potentiates GABA-activated currents in these neurons and blocks NMDA-activated potentiation in cortical and hippocampal neurons. The peptide modulates activity of presynaptic NMDA receptors, which is seen from changes in (45)Ca(2+) uptake into synaptosomes of the brain cortex after uptake stimulation with glutamate and NMDA.

Specificity of glutamate receptors in the P2 synaptosomal fraction from the cerebral cortex of newborn rats was studied by measuring (45)Ca(2+) uptake by synaptosomes in the presence of agonists of ionotropic and metabotropic glutamate receptors. It was shown that P2 synaptosomal fraction from rat cortex contains NMDA receptors, kainate receptors, and group 1 metabotropic receptors.

Malaria was eradicated on the territory of Moscow in 1960; its imported cases mainly from the countries of Asia and Africa and sporadic cases secondary to the imported ones were recorded during the following 40 years. Mass migration of the population (businessmen, seasonal workers, etc.) from the endemic CIS countries in the late 1990s, particularly to the Moscow Region, increased cases of tertian malaria, resulting from the transmission of the infection by the mosquitoes Anopheles. A total of 793 cases of malaria, including 27 parasitic carriers, were recorded from 2000 to 2004. The proportion of Muscovites was 24.1% of the total cases of malaria in 2002 and increased up to 50% in 2004. The causative agents of tertian malaria (Plasmodium vivax) were detected in 74.8% of the total number of cases. The ongoing importation of malaria from Tadjikistan and Azerbaijan, the late establishment of final diagnosis, the shortage of antimalarial drugs make the malaria situation worse in Moscow, which requires that antiepidemic measures should be intensified in the coming 3 years.

Different frequencies of chromosomal alterations in salivary gland polytene chromosomes AB, CD and EF were described in larvae of Chironomus riparius (syn. Chironomus thummi) from the trace metal-polluted station of Santena on the river Banna, near Turin, and from the unpolluted station of Corio (40 Km from Turin) which was taken as a reference area. In a sample of 56 larvae from Santena, no specimen with the standard karyotype in all cells of the salivary glands was found. Different types of aberrations were found: 33 paracentric and five pericentric inversions, three deficiencies, four amplified sections and one chromatid break. Fifteen out of the 38 inversions and two amplified sections appeared to be inherited, while all the other aberrations were somatic. Most of the aberrations' breakpoints were located on both sides of the centromere regions, where constitutive heterochromatin is present. Also functional alterations were observed (mainly telomere and centromere decondensations and nine novel puffs). In a sample of 49 larvae of a population from the well-preserved area of Corio only six somatic and one inherited paracentric inversions were found. These results suggest that the strong destabilization of the genomes of C. riparius larvae from Santena could be a reaction to the activity of the toxic substances present in the polluted sediments of the river Banna.

Potential immunodominant epitopes were predicted on the basis of a theoretical analysis of the antigenic structure of the VP1 protein of the type Asia-1 foot-and-mouth disease virus. Peptides corresponding to the 140-153, 136-153, 132-153, 143-157, 137-157, and 193-208 fragments of the VP1 protein sequence were synthesized by the solid phase method, and the immunogenic properties of the peptides were studied on guinea pigs. The shortest peptide exhibiting the protective effect was found to correspond to the, 140-153 fragment of the VP1 sequence. The Plm-(Gly)3-(140-153)-(Gly)2-Lys(Plm)-Leu and [Ac-(140-153)-(Gly)3]8-(Lys)7-Gly synthetic constructions in combination with adjuvants provided up to 80% protection of immunized animals against infection with the foot-and-mouth disease virus.

The effect of interleukin-1beta on presynaptic NMDA receptors was evaluated by studying NMDA-induced 45Ca2+ uptake by synaptosomes from rat brain cortex. Interleukin-1beta inhibited 45Ca2+ uptake by synaptosomes. Our results indicate that interleukin-1beta modulates presynaptic NMDA receptors and is probably involved in the regulation of synaptic transmission in the central nervous system.

Peptides were synthesized, which, according to theoretical analysis of the antigenic structure of protein VP1 of foot-and-mouth disease (FMD) virus types A, 0, and Asia 1, corresponded to potential immunodominant protein sites. Activities of the peptides were studied by solid-phase indirect radioimmunoassay on polyethylene film with purified immunoglobulins against intact FMD virus. Virtually no cross reactions were observed. Blood sera of cattle convalescent after FMD were tested with the FMD virus and peptides containing VP1 fragments 141-160 (A22 No. 550), 140-160 (O1 No. 194), and 140-153 (Asia 1 No. 48). The specificity of interactions between the sera and peptides and the virus was uniform, this permitting the identification of the virus type which caused the disease.

The reaction of chicken myoblasts in competent primary culture during their contact with activated rat neutrophils was evaluated. In the incubation medium dynamics of activities were defined for proteinases and creatine kinase, protein content, as well as for DNA and protein marked precursor increase. An increase in myoblast membrane permeability has been revealed. It is shown that the culture is a sensitive model for investigation of physiological interaction of neutrophils with muscle tissue cells.

A cytogenetic analysis was carried out on a population of Chironomus balatonicus (Chironomidae, Diptera) from Chernobyl, a highly radioactive area of the Kiev region. Several chromosomal aberrations were established unique to a population of Chironomus balatonicus living in an area contaminated by radioactive waste. Five new heterozygous inversions, deficiencies in arms C, D, E, F and chromatid breaks were found in the irradiated population but not in nonirradiated populations. A pericentric inversion in chromosome AB occurred at a relatively high frequency. Genome aberrations expressed by a heterochromatized 'B' chromosome were evident. In the irradiated and nonirradiated populations common inversions occurred showing variation in their frequency depending on specific environmental conditions. The somatic and also the germ cells were characterized by a number of heteropycnotic nuclei and vacuolized chromosomes. Both the somatic and germ cells showed changes in the structural and functional organization of heterochromatin and this was particularly marked in the telomeric sectors of the chromosomes. The heterochromatin which is extremely sensitive to radioactivity appears to protect euchromatin from adverse environmental conditions.

Neutrophil proteinases have been studied for their activity in the rat skeletal muscles after muscular activity using antiserum to neutrophils. The increase of proteolytic activity after physical load partially determined by neutrophil proteinases in the skeletal muscles is shown.

Abdominal neutrophils effect on rat skeletal muscle m. soleus was investigated in vitro. The incubation was carried out in Hanks balanced solution within 24 hrs. It was a release of proteins from m. soleus 1 hr later. Creatine kinase (CK) and aspartate aminotransferase (AAT) activities increase was detected in incubation medium. The neutrophils released their proteins quicker than muscles. A dramatic inhibition of CK and AAT activities took place during coincubation of m. soleus and neutrophils. Zymosan-activated cells had a higher inhibition potency in comparison to nonactivated neutrophils. Analysis of proteinase and myeloperoxidase activities in incubation medium has given evidence that CK and AAT inhibition by non-activated neutrophils mainly depends on cell-secreted proteinases. Zymosan-activated neutrophil inhibition of CK and AAT consists of proteinases and myeloperoxidase effects. AAT appeared to be more resistant than CK to the damage by neutrophils. The used approach failed to demonstrate the direct damage effect of neutrophils on m. soleus, but the described enzyme inhibition mechanism can take place in vivo during leukocyte infiltration of skeletal muscles after intensive muscular activity.

A study was made of the influence of creatine phosphate (CP) on creatine kinase activity and the total protein status of the primary chicken myoblast culture. It has been established that the two-hour exposure of the developing culture in the presence of 2 and 4 mM CP is accompanied by the lowering of the content of water extractable protein and a considerable increase of creatine kinase activity. Meanwhile the total activity of the enzyme rises in the non-differentiated and falls in the differentiated one. Electrophoretic analysis has shown CP to produce not only quantitative but also deep qualitative changes in the myoblast protein spectrum. It is assumed that the changes revealed are nonspecific.

The information on the initial step of enzymatic reaction of 2.5S RNA isolated from muscle 1,4-alpha-glucane branching enzyme (EC 2.4.1.18), isomerase amylose, has been reported for the first time. The 2.5S RNA-polysaccharide (starch) complex was isolated and partly characterized. It was shown that the complex retains its integrity during precipitation with ethanol, gel-filtration on a Biogel P-150 column and electrophoresis but dissociates into RNA and starch upon fractionation on a DEAE-52 cellulose column. Treatment of the original preparation with RNAase fully reflects the complex formation: with a decrease in the RNA content in the original preparation that of the synthesized complex diminishes. The RNA-polysaccharide complex exhibits the properties of the branching enzyme; its enzymatic activity markedly exceeds that of the free RNA.

Possibility of ortho-, para-, meta-methylphenyl and methoxyphenyl-derivates of MPTP to produce parkinsonism was investigated. Only ortho-methylphenyl- and ortho-methoxyphenyl-derivates of MPTP cause a persistent loss in dopamine content in the brain and produced the clinical symptoms of parkinsonism. All substances produced Parkinsonian-like syndrome gives the symptoms of activation of nervous system during 0.5-1 h after injection and symptoms of depression in following 3 h of observations.

It has been stated that separate and simultaneous administration into the animal organism of low doses of copper, lead and nickel with drinking water is accompanied by dose-dependent change in the level of not only administered but also endogenic elements in animals' organs. A supposition has been made that the character of changes in the content of the administered and endogenic metals is associated in the organism of animals with the depth of disturbances in metabolic processes.

The spontaneous mutation rate was studied in cultured human and Chinese hamster cells. Mutations to 8-azaguanine (8-AG) and 6-mercaptopurine (6-M) resistance and mutations controlling the ability to proliferate in a medium with low glucose concentration (“glucose-independence”) were investigated. The following cell lines were used: a hypodiploid clone 237 1 and a hyperdiploid clone 431-8 of Chinese hamster cells derived from line BIId-ii-FAF28; diploid lung cell strains of human male embryos L-53 and L-54 and of female embryo WI-38; an aneuploid line Glen, obtained from strain WI-38 by transformation. Using the fluctuation test, it was shown that the mutations for resistance to purine analogues and glucose-independence arose spontaneously. Karyological analysis of independent mutants (10 clones of Chinese hamster cells resistant to purine analogues and 5 glucose-independent clones) did not reveal any correlation between the acqsisition of the mutant phenotype and the appearance of some specific chromosome change. It is suggested that the characters studied are due to gene mutations. The rates of spontaneous mutation to 8-AG and 6-M resistance in Chinese hamster cells are respectively 1.5·10 −5 and 0.4·10 −5 . The spontaneous rate to glucose-independence mutations is as high as 10 −4 . The latter character may be due to mutations affecting not a single but several different loci. Consequently the mutation rate obtained could be the sum of the rates of mutations in all these loci. No difference was observed between the rates of mutation to 8-AG resistance in the human diploid strain L-54 and the aneuploid line Glen (about 7·10 −5 . The relative mutability of normal diploid and malignant aneuploid somatic cells and the levels of mutability in somatic and germ cells are discussed.

The primary structure of the nucleic acid from the branching enzyme 1,4-alpha-D-glucan: 1,4-alpha-D-glucan 6-alpha-(1,4-alpha-glucano)-transferase (2.5-S RNA) isolated from rabbit muscles has been elucidated. The polyribonucleotide consists of 31 nucleotides; the unique features of the polyribonucleotide are the unusually high content of modified nucleotides (32%) and guanine residues (40%). Apparently 2.5-S RNA belongs to a class of nucleic acids unknown up to now. It is the first time that the structure of a nucleic acid component from a ribonucleoenzyme has been defined. This work is a preprequisite for gaining insight into the intimate activating effect of the poly-ribonucleotide on the enzyme action.

After polyethylene glycol treatment of GPRT- mutant cells, 12 clones were isolated on the ATG medium showing intragenic complementation. Karyological analysis confirmed the hybrid nature of the clones isolated. The GPRT activity in the hybrid clones, as assessed in vitro, exceeded the sum of parental activities. In vivo incorporation of 14C-hypoxanthine showed the GPRT activity in the hybrids to be an order of magnitude higher than in the mutant parental cells. Moreover, the GPRT activity in the hybrid clones was found to increase considerably during cultivation on the ATG medium; hence, their ability to multiply on this selective medium. All hybrid cells surviving and multiplying on the ATG medium were shown to maintain a high enough resistance to 8-AG, 6-MP and, to somewhat less extent, to 6-TG. The frequency of complementation was determined for the cells of mutant clones selected on media with different purine base analogues. The complementation map for the GPRT locus was constructed and proved to be linear. Five groups of complementation were specified.