[Potential for infusion correction of COVID-19-associated endotheliopathy].

AIM: To evaluate the relationship between the systemic inflammatory response and the severity of COVID-19-associated endotheliopathy and the effect of succinate-containing crystalloid solution (sodium meglumine succinate) on it in patients with severe COVID-19. MATERIALS AND METHODS: Clinical and laboratory parameters of 53 intensive care unit's patients with COVID-19 complicated by community-acquired bilateral multisegmental pneumonia were analyzed. Intensive therapy complex of 27 patients (study group) included daily infusion of 1.5% solution of sodium meglumine succinate (Reamberin) in the daily dose of 10 ml/kg for at least 11 days (or during the whole stay in the unit). A similar volume of Ringer's solution was present in the control group of 26 patients. The levels of endotheliocytosis, homocysteine, and systemic inflammatory response were determined at all stages of the study. RESULTS: The evaluation of endotheliopathy degree in the meglumine succinate group showed a significant reduction of initially elevated levels of endotheliemia and homocysteinemia at all study stages. The pattern of changes in the study group was highly correlated (r=0.90-0.96) with the dynamics of systemic inflammatory response parameters-fibrinogenemia, C-reactive protein and interleukin-6 levels. As normalization of the immune imbalance, we regarded the termination of lymphopenia in the Reamberin group. CONCLUSION: Early inclusion of Reamberin infusion into intensive therapy of severe COVID-19, in comparison with Ringer's solution, leads to significant and stable correction of the severity of systemic inflammatory response, which in turn is naturally reflected in the severity of endothelial dysfunction, multiple organ failure, and also leads to a decrease in 28-day mortality.

Systemic Amyloidosis Model on Young Mice.

Subcutaneous daily injection (with neglect of aseptics) of 0.5 ml solution of soybean cream substitute (10% volume in distilled water) during 30 days caused systemic amyloidosis in 30-day-old mice. All the known methods for induction of systemic amyloidosis are based on the use of old animals, as senile tissue bradytrophy allows effective simulation of amyloidosis.

First Report of Beet severe curly top virus in Jalapeño Pepper in Chihuahua, Mexico.

Curly top is a serious problem in many irrigated crops in the semiarid areas in the western United States. The disease is caused by a complex of leafhopper-transmitted curtoviruses, one of which, Beet mild curly top virus (BMCTV), was previously found in chili pepper in Zacatecas and Aguascalientes, Mexico (3). During the past few years, sporadic symptoms similar to curly top disease were observed in jalapeño pepper in the south-central area of Chihuahua State. Symptomatic plants were scattered in otherwise healthy looking pepper stands and displayed stunting and yellowing. Affected leaves were brittle, showed upward curling, and a distinct green vein pattern with interveinal yellowing. In June and August of 2010, field surveys were conducted in Cordillera-Escuadra, Meoqui-Estacion Consuelo, Meoqui-Lomas del Consuelo, and Delicias-Presa Francisco I Madero. Ninety-four leaf samples were collected from symptomatic jalapeño pepper plants and subjected to ELISA and PCR testing for curly top. Of the 94 samples, 11 were found to be positive by triple-antibody sandwich-ELISA with polyclonal antibodies against curly top (2). To confirm the identification of curly top and type the specific curtovirus identified, four ELISA-positive samples were subjected to a PCR analysis using a virus-specific primer set for curtovirus typing designed by Chen et al. (1). All four samples tested produced a single 720-bp band with primers BSCTVv2688 and BGc396 (1) characteristic of the Beet severe curly top virus (BSCTV). These curly top-specific PCR amplicons were sequenced and found to be 99% similar to the BSCTV nucleotide sequence in the C1 gene region (GenBank Accession No. X97203); corresponding sequences were deposited in GenBank under Accession Nos. JF437870 to JF437873. To our knowledge, this is the first report of the curly top virus in the State of Chihuahua, demonstrating that curly top is established and common in jalapeño pepper here and will need surveillance in other vegetable crops under irrigation. References: (1) L. F. Chen et al. Plant Dis. 94:99, 2010. (2) J. Durrin et al. Plant Dis. 94:972, 2010. (3) R. Velásquez-Valle et al. Plant Dis. 92:650, 2008.

First Identification of an Unusual Recombinant Potato virus Y Strain in Potato in Mexico.

Potato virus Y (PVY) has been reported in potato crops in Mexico (3), with tobacco necrotic variants found in the central State of Mexico (4). Nevertheless, many individual states are currently declared PVY free and distribution of individual strains of PVY in potato in different states of Mexico and in different solanaceous crops had not yet been studied. A limited field PVY survey was conducted on potato in the State of Chihuahua in August 2009. More than 900 random potato leaf samples were collected from cvs. Snowden, Atlantic, FL1867, Felsina, Fianna, Gigant, and Alpha. Seven were found to be PVY-positive and had been collected from cvs. Fianna, Snowden, and FL1867. The PVY status of the collected samples was initially determined with the PVY-specific Immunostrips (Bioreba, Reinach, Switzerland) and by double-antibody sandwich-ELISA using the polyclonal PVY detection kit (Agdia, Elkhart, IN). To determine the strain specificity of these PVY isolates following ELISA tests, the infected original samples were inoculated onto tobacco plants at the four-leaf stage and symptom appearance and development were observed for 8 weeks side-by-side with control isolates PB-Oz (PVYO), N4 (PVYNTN), and Mont (PVYN) (1), followed by the standard PVY strain typing by reverse transcription (RT)-PCR (2). Only one of the PVY-positive samples, originally from symptomless potato cv. Fianna, induced systemic PVY infection in tobacco by producing stunting, mosaic, and vein clearing. No systemic vein necrosis, characteristic of isolates Mont and N4, was observed in Nicotiana tabacum cvs. Burley, Xanthi, or Samsun after inoculation with this isolate during all 8 weeks of observation. This isolate, PVY-M3, was typed as a PVY recombinant by RT-PCR, with two recombinant junctions characteristic of European PVYNTN strains (2). It was further analyzed by triple-antibody sandwich-ELISA using four PVYO and PVYN strain-specific monoclonal antibodies. Monoclonals 1F5 (Agdia) and SASA-N (Scottish Agriculture Science Agency [SASA], Edinburgh) reacted to this isolate and identified PVY-M3 serologically as PVYN serotype, characteristic of other PVYNTN recombinants. Monoclonals MAb2 (Agdia) and SASA-O (SASA), specific to PVYO and PVYC strains, did not react to PVY-M3. Taken together, the combination of biological, serological, and molecular characteristics define this recombinant isolate from Mexico as belonging to the same PVY strain group represented by the isolate PVY-L26 (1). To our knowledge, this is the first report of such an unusual PVYNTN recombinant strain from Mexico. Presence of this isolate, with no vein necrotic symptoms induced on tobacco and with PVYNTN genome, will necessitate development of new detection methods for the seed potato industry in Mexico. References: (1) X. Hu et al. Virus Res. 143:68, 2009. (2) J. L. Lorenzen et al. Plant Dis. 90:935, 2006. (3) L. P. Moreno et al. Rev. Mex. Fitopatol. 22:187, 2004. (4) V. R. Ramirez-Rodriguez et al. Virol. J. 6:48, 2009.

[Alcoholic hepatosis in pathogenesis of hepatic encephalopathy].

The problem of chronic alcoholism has a great social importance. In the literature there are not methodological grounds for verification of causes and mechanisms of unexpected death caused by chronic alcoholic intoxication. Given literature data show the reasonability of study of hepatic encephalopathy in combination with pathomorphological, histochemical and biochemical analyses in comparison with forensic chemical analysis of alcohol concentration in human organism. That will make possible to determine the mechanisms of tanatogenesis of alcoholic intoxication.

Characterization of the beet yellow stunt virus coat protein gene.

ABSTRACT The beet yellow stunt virus (BYSV) genome contains at least nine open reading frames (ORFs) that code for proteins ranging from 6 to 66 kDa. Based on amino acid sequence comparisons, the coat protein (CP) was previously identified as the product of ORF7. We expressed the product of ORF7 in bacteria and confirmed that ORF7 codes for the BYSV CP by immunoblotting. BYSV is a phloem-limited virus, and virus CP antigen of a quality sufficient for diagnostic antisera production has not been available. To produce BYSV antigen free of plant host contaminants, ORF7 was cloned into a pMAL bacterial expression vector. The resulting fusion protein was affinity-purified and used as an antigen to raise anti-BYSV CP antisera in rabbits and guinea pigs. Using these antisera, an indirect double-antibody sandwich (DAS) enzyme-linked immunosorbent assay (ELISA)-based diagnostic system was developed. This indirect DAS-ELISA format enabled reliable detection of BYSV in tissue extracts from virus-infected lettuce diluted up to 5,000 times. The diagnostic system developed may enable large-scale epidemiological studies of BYSV using simple serological techniques. The antisera raised had a titer exceeding 1 x 10(5) in immunoblots and easily detected the 23.7-kDa BYSV CP in virus-infected lettuce and sowthistle plants. In these two plant species, BYSV CP was detected as two closely migrating bands during electrophoresis, which may suggest posttranslational CP modifications. To further characterize the BYSV CP gene, the 5'-untranslated region (UTR) of the BYSV CP subgenomic RNA (sgRNA) was cloned and sequenced. The CP-encoding, approximately 1.9-kb sgRNA has an AT-rich, 66-nucleotide-long 5'-UTR colinear to the genomic sequence upstream of ORF7.

Modulation of the antigenic reactivity of the citrus tristeza virus coat protein.

Trapping properties of a panel of monoclonal antibodies (Mabs) raised against citrus tristeza virus (CTV) were analyzed in an indirect double-antibody sandwich ELISA (I-DAS-ELISA). These antibodies had been previously assigned by serological specificity into five groups (I to V). Mabs from group V, which are directed to conformational epitopes, trapped significant amounts of virus antigen from CTV-infected plant tissue at IgG concentration above 10 ng/ml. Mabs from groups I to IV, which are directed to linear, continuous epitopes, performed poorly as coating antibodies, even at a 1 microgram/ml concentration of the IgG's, indicating that the respective linear epitopes were inaccessible. However, when Mabs from groups I to IV were combined with a small amount of Mabs from group V, a substantial increase in trapping of the CTV antigen was recorded. In this 'two antibody-binding assay' previously cryptic, linear epitopes of the CTV CP apparently became accessible to the Mabs from groups I to IV. Modulation of the antigenic reactivity of the CTV CP was also recorded upon binding of the Mabs directed to the conformational epitopes in solution. Induced exposure of the linear epitopes of the CTV CP was revealed in 'two antibody-binding assays' with pairwise combinations of different mouse Mabs and several rabbit and chicken polyclonal antisera with different serological specificities, including antisera to bacterially expressed CP fragments. This mixed coating in I-DAS-ELISA resulted in substantially increased efficiency of the virus antigen trapping by antisera produced against bacterially expressed protein fragments and an increased sensitivity of the CTV detection after optimization of the ratio between conformational and linear antibodies.

Organization of the 3'-terminal half of beet yellow stunt virus genome and implications for the evolution of closteroviruses.

The 3'-terminal half of the beet yellow stunt virus (BYSV) genome 10,545 nt, has been cloned and sequenced. The sequenced portion of the BYSV genome encompasses 10 open reading frames (ORFs) and 241 nt of the 3' untranslated region. The sequence spans, in the 5' to 3' direction, the C-terminal region of the replication-associated polyprotein gene (ORF 1a) which includes the set of motifs typical of helicases (HEL), the entire 53-kDa polymerase (RdRp) gene (ORF 1b), and genes encoding 30-kDa (ORF 2), 6-kDa (ORF 3), 66-kDa (ORF 4), 61-kDa (ORF 5), 25-kDa (ORF 6), 23.7-kDa (coat protein, CP) (ORF 7), 18-kDa (ORF 8), and 22-kDa (ORF 9) proteins. The double-stranded RNA "replicative form" of the BYSV was demonstrated to have a nontemplate G residue at the 3' terminus of the (+) strand. The RdRp of BYSV is presumably expressed via a +1 ribosomal frameshift. The five-gene module conserved among closteroviruses was identified in BYSV; it includes a gene array coding for a 6-kDa small hydrophobic protein, a 66-kDa homolog of the cellular HSP70 heat shock proteins, a 61-kDa protein, and a 25-kDa diverged copy of the CP followed by the CP gene itself. Phylogenetic analysis of the replication-associated HEL and RdRp domains as well as proteins from the five-gene module demonstrated the closest relationship between BYSV and two other closteroviruses, beet yellows (BYV) and citrus tristeza (CTV) viruses. Like CTV, the BYSV genome contains a 30-kDa protein gene between the RdRp and the 6-kDa protein genes, and like BYV it has only two genes downstream of the CP gene. The organization of the BYSV genome appears to be intermediate between BYV and CTV, which suggests that these three viruses might represent three distinct but probably close stages in the closterovirus evolution.

Complete sequence of the citrus tristeza virus RNA genome.

The sequence of the entire genome of citrus tristeza virus (CTV), Florida isolate T36, was completed. The 19,296-nt CTV genome encodes 12 open reading frames (ORFs) potentially coding for at least 17 protein products. The 5'-proximal ORF 1a starts at nucleotide 108 and encodes a large polyprotein with calculated MW of 349 kDa containing domains characteristic of (from 5' to 3') two papain-like proteases (P-PRO), a methyltransferase (MT), and a helicase (HEL). Alignment of the putative P-PRO sequences of CTV with the related proteases of beet yellows closterovirus (BYV) and potyviruses allowed the prediction of catalytic cysteine and histidine residues as well as two cleavage sites, namely Val-Gly/Gly for the 5' proximal P-PRO domain and Met-Gly/Gly for the 5' distal P-PRO domain. The autoproteolytic cleavage of the polyprotein at these sites would release two N-terminal leader proteins of 54 and 55 kDa, respectively, and a 240-kDa C-terminal fragment containing MT and HEL domains. The apparent duplication of the leader domain distinguishes CTV from BYV and accounts for most of the size increase in the ORF 1a product of CTV. The downstream ORF 1b encodes a 57-kDa putative RNA-dependent RNA polymerase (RdRp), which is probably expressed via a +1 ribosomal frameshift. Sequence analysis of the frameshift region suggests that this +1 frameshift probably occurs at a rare arginine codon CGG and that elements of the RNA secondary structure are unlikely to be involved in this process. The complete polyprotein resulting from this frameshift event has a calculated MW of 401 kDa and after cleavage of the two N-terminal leaders would yield a 292-kDa protein containing the MT, HEL, and RdRp domains. Phylogenetic analysis of the three replication-associated domains, MT, HEL, and RdRp, indicates that CTV and BYV form a separate closterovirus lineage within the alpha-like supergroup of positive-strand RNA viruses. Two gene blocks or modules can be easily identified in the CTV genome. The first includes the replicative MT, HEL, and RdRp genes and is conserved throughout the entire alpha-like superfamily. The second block consists of five ORFs, 3 to 7, conserved among closteroviruses, including genes for the CTV homolog of HSP70 proteins and a duplicate of the coat protein gene. The 3'-terminal ORFs 8 to 11 encode a putative RNA-binding protein (ORF 11), and three proteins with unknown functions; this gene array is poorly conserved among closteroviruses.(ABSTRACT TRUNCATED AT 400 WORDS)

Screening of the closterovirus genome by degenerate primer-mediated polymerase chain reaction.

The genome of beet yellows virus (BYV), the type representative of the closterovirus group, encodes a homologue of the cellular heat-shock protein (HSP) 70 family. A pair of degenerate primers targeted to motifs A and E, which are highly conserved in HSP70s, was synthesized. Genomes of several definite and possible members of the closterovirus group were screened for the presence of the HSP70 gene with PCR using these degenerate primers. BYV, citrus tristeza virus (CTV), beet yellow stunt virus (BYSV) and carnation necrotic fleck virus templates produced 1 kb amplification products, which were shown by sequencing to represent fragments of the respective HSP70 genes. Further screening was performed with an additional degenerate primer targeted to the motif IV of the putative viral polymerase. This degenerate primer and specific primers complementary to the 5' region of the HSP70 genes of the respective viruses were used to estimate the distance between polymerase motif IV and the start point of the HSP70 gene for BYV (approximately 1.1 kb), CTV and BYSV (around 2.0 kb) by PCR. The amplified genome regions of CTV (3026 nucleotides) and BYSV (2837 nucleotides) were cloned and sequenced. CTV and BYSV were found to encode the gene for an additional 30K (BYSV) or 33K (CTV) protein between the polymerase and the small hydrophobic protein genes, which was absent in BYV. These two 30K proteins displayed very weak similarity to each other, unlike the highly conserved polymerases, hydrophobic proteins and HSP70s of BYV, CTV and BYSV. Degenerate primer-mediated PCR proved to be an efficient tool for rapid screening and subsequent cloning of the viral genomes.

[Secretory immunoglobulin of the stomach in patients with chronic gastritis]. / Sekretornyi immunoglobulin zheludka u bol'nykh khronicheskim gastritom.

The determination of the level of secretory IgA by a method of radial immunodiffusion after Mancini in the gastric juice of 48 patients with chronic gastritis in correlation with the status of the gastric mucosa, the level of acidification and the phase of exacerbation has shown diagnostic potentialities of the method. The highest IgA level was detected in patients with "rearrangement" gastritis and in patients with sharply suppressed gastric secretion. In marked atrophy of the gastric mucosa IgA secretion was significantly lowered. The period of remission was attended by a decrease in IgA secretion as compared with the phase of exacerbation.

[Interferon-inducing and antiviral activity of dsRNA of yeast origin]. / Interferonindutsiruiushchaia i protivovirusnaia aktivnost' dsRNK drozhzhevogo proiskhozhdeniia.

The paper describes derivation of double-stranded RNA from K plasmid of Sachcaromyces cerevisiae yeast by means of electrophoresis in agar gel and by differential salting out with 4 M lithium chloride. Studies in vitro and in vivo demonstrated a high interferon-inducing and antiviral activity of dsRNA preparations from the yeast plasmid.

Venera 13 and venera 14: sedimentary rocks on venus?

Venera 13 and Venera 14 transmitted almost complete panoramic views of their landing sites. Analyses of the photographs show the presence of rock formations undergoing geomorphic degradation. The formations display ripple marks, thin layering, differential erosion, and curvilinear fracturings. Some of them are interpreted as lithified clastic sediments. The lithification could have taken place at depth or at the surface, resulting in a type of duricrust. The origin of the sediments is unknown but could be aeolian, volcanic, or related to impacts or to turbidity currents.

[Interferon-inducing and antiviral activity of levamisole]. / Interferonindutsiruiushchaia i protivovirusnaia aktivnost' levamizola.

It was shown that levamisol administered orally to mice induced production of interferon with its maximum level in 4-6 hours and prolonged subsequent circulation in the host (the observation period of 5 days). Antiviral activity of levamisol in experimental forest-spring encephalitis was shown (protection of 35-40 per cent). When levamisol were used in combination with polyguacyl, an additive effect was recorded.

[Combined use of isoprinosine and an interferon inducer in experimental viral infection]. / Kombinirovannoe primenenie izoprinozina i induktora interferona pri éksperimental'noi virusnoi infektsii.

It was shown for the first time that isoprinosine has an antiviral effect in treatment of experimental forest-spring encephalitis. The combined use of isoprinosine with mouse interferon in treatment of the infected albino mice potentiated antiviral effect as compared to the effect of substances used alone. In addition, the combined use of isoprinosine and interferon inductor in minimum nontoxic amounts resulted in a 4--8 fold increase in the titers of the serum interferon and a statistically significant increase in the mouse resistance to the viral infection. It was found that the efficacy of the combined use of isoprinosine and mouse interferon or interferon inductor increased, when the interval between the drug administrations was prolonged up to 24 hours. The maximum effect providing 75 per cent protection of the animals from the viral infection and an increase in the average life span of the mice from 7.9 to 17.4 days was observed with the combined use of isoprinosine and interferon inductor.

[Biological activity of a new Soviet natural inducer, double-stranded RNA]. / Biologicheskaia aktivnost' novogo otechestvennogo prirodnogo induktora--dvunitevoi RNK.

Antiviral activity of a two-spiral RNA (ts RNA), a new natural interferon inductor was studied. It was shown that ts RNA extracted from a phage infected E. coli culture was an active inductor of interferon and resistance to infection with the forestspring encephalitis virus experimental animals. In experiments on 10-12 g mice ts RNA administered in a dose of 50 micrograms/mouse 6 hours after the infection induced up to 1280 units/ml of the serum interferon. When the inductor was administered repeatedly, the experimental animals developed hyporeactivity resulting in a marked decrease in interferon production after the 3rd subsequent injection. The most pronounced effect with respect to the forest-spring encephalitis virus was observed when the inductor was administered intraperitoneally in a dose of 20 micrograms/mouse 4 hours before the infection. The protective effect was less pronounced when the inductor was administered 24 and 48 hours before the infection. A two-fold administration of the inductor did not increase the antiviral effect. When the inductor was administered in a dose of 100 micrograms 14 days before the infection, the animals showed an increase in the nonspecific resistance to the infection resulting in a marked antiviral effect.

[Effect of prolonged storage times on the infectivity of lyophilized arboviruses]. / Vliianie dlitel'nykh srokov khraneniia na infektsionnost' liofilizovannykh arbnovirusov.

Lyophilized arboviruses of the following taxonomic groups: alfaviruses, flaviviruses, Bunyamwera supergroup viruses, as well as arenaviruses were used in the study. These viruses had been stored for a long time at --20 degree C. Determinations of the infectious titers in suckling mice showed that in a period of over 10 years (up to 21 years) no complete loss of infectivity occurred in any virus groups.

Comparison of interferon-inducing activities and antiviral effects of tobacco mosaic virus, tilorone and sodium nucleinate.

Endogenous interferon was produced in animals in response to the administration of tobacco mosaic virus (TMV), tilorone and sodium nucleinate. The relationship between interferon production and the kind of inducer and the route of its administration was studied. TMV was completely innocuous for Macaca rhesus monkeys and mice and caused no untoward effects in humans upon peroral administration. TMV, tilorone and sodium nucleinate given per os exerted a marked protective effect in mice against tick-borne encephalitis, eastern and western equine encephalomyelitis and influenza virus infections.