Exergames： neuroplastic hypothesis about cognitive improvement and biological effects on physical function of institutionalized older persons

Exergames can be considered a dual task because the games are performed by a man-videogame interface, requiring cognitive and motor functions simultaneously. Although the literature has shown improvements of cognitive and physical functions due to exergames, the intrinsic mechanisms involved in these functional changes have still not been elucidated. The aims of the present study were（1） to demonstrate the known biological mechanisms of physical exercise regarding muscle adaptation and establish a relationship with exergames; and（2） to present a neurobiological hypothesis about the neuroplastic effects of exergames on the cognitive function of institutionalized older persons. These hypotheses are discussed.

Interleukin 28B-related polymorphisms: A pathway for understanding hepatitis C virus infection?

AIM:To analyze the role of rs12979860 and rs8099917polymorphisms in hepatitis C virus(HCV)genotype 1infection of Brazilians.METHODS:A total of 145 adult patients diagnosed with genotype 1 chronic hepatitis C(CHC)who had completed a 48-wk regimen of pegylated-interferonα-2a or-2b plus ribavirin combination therapy were recruited from six large urban healthcare centers and199 healthy blood donors(controls)from a single site between January 2010 and January 2012.Data on the patients’response to treatment was collected.Polymerase chain reaction-restriction fragment length polymorphism genotyping of the interleukin(IL)28B gene fragment encompassing the single nucleotide polymorphisms(SNPs)rs12979860(C/T)and rs8099917(T/G)was carried out for 79 of the CHC patients and 199 of the controls.Bi-directional amplicon sequencing of the two SNPs was carried out for the remaining 66 CHC patients.RESULTS:SNP rs12979860 genotyping was successful in 99.5%of the controls and 97.2%of the CHC patients,whereas the SNP rs8099917 genotyping was successful in 95.5%of the controls and 100%of the CHC patients.The genotype and allele distributions for both rs12979860 and rs8099917 were significantly different between the control and CHC patient groups,with significantly higher genotype frequencies of CC and TT in the controls(P=0.037 and 0.046,respectively)and of TT and GG in the CHC patients(P=0.0009and 0.0001,respectively).Analysis of the CHC patients who achieved sustained virological response(SVR)to treatment(n=55)indicated that the rs12979860 C allele and CC genotype were predictors of SVR(P=0.02).No significant correlation was found between rs8099917 genotypes and treatment response,but carriers of the T allele showed significantly higher rates of SVR(P=0.02).Linkage disequilibrium analysis of the group that achieved SVR showed a significant association between rs12979860 and rs8099917(P=0.07).CONCLUSION:The higher allele frequency of rs12979860 C and rs8099917 T observed in non-HCVinfected individuals may indicate a potential protective role for these IL28B-related polymorphisms.

Physiological Performance of Rice Seeds Treated to Thiamethoxam and Placed under Storage

High-yielding crops can only achieve their potential when the appropriate technological tools are applied and weather conditions do not impose restrictions. One such technological tool is the treatment of seeds, in which different products are used and, in some cases, cause little-known effects capable of modifying the plant’s metabolism and/or its morphology. The insecticide thiamethoxam alters the plant’s physiology and morphology, accelerating its development, thus enhancing its vigor. The objective of this study was to evaluate the influence of thiamethoxam on the physiological performance of rice seeds during storage. The research was conducted in a green-house and at the laboratory of the Department of Plant Science, Faculty of Agronomy Eliseu Maciel —UFPel (Federal University of Pelotas). Rice seeds from cultivars INIA Olimar and El Paso L144 were treated to a commercial product containing 350 grams of active ingredient thiamethoxam at doses of 0, 100, 200, 300, 400 and 500 g per 100 kg of seed. Seeds were tested for physiological quality immediately after being treated (check treatment) and at 3, 6, 9 and 12 months under storage, through germination test, cold test, accelerated aging and greenhouse emergence test. The concentration range from 200 to 400 g a.i. 100 kg-1 seed yielded the highest germination and seed vigor values for both rice cultivars during the storage period.

Recycling process and proto-kimberlite melt metasomatism in the lithosphere-asthenosphere boundary beneath the Amazonian Craton recorded by garnet xenocrysts and mantle xenoliths from the Carolina kimberlite

Here we present new data on the major and trace element compositions of silicate and oxide minerals from mantle xenoliths brought to the surface by the Carolina kimberlite,Pimenta Bueno Kimberlitic Field,which is located on the southwestern border of the Amazonian Craton.We also present Sr-Nd isotopic data of garnet xenocrysts and whole-rocks from the Carolina kimberlite.Mantle xenoliths are mainly clinopyroxenites and garnetites.Some of the clinopyroxenites were classified as GPP–PP–PKP(garnet-phlogopite peridotite,phlogopite-peridotite,phlogopite-K-richterite peridotite)suites,and two clinopyroxenites(eclogites)and two garnetites are relicts of an ancient subducted slab.Temperature and pressure estimates yield 855–1102℃ and 3.6–7.0 GPa,respectively.Clinopyroxenes are enriched in light rare earth elements(LREE)(La_(N)/Yb_(N)=5–62;Ce_(N)/Sm_(N)=1–3;where N=primitive mantle normalized values),they have high Ca/Al ratios(10–410),low to medium Ti/Eu ratios(742–2840),and low Zr/Hf ratios(13–26),which suggest they were formed by metasomatic reactions with CO_(2)-rich silicate melts.Phlogopite with high TiO_(2)(>2.0 wt.%),Al_(2)O_(3)(>12.0 wt.%),and FeOt(5.0–13.0 wt.%)resemble those found in the groundmass of kimberlites,lamproites and lamprophyres.Conversely,phlogopite with low TiO_(2)(<1.0 wt.%)and lower Al_(2)O_(3)(<12.0 wt.%)are similar to those present in GPP-PP-PKP,and in MARID(mica-amphibole-rutile-ilmenite-diopside)and PIC(phlogopite-ilmenite-clinopyorxene)xenoliths.The GPP-PP-PKP suite of xenoliths,together with the clinopyroxene and phlogopite major and trace element signatures suggests that an intense proto-kimberlite melt metasomatism occurred in the deep cratonic lithosphere beneath the Amazonian Craton.The Sr-Nd isotopic ratios of pyrope xenocrysts(G3,G9 and G11)from the Carolina kimberlite are characterized by high ^(143)Nd/^(144)Nd(0.51287–0.51371)and eNd(+4.55 to+20.85)accompanied with enriched ^(87)Sr/^(86)Sr(0.70405–0.71098).These results suggest interaction with a proto-kimberlite melt compositionally similar with worldwide kimberlites.Based on Sr-Nd whole-rock compositions,the Carolina kimberlite has affinity with Group 1 kimberlites.The Sm-Nd isochron age calculated with selected eclogitic garnets yielded an age of 291.9±5.4 Ma(2σ),which represents the cooling age after the proto-kimberlite melt metasomatism.Therefore,we propose that the lithospheric mantle beneath the Amazonian Craton records the Paleozoic subduction with the attachment of an eclogitic slab into the cratonic mantle(garnetites and eclogites);with a later metasomatic event caused by proto-kimberlite melts shortly before the Carolina kimberlite erupted.

Hybrid Rice Seed Treatment with Pesticides Improves Its Physiological Quality and Performance

Pathogens in the soil or transmitted by seeds can reduce the stand of plants, affecting the yield of the crop. Furthermore, in the case of rice, the low temperature is a limiting factor in culture. The treatment of seeds with insecticides is an alternative to reduce the damage caused by pests incidents during the early stages of plant development and in some cases, may assist in the germination of seeds exposed to low temperatures. In this sense, the objective of this study was to evaluate the influence of seed treatment with insecticides in the physiological quality and yield of hybrid rice seeds, cv. Inov CL. For this, three experiments were conducted with treated seeds in different doses of thiamethoxam + lambda-cyhalothrin and fipronil totaling seven treatments, including the control (no product). In the first experiment, it was determined seed germination to 10°C, 13°C, 16°C, 20°C and 25°C. In the second experiment, it was assessed seed vigor through accelerated aging, cold test and emergency field. The third experiment was carried out into the field to observe the number of panicles m-2 and crop yield. It is concluded that treatment of hybrid rice seeds, cv. Inov CL with thiamethoxam + lambda-cyhalothrin and fipronil increases the germination percentage of seed and early seedling performance. Treatment of hybrid rice seeds, cv. Inov CL with thiamethoxam + lambda-cyhalothrin provides greater germination and early seedling performance compared to treatment with fipronil. Treatment of hybrid rice seeds, cv. Inov CL with thiamethoxam + lambda-cyhalothrin in dosages of 500 mL 100 kg-11 of seeds or 500 mL 45 kg-1 of seeds increases grain yield.

Excessive Moderator Band in a Female Cat’s Left Ventricle

This report describes a case of cardiomyopathy associated with excessive moderator bands in a 7 years old cat. In the cat echocardiography examination was observed cardiomegaly, decreased systolic function and thinning of the left ventricle wall. The electrocardiogram showed the occurrence of supraventricular tachycardia. The treatment was initiated with digoxin, furosemide and taurine supplementation, with an improvement of the clinical signs, however the animal died in a fall. The diagnosis of cardiomyopathy associated with excessive moderator band, was based on the multiple cardiac bunds which connect the septum to the free wall of the left ventricle observed in the cat necropsy.

Argentina and Brazil： Laws as Mediators of Social Identities

The research is a brief analysis of recent legislation in Argentina about the intangible heritage and its relationship with Brazilian instruments. This is a study about links between the two laws and their touch points and their differences. It will be seen that the legislative actions follow a common sense but are targeted for different acts of management of cultural heritage. We will be able to observe how legislation, whether it is Argentine or Brazilian, works as a mediator between the reclaiming of heritage （as in the acknowledgement of cultural practices） and the government actions towards cultural heritage. This shows that Argentina, despite not creating specific federal legislation regarding intangible cultural heritage, is aware of the need to preserve the cultural events that belong in this category of heritage

Mycobacterium bovis BCG as a Delivery System for the dtb Gene Antigen from Diphtheria Toxin

Diphtheria is a fulminant bacterial disease caused by toxigenic strains of Corynebacterium diphtheriae whose local and systemic manifestations are due to the action of the diphtheria toxin (DT). The vaccine which is used to prevent diphtheria worldwide is a toxoid obtained by detoxifying DT. Although associated with high efficacy in the prevention of disease, the current anti-diphtheria vaccine, one of the components of DTP (diphtheria, tetanus and pertussis triple vaccine), may present post vaccination effects such as toxicity and reactogenicity resulting from the presence of contaminants in the vaccine that originated during the process of production and/or detoxification. Therefore, strategies to develop a less toxic and at the same time economically viable vaccine alternatives are needed to improve existing vaccines in use worldwide. In this study, the Moreau substrain of BCG which is used in Brazil as a live vaccine against human tuberculosis was genetically modified to carry and express the gene encoding for the diphtheria toxin fragment B (DTB). As such, the DNA sequence encoding the dtb gene was cloned into the pUS977 shuttle vector for cytoplasmic expression and successfully introduced into BCG cells by electroporation. Mice immunized with recombinant BCG expressing DTB showed seroconversion with the detection of specific antibodies against DTB. Also, rBCGs stably expressing DTB persisted up to 60 days in the absence of selective pressure in mice and cell viability did not change significantly during the period tested. Finally, immune sera from BALB/c mice vaccinated with rBCGpUS977dtbPW8 were preliminarily tested for their capacity of neutralizing the diphtheria toxin in the Vero Cells assay.

Essential amino acid ratios and mTOR affect lipogenic gene networks and miRNA expression in bovine mammary epithelial cells

Background： The objective of this study was to study how changing the ratio of Lys to Thr, Lys to His, and Lys to Val affects the expression of lipogenic genes and microRNA （miRNA） in bovine mammary epithelial cells. Results： Triplicate cultures with the respective ＂optimal＂ amino acid （AA） ratio （OPAA = Lys：Met 2.9：1; Thr：Phe 1.05：1; Lys：Thr 1.8：1; Lys：His 2.38：1; Lys：Val 1.23：1） plus rapamycin （OPAARMC; positive control）, OPAA, Lys：Thr 2.1：1 （LT2.1）, Lys：Thr 1.3：1 （LT1.3）, Lys：His 3.05：1 （LH3.0）, or Lys：Val 1.62：1 （LV1.6） were incubated in lactogenic medium for 12 h. The expression of 15 lipogenic genes and 7 miRNA were evaluated. Responses to LT2.1, LT1.3, LH3.0, and LV1.6 relative to the control （OPAARMC） included up-regulated expression ofACSS2, FABP3, ACACA, FASN, SCD, LPIN1, INSIG1, SREBF1, PPARD, and NR1H3 （commonly known as LXR-a）. Furthermore, LV1.6 up-regulated expression of ACSL1, DGAT1, and RXRA and down-regulated PPARG expression. Although no effect of OPAA on expression of PPARG was observed, compared with the control, OPAA up-regulated expression of the PPAR targets ACSS2, FABP3, ACACA, FASN, SCD, LPIN1, INSIG1, and SREBF1. Compared with the control, the expression of the anti-lipogenic MIR27AB was down-regulated by OPAA, LT2.1, LT1.3 and LH3.0. In contrast, compared with the control, the expression of the pro-lipogenic MIR21 was up-regulated by LT2.1, LT1.3, LH3.0, and LV1.6. Conclusions： The observed up-regulation of lipogenic gene networks and the changes in expression of key miRNA involved in the control of lipogenic balance are indicative of a potentially important role of EAA ratios and mTOR signaling in the regulation of milk fat synthesis.

Morphophysiological Responses of Olive Plants of the Arbequina Cultivar in Acid Soils

The current study aimed to investigate the morphophysiological responses with determinations of the plant height, stem diameter, chlorophyll content, and leaf nutrients of ‘Arbequina’ olive plant in acid soils. For evaluations of plant height, stem diameter, chlorophyll content, the experimental design was completely randomized arranged in split-plot design. The factor allocated to the main plots was consisted of the time after transplant (0, 30, 60, 90, 120 and 150 days after transplant—DAT) and, the factor arranged in the subplots was composed by pH with six levels 2.9;3.1;3.9;4.3;5.0;and, 6.3 (witness). In determination of leaf nutrient content (nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, copper, zinc, iron, manganese, aluminium and boron) was followed the same experimental design, however, only pH was tested. Plant height, stem diameter and chlorophyll content (SPAD) are not prejudiced by acidic pH up to 150 DAT. For the different pH levels tested, the nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, copper, zinc, iron, manganese and boron foliar contents are adequate for the olive crop, except nitrogen at pH 2.9. The ‘Arbequina’ olive plants adequately support acidic soils even with accentuated additions in the foliar aluminium content.

Reiterated homogenization of a laminate with imperfect contact:gain-enhancement of effective properties

A family of one-dimensional(1D) elliptic boundary-value problems with periodic and rapidly-oscillating piecewise-smooth coefficients is considered. The coefficients depend on the local or fast variables corresponding to two different structural scales. A finite number of imperfect contact conditions are analyzed at each of the scales. The reiterated homogenization method(RHM) is used to construct a formal asymptotic solution. The homogenized problem, the local problems, and the corresponding effective coefficients are obtained. A variational formulation is derived to obtain an estimate to prove the proximity between the solutions of the original problem and the homogenized problem. Numerical computations are used to illustrate both the convergence of the solutions and the gain of the effective properties of a three-scale heterogeneous 1D laminate with respect to their two-scale counterparts. The theoretical and practical ideas exposed here could be used to mathematically model multidimensional problems involving multiscale composite materials with imperfect contact at the interfaces.

Physiological Performance of Common Bean Seeds Treated with Bioactivator with and without Moisture Restriction

The commonbean (Phaseolus vulgaris L.) is an important agricultural crop due to its nutritional properties. Insecticides and fungicides employed to control pests and diseases in the common bean may cause as yet unknown effects on the plant’s morphology and metabolism. Bioactivators are complex organic substances, capable of exerting modification upon growth patterns, plant DNA transcription, gene expression, metabolic enzymes and mineral nutrition. Thiamethoxam insecticide has shown positive action on the enhancement of vigor expression, phytomass accretion and the promotion of deeper root systems. The aim of this work was to evaluate the effect of thiamethoxam on the physiological performance of common bean seeds. Seeds from cultivars Pérola and IAPAR Siriri were treated to thiamethoxam concentrations of 0, 100, 200, 300, 400 and 500 mL&middotkg-1 seed. Physiological quality was measured through tests for germination value, first count, cold test, accelerated aging and seedling shoot and root lengths, all undertaken in the presence and absence of moisture stress. Thiamethoxam stimulated the physiological performance of common bean seeds under moisture stress or in the absence of it, with levels varying according to cultivar and seed lot. The best physiological seed performance was obtained within the range of 200 mL&middotkg-1 seed to 400 mL&middotkg-1 seed.

Isoenzyme Expression in Bean Seed Germination Treated with Thiamethoxam with and without Drought Stress

The thiamethoxam acts as enhancer, allowing maximizing the expression of seed vigor. Isozymes are products of gene expression, highly influenced by the environment, because genes that control its expression are manifested in certain stages of development and in specific organs and tissues, or even under the action of certain stimulus. This study evaluated the isoenzyme expression in bean seedlings from seeds treated with thiamethoxam. Bean seeds were used, cultivar Pérola and IAPAR Siriri, submitted or not to drought stress. Seeds were treated at doses of thiamethoxam: 0, 1, 2, 3, 4 and 5 mL·kg-1 of seed. Isozymes were extracted from seedlings collected five days after sowing. We evaluated the expression of isoenzymes: Glutamate Oxalacetate transaminase (GOT), acid phosphatase (FAC), peroxidase (PO), Esterase (EST), malate dehydrogenase (MDH) and glutamate dehydrogenase (GTDH). Interpretation of the results was based on visual analysis of electrophoresis gels, considering the presence/absence and the intensity of each electrophoretic band. Bean seeds treated with thiamethoxam with doses up 3 mL·kg-1 of seed are higher germination untreated, even when submitted to drought stress. The expression of isozymes GOT, FAC, PO, EST, MDH and GTDH is not affected significantly in bean seedlings of Pérola and IPR Siriri under different doses of thiamethoxam with and without drought stress.

Yield and quality of maize following the foliar application of a fertilizer based on the byproduct “shale water”

The water extracted from the shale rock (shale water) through the pyrolysis process to obtain fuel oil and other products shows a composition based on organic compounds and a wide range of minerals and trace elements with an important role in plant nutrition, suggesting its use as a fertilizer. Thus, the influence of foliar application of shale water (SW), with or without the micronutrients zinc (Zn), manganese (Mn), copper (Cu), boro (B) and molybdenum (Mo), was evaluated regarding yield and quality of maize grains. The yield, the total antioxidant activity, and the content of starch, phenolic compounds and carotenoids were improved in maize grains following the application of three doses of 7 L/ha of SW, which indicates that SW may influence the primary and secondary metabolisms. The application of SW with micronutrients resulted in the increase of grain yield;however, did not result in the improvement of grain quality. The foliar fertilizer formulations also had an influence in the content of minerals and aminoacids of the grain. The results indicate that SW has potential to be used in agriculture to improve yield and quality of maize.

Eucalyptus Oil-Mediated Synthesis of Gold Nanoparticles

The search for efficient methods for the synthesis of metal nanoparticles has been widely explored.Within this context,the use of biological materials such as plants,algae,bacteria and fungi has been reported to various metal nanoparticles as an efficient,low-cost and environmental friendly approach.In this paper we present a single-step method for synthesizing gold nanoparticles,by using essential oils from Brazilian Eucalyptus leaves(urograndis and dunnii).The results strongly suggest that the presence of a stronger bioreducing agent,α-Terpinyl acetate,in the Eucalyptus urograndis,results in higher antioxidant capacity and hence in the synthesis of gold nanoparticles with more controlled size distribution.

Bioactive Compounds and Antifungal Activities of Extracts of Lamiaceae Species

Origanum vulgare L. (oregano) and Rosmarinus officinalis L. (rosemary) are vegetal species belonging to the family Lamiaceae, popularly known as oregano and rosemary. Aromatic plants are used in the treatment and prevention of diseases and in the culinary as functional food in the preparation and conservation of foods. In the chemical composition of oregano and rosemary are present bioactive compounds with antimicrobial, antioxidant and flavoring effect. Several reports in the literature have presented the chemical composition and biological activity of the essential oils of oregano and rosemary. However, few studies have been carried out regarding the chemical composition and biological potential of the aqueous and ethanolic extracts of Origanum vulgare L. and Rosmarinus officinalis L. Evidencing a need to investigate the chemical composition and antifungal activity of these extracts. The objective of the study was to evaluate the bioactive compounds and antifungal activity of the aqueous and ethanolic extract of Origanum vulgare L. and Rosmarinus officinalis L. The aqueous and ethanolic extracts of Origanum vulgare L. and Rosmarinus officinalis L. present in the chemical composition phenolic acids and flavonoids. The antifungal test of the aqueous and ethanolic extract of Origanum vulgare L. and Rosmarinus officinalis L. presented antifungal potential against Candida globosa, Cryptococcus laurentii, Trichosporum assai, Rhodotorula sp., Candida albicans, Kodamaea ohmeri, Saccharomyces and Geotrichum. According to the results obtained in this study, it was concluded that the ethanolic extract of oregano and rosemary present antifungal activity against several yeasts tested, thus proving that these plant species must be carefully evaluated, aiming at a potential for use as an antimicrobial agent.

Impact of Cluster Thinning on Quality of“Malbec”Grapes in Encruzilhada do Sul-RS

The cultivar Malbec has been present’s good performance and quality in the South Half of Rio Grande do Sul (RS). In this context, we aimed to evaluate the quality of “Malbec” grapes in relation to the phytotechnical, physicochemical parameters and bioactive compounds under different intensities of cluster thinning in Encruzilhada do Sul-RS. The experiment was carried out in a commercial vineyard in the 2015/16 with the cultivar Malbec (Vitis vinifera L.) under rootstock “1103 Paulsen”, with spacing of 2.2 m between rows and 1.1 m between plants. The experimental design was of randomized complete blocks with five replications. The treatment factor tested was the cluster thinning, with four levels (0%, 10%, 30% and 50%), performed in véraison. The evaluated phytotechnical variables were cluster length, cluster fresh weight, berries fresh weight, number of berries, rachis fresh weight. Also what were determined are the number of clusters, production and Ravaz index. The physicochemical variables verified were pH, soluble solids (SS), titratable acidity (TA) and SS/TA ratio. The bioactive compounds were total polyphenols index and total anthocyanins. The appropriate balance between source and sink, by the Ravaz index, occurs to zero and 10% of cluster thinning. The “Malbec” grapes present an increase of the maturation index (SS/TA) at 50% of cluster thinning, due to the low titratable acidity at this level. The total polyphenols index has a larger increment when the plants are submitted to the cluster thinning of 10%, providing a larger quality to the fruits. At all the levels of cluster thinning tested, when compared to the control, there was a greater contribution of total anthocyanins in the berries.

Accelerated Aging in Tomato Seeds

The tomato plant (Lycopersicum esculentum L.) is a perennial, shrubby-like vegetable with high reproductive ability, which behaves like a typical annual crop. The aim of this work was to select suitable tests to assess the quality of different lots of tomato seeds. Six batches of tomato seeds were assessed for parameter values: moisture content, germination, first count, accelerated aging (standard: 40 mL of water;saline solution: 11 g NaCl/100mL of water and saturated saline solution: 40 g NaCl/100mL water) and seedling emergence in soil. Accelerated aging through the use of saturated salt solution for 48 h or 72 h at 41°C resulted in an alternative and efficient way to evaluate the physiological quality of tomato seeds. The standard accelerated aging test was less efficient and exhibited higher moisture variation.

Chemical Composition and Antiviral Effect of Extracts of Origanum vulgare

This study aimed determine the activity of aqueous and ethanolic extracts of Origanum vulgare against some viruses of veterinary importance (bovine viral diarrhea virus (BVDV), equine arteritis virus (EAV), equine influenza virus (EIV), feline calicivirus (FCV), canine distemper virus (CDV), canine adenovirus (CAV), and canine cororavirus (CCoV) by evaluating the possibility of inhibition of viral particles production. The aqueous extract from 1600 μg/mL did not show cytotoxicity for all cellular lineages evaluated, Madin Darby bovine kidney cells (MDBK), Rabbit kidney cells (RK 13), Madin Darby canine kidney cells (MDCK) and Crandell feline kidney cells (CRFK), and the ethanolic extract of Origanum vulgare was not toxic at 600 μg/mL. The addition of aqueous extract of Origanum vulgare in media resulted in a significant reduction of the EAV titer from 105.42 infecting dose for cellular culture at 50% (TCID50) to 102.09 TCID50/100 μL while in the presence of the ethanolic extract of Origanum vulgare in media resulted in a significant reduction of the EAV titer from 105.42 TCID50 to 100.79 TCID50/100 μL. To CDV the addition of aqueous extract resulted in a reduction from 102.00 TCID50 to 100.00 TCID50/100 μL while in the presence of the ethanolic extract titers were reduced from 102.00 TCID50 to 101.50 TCID50/100 μL. No significant differences in titers regarding the others analyzed viruses were detected. With respect to chemical analysis of the extracts of Origanum vulgare, were identified in the ethanol extract phenolics rosmarinic acid, caffeic acid, carnosol, p-coumaric acid, carnosic acid, luteolin, apigenin, kaempferol and quercetin. In aqueous extracts of Origanum vulgare were detected rosmarinic acid, p-coumaric acid carnosic acid, luteolin, apigenin, kaempferol and quercetin. The data obtained stimulate other biological assays in order to determine which compounds are responsible for the antiviral activity as well as which are the mechanisms involved. The results presented and the considerations we were able to draw from them allowed us to conclude that the ethanolic extract of Origanum vulgare demonstrated lower cell viability than the aqueous extract and has significant antiviral activity against EAV and the both aqueous and ethanolic extracts have antiviral action against CDV.

Plasmid instability when the hsp60 gene promoter is used to express the protective non-toxic fragment B of the diphtheria toxin in recombinant BCG

The genetic modification of the live attenuated Mycobacterium bovis BCG to deliver a protective Corynebacterium diphtheriae antigen in vivo could be a safer and less costly alternative to the new and more expensive DTP vaccines available today, in particular to third world-countries. The stability of expression of heterologous antigens in BCG, however, is a major challenge to the use of live recombinant bacteria in vaccine development and appears to be dependent to a certain extent, on a genetic compatibility between the expression cassette within the plasmid construct and the mycobacterium host. In the quest for the best recombinant BCG transformant to express the dtb gene of C. diphtheriae we generated two new rBCG strains by transforming the Moreau substrain of BCG with the mycobacterial expression vectors pUS973 and pUS977, each one carrying a different promoter to drive the expression of the target antigen. After transformation recombinant BCG clones were selected on Middlebrook 7H10 kanamycin Agar plates, expanded in Middlebrook 7H9 kanamycin Broth and analyzed by agarose gel electrophoresis and immunoblotting. rBCGs transformed with the construct carrying the weak PAN promoter from M. paratuberculosis stably expressed the dtb gene. Conversely, rBCGs transformed with the construct carrying the strong mycobacterium hsp60 promoter were unstable and consequently unfit for the expression of the C. diphtheriae gene.