Constitutional genomic imbalances are known to cause malformations, disabilities, neurodevelopmental delay, and dysmorphia and can lead to dysfunctions in the cell cycle. In extremely rare genetic conditions such as small supernumerary marker chromosomes (sSMC), it is important to understand the cellular consequences of this extra marker, as well the factors that contribute to their maintenance or elimination through successive cell cycles and phenotypic impact. The study of chromosomal mosaicism provides a natural model to characterize the effect of aneuploidy on genome stability and compare cells with the same genetic background and environment exposure, but differing in the presence of sSMC. Here, we report the functional characterization of different cell lines from two familial patients with mosaic sSMC derived from chromosome 12. We performed studies of proliferation dynamics, stability, and variability of these cells using fluorescent in situ hybridization (FISH), sister chromatid exchanges (SCE), and conventional staining. We also quantified the telomere-related genomic instability of sSMC cells using 3D telomeric profile analysis by quantitative-FISH. sSMC cells exhibited differences in the cell cycle dynamics compared to normal cells. First, the sSMC cells exhibited lower proliferation index and higher frequency of SCE than normal cells, associated with a higher level of chromosomal instability. Second, sSMC cells exhibited more telomeric-related genomic instability. Lastly, the differences of sSMC cells distribution among tissues could explain different phenotypic repercussions observed in patients. These results will help in our understanding of the sSMC stability, maintenance during cell cycle, and the cell cycle variables involved in the different phenotypic manifestations.

RESUMO: Este estudo analisou a eficácia do uso de diferentes níveis de um produto à base de cipermetrina a 6% e citronela a 2%, na produção de frangos de corte e no controle do cascudinho (Alphitobius diaperinus). Um total de 648 pintos de um dia de idade (Cobb®) foram distribuídos em três câmaras com os tratamentos (sem aplicação do produto comercial - controle, e duas concentrações do produto comercial - 2,00 e 3,33g/m2) em oito repetições com 27 aves cada. O produto comercial a base de cipermetrina foi aplicado nas câmaras com um atomizador. A população de cascudinho foi amostrada semanalmente (1, 7, 14, 21, 28, 35 e 42 dias do experimento) usando armadilhas, em três pontos (frente, meio e fundo) de cada parcela. Foram avaliados o desempenho, rendimentos de carcaça e cortes. Aos 21, 28, 35 e 42 dias do experimento, o número de adultos e larvas de cascudinho foi menor nos grupos de tratamento envolvendo 2,00 e 3,33g/m2 do produto quando comparado ao grupo controle. No primeiro e no 14º dia do estudo, o número de cascudinhos adultos foi menor no grupo tratado com 3,33g/m2 do produto em comparação ao grupo controle, embora os resultados não tenham diferido estatisticamente daqueles obtidos com 2,00g/m2 do produto. A conversão alimentar de 1 a 21 dias de idade foi significativa, com os melhores resultados para essa variável quando aplicado 3,33g do produto por metro quadrado e no tratamento controle. Em conclusão, a aplicação do produto comercial em ambas as concentrações (2,00 e 3,33g/m2) foi eficaz no combate a adultos e larvas de Alphitobius diaperinus e, não influenciou o desempenho.

ABSTRACT: This study analysed the effectiveness of using different levels of a product based on 6% cypermethrin and 2% citronella on broiler production and control the lesser mealworm (Alphitobius diaperinus). A total of 648 one-day-old chicks (Cobb®) which were allocated to one of three treatments (no application of the commercial product - control, and two concentrations of the product 2.00 and 3.33g/m2) in eight replicates with 27 birds each. The commercial product, which was based on cypermethrin was applied in the chambers with an atomizer. The lesser mealworm population was sampled weekly (1, 7, 14, 21, 28, 35 and 42 days of the experiment) using traps, at three points (front, middle and back) inside the pen. The performance, carcass and cut yields were evaluated. At 21, 28, 35 and 42 days of the experiment, the number of lesser mealworm adults and larvae was lower in the treatment groups involving 2.00 and 3.33g/m2 of the product than in the control group. On the first and 14th days of the study, the number of lesser mealworms adults was lower in the group treated with 3.33g/m2 of the product compared to the control group, although the results did not differ statistically from those obtained with 2.00g/m2 of the product. In conclusion, the application of the commercial product at both concentrations (2.00 and 3.33g/m2) was effective in combating Alphitobius diaperinus adults and larvae, and the performance was not influenced by treatments.

RESUMO O objetivo foi avaliar os efeitos de uma subdose de hCG aplicada no acuponto Hou Hai na qualidade do corpo lúteo (CL) e na indução da ovulação em éguas. Quinze éguas mestiças foram distribuídas em blocos ao acaso, sendo o período utilizado como fator de blocagem, em: T1 = 1500 UI de hCG por via intravenosa (IV); T2 = 450 UI de hCG aplicado no falso acuponto (IV) e T3 = 450 UI de hCG aplicada no acuponto Hou Hai. Avaliou-se diâmetro médio do CL, concentração sérica de progesterona (P4), vascularização do folículo pré-ovulatório e do CL. As fêmeas que receberam 450 UI de hCG no acuponto Hou Hai apresentaram maiores taxas de ovulação (33,33%) 48h após a indução. O número de pixels coloridos (NPC) mínimo do folículo pré-ovulatório das fêmeas do grupo controle foi superior (40,33) ao das éguas que receberam 450 UI de hCG IV no falso acuponto (36,84) e semelhante ao das éguas que receberam hCG no acuponto Hou Hai (39,31); correlações moderadamente positivas foram encontradas entre o diâmetro do CL e a concentração de P4, ambos no D8 (P <0,05). A administração IV de 450 UI de hCG ou no acuponto Hou Hai foi eficiente na indução da ovulação e na garantia da qualidade do CL nas éguas.

ABSTRACT The objective of this study was to evaluate the effects of an hCG sub dose applied at the Hou Hai acupoint on corpus luteum (CL) quality and ovulation induction in mares. Fifteen crossbred mares were distributed in randomized blocks and used in three periods with each period employed as the blocking factor in three treatments: T1 = 1500 IU of hCG via intravenous (IV); T2 = 450 IU of hCG applied at the false acupoint (IV); and T3 = 450 IU of hCG applied at the Hou Hai acupoint. Mean diameter of the CL, serum concentration of progesterone (P4), vascularization of the pre-ovulatory follicle and CL were evaluated. Females administered 450 IU of hCG at the Hou Hai acupoint exhibited greater ovulation rates (33.33%) 48h after induction; The minimum number of colored pixel (NCP) of the pre-ovulatory follicle of control females was superior (40.33) to that of mares administered 450 IU of hCG IV at the false acupoint (36.84) and similar to that of those administered hCG at the Hou Hai acupoint (39.31). Further, moderately positive correlations were found between the CL diameter and the P4 concentration on D8 (P<0.05). IV administration of 450 IU of hCG or at the Hou Hai acupoint was efficient at inducing ovulation and ensuring the quality of CL in mares.

RESUMO O objetivo deste trabalho foi avaliar a influência da inclusão da rama de mandioca na dieta de cordeiros confinados e semiconfinados, em substituição parcial de alimentos concentrados, como forma de redução de custos de produção. O experimento foi conduzido no Centro de Tecnologia de Ovinos (CTO), município de Campo Grande, MS. Foram utilizados 56 cordeiros do grupo genético Pantaneiro, sendo 28 machos e 28 fêmeas, com pesos médios de 18,84kg±2,02kg. As dietas recebidas foram: tratamento controle com ração comercial e tratamentos contendo a parte aérea de mandioca (PAM) em substituição de 10% dessa ração. Os machos confinados obtiveram ganho de peso médio diário de 210g e os semiconfinados, de 178g. No semiconfinamento, as fêmeas controle tiveram GMD de 120g e menor desempenho entre os grupos experimentais. A inclusão PAM acarretou uma redução de 31,29% para a mesma categoria animal. Entre os sistemas de produção, o semiconfinamento apresentou maior lucratividade média, 6,4%, e uma rentabilidade de 7,99% em relação ao sistema de confinamento, que obteve 0,04% de lucratividade e 0,87% de rentabilidade. Conclui-se que a inclusão da parte aérea da mandioca aumenta a eficiência econômica dos sistemas produtivos sem afetar o desenvolvimento dos animais em terminação.

ABSTRACT The scope of this study was to evaluate the inclusion of raw cassava aerial parts in the diet of lambs in partial substitution of rations, in order to reduce production costs. The experiment was conducted at Sheep Technology Centre (CTO) in Campo Grande City, MS. A sample of 56 lambs from Pantaneiro genetic group was used, 28 males and 28 females, average weight of 18.84kg±2.02kg. The inclusion of PAM decreased the costs by 31.29% for the same category of animal. Among the production systems the semi-feedlot, showed the highest average profitability of 6.4% and a return of 7.99%, compared to the feedlot system, which obtained 0.04% 0.87% profitability and return. In conclusion, the inclusion of the aerial part of cassava increases the economic efficiency of production systems without affecting the development of finishing animals.

Extratos aquosos da planta medicinal Achillea millefolium contêm macromoléculas de interesse para desenvolver fitodefensivos para a agricultura. Duas frações de mil folhas foram obtidas por ultrafiltração, E1 (contendo moléculas maiores que 30 kDa), e E3 (peptídeos entre 1 e 10 kDa) que inibiram o crescimento das bactérias fitopatogênicas Ralstonia solanacearum, gram-negativa, e Clavibacter michiganensis subsp. michiganensis, gram-positiva, com dependência de concentração. Os valores de concentração inibitória mínima (CIM) para ambos os extratos e bactérias foram baixos, entre 20 e 80µM. A CIM relativa à proteína total evidenciou a presença de macromoléculas muito ativas em E3, embora com baixa concentração proteica. E3 se aplica à prospecção de peptídeos antimicrobianos. Estimar a CIM relativa à quantidade de amostra vegetal valorizou o potencial antimicrobiano natural de E1, que contém alta concentração proteica. E1e E3 se aplicam ao desenvolvimento de fitodefensivos para uso biotecnológico. A ultrafiltração fracionou as amostras de forma nativa, rápida, e com baixo custo; além de dessalinizar, clarificar, purificar, e concentrar E1 e E3. Esse estudo inédito sobre a separômica e a ação antimicrobiana de extratos macromoleculares aquosos de mil folhas sugere que plantas cicatrizantes podem apresentar grande potencial para desenvolver fitodefensivos agrícolas naturais não danosos, à semelhança de medicamentos fitoterápicos.

Aqueous extracts from the medicinal plant Achillea millefolium contain macromolecules of interest to develop agrochemicals for agriculture. Two fractions of "mil folhas" were obtained by ultrafiltration, E1 (containing molecules larger than 30 kDa) and E3 (peptides between 1 and 10 kDa), which inhibited the growth of phytopathogenic bacteria Ralstonia solanacearum, gram-negative, and Clavibacter michiganensis subsp. michiganensis, gram-positive, concentration-dependent. The values of minimum inhibitory concentration (MIC) for both extracts and both bacteria were low, ranging from 20 to 80µM. The MIC relative to total protein evidenced the presence of very active macromolecules in E3, although showing low protein concentration. E3 applies to the prospection of antimicrobial peptides. The estimated MIC relative to the amount of plant sample valued the natural antimicrobial potential of E1, which contains high protein concentration. E1 and E3 can be used in the development of agrochemicals for biotechnological purposes. The ultrafiltration procedure fractionated the samples in a rapid and native way and at a low cost; it also desalted, clarified, concentrated and purified both E1 and E3. This pioneering study on the separomics and on the antimicrobial activity of macromolecular aqueous extracts from "mil folhas" suggests that healing plants have great potential to develop non-harmful agricultural natural agrochemicals, similarly to the available phytotherapic drugs.

Implantation of Walker 256 tumor decreases acute systemic inflammation in rats. Inflammatory hyperalgesia is one of the most important events of acute inflammation. The L-arginine/NO/cGMP/K+ATP pathway has been proposed as the mechanism of peripheral antinociception mediated by several drugs and physical exercise. The objective of this study was to investigate a possible involvement of the NO/cGMP/K+ATP pathway in antinociception induced in Walker 256 tumor-bearing male Wistar rats (180-220 g). The groups consisted of 5-6 animals. Mechanical inflammatory hypernociception was evaluated using an electronic version of the von Frey test. Walker tumor (4th and 7th day post-implantation) reduced prostaglandin E2- (PGE2, 400 ng/paw; 50 µL; intraplantar injection) and carrageenan-induced hypernociception (500 µg/paw; 100 µL; intraplantar injection). Walker tumor-induced analgesia was reversed (99.3% for carrageenan and 77.2% for PGE2) by a selective inhibitor of nitric oxide synthase (L-NAME; 90 mg/kg, ip) and L-arginine (200 mg/kg, ip), which prevented (80% for carrageenan and 65% for PGE2) the effect of L-NAME. Treatment with the soluble guanylyl cyclase inhibitor ODQ (100% for carrageenan and 95% for PGE2; 8 µg/paw) and the ATP-sensitive K+ channel (KATP) blocker glibenclamide (87.5% for carrageenan and 100% for PGE2; 160 µg/paw) reversed the antinociceptive effect of tumor bearing in a statistically significant manner (P < 0.05). The present study confirmed an intrinsic peripheral antinociceptive effect of Walker tumor bearing in rats. This antinociceptive effect seemed to be mediated by activation of the NO/cGMP pathway followed by the opening of KATP channels.

The effect of substitution of equine chorionic gonadotropin (eCG) by follicle stimulating hormone (FSH) in synchronization protocols of ovulation in dairy goats was evaluated. Twelve goats received intravaginal sponges impregnated with 60mg of medroxyprogesterone acetate (MAP) for 10 days. The sponges were removed and the animals were distributed into two groups (G): G1 (n=6) treated with 0.5mL of a synthetic analogue of PGF2 α and 100 IU of eCG for each 10kg weight, intramuscular injection (IM); and G2 (n=6) treated with 0.5mL of a synthetic analogue of PGF2α and 20mg of FSH (IM). All animals were monitored for estrus detection with aid of a ruffian after sponge removal. The ovarian dynamics were analyzed by ultrasound, since six hours after sponge removed. Each animal was analyzed in time elapsed of six hours until 12 hours after ovulation detection. For data analyses, the Wilcoxon test and variance analyses were used. There was not difference between the analyzed parameters (P>0.05). In this way, eCG can be replaced by FSH in synchronization protocols of ovulation in dairy goats.

O papel da bactéria proteolítica na exacerbação do vírus influenza tem sido demonstrado em hospedeiros naturais como porcos e humanos. Foram coletadas 407 amostras do trato respiratório de indivíduos apresentando manifestações clínicas, durante a estação da influenza (2003-2005) na cidade de São Paulo. Este trabalho teve como objetivo avaliar a incidência de determinadas bactérias que junto com vírus co-infectarem o trato respiratório humano. Testes bacteriológicos, e virológicos como imunofluorescência (IF), RT/PCR e hemaglutinação (HA) foram usados nas investigações viral e bacteriana. Pelo teste de IF foram selecionadas trinta e sete (9,09%) amostras positivas para o vírus influenza. A presença do vírus influenza foi confirmada pela técnica de RT/PCR. Pelos testes bacteriológicos e do agar caseina, verificou-se que 18 (48,64%) dos indivíduos foram infectados com bactérias proteolíticas tais como Staphylococcus spp., Streptococcus spp. e Pseudomonas spp. Destas amostras, 13 (35,13%) foram co-infectadas com vírus influenza tipo A, e 5 (13,51%) com influenza tipo B. No experimento in vitro com influenza e S. aureus, detectou-se aumento do título hemaglutinante deste vírus, após contacto de 30 min a 25 ºC. Os resultados obtidos revelaram a ocorrência de co-infecção com bactéria proteolítica e vírus influenza nos indivíduos avaliados. Estes achados corroboram com a investigação do sinergismo, entre bactéria e vírus, que poderia ser capaz de potencializar infecção respiratória, aumentando os riscos aos hospedeiros.

A role for proteolytic bacteria in the exacerbation of influenza virus has been shown in natural hosts such as pigs and humans. Four hundred seven samples were collected from the respiratory tract of individuals presenting clinical manifestations, during influenza season (2003-2005) in São Paulo City. The aim of this study was to evaluate the incidence of determined bacteria co-infecting virus in human respiratory tract. Tests, such as bacteriological, immunofluorescence (IF), RT/PCR and hemagglutination (HA) were used for bacterial and viral investigation. Thirty seven (9.09%) positive for influenza virus were screened by IF. The RT/PCR confirmed the presence of influenza virus in these samples. Bacterial and agar casein tests demonstrated that 18 (48.64%) individuals were infected with proteolytic bacteria such as Staphylococcus spp., Streptococcus spp. and Pseudomonas spp. Among these samples, 13 (35.13%) were co-infected with influenza A virus. Influenza type B, co-infecting bacteria were found in five (13.51%) samples. In vitro the S. aureus protease increased the influenza HA titer after contact for 30 min at 25 ºC. Results revealed the occurrence of co-infection with proteolytic bacteria and influenza in the evaluated individuals. This finding corroborates that virus versus bacteria synergism could be able to potentiate respiratory infection, increasing damage to hosts.

In the present investigation we studied the fusogenic process developed by influenza A, B and C viruses on cell surfaces and different factors associated with virus and cell membrane structures. The biological activity of purified virus strains was evaluated in hemagglutination, sialidase and fusion assays. Hemolysis by influenza A, B and C viruses ranging from 77.4 to 97.2%, from 20.0 to 65.0%, from 0.2 to 93.7% and from 9.0 to 76.1% was observed when human, chicken, rabbit and monkey erythrocytes, respectively, were tested at pH 5.5. At this pH, low hemolysis indexes for influenza A, B and C viruses were observed if horse erythrocytes were used as target cells for the fusion process, which could be explained by an inefficient receptor binding activity of influenza on N-glycolyl sialic acids. Differences in hemagglutinin receptor binding activity due to its specificity to N-acetyl or N-glycolyl cell surface oligosaccharides, density of these cellular receptors and level of negative charges on the cell surface may possibly explain these results, showing influence on the sialidase activity and the fusogenic process. Comparative analysis showed a lack of dependence between the sialidase and fusion activities developed by influenza B viruses. Influenza A viruses at low sialidase titers (<2) also exhibited clearly low hemolysis at pH 5.5 (15.8%), while influenza B viruses with similarly low sialidase titers showed highly variable hemolysis indexes (0.2 to 78.0%). These results support the idea that different virus and cell-associated factors such as those presented above have a significant effect on the multifactorial fusion process