The growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities in two rearing systems.

This study evaluated the growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities using two rearing systems. The experiment was conducted in a factorial design (4x2) with four stocking densities (0.1, 0.4, 0.7, and 1.0 g/L), two type of aquarium tanks (glass and ceramic aquariums), and four replicates. The experiment lasted 60 days using 148 juvenile fish (3.05 ± 0.09 g) randomly placed in 32 aquariums (50 L) equipped with filters and aeration. All fish received two meals a day ad libitum (8:00 and 16:00). Water quality parameters such as temperature, dissolved oxygen, pH, and total ammonia were measured. At the end of the experiment, all fish were measured and weighed to determine growth performance and then subjected to parasitological analysis. The data were analyzed with a two-way ANOVA with post-hoc Tukey test (p<0.05). No effects on growth performance at different stocking densities were observed. However, there was an increase in Capillaria pterophylli infestation in the high stocking density within ceramic aquariums. Thus, this study recommends the use of 1.0 g/L for the intensive aquaculture system of freshwater angelfish, and applying cleaning management to avoid parasite infestation, particularly in ceramic aquariums.

Porcine respiratory disease complex after the introduction of H1N1/2009 influenza virus in Brazil.

From 2009 to 2015, 74 lungs from suckling (6.8%), nursing (70.3%), fattening (20.3%) pigs and pregnant sows (2.7%) with respiratory signs from pig farms in Southern Brazil were submitted to a diagnostic laboratory for necropsy and/or histologic examination and screening for respiratory agents by RT-qPCR, immunohistochemistry (IHC), virus isolation (VI) and subtyping for influenza A virus (IAV), IHC and nested PCR for Mycoplasma hyopneumoniae (Mhyo), PCR for porcine circovirus 2 (PCV2), RT-qPCR for porcine reproductive and respiratory syndrome virus (PRRSV) and bacterial culture. All lung samples were positive for IAV using RT-qPCR. Seventy-two lungs had histologic lesions associated with acute to subacute IAV infection characterized by necrotizing bronchiolitis/bronchitis or bronchointerstitial pneumonia with lymphocytic peribronchiolitis and bronchiolar/bronchial hyperplasia, respectively. Forty-nine lungs (66.2%) were positive by IHC for IAV nucleoprotein. The H1N1/2009 was the most common subtype and the only IAV detected in 58.1% of lungs, followed by H1N2 (9.5%) and H3N2 (6.8%). Coinfection of IAV and Mhyo was seen in 23 (31%) cases. Although 14.9% of the lungs were positive for PCV2 using PCR, no suggestive lesions of PCV2 disease were observed. Porcine reproductive and respiratory syndrome virus (PRRSV) was not detected, consistent with the PRRS-free status of Brazil. Secondary bacterial infections (8/38) were associated with suppurative bronchopneumonia and/or pleuritis. Primary IAV infection with Mhyo coinfection was the most common agents found in porcine respiratory disease complex (PRDC) in pigs in Southern Brazil.

Genome-wide association study of periweaning failure-to-thrive syndrome (PFTS) in pigs.

Porcine periweaning-failure-to-thrive syndrome (PFTS) is a condition that affects newly weaned piglets. It is characterised by a progressive debilitation leading to death, in the absence of infectious, nutritional, management or environmental factors. In this study, we present the first report of PFTS in South America and the results of a genome-wide association study to identify the genetic markers associated with the appearance of this condition in a crossbred swine population. Four chromosomal regions were associated with PFTS predisposition, one located on SSCX, one on SSC8, and the two other regions on SSC14. Regions on SSC8 and SSC14 harbour important functional candidate genes involved in human depression and might have an important role in PFTS. Our findings contribute to the increasing knowledge about this syndrome, which has been investigated since 2007, and to the identification of the aetiology of this disease.

Avaliação histopatológica de órgãos reprodutivos e bexiga de fêmeas suínas descartadas / Histopathology of reproductive organs and bladder of culled sows

Em um sistema intensivo de produção de suínos, as falhas reprodutivas são uma das principais razões de descarte de matrizes e queda nos índices produtivos. A infecção urinária (cistite) e as endometrites são consideradas importantes causas de descarte em fêmeas suínas, por terem consequências reprodutivas relevantes e elevarem a taxa de reposição do plantel. O presente estudo teve o objetivo de avaliar o aparelho reprodutivo e a bexiga de fêmeas suínas de descarte normal de granjas, bem como investigar a existência de relação entre as patologias encontradas. Foram examinadas 79 matrizes suínas oriundas de 20 rebanhos localizados no Estado de Santa Catarina. De cada fêmea foram coletados os ovários, fragmentos de útero e bexiga. Dentre as fêmeas avaliadas, 32 (40,5%) tinham diferentes graduações de cistite, 24 (30,4%) tinham algum tipo de inflamação uterina, e 9 (11,4%) estavam em anestro, com ovários inativos. Contudo, não foi observada dependência significativa entre cistite e endometrite nas amostras analisadas.

Reproductive failures are the major reasons for removal of sows and decrease of production rates in an intensive swine production system. Urinary infection and endometritis are considered important causes for culling of sows, due to relevant reproductive consequences and increase of the replacement rates. The present study aimed to evaluate the reproductive and urinary system of culled sows, as well as investigate the occurrence of cystitis and endometritis in analyzed sows. Samples, such as ovaries, uterus fragments and bladder were collected from 79 sows originated from 20 farms of Santa Catarina State. Results showed that, 32 (40,5%) analyzed sows presented cystitis in different levels, 24 (30,4%) had some class of uterine inflammation, and 9 (11,4%) were in anestrous, with inactive ovaries. However, unsignificative dependence between cystitis and endometritis in analyzed samples was observed.

Review of influenza A virus in swine worldwide: a call for increased surveillance and research.

Pigs and humans have shared influenza A viruses (IAV) since at least 1918, and many interspecies transmission events have been documented since that time. However, despite this interplay, relatively little is known regarding IAV circulating in swine around the world compared with the avian and human knowledge base. This gap in knowledge impedes our understanding of how viruses adapted to swine or man impacts the ecology and evolution of IAV as a whole and the true impact of swine IAV on human health. The pandemic H1N1 that emerged in 2009 underscored the need for greater surveillance and sharing of data on IAV in swine. In this paper, we review the current state of IAV in swine around the world, highlight the collaboration between international organizations and a network of laboratories engaged in human and animal IAV surveillance and research, and emphasize the need to increase information in high-priority regions. The need for global integration and rapid sharing of data and resources to fight IAV in swine and other animal species is apparent, but this effort requires grassroots support from governments, practicing veterinarians and the swine industry and, ultimately, requires significant increases in funding and infrastructure.

Culturing and molecular methods to assess the infectivity of porcine circovirus from treated effluent of swine manure.

Samples were collected at the effluent of two swine manure treatment systems and were analyzed by qPCR to determine the presence and amounts of porcine circovirus (PCV2) genetic material. ST cells were inoculated with the positive samples to evaluate virus viability and for viral genotyping. Twenty-five water samples were collected monthly from treated effluent (March 2009 to December 2010). The PCV2 genome was identified by qPCR in 60% of the samples, and all of the positive samples were able to infect ST cells in vitro. Positive samples were genotyped and 60% of them were positive for both PCV2a and PCV2b, 20% were positive for genotype 2a, and 20% were positive for genotype 2b. Our results suggest that these viruses were able to resist the regular wastewater treatment, and this finding demonstrates the necessity of adding a virus inactivation step to the treatment system to guarantee the safety of water reuse.

Phylogenetic comparison of the carboxy-terminal region of glycoprotein C (gC) of bovine herpesviruses (BoHV) 1.1, 1.2 and 5 from South America (SA).

Different types and subtypes of bovine herpesvirus 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, in such a way that type/subtype differentiation has become an essential tool for understanding the pathogenesis and epidemiology of BoHV infections. In search for a genomic region that would allow a clear distinction between BoHV-1 and BoHV-5, the carboxy-terminal portion of glycoprotein C (gC), corresponding to residues 321-450 (BoHV-1) and 301-429 (BoHV-5) of 23 South American (SA) isolates (Brazil mostly) was amplified and sequenced. The nucleotide sequence alignments revealed levels of genomic similarity ranging from 98.7 to 99.8% among BoHV-1 isolates, 88.3 to 92% between BoHV-1/BoHV-5 and 96 to 99.7% among BoHV-5 isolates. At the amino acid level, sequence similarity varied ranging from 97.5 to 99.5% among BoHV-1, 77.5 to 84.4% between BoHV-1/BoHV-5 and 92.1 to 99.5% (BoHV-5/BoHV-5). The isolates could be clearly separated into BoHV-1.1, BoHV-1.2 and BoHV-5 after phylogenetic analysis. The results suggest that the phylogenetic analysis performed here can be used as a potential molecular epidemiological tool for herpesviruses.

Disseminação do vírus da doença de Aujeszky, envolvendo o comércio de reprodutores suínos de reposição / Spread of virus of Aujeszky's disease by the trade of replacement swine breeders

Relata-se a disseminação da infeção pelo vírus da doença de Aujeszky (VDA) a partir da comercialização de reprodutores suínos oriundos de duas granjas de reprodutores suídeos certificada (GRSC) que tiveram surto da DA. Após a confirmação do diagnóstico de DA, os suínos comercializados 37 a 45 dias anteriores aos surtos foram recolhidos, submetidos ao exame sorológico para o VDA e enviados ao abate. Nos rebanhos-destino foram realizados exames sorológicos para o VDA 17 a 37 dias após o recolhimento, naqueles que estavam lojados nas baias vizinhas àquelas onde haviam sido alojados os suínos comprados; seis meses mais tarde, foi realizada outra sorologia por amostragem dos reprodutores. No total, 52 rebanhos compraram suínos das duas granjas, e, destes, 37 (69,8 por cento) receberam, pelo menos, um animal com sorologia positiva para o VDA. Somente sete (18,9 por cento) deles apresentaram sorologia positiva para o VDA, e em 30 (81,1 por cento), não houve indícios de disseminação da infecção. A contaminação pelo VDA de granjas GRSC representa enorme potencial para a disseminação da infecção, por meio do comércio de suínos de reposição. A rastreabilidade dos animais comercializados em um período anterior ao diagnóstico, com imediata remoção dos lotes de suínos dos rebanhos-destino, evitou a disseminação da infecção

It was reported the spreading of the infection caused by the virus of Aujesky's disease (VAD) by the commercialization of breeders originated from two pigs farms GRSC (Farms of Certified Swine Breeders - FCSB) which had an outbreak of Aujeszky's disease. After the positive diagnosis of Aujesky's disease, the pigs traded from 37 to 45 day before the outbreaks were removed from the herd, bled for serological exams for the VAD and sent to slaughterhouses. The herds which received these pigs were serologically tested for the VAD, 17 to 37 days after the removal of the animals. Serological tests were also performed in pigs lodged at neighboring cages to those which had been lodged with the pigs bought and a sampling test was done six months later. Thus, 52 flocks bought pigs from the farms 1 and 2. From those, 37 (69.8 percent) received at least one serum-positive pig for the VAD. Only seven (18.9 percent) of them were infected and 30 (81.1 percent) pigs showed no indications of infection by the VAD. The occurrence of VAD in FCSB farms represents huge potential for spreading of the VAD, by the trade of replacement pigs. The traceability investigation and removal of the animals traded before the diagnosis, with immediate removal of the positive lots of pigs at the destiny herd, avoided the spread of the infection

Uso do plasma suíno ultrafiltrado na recuperação de leitões com sinais clínicos de circovirose / The use of ultra-filtered swine plasma in the recovery of pigs with clinical signs of porcine circovirus infection

Testou-se o efeito do plasma suíno ultrafiltrado spray-dried, associado a um acidificante comercial na água de bebida para a recuperação de leitões com sinais clínicos da síndrome multissistêmica do definhamento dos suínos (SMDS). Utilizaram-se 40 leitões com sinais clínicos da SMDS, selecionados 20 dias após o alojamento em uma unidade de terminação, distribuídos em quatro tratamentos (T) de 10 leitões cada. No T1, os animais receberam água clorada à vontade (controle); no T2, solução com 2,5 por cento do plasma sangüíneo diluído em água; no T3, acidificante (Selko®) diluído em água na dosagem de 12ml/10l e, no T4, solução com 2,5 por cento do plasma sangüíneo e o acidificante na dose de 12ml/10l, diluídos em água. Os leitões não foram medicados e foram sacrificados aos 28 ou 40 dias de experimento para avaliação sorológica e patológica. Não houve diferença no ganho de peso e na situação clínica-patológica entre os tratamentos. Entretanto, os leitões do T4 estavam em melhor situação clínica-patológica. Os leitões dos quatro tratamentos tiveram boa recuperação, sem terem sido medicados. Observou-se alta freqüência de lesões compatíveis com a SMDS nos pulmões, rins e linfonodos. Concluiu-se que o plasma spray dried associado ao ácido não melhoraram o desempenho e a situação clínica-patológica de leitões com sintomas da SMDS

The effect of the ultra-filtered spray-dried porcine plasma, associated to a commercial acid in the drinking water was tested for recovering pigs with clinical signs of the porcine postweaning multisystemic wasting syndrome (PMWS). Forty piglets with clinical signs of the PMWS were used following a selection at 20 days after their housing in one finishing facility. They were divided in four treatment groups (T) of 10 pigs each: T1 - chlorine treated water ad libitum (control); T2 - solution prepared with 2.5 percent of plasma diluted in water; T3 - acid (Selko® ) diluted in water at the concentration of 12 ml/10l; T4 - solution prepared with 2.5 percent of plasma diluted and the acid (Selko® ) diluted in water at the concentration of 12 ml/10l. The pigs received no medication and were euthanized at 28 or 40 days after the beginning of the experiment for serological and pathological tests. Differences at the weight gain and in the clinical-pathological situation were not observed among the treatments. However, pigs from T4 were in better clinical-pathological situation. The pigs of all four treatments showed a good recovery, although they were not medicated. Even though, it was observed a high frequency of lesions compatible to PMWS in the lungs, kidneys and lymph nodes. It was concluded that the plasma spray-dried associated to the acid did not improve the performance and the clinical-pathological situation of pigs with clinical signs of PMWS

Comparing ectopic bone growth induced by rhBMP-2 on an absorbable collagen sponge in rat and rabbit models.

Recombinant human Bone Morphogenetic Protein-2 (rhBMP-2) is currently employed as an autograft replacement for spinal fusion. The morphogen is incorporated onto its carrier, an absorbable collagen sponge (ACS), in the operating room. Although the effectiveness of the rhBMP-2/ACS implant in stimulating bone formation in human subjects has now been well established, further investigations of its use are necessary to deepen our understanding of its performance. The objective of the present study was to determine whether fluid released from the rhBMP-2/ACS implant could induce bone growth in tissue sites away from the implant site. We first measured the amount of protein in the fluid released from the rhBMP-2-soaked ACS during intraoperative handling. Variables included soak time and degree of compression. In the compression group that most closely approximated intraoperative conditions, more than 95% of the rhBMP-2 protein was retained by the ACS following a 15-min. soak time. This in vitro study was followed by an in vivo ectopic implant experiment using rat and rabbit models. The animal investigation compared the amount of bone induced by rhBMP-2 solution alone versus the de novo bone formation induced by rhBMP-2/ACS implants with varying concentrations of rhBMP-2. No ossicles were found at the sites where rhBMP-2 solution was injected in either animal species. Twenty-two of the 24 subcutaneous sites in the rats implanted with the rhBMP-2/ACS constructs displayed the presence of the typical 4- and 12-week ossicle. There were no noticeable differences in the size and shape of the ossicles after 4 and 12 weeks. There was a greater percentage of implant sites without ossicles in the rabbits, compared to the rats.

Coinfecção experimental de circovírus suíno tipo 2 isolado no Brasil e parvovírus suíno em suínos SPF / Experimental coinfection of SPF pigs with porcine circovirus type 2 isolated in Brazil and porcine parvovirus

Avaliou-se a patogenicidade do circovírus suíno tipo 2 (PCV2) isolado no estado de Santa Catarina mediante coinfecção experimental com parvovírus suíno (PPV). Foram utilizados 24 leitões specific pathogen free (SPF) com cinco dias de idade, distribuídos em quatro grupos (G), alojados em salas independentes e inoculados por via intranasal: G1 - controle (n=4); G2 - inoculados com PCV2 (n=7); G3 - inoculados com PPV (n=6); G4 - inoculados com PCV2 e PPV (n=7). Os animais foram monitorados diariamente para avaliação clínica e necropsiados 48 dias após a infecção. As principais lesões anatomopatológicas observadas nos suínos do G2 e G4 foram: aumento do volume dos linfonodos, depleção linfocitária com redução dos folículos linfóides nos órgãos linfocitários e presença de infiltrado eosinofílico nos linfonodos. A técnica de nested-PCR para PCV2 foi utilizada detectando DNA viral em órgãos de todos os animais do G2 e G4. O PCV2 infectou suínos SPF por via intranasal e foi detectado em outros órgãos, com mais lesões histopatológicas e em maior proporção nos animais coinfectados com PPV (G4), quando comparados aos infectados somente com PCV2 (G2).

The virulence of porcine circovirus type 2 (PCV2) isolated in Santa Catarina State by coinfection with porcine parvovirus (PPV) was investigated. Twenty-four, 5-day-old SPF pigs were distributed into four groups, housed in separate rooms and inoculated by intranasal route: G1 - control (n=4); G2 - inoculated with PCV2 (n=7); G3 - inoculated with PPV (n=6); G4 - inoculated with PCV2 and PPV (n=7). The animals were monitored daily for clinical evaluation and were necropsied 48 days after the infection. The pathological lesions seen in G2 and G4 pigs were: enlargement of lymph nodes, mild to moderate lymphoid cell depletion, affecting lymphoid follicles in lymphoid organs and presence of infiltration by eosinophils in lymph nodes. PCV2 DNA was detected by a nested-PCR in all pigs of G2 and G4. These findings confirmed that pigs were successfully infected intranasally with PCV2. The presence of PCV2 DNA in tissue samples and the pathological lesions were more evident in pigs infected with both PCV2 and PPV than in pigs infected with PCV2 alone.

Diagnosis of post-weaning multisystemic wasting syndrome in pigs in Brazil caused by porcine circovirus type 2

Este trabalho descreve a primeira caracterização preliminar de isolados de circovírus suíno tipo 2 (PCV2) a partir de órgãos de suínos acometidos pela síndrome da refugagem multissistêmica (SRM) no Brasil. Leitões doentes foram examinados à necropsia e por histopatologia. Análises macroscópicas e microscópicas demonstraram lesões típicas de SRM, respectivamente, emagrecimento, aumento do volume dos linfonodos, atrofia de timo e pneumonia intersticial, e linfadenite granulomatosa com células sinciciais, entre outras. A presença de antígeno ou DNA do PCV2 foi demonstrada por imunoperoxidase ou reação da polimerase em cadeia nested (nested PCR), respectivamente. Foi possível diferenciar PCV1 e PCV2 por análises de polimorfismo de fragmento de restrição (RFLP) do fragmento amplificado da PCR. O DNA do PCV2 foi detectado em 70 por cento (14/20) das amostras obtidas de suíno com sintomatologia clínica e lesões associadas com SRM. Este estudo apresenta a associação de PCV2 com lesões e sinais clínicos de SRM em suínos e também indica que os isolados do PCV2 brasileiros podem apresentar variações genômicas.

Analysis of fumonisin B1-induced apoptosis.

Fumonisins are mycotoxins produced by Fusarium moniliforme, a prevalent fungus that infects corn and other cereal grains. Fumonisin B1(FB1 is the most common mycotoxin produced by F. moniliforme, suggesting it has toxicologic significance. The structure of FB1 resembles sphingoid bases, and it inhibits ceramide synthase. Because sphingoid bases regulate cell growth, differentiation, transformation, and apoptosis, it is not surprising to find that FB1 can alter growth of certain mammalian cells. Previous studies concluded FB1-induced apoptosis, or cell cycle arrest, in African green monkey kidney fibroblasts (CV-1). In this study we have identified genes that inhibit FB1 induced apoptosis in CV-1 cells and two mouse embryo fibroblasts (MEF). A baculovirus gene, inhibitor of apoptosis (CpIAP), protected these cells from apoptosis. CpIAP blocks apoptosis induced by the tumor necrosis factor (TNF) pathway as well as other mechanisms. Further support for the involvement of the TNF signal transduction pathway in FB1 induced apoptosis was the cleavage of caspase 8. Inhibition of caspases by the baculovirus gene (italic)p35 also inhibited FB1-induced apoptosis. The tumor suppressor gene p53 was not required for FB1 induced apoptosis because p53-/- MEF undergo apoptosis following FB1 treatment. Furthermore, Bcl-2 was not an effective inhibitor of FB1-induced apoptosis in CV-1 cells or p53+/+ MEF. In summary, these results provide new information to help understand the mechanism by which FB1 induces apoptosis.

Analysis of latency in cattle after inoculation with a temperature sensitive mutant of bovine herpesvirus 1 (RLB106).

Calves were inoculated with the bovine herpes virus 1 (BHV-1) vaccine strain (RLB 106), which is a temperature sensitive mutant. The route of inoculation was intranasal instillation or intramuscular (i.m.) injection (flank or neck). As a control, five calves were given placebo by i.m. injection of the neck. Regardless of the infection route, clinical symptoms did not occur. However, BHV-1 neutralizing antibodies were detected after inoculation demonstrating that sero-conversion occurred. At 60 days post-inoculation, dexamethasone was given by i.m. injection to attempt reactivation of RLB 106. Only those calves inoculated by the intranasal route shed virus leading to an increase in BHV-1 specific antibodies. As expected, viral DNA and the latency related-RNA were detected in trigeminal ganglia (TG) of calves inoculated by the intranasal route. In contrast, viral nucleic acid was not detected in TG of calves inoculated by the i.m. route or in calves inoculated with placebo. In cervical ganglia or sacral dorsal root ganglia, viral nucleic acid was not consistently detected. This study provides evidence that efficient latency and reactivation does not occur following i.m. inoculation. Since serum-neutralizing antibodies were detected in all inoculated calves, i.m. inoculation led to sero-conversion.

Virus-induced neuronal apoptosis blocked by the herpes simplex virus latency-associated transcript.

Latent infections with periodic reactivation are a common outcome after acute infection with many viruses. The latency-associated transcript (LAT) gene is required for wild-type reactivation of herpes simplex virus (HSV). However, the underlying mechanisms remain unclear. In rabbit trigeminal ganglia, extensive apoptosis occurred with LAT(-) virus but not with LAT(+) viruses. In addition, a plasmid expressing LAT blocked apoptosis in cultured cells. Thus, LAT promotes neuronal survival after HSV-1 infection by reducing apoptosis.

The latency-related gene of bovine herpesvirus 1 inhibits programmed cell death.

Although viral gene expression occurs in the peripheral nervous system during acute infection, bovine herpesvirus 1 (BHV-1) gene expression is extinguished, many neurons survive, and latency ensues. The only abundant viral transcript expressed during latency is the latency-related (LR) RNA, which is alternatively spliced in trigeminal ganglia during acute infection (L. Devireddy and C. Jones, J. Virol. 72:7294-7301, 1998). A subset of neurons express a protein encoded by the LR gene and the LR protein (LRP) is associated with cyclin-dependent kinase 2 (Cdk2)/cyclin complexes during productive infection (Y. Jiang, A. Hossain, M. T. Winkler, T. Holt, A. Doster, and C. Jones, J. Virol. 72:8133-8142, 1998). LR gene products inhibit cell cycle progression, perhaps as a result of LRP interacting with Cdk2/cyclin complexes. During acute infection, expression of cyclin A occurs in trigeminal ganglionic neurons (L. M. Schang, A. Hossain, and C. Jones, J. Virol. 70:3807-3814, 1996). Inappropriate expression of G(1)- and S-phase cyclins can initiate programmed cell death (PCD), apoptosis, in neurons, suggesting that LR gene products inhibit PCD. To test this hypothesis, we modified an assay to measure PCD frequency in transiently transfected cells. C(6)-ceramide, fumonisin B(1) (FB(1)), or etoposide was used to initiate PCD following transfection of cells with plasmids expressing LR gene products and the beta-galactosidase gene. Transfected cells that survived were quantified by counting beta-galactosidase-positive cells. Plasmids that expressed LR gene products promoted survival of monkey kidney (CV-1), human lung (IMR-90), or mouse neuroblastoma (neuro-2A) cells after induction of PCD. Plasmids with termination codons at the beginning of LR open reading frames or deletion of sequences that mediate splicing of LR RNA did not promote cell survival following PCD induction. We hypothesize that LR gene products play a role in promoting survival of postmitotic neurons during acute infection or reactivation.

Fumonisin B1, a mycotoxin contaminant of cereal grains, and inducer of apoptosis via the tumour necrosis factor pathway and caspase activation.

Fumonisins are mycotoxins produced by Fusarium moniliforme, a prevalent fungus which infects corn or other cereal grains. Fumonisin B1 (FB1) is the most common mycotoxin produced by F. moniliforme, suggesting that it has toxicological significance. The structure of FB1 resembles sphingoid bases and it inhibits ceramide synthase. As sphingoid bases regulate cell growth, differentiation, transformation and apoptosis, it is reasonable to hypothesize that FB1 can also regulate these activities. Previous studies concluded that FB1 induced apoptosis or cell-cycle arrest in CV-1 cells (African green monkey kidney fibroblasts). In this study, we have identified genes that inhibit FB1-induced apoptosis in CV-1 cells and in two primary human cell types (lung fibroblasts and neonatal kidney cells). A baculovirus gene. inhibitor of apoptosis (IAP), protected CV-1 and the human cells from apoptosis. IAP blocks apoptosis which is induced by the tumour necrosis factor (TNF) pathway. Inhibition of interleukin converting enzymes (ICE proteases or caspases) by the baculovirus gene p35 also inhibited FB1-induced apoptosis. FB1 treatment led to cleavage of Rb (retinoblastoma protein) at its C-terminus in CV-1 or human lung cells. As the C-terminus of Rb is cleaved by ICE proteases during apoptosis, this supports an active role for ICE proteases in FB1-induced apoptosis. The tumour suppressor gene p53 was not required for FB1-induced apoptosis because p53-/- primary mouse embryo fibroblasts underwent apoptosis following FB1 treatment. Furthermore, Bcl-2 was not an effective inhibitor of FB1-induced apoptosis in CV-1 or IMR-90 cells. In summary, these results demonstrate that the TNF pathway and caspases plays an important role in FB1-induced apoptosis.

Characterization of cell-cycle arrest by fumonisin B1 in CV-1 cells.

Fusarium moniliforme is a widespread fungal pathogen which primarily infects corn, but can also infect rice or wheat. Fusarium moniliforme produce several mycotoxins, the most prominent of which is called fumonisin B1 (FB1). Epidemiological studies have indicated that ingestion of fumonisins correlates with a higher incidence of oesophageal cancer in Africa and China. Fumonisins also cause a neurodegenerative disease in horses, induce hepatic cancer in rats, are nephrotoxic in rats, or cause pulmonary oedema in swine. Structurally, fumonisins resemble sphingolipids and can alter sphingolipid biosynthesis. suggesting that sphingolipid alterations play a role in disease and carcinogenesis. Previous studies determined that FB1 blocked cell-cycle progression in CV-1 cells but not COS-7 cells. Herein, we have examined the effects that FB1 treatment has on cell-cycle regulatory proteins. Our studies established that FB1 treatment of CV-1 cells, but not COS-7 cells, leads to dephosphorylation of the retinoblastoma (Rb) protein. Cyclin dependent kinase 2 (CDK2) activity was repressed five- to 10-fold and cyclin E protein levels were lower in CV-1 cells after fumonisin treatment. Two CDK inhibitors, Kip1 and Kip2, were induced within 3 hours after fumonisin treatment of CV-1 cells, suggesting these two proteins mediate cell-cycle arrest induced by FB1. This mycotoxin caused large increases in sphinganine within 3 hours after addition of FB1. As sphingoid bases are known to induce Rb phosphorylation, this increase in sphinganinie might be the stimulus for the suppression of cyclin dependent kinase activities via Kip1 and Kip2. The ability of FB1 to accumulate sphingosine or sphinganine and arrest the cell cycle in some cells but not others may play an important role in carcinogenesis or disease.

Analysis of cyclin-dependent kinase activity after herpes simplex virus type 2 infection.

Small DNA viruses (adenoviruses, simian virus 40, or human papillomaviruses) induce S-phase progression but prevent cell division to provide precursors for viral DNA replication. Herpes simplex viruses types 1 or 2 (HSV-1 or HSV-2) contain genes which encode DNA-metabolizing enzymes, for example, ribonucleotide reductase, thymidine kinase and dUTPase, suggesting that S-phase factors are not required for an efficient infection. However, several studies indicated that HSV induces some events that occur during cell-cycle progression. To determine if HSV-2 induces S-phase entry, we examined serum-arrested African green monkey kidney cells (CV-1) after infection. Two hours after infection steady-state levels of the S-phase-specific cyclin, cyclin A, increased. S-phase cyclin-dependent kinase activity (CDK2) was stimulated 10-fold 8 h after infection but decreased at 16 or 24 h after infection. Mitotic CDK activity (CDC2) was not activated after infection, in part due to decreases in CDC2 protein levels and inactivation of enzymatic activity resulting from tyrosine phosphorylation of CDC2. Furthermore, CDK4 activity was not dramatically affected by infection. These studies indicate that HSV-2 infection selectively activates CDK2 after infection but cell-cycle progression does not occur. We hypothesize that infection activates certain components of the cell cycle which enhance viral gene expression and DNA replication.

Validation of an automated arterial tonometry monitor using Association for the Advancement of Medical Instrumentation standards.

BACKGROUND: Arterial tonometry is a noninvasive technique for monitoring the arterial blood pressure in a continuous manner. Real-time arterial blood pressure measurements are advantageous in many clinical settings, but arterial tonometric devices must be validated prior to clinical acceptance. Guidelines for accuracy for noninvasive blood pressure monitors have been established by the Association for the Advancement of Medical Instrumentation. OBJECTIVE: To test the Colin Pilot 9200 configured with an arterial tonometry module with 20 patients and to compare tonometric blood pressure measurements with intra-arterial blood pressure measurements. METHODS: All of the patients in the study were aged over 14 years and weighed over 35 kg; testing was performed in the operating room or in the intensive care unit. Data from each patient consisted of multiple simultaneous recordings of tonometric and intra-arterial blood pressure values. The data were then compared; the mean and SD of the difference between the two measurement devices were then calculated. RESULTS: Tonometric values were slightly less than the intra-arterial pressure measurements; the mean difference for systolic blood pressure was 2.24 +/- 8.7 mmHg and for diastolic pressure was 0.26 +/- 8.88 mmHg. CONCLUSION: The arterial tonometry module incorporated into the Colin Pilot 9200 was investigated for use with selected adult and pediatric populations. With our patients, it generated accurate data throughout a wide blood pressure range. It satisfied Association for the Advancement of Medical Instrumentation standards for mean systolic and diastolic blood pressure measurements and only minimally exceeded the allowable SD. This technology should prove to be a valuable tool for noninvasive blood pressure monitoring of various patient populations.