Orientador: Mário Massayuki Inomoto / Resumo: Amostras de solo e raízes foram coletadas em plantações de café no estado de São Paulo, com o objetivo de determinar a importância e a ocorrência dos nematóides das lesões na cultura do café. A espécie de Pratylenchus mais freqüente foi P. brachyurus (solo: 13,2 %; raízes: 18,3 %), mas geralmente em baixas densidades. O nematóide das lesões do café, P. coffeae, ocorreu em 5,1 % das amostras de raízes, mas em altas densidades e causando mais danos do que a primeira espécie. Outra espécie, P. vulnus, foi encontrada somente em uma localidade. Este é o primeiro relato de P. vulnus em café. Outros fitonematóides identificados nas amostras foram: Xiphinema brevicolle, Xiphinema sp., Paratrichodorus minor, Paratrichodorus sp., ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Soil and root samples were collected from coffee plantations in the state of São Paulo, in order to determine the occurrence and importance of lesion nematodes in coffee culture. The most frequent species of Pratylenchus was P. brachyurus (soil: 13,2 %; roots: 18,3 %), but generally in low densities. The coffee lesion nematode, P. coffeae, occurred in 5,1 % of root samples, but in higher densities and causing more damage than the former species. Other species, P. vulnus, was found in one single locality. This is the first report of P. vulnus on coffee. Other phytonematodes identified in the survey were: Xiphinema brevicolle, Xiphinema sp., Paratrichodorus minor, Paratrichodorus sp., Tylenchorhynchus sp., Helicotylenchus dihystera, H. californicus, H. erythrinae, Helicotylenchus sp., Scutellonema sp., Rotylenchulus reniformis, Meloidogyne exigua, M. incognita, M. coffeicola, Meloidogyne sp, Criconemella onoensis, C. ornata, C. sphaerocephala, Criconemella sp., Hemicriconemoides strictathecatus, and Paratylenchus sp. ...(Complete abstract, click electronic access below) / Doutor

HIFU is the upcoming technology for noninvasive or minimally invasive tumor ablation via the localized acoustic energy deposition at the focal region within the tumor target. The presence of cavitation bubbles had been shown to improve the therapeutic effect of HIFU. In this study, we have investigated the effect of HIFU on temperature rise and cavitation bubble activity in ethanol-treated porcine liver and kidney tissues. We have also explored changes in the viability and proliferation rate of HepG2, SW1376, and FB1 cancer cells with their exposure to ethanol and HIFU. Tissues were submerged in 95% ethanol for five hours and then exposed to HIFU generated by a 1.1 MHz transducer or injected into focal spot before HIFU exposure. Cavitation events were measured by a passive cavitation detection technique for a range of acoustic power from 1.17 W to 20.52 W. The temperature around the focal zone was measured by type K or type E thermocouples embedded in the samples. In experiments with cancer cells, 2.7 millions cells were treated with concentration of ethanol at concentration 2%, 4%, 10%, 25%, and 50% and the cell were exposed to HIFU with power of 2.73 W, 8.72 W, and 12.0 W for 30 seconds. Our data show that the treatment of tissues with ethanol reduces the threshold power for inertial cavitation and increases the temperature rise. The exposure of cancer cells to various HIFU power only showed a higher number of viable cells 24 to 72 hours after HIFU exposure. On the other hand, both the viability and proliferation rate were significantly decreased in cells treated with ethanol and then HIFU at 8.7 W and 12.0 W even at ethanol concentration of 2 and 4 percent. In conclusion, the results of our study indicate that percutaneous ethanol injection (PEI) and HIFU have a synergistic effect on cancer cells ablation. / acase@tulane.edu

Executive functioning is a multidimensional concept encompassing higher-order adaptive abilities, such as judgment, decision-making, self-monitoring, planning, and emotional regulation. Disruption in executive functioning often results in devastating impairments in vitally-important areas of life, such as one's ability to hold employment and maintain social relationships. Executive functions have been associated primarily with the prefrontal cortex. However, the nature and degree of the association between frontal lobe damage and performance on executive functioning tests remains controversial. Research suggests that the association may vary based on the specific location of damage within the prefrontal cortex, as well as the used measure of executive functioning. Few investigations have systematically addressed these variables. The current study employed the lesion method to investigate the relationship between performance on a battery of executive functioning tests and damage to specific regions of the prefrontal cortex. Three groups of participants with lesions in one of the locations of interest [ventromedial prefrontal (VMPC, n = 14), dorsolateral prefrontal (DLPC, n = 14), and non-frontal (n = 18)] were administered the Delis-Kaplan Executive Function System (D-KEFS, 2001), a comprehensive battery of executive functioning tests. Results revealed no statistically-significant differences between group performances on the D-KEFS primary measures. However, a qualitative analysis of the results revealed several meaningful group differences. It appears that some relationship exists between frontal lobe damage, particularly in the DLPC, and decreased performance on several executive functioning tests but further research overcoming the methodological limitations of most existing literature on this topic is needed to clearly resolve this issue.

Phakopsora meliosmae-myrianthae, a causal agent of Asian grapevine leaf rust, and Phakopsora pachyrhizi, a causal agent of Asian soybean rust, cause severe epidemics on their crop hosts. Both Phakopsora spp. seem to behave differently to other rusts, i.e. showing a high frequency of pustules on leaves concomitantly with host tissue necrosis and leading to premature defoliation. To shed light on the epidemiology of these rusts, this study aimed to: (i) compare the colonization progress of P. meliosmae-myrianthae and P. pachyrhizi on their hosts, by determination of fungal biomass via histological staining and quantitative polymerase chain reaction (qPCR); (ii) compare the effects of Phakopsora spp. on host photosynthesis and the monocyclic components: incubation, latent, and infectious periods, number of uredinia, and disease severity over time; and (iii) estimate the relative defoliation rate caused by Asian grapevine and soybean rusts and its relationship to a range of disease severity. All pathosystems showed lesion expansion. Mycelial colonization did not extend beyond the lesion border. No increase in the number of lesions was observed over time, but formation of new uredinia of P. pachyrhizi and P. meliosmae-myrianthae within an existing lesion, without the need for a new infection site, increased by 9- to 19-fold, respectively. Incubation and latent periods were coincident for 8 days in Asian grapevine leaf rust and 13 days in Asian soybean rust. Minimum infectious periods were 21 days for P. meliosmae-myrianthae and 13 days for P. pachyrhizi, and both pathogens presented several sporulation peaks. Both Phakopsora rusts showed an increase in disease severity during monocycle, with similar progress rates that were estimated with the monomolecular model as 0.06 and 0.05 day-1 for grapevine and soybean rusts, respectively. P. meliosmae-myrianthae and P. pachyrhizi infection reduced relative photosynthetic rates by 22% and 5%, respectively, before the onset of symptoms. Defoliation rates of grapevine and soybean rusts were positively correlated with mean disease severity, according to a logarithmic model. On symptomless grapevine and soybean leaves, defoliation rates were 0.05 and 0.06 day-1, respectively. On diseased grapevine leaves, defoliation rate was 0.13 day-1 for leaves with disease severity between 12.1% and 25%. The rate of defoliation on soybean leaflets was 0.12 day-1 when disease severity was between 25% and 60%. Our findings showed that the epidemiological behaviour of P. meliosmae-myrianthae is similar to that of P. pachyrhizi. The continuous increase in the number of uredinia within lesions ensures an infectious period with several urediniospore production peaks, which is an efficient survival mechanism for these pathogens in the tropics. This might be directly related to the frequent epidemics caused by Phakopsora spp. / Phakopsora meliosmae-myrianthae, agente causal da ferrugem asiática da videira, e Phakopsora pachyrhizi, agente causal da ferrugem asiática da soja, ocasionam epidemias severas em seus hospedeiros. O comportamento dessas Phakopsora spp. parece não seguir o padrão de outras ferrugens, por exemplo apresentam elevada frequência de pústulas no limbo foliar concomitantemente à necrose foliar e desfolha precoce. Para elucidar a epidemiologia dessas ferrugens, este estudo teve como objetivos: (i) comparar o progresso da colonização de P. meliosmae-myrianthae e de P. pachyrhizi, em seus hospedeiros, pela determinação da biomassa fúngica via coloração histológica e via reação em cadeia da polimerase em tempo real (qPCR); (ii) comparar os efeitos de Phakopsora spp. na fotossíntese dos seus hospedeiros e os componentes monocíclicos: períodos de incubação, latente e infeccioso, número de uredínios e severidade da doença ao longo do tempo; e (iii) estimar as taxas relativas de desfolha ocasionadas pelas ferrugens da videira e da soja em função da severidade das doenças. Todos os patossistemas apresentaram crescimento da lesão. A colonização micelial não se estendeu além da borda da lesão. Não foi observado aumento no número de lesões ao longo do tempo, mas o número de novos uredínios de P. pachyrhizi e de P. meliosmae- myrianthae nas lesões aumentou em 9 e 19 vezes, respectivamente. Os períodos de incubação e latente foram coincidentes: 8 dias para ferrugem da videira e 13 dias para a ferrugem da soja. Os períodos infecciosos foram de, no mínimo, 21 dias para P. meliosmae-myrianthae e de 13 dias para P. pachyrhizi e foram compostos de vários picos de esporulação para ambas as ferrugens. Durante o monociclo, as duas ferrugens apresentaram aumento da severidade, com similar taxa de progresso, determinada pelo modelo monomolecular, de 0,06 dia-1 e 0,05 dia-1, para as ferrugens da videira e da soja, respectivamente. Phakopsora meliosmae-myrianthae e P. pachyrhizi reduziram, em média, 22% e 5% da taxa fotossintética líquida nas folhas infectadas antes do aparecimento dos sintomas, respectivamente. As taxas de desfolha da ferrugem da videira e da soja foram positivamente correlacionadas com a severidade média das doenças, de acordo com um modelo logarítmico. Nas folhas de videira e de soja sem sintomas, as taxas de desfolha foram de 0,05 dia-1 e 0,06 dia-1, respectivamente. Nas folhas de videira com severidade da doença entre 12,1 a 25%, a taxa de desfolha foi de 0,13 dia-1 e nos folíolos de soja com severidade da doença entre 25 a 60%, foi de 0,12 dia-1. Nossos resultados mostram que o comportamento epidemiológico de P. meliosmae-myrianthae é semelhante ao de P. pachyrhizi. O contínuo aumento do número de uredínios na lesão favorece um período infeccioso com vários picos de produção de urediniósporos, sendo um eficiente mecanismo de sobrevivência desses patógenos nos trópicos. Essas características podem estar diretamente relacionadas às frequentes epidemias ocasionadas por Phakopsora spp.

No description available.

The incidence of Squamous Cell Carcinoma is growing in certain populations to the extent that it is now the most common skin lesion in young men and women in high ultraviolet exposure regions such as Queensland. In terms of incidence up to 45% of the Australian population over 40 years of age is thought to possess the precancerous Solar Keratosis lesion and with a small but significant chance of progression into SCC, understanding the genetic events that play a role in this process is essential. The major aims of this study were to analyse whole blood derived samples for DNA aberrations in genes associated with tumour development and cellular maintenance, with the ultimate aim of identifying genes associated with non-melanoma skin cancer development. This study had an explicit emphasis on the mitochondrial genome and nuclear genes that encode for subunits in the mitochondrial regulated energy transducing oxidative phosphorylation pathways. More specifically the first aim of this project was to analyse the NDUFA8, PTCH, NDUFAS, SMOH, SDHD, MMPI2, NDUFV1, EMSI, COXVIIc, and RASAI genes via non-specific fluorophoric Real-Time PCR for genetic aberrations in an affected Solar Keratosis and control cohort. The second aim was to analyse two specific genes, SDHD and MMPI2, for copy number aberrations via Dual-Labelled Probe Real-Time PCR in the same affected Solar Keratosis and control cohort. The third aim was to analyse Mitochondrial DNA Depletion syndrome (MDS) in a chemically exposed RAAF personnel cohort via Dual-Labelled Probe Real-Time PCR. The significance of these studies is in their contribution to the knowledge of the genetic pathways that are malformed in the progression and development of the pre-cancerous skin lesion Solar Keratosis. Furthermore, it would determine whether the genes analysed in this study exist in greater prevalence in the affected Solar Keratosis population compared to the control cohort. With regard to the MDS component, identifying the presence of this disease in these individuals was initially undertaken as part of a study to provide evidence in compensation claims. The diagnosis may assist in their medical therapy, insofar as some of them were now suffering from liver malfunctions and atypical male breast cancer. Another application of this effective and low cost method of diagnosing MDS is in populations with high HTV incidences. This is due to the fact that the most common drug used to treat this disease can give rise to the expression of MDS, thus further complicating the health status of HIV infected individuals. The analysis of this research was accomplished via the Real-Time PCR technique, with a non-specific fluorophore component in addition to specific Dual-Labelled Probe components, to ascertain the general nature of any aberration identified in the sample cohort. This project also employed additional methods of analysis such as DHPLC and DNA sequencing to assist in determining the veracity of its aims, particularly in terms of the precise detection of genetic aberrations via Real-Time PCR. Patients exhibiting male breast cancer and liver malftinctions were also analysed via Dual-Labelled Probe RealTime PCR to ascertain the presence of Mitochondrial DNA Depletion syndrome, a disorder characterised by lactic acidosis, liver failure, seizures, and congestive heart failure. Determining the presence of this syndrome in these patients would assist in their medical treatment, and contribute to the analytical methods available to diagnose this syndrome, which is known to occur in HIV sufferers due to the nucleoside drugs used to combat the disease. Real-Time PCR can adequately gauge the integrity of a genetic area in terms of amplicon malformities (non-specific-fluorophoric) and DNA copy number aberrations (Dual-Labelled Probe) via fluorophore signal differentials compared to wild-type samples and housekeeper profiles. The results of the first component of this project, namely the analysis of five gene pairs by non-specific fluorophoric Real-Time PCR, highlighted that a significantly higher incidence of putative aberrants is evident in the affected population when compared to the control cohort. The genes analysed were NDUFA8, PTCH, NDUFA5, SMOH, SDHD, MMP 12, NDUFVI, EMS 1, COXVIIc, and RASA 1. These ten genes were subdivided into five pairs; one of the pair being a gene associated with the development of a non-melanotic skin cancer (NMSC), the other a gene encoding for a subunit of the Electron Transport Chain (ETC). Each of these pairs exists in close proximity to one another on a particular chromosomal locale. Differences were highlighted in the single gene triplicate run population. The ETC genes (NDUFA8, NDUFA5, SDHD, NIDUFVI, COXVIIc) exhibited 10 / 720 (1.37%) as being putative mutants in the control population, compared to 117 / 675 (17.3%) for the affected population (p value less than 0.0001). The NMSC gene analysis (PTCH, SMOH, MMPI2, EMSI, RASA1) produced a 16 / 720 (2.22%) ratio for the control population, with the affected population having an incidence of 97 / 675 (14.4 %) for putative mutants (p value less than 0.0001). The observance of putative aberrants in the NDUFVI (p less than 0.018), EMS1 (p less than 0.003), COXVTIc (p less than 0.001), and RASA I (p less than 0.009) genes in the affected Solar Keratosis (SK) population was significantly higher than that observed in the control population. The majority of aberrations detected via the non-specific fluorophoric Real-Time PCR technique were small nucleotide base insertions and deletions. The analysis of the SK affected and control cohort via Real-Time PCR proved a cost-effective and reliable method in identifying the presence of DNA aberrations such as non-instructional sites. The results of the second component extended the findings of the non-specific fluorophoric analysis. The SDHD and MMPI 2 genes were analysed for copy number aberrations via Dual-Labelled Probe Real-Time PCR for genetic aberrations the same affected and control Solar Keratosis cohort. It was found that 12 of 279 samples had identifiable copy-number aberrations in either the SDHD or MMPI2 gene (this means that a genetic section of either of these two genes is aberrantly amplified or deleted), with five of the samples exhibiting aberrations in both genes. The MMPI2 gene also had nine samples identified as possessing an intronic heterozygous base-pair substitution anomaly via DNA sequencing. The NDUFA8 gene had 12 samples identified as anomalous via the DHPLC technique, 11 of which were identified via non-specific fluorophoric Real-Time PCR, with the analysis performed to verify the accuracy of the Real-Time technique in identifying DNA aberrations. This study identified DNA aberrations in an affected Solar Keratosis and control cohort and ascertained several particular genomic abnomialities in the SDHD, MMPI2 and NDUFA8 genes, with an emphasis on copy-number aberrations and amplicon abnormalities. In the third component of this study, namely the analysis of Mitochondrial DNA Depletion syndrome (MDS) in a jet-fuel exposed RAAF personnel cohort via Dual-Labelled Probe Real-Time PCR, the results indicated that four of the seven patients were expressing MDS. Of the four patients who exhibited a reduction in mitochondrial copy-number the average decrease was of a four-fold level, or approximately a depletion of mitochondrial copies from 200 plus to ~ 54 (74 % reduction in MtDNA). The patients who contributed DNA for investigation into the presence of MDS were suffering from liver malfunction and atypical male breast cancer. The Dual-Labelled Probe technique proved a reliable and cost effective method in identifying the presence of MDS in these patients, with the DNA extracted from fresh white blood cells that had been isolated using the Ficoll-Hypaque method. The importance of this is that accurate levels of Mitochondrial DNA copy numbers can be ascertained in white blood cells as it removes the presence of platelets, which also contain mitochondria but no nucleus. The analysis of ETC and NMSC associated genes in addition to mitochondrial copy number integrity means that this study investigated two aspects of the carcinogenetic pathway i.e. abnormal energy regulation and the regulation of micromolecular and macromolecular cellular homeostatic mechanisms. The mechanism of programmed cell death or apoptosis is regulated by the mitochondria and the ability of a genetically damaged cell to evade the apoptotic process is directly linked to a cell becoming cancerous. It is only after the evasion of apoptosis and the replication of the damaged cells' DNA into daughter cells that neoplastic events can occur. Thus, this study contributed to the understanding of how neo-plastic lesions may develop and progress into invasive tumours. It additionally assisted in proving the effectiveness of the RealTime PCR technique in detecting DNA aberrations and mitochondrial copy number anomalies.

<p>Consequences of atherosclerosis may result in a number of diseases of the cardiovascular system that represent serious health problems and major causes of morbidity and mortality worldwide. Although it is initially considered as disease of fibro-lipid and thrombus deposition in the arterial wall, it also involves an ongoing inflammatory response.</p><p>Normally, the inflammatory response is considered as a protective defence mechanism of the body. However, if the inflammation gets out of proportion to the threat it is dealing with, it may then result in a sustained chronic disorder and thus may underlie the initial stage of atherogenesis. The work of this thesis focuses on the expression of cytokines/chemokines and the vascular transcriptional response to inflammation, i.e. LPS in atherosclerosis. This has mainly been studied in animal models of atherosclerosis; consequently, we set out to investigate these events using human material in vitro (human carotid lesions).</p><p>Employing quantitative analysis, we were able to detect a significant induction of protein and mRNA of the cytokines IL-1β, IL-6, IL-10 and TNF-α and the chemokines IL-8 and MCP-1 by LPS in both atherosclerotic and non-atherosclerotic vessels. In contrast, LPS induction of TNF-α, IL-1β and IL-10 was solely observed in the lesions, but not in normal arteries. In addition, the impact of IL-1 gene polymorphism on the risk of myocardial infarction (MI) was estimated by DNA genotyping of 387 survivors of a first MI and 387 sex and age-matched control subjects. We found no statistically significant differences in either genotypic distribution or allelic frequencies of IL-1β (-511) or IL-1Ra (VNTR) polymorphisms between first-time survivors of myocardial infarction and their age-matched healthy controls. Incontrast, our results demonstrated a strong association between the IL-1Ra genotype and severity of angiographically determined coronary artery disease in post-MI patients. To further investigate the vascular response to inflammation, we used gene array analysis to evaluate the human vascular transcriptional response to LPS of non-atherosclerotic human renal arteries compared to carotid lesions. In LPS treated renal arteries, 54% of the transcripts gave a detectable signal, where 4% were upregulated and 3.8% down-regulated. In the LPS stimulated carotid lesions, 44% of transcripts were detected. In this latter group, 5.1% of transcripts were increased and 3.3% decreased. Interestingly, a newly identified virus-inducible antiviral protein, CMV inducible gene ₅/viperin (Cig₅), was among the most strongly induced gene in both normal and atherosclerotic biopsies. Single gene analysis revealed viperin in the endothelium of human atherosclerotic lesions. Further, viperin was induced in vascular cells by inflammatory stimuli and CMV infection.</p><p>In conclusion we show that atherosclerotic vessels produce more proinflammatory cytokines/chemokines than normal vessels. Interestingly, our results indicate that LPS enhances the expression of cytokines/chemokines in a similar pattern both in lesions and normal arteries. However, the response is stronger in atherosclerotic lesions. Furthermore, our results suggest that genetic polymorphisms within the IL-1Ra loci may influence the severity of CAD. Finally, the CMV inducible gene ₅/viperin have been identified as a putative culprit molecule in vascular inflammation and atherosclerosis.</p>

The overall aim of this thesis is to evaluate the risk for increased morbidity and mortality due to long-term arsenic exposure via drinking water by use of epidemiological and spatial approaches in studies performed in Matlab, Bangladesh. A total of 166,934 individuals aged 4 years and above were screened for skin lesions in 2002-2003. Another sample of 115,903 adults aged 15 years or more and a third sample of 26,972 pregnancies in 1991-2000 were considered in a historical cohort and an ecological study, respectively, where risk of adult mortality and spatial clusters of foetal loss and infant death were analysed in relation to arsenic exposure. More than 70% of the tube-wells in the study area exceeded the threshold for arsenic contamination according to the WHO guideline (10 µg/L). The prevalence of arsenic-induced skin lesions was 3/1000 and men had significantly higher prevalence of skin lesions (SMR 158, 95% CI: 133-188) compared to women. There was an increased risk for death in adulthood due to all non-accidental causes (hazards ratio = 1.16, [95% CI 1.06-1.26]) even at a low level of arsenic contamination (10-49 µg/L). Slightly lower risks were observed for death in cancers (1.44 [1.06-1.95]) and infectious diseases (1.30 [1.13-1.49]) at 50-149 µg/L, but for cardiovascular diseases, it was evident (1.23 [1.01-1.51]) from the level 150-299 µg/L. A dose-response relationship was observed for each of these causes. We identified high and low risk clusters of foetal loss and infant death that coincided with identified high and low clusters of arsenic exposure. Water arsenic concentration of the reported main water source was significantly correlated with arsenic concentration in urine, which reflects current arsenic intake from all sources (R2=0.41, ρ&lt; 0.0001), and the influence of neighbouring water sources was minimal. The study findings underlines that the ongoing arsenic exposure has resulted in a series of severe public health consequences in Bangladesh that call for reinforcement in the mitigation efforts. Knowledge about the spatial distribution of exposure and health effects may be of value in that process.

No description available.

In Syrian hamsters, social behavior is mediated exclusively by chemosensory cues and circulating gonadal steroid hormones. Where these two signals are processed in the brain is unknown, but the posterior bed nucleus of the stria terminalis (pBNST) has been suggested as a candidate site. Therefore, we tested male hamsters’ preference for opposite-sex odors following excitotoxic lesions of the pBNST. Lesions of the pBNST (pBNST-X) eliminated male hamsters’ preference for opposite-sex odors. Furthermore, pBNST-X males spent significantly less time investigating female odors than clean odors and significantly less time investigating female odors than control males did. Lesions of the pBNST did not change male hamsters’ investigation of male odors. The deficits observed in pBNST-X males were not due to a failure to discriminate between odors, as pBNST-X males were able to distinguish between odors. Together, these data suggest the pBNST is critical for opposite-sex odor preference in male hamsters.