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Find video protocols related to scientific articles indexed in Pubmed.
Cell wall bound anionic peroxidases from asparagus byproducts.
J. Agric. Food Chem.
PUBLISHED: 09-22-2014
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Asparagus byproducts are a good source of cationic soluble peroxidases (CAP) useful for the bioremediation of phenol-contaminated wastewaters. In this study, cell wall bound peroxidases (POD) from the same byproducts have been purified and characterized. The covalent forms of POD represent >90% of the total cell wall bound POD. Isoelectric focusing showed that whereas the covalent fraction is constituted primarily by anionic isoenzymes, the ionic fraction is a mixture of anionic, neutral, and cationic isoenzymes. Covalently bound peroxidases were purified by means of ion exchange chromatography and affinity chromatography. In vitro detoxification studies showed that although CAP are more effective for the removal of 4-CP and 2,4-DCP, anionic asparagus peroxidase (AAP) is a better option for the removal of hydroxytyrosol (HT), the main phenol present in olive mill wastewaters.
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Epidural anesthesia in capuchin monkeys (Sapajus libidinosus).
J. Med. Primatol.
PUBLISHED: 09-11-2014
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Many primates are kept in breeding for scientific or conservation purposes, but much of the clinical care is represented by trauma. To provide more effective interventions in some of these cases, the present study aimed to evaluate the technique of local anesthesia through the epidural space in capuchin monkeys (Sapajus libidinosus).
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Combined impact of hepatitis C virus genotype 1 and interleukin-6 and tumor necrosis factor-? polymorphisms on serum levels of pro-inflammatory cytokines in Brazilian HCV-infected patients.
Hum. Immunol.
PUBLISHED: 09-03-2014
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We investigated the association between hepatitis C virus (HCV) genotypes and host cytokine gene polymorphisms and serum cytokine levels in patients with chronic hepatitis C. Serum IL-6, TNF-?, IL-2, IFN-?, IL-4, IL-10, and IL-17A levels were measured in 67 HCV patients (68.2% genotype 1 [G1]) and 47 healthy controls. The HCV patients had higher IL-6, IL-2, IFN-?, IL-10, and IL-17A levels than the controls. HCV G1 patients had higher IL-2 and IFN-? levels than G2 patients. The -174IL6G>C, -308TNF?G>A, and -1082IL10A>G variants were similarly distributed in both groups. However, HCV patients with the -174IL6GC variant had higher IL-2 and IFN-? levels than patients with the GG and CC variants. Additionally, HCV patients with the -308TNF?GG genotype had higher IL-17A levels than patients with the AG genotype, whereas patients with the -1082IL10GG variant had higher IL-6 levels than patients with the AA and AG variants. A significant proportion of HCV patients had high levels of both IL-2 and IFN-?. The subgroup of HCV patients with the G1/IL6CG/TNF?GG association displayed the highest proportions of high producers of IL-2 and IFN-? whereas the subgroup with the G1/TNF?GG profile showed high proportions of high producers of IL-6 and IL-17A. HCV patients with other HCV/cytokine genotype associations showed no particular cytokine profile. Our results suggest that HCV genotype G1 and IL-6 and TNF-? polymorphisms have a clinically relevant influence on serum pro-inflammatory cytokine profile (IL-2 and IFN-?) in HCV patients.
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Caffeic acid protects mice from memory deficits induced by focal cerebral ischemia.
Behav Pharmacol
PUBLISHED: 08-30-2014
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Brain ischemia pathophysiology involves a complex cascade of events such as inflammation and oxidative stress that lead to neuronal loss and cognitive deficits. Caffeic acid (CA) is a natural phenolic compound with antioxidant and anti-inflammatory properties. To evaluate the neuroprotective efficacy of this compound in mice subjected to a permanent middle cerebral artery occlusion, animals were pretreated and post-treated with CA, 2, 20, and 60?mg/kg/day, intraperitoneally, at 24, 48, 72, 96, or 120?h after ischemia. Animals were evaluated at 24?h after the permanent middle cerebral artery occlusion for brain infarction and neurological deficit score. At 72?h after the occlusion, animals were evaluated for locomotor activity, working memory, and short-term aversive memory; long-term aversive memory was evaluated 24?h after the evaluation of short-term aversive memory. Finally, at 120?h after the event, spatial memory and the expression levels of synaptophysin (SYP), SNAP-25, and caspase 3 were evaluated. The treatment with CA reduced the infarcted area and improved neurological deficit scores. There was no difference in locomotor activity between groups. The working, spatial, and long-term aversive memory deficits improved with CA. Furthermore, western blotting data showed that the expression of SYP, which correlates with synaptic formation and function, decreased after ischemic insult, and CA inhibited the reduction of SYP expression. Ischemia also increased, and CA treatment decreased, caspase 3 expression. These results suggest that CA exerts neuroprotective and antidementia effects, at least in part, by preventing the loss of neural cells and synapses in ischemic brain injury.
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Deconstructing the DGAT1 enzyme: Binding sites and substrate interactions.
Biochim. Biophys. Acta
PUBLISHED: 08-23-2014
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Diacylglycerol acyltransferase 1 (DGAT1) is a microsomal membrane enzyme responsible for the final step in the synthesis of triacylglycerides. Although DGATs from a wide range of organisms have nearly identical sequences, there is little structural information available for these enzymes. The substrate binding sites of DGAT1 are predicted to be in its large luminal extramembranous loop and to include common motifs with acyl-CoA cholesterol acyltransferase enzymes and the diacylglycerol binding domain found in protein kinases. In this study, synthetic peptides corresponding to the predicted binding sites of DGAT1 enzyme were examined using synchrotron radiation circular dichroism spectroscopy, fluorescence emission and adsorption onto lipid monolayers to determine their interactions with substrates associated with triacylglyceride synthesis (oleoyl-CoA and dioleoylglycerol). One of the peptides, Sit1, which includes the FYxDWWN motif common to both DGAT1 and acyl-CoA cholesterol acyltransferase, changes its conformation in the presence of both substrates, suggesting its capability to bind their acyl chains. The other peptide (Sit2), which includes the putative diacylglycerol binding domain HKWCIRHFYKP found in protein kinase C and diacylglycerol kinases, appears to interact with the charged headgroup region of the substrates. Moreover, in an extended-peptide which contains Sit1 and Sit2 sequences separated by a flexible linker, larger conformational changes were induced by both substrates, suggesting that the two binding sites may bring the substrates into close proximity within the membrane, thus catalyzing the formation of the triacylglyceride product.
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Linear B-cell epitope mapping of MAPK3 and MAPK4 from Leishmania braziliensis: implications for the serodiagnosis of human and canine leishmaniasis.
Appl. Microbiol. Biotechnol.
PUBLISHED: 08-18-2014
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The correct and early identification of humans and dogs infected with Leishmania are key steps in the control of leishmaniasis. Additionally, a method with high sensitivity and specificity at low cost that allows the screening of a large number of samples would be extremely valuable. In this study, we analyzed the potential of mitogen-activated protein kinase 3 (MAPK3) and mitogen-activated protein kinase 4 (MAPK4) proteins from Leishmania braziliensis to serve as antigen candidates for the serodiagnosis of human visceral and tegumentary leishmaniasis, as well as canine visceral disease. Moreover, we mapped linear B-cell epitopes in these proteins and selected those epitopes with sequences that were divergent in the corresponding orthologs in Homo sapiens, in Canis familiaris, and in Trypanosoma cruzi. We compared the performance of these peptides with the recombinant protein using ELISA. Both MAPK3 and MAPK4 recombinant proteins showed better specificity in the immunodiagnosis of human and canine leishmaniasis than soluble parasite antigens and the EIE-leishmaniose-visceral-canina-bio-manguinhos (EIE-LVC) kit. Furthermore, the performance of this serodiagnosis assay was improved using synthetic peptides corresponding to B-cell epitopes derived from both proteins.
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Role of gastric acid inhibition, prostaglandins and endogenous-free thiol groups on the gastroprotective effect of a proteolytic fraction from Vasconcellea cundinamarcensis latex.
J. Pharm. Pharmacol.
PUBLISHED: 07-27-2014
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The aim of this study was to extend our knowledge about the mechanism involved in the gastroprotective effect of P1G10, a proteolytic fraction rich in cysteine proteinases from Vasconcellea cundinamarcensis (syn. Carica candamarcensis) latex, which demonstrated gastric healing and protection activities in rats.
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IL-4/CCL22/CCR4 Axis Controls Regulatory T Cell Migration That Suppress Inflammatory Bone Loss in Murine Experimental Periodontitis.
J. Bone Miner. Res.
PUBLISHED: 07-04-2014
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Inflammatory bone resorption is a hallmark of periodontitis, being Treg and Th2 cells independently associated with disease progression attenuation. In this study, we employed an infection-triggered inflammatory osteolysis model to investigate the mechanisms underlying Treg and Th2 cell migration and impact on disease outcome. A. actinomycetemcomitans-infected C57Bl/6 (WT) mice develop an intense inflammatory reaction and alveolar bone resorption, being Tregs of Th2 cells migration temporally associated with disease progression attenuation. Tregs extracted from the lesions preferentially express CCR4 and CCR8, while Th2 cells express CCR3, CCR4 and CCR8. The absence of CCR5 and CCR8 did not impact Th2 and Tregs migration or disease outcome in a significant manner. CCR4KO mice presented a minor reduction in Th2 in parallel with major impairment of Tregs migration, associated with increased inflammatory bone loss and higher pro-inflammatory and osteoclastogenic cytokines levels. The blockade of the CCR4 ligand CCL22 in WT mice resulted in increased inflammatory bone loss phenotype similarly to CCR4KO strain. Adoptive transfer of CCR4(+) Tregs to CCR4KO strain revert the increased disease phenotype to WT mice-like levels, being the production of CCL22 in the lesions mandatory for Tregs migration and the consequent bone loss arrest. The local release of exogenous CCL22 provided by PLGA-microparticles promote Tregs migration and disease arrest in the absence of endogenous CCL22 IL-4KO strain, characterized by the lack of endogenous CCL22 production, defective Tregs migration and exacerbated bone loss. In summary, our results demonstrate that the involvement of IL-4/CCL22/CCR4 axis in the migration of Tregs to osteolytic lesions sites, and attenuates development of lesions by inhibiting inflammatory migration and the production of pro-inflammatory and osteoclastogenic mediators. © 2014 American Society for Bone and Mineral Research.
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Tyms double (2R) and triple repeat (3R) confers risk for human oral squamous cell carcinoma.
Mol. Biol. Rep.
PUBLISHED: 06-17-2014
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The oral cancer is responsible for approximately 3 % of cases of cancer in Brazil. Epidemiological studies have associated low folate intake with an increased risk of epithelial cancers, including oral cancer. Folic acid has a key role in DNA synthesis, repair, methylation and this is the basis of explanations for a putative role for folic acid in cancer prevention. The role of folic acid in carcinogenesis may be modulated by polymorphism C677T in MTHFR and tandem repeats 2R/3R in the promoter site of TYMS gene that are related to decreased enzymatic activity and quantity and availability of the enzyme, respectively. These events cause a decrease in the synthesis, repair and DNA methylation, which can lead to a disruption in the expression of tumor suppressor genes as TP53. The objective of this study was investigate the distribution of polymorphisms C677T and tandem repeats 2R/3R associated with the development of oral squamous cell carcinoma (OSCC). 53 paraffin-embedded samples from patients who underwent surgery but are no longer at the institution and 43 samples collected by method of oral exfoliation by cytobrush were selected. 132 healthy subjects were selected by specialists at the dental clinics of the Faculdade de Odontologia de Pernambuco-FOP. The MTHFR genotyping was performed by PCR-RFLP, and the TYMS genotyping was performed by conventional PCR. Fisher's Exact test at significant level of 5 %. Odds ratios (ORs) and 95 % confidence intervals (CIs) were used to measure the strength of association between genotype frequency and OSCC development. The results were statistically significant for the tandem repeats of the TYMS gene (p = 0.015). The TYMS 2R3R genotype was significantly associated with the development of OSCC (OR = 3.582; 95 % CI  1.240-10.348; p = 0.0262) and also the genotype 3R3R (OR = 3.553; 95 % CI  1.293-9.760; p = 0.0345). When analyzed together, the TYMS 2R3R + 3R3R genotypes also showed association (OR = 3.518; 95 % CI  11.188-10.348; p = 0.0177). No differences for the MTHFR C677T polymorphisms distribution were found between the oral cancer patients and controls subjects in our study (p = 0.499). Therefore, these data suggest that determination of TYMS tandem repeats could provide information on the comprehension of the risk factors and prevention of the OSCC.
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Streptococcus agalactiae in Brazil: serotype distribution, virulence determinants and antimicrobial susceptibility.
BMC Infect. Dis.
PUBLISHED: 06-06-2014
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Group B Streptococcus (GBS) remains a major cause of neonatal sepsis and is also associated with invasive and noninvasive infections in pregnant women and non-pregnant adults, elderly and patients with underlying medical conditions. Ten capsular serotypes have been recognized, and determination of their distribution within a specific population or geographical region is important as they are major targets for the development of vaccine strategies. We have evaluated the characteristics of GBS isolates recovered from individuals with infections or colonization by this microorganism, living in different geographic regions of Brazil.
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Protective effect of prolactin against methylmercury-induced mutagenicity and cytotoxicity on human lymphocytes.
Int J Environ Res Public Health
PUBLISHED: 06-03-2014
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Mercury exhibits cytotoxic and mutagenic properties as a result of its effect on tubulin. This toxicity mechanism is related to the production of free radicals that can cause DNA damage. Methylmercury (MeHg) is one of the most toxic of the mercury compounds. It accumulates in the aquatic food chain, eventually reaching the human diet. Several studies have demonstrated that prolactin (PRL) may be differently affected by inorganic and organic mercury based on interference with various neurotransmitters involved in the regulation of PRL secretion. This study evaluated the cytoprotective effect of PRL on human lymphocytes exposed to MeHg in vitro, including observation of the kinetics of HL-60 cells (an acute myeloid leukemia lineage) treated with MeHg and PRL at different concentrations, with both treatments with the individual compounds and combined treatments. All treatments with MeHg produced a significant increase in the frequency of chromatid gaps, however, no significant difference was observed in the chromosomal breaks with any treatment. A dose-dependent increase in the mitotic index was observed for treatments with PRL, which also acts as a co-mitogenic factor, regulating proliferation by modulating the expression of genes that are essential for cell cycle progression and cytoskeleton organization. These properties contribute to the protective action of PRL against the cytotoxic and mutagenic effects of MeHg.
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Physical Stress Echocardiography: Prediction of Mortality and Cardiac Events in Patients with Exercise Test showing Ischemia.
Arq. Bras. Cardiol.
PUBLISHED: 06-02-2014
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Background: Studies have demonstrated the diagnostic accuracy and prognostic value of physical stress echocardiography in coronary artery disease. However, the prediction of mortality and major cardiac events in patients with exercise test positive for myocardial ischemia is limited. Objective: To evaluate the effectiveness of physical stress echocardiography in the prediction of mortality and major cardiac events in patients with exercise test positive for myocardial ischemia. Methods: This is a retrospective cohort in which 866 consecutive patients with exercise test positive for myocardial ischemia, and who underwent physical stress echocardiography were studied. Patients were divided into two groups: with physical stress echocardiography negative (G1) or positive (G2) for myocardial ischemia. The endpoints analyzed were all-cause mortality and major cardiac events, defined as cardiac death and non-fatal acute myocardial infarction. Results: G2 comprised 205 patients (23.7%). During the mean 85.6 ± 15.0-month follow-up, there were 26 deaths, of which six were cardiac deaths, and 25 non-fatal myocardial infarction cases. The independent predictors of mortality were: age, diabetes mellitus, and positive physical stress echocardiography (hazard ratio: 2.69; 95% confidence interval: 1.20 - 6.01; p = 0.016). The independent predictors of major cardiac events were: age, previous coronary artery disease, positive physical stress echocardiography (hazard ratio: 2.75; 95% confidence interval: 1.15 - 6.53; p = 0.022) and absence of a 10% increase in ejection fraction. All-cause mortality and the incidence of major cardiac events were significantly higher in G2 (p < 0. 001 and p = 0.001, respectively). Conclusion: Physical stress echocardiography provides additional prognostic information in patients with exercise test positive for myocardial ischemia.Fundamento: Estudos têm demonstrado a acurácia diagnóstica e o valor prognóstico da ecocardiografia com estresse físico na doença arterial coronária, mas a predição de mortalidade e de eventos cardíacos maiores, em pacientes com teste ergométrico positivo para isquemia miocárdica, é limitada. Objetivo: Avaliar a predição de mortalidade e de eventos cardíacos maiores pela ecocardiografia com estresse físico em pacientes com teste ergométrico positivo para isquemia miocárdica. Métodos: Trata-se de uma coorte retrospectiva em que foram estudados 866 pacientes consecutivos, com teste ergométrico positivo para isquemia miocárdica, submetidos à ecocardiografia com estresse físico. Os pacientes foram divididos em dois grupos: ecocardiografia com estresse físico negativa (G1) ou positiva (G2) para isquemia miocárdica. Os desfechos avaliados foram mortalidade por qualquer causa e eventos cardíacos maiores, definidos como óbito cardíaco e infarto agudo do miocárdio não fatal. Resultados: O G2 constituiu-se de 205 (23,7%) pacientes. Durante o seguimento médio de 85,6 ± 15,0 meses, ocorreram 26 óbitos, sendo seis por causa cardíaca, e 25 casos de infarto agudo do miocárdio não fatais. Os preditores independentes de mortalidade foram idade, diabetes melito e a ecocardiografia com estresse físico + (hazard ratio: 2,69; intervalo de confiança de 95%: 1,20 - 6,01; p = 0,016), com os seguintes eventos cardíacos maiores: idade, doença arterial coronária prévia, ecocardiografia com estresse físico + (hazard ratio: 2,75; intervalo de confiança de 95%: 1,15 - 6,53; p = 0,022) e ausência do incremento de 10% na fração de ejeção. A mortalidade por qualquer causa e os eventos cardíacos maiores foram significativamente superiores no G2 (p < 0, 001 e p = 0,001, respectivamente). Conclusão: A ecocardiografia com estresse físico oferece informações prognósticas adicionais em pacientes com teste ergométrico positivo para isquemia miocárdica.
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Factors associated with neural alterations and physical disabilities in patients with leprosy in São Luis, State of Maranhão, Brazil.
Rev. Soc. Bras. Med. Trop.
PUBLISHED: 05-28-2014
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Leprosy is a chronic infectious disease that is caused by Mycobacterium leprae. The objective of this study was to evaluate the risk factors that are associated with neural alterations and physical disabilities in leprosy patients at the time of diagnosis.
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Expression of heat shock proteins in periapical granulomas.
J Endod
PUBLISHED: 05-28-2014
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Cells from virtually all organisms respond to a variety of stresses by the rapid synthesis of a highly conserved set of polypeptides termed heat shock proteins (HSPs). HSPs protect cells under adverse conditions such as infection, inflammation, and disease. We hypothesize that endodontic infection might result in an imbalance in the expression of heat shock genes, accounting for different clinical outcomes in periapical lesions.
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Temporal bone dissection practice using a chicken egg.
Otol. Neurotol.
PUBLISHED: 05-21-2014
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Temporal bone drilling practice constitutes an essential stage in training for the surgical approach to this complex anatomic structure. To facilitate adaptation and surgical skills in otologic surgery, we recall the easy cost-effective practice of drilling a chicken egg.
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Raising awareness of new psychoactive substances: chemical analysis and in vitro toxicity screening of 'legal high' packages containing synthetic cathinones.
Arch. Toxicol.
PUBLISHED: 04-07-2014
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The world's status quo on recreational drugs has dramatically changed in recent years due to the rapid emergence of new psychoactive substances (NPS), represented by new narcotic or psychotropic drugs, in pure form or in preparation, which are not controlled by international conventions, but that may pose a public health threat comparable with that posed by substances listed in these conventions. These NPS, also known as 'legal highs' or 'smart drugs', are typically sold via Internet or 'smartshops' as legal alternatives to controlled substances, being announced as 'bath salts' and 'plant feeders' and is often sought after for consumption especially among young people. Although NPS have the biased reputation of being safe, the vast majority has hitherto not been tested and several fatal cases have been reported, namely for synthetic cathinones, with pathological patterns comparable with amphetamines. Additionally, the unprecedented speed of appearance and distribution of the NPS worldwide brings technical difficulties in the development of analytical procedures and risk assessment in real time. In this study, 27 products commercialized as 'plant feeders' were chemically characterized by gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy. It was also evaluated, for the first time, the in vitro hepatotoxic effects of individual synthetic cathinones, namely methylone, pentedrone, 4-methylethcathinone (4-MEC) and 3,4-methylenedioxypyrovalerone (MDPV). Two commercial mixtures ('Bloom' and 'Blow') containing mainly cathinone derivatives were also tested, and 3,4-methylenedioxymethamphetamine (MDMA) was used as the reference drug. The study allowed the identification of 19 compounds, showing that synthetic cathinones are the main active compounds present in these products. Qualitative and quantitative variability was found in products sold with the same trade name in matching or different 'smartshops'. In the toxicity studies performed in primary cultured rat hepatocytes, pentedrone and MDPV proved to be the most potent individual agents, with EC50 values of 0.664 and 0.742 mM, respectively, followed by MDMA (EC50 = 0.754 mM). 4-MEC and methylone were the least potent substances, with EC50 values significantly higher (1.29 and 1.18 mM, respectively; p < 0.05 vs. MDMA). 'Bloom' and 'Blow' showed hepatotoxic effects similar to MDMA (EC50 = 0.788 and 0.870 mM, respectively), with cathinones present in these mixtures contributing additively to the overall toxicological effect. Our results show a miscellany of psychoactive compounds present in 'legal high' products with evident hepatotoxic effects. These data contribute to increase the awareness on the real composition of 'legal high' packages and unveil the health risks posed by NPS.
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Simultaneous analysis of T helper subsets (Th1, Th2, Th9, Th17, Th22, Tfh, Tr1 and Tregs) markers expression in periapical lesions reveals multiple cytokine clusters accountable for lesions activity and inactivity status.
J Appl Oral Sci
PUBLISHED: 03-28-2014
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Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas.
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Valorization of pomegranate peel from 12 cultivars: dietary fibre composition, antioxidant capacity and functional properties.
Food Chem
PUBLISHED: 03-14-2014
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The dried powdered fruit peels of pomegranate (Punica granatum L.) (PomP) from 12 cultivars were used to extract and characterise their dietary fibre (DF) and to assess their functional and antioxidant properties. The total DF content varied between 33.10 and 62/100 g. The cellulose, Klason lignin, uronic acid and total neutral sugars (NS) composition of DF was: 16.53-22.71, 20.59-41.86, 13.98-23.31 and 16.88-19.66/100g, respectively. Arabinose and xylose were the most present NS with more than 60% of total NS content. The ratio of insoluble to soluble DF was around 1, reflecting the balanced composition of PomP's DF. Besides, PomP powder showed intermediate values for water- and oil-holding capacities: 2.31-3.53 and 2.80-4.05 mL/g, respectively, and strong retardation effect on the dialysis of glucose, reaching ?60%. Also, it has been shown that most of the antioxidants can be extracted, based on the strong soluble antioxidant activity (2018-2649 ?mol Trolox/g) compared to the insoluble one (13-23 ?mol Trolox/g).
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Unraveling vitamin B12-responsive gene regulation in algae.
Plant Physiol.
PUBLISHED: 03-13-2014
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Photosynthetic microalgae play a vital role in primary productivity and biogeochemical cycling in both marine and freshwater systems across the globe. However, the growth of these cosmopolitan organisms depends on the bioavailability of nutrients such as vitamins. Approximately one-half of all microalgal species requires vitamin B12 as a growth supplement. The major determinant of algal B12 requirements is defined by the isoform of methionine synthase possessed by an alga, such that the presence of the B12-independent methionine synthase (METE) enables growth without this vitamin. Moreover, the widespread but phylogenetically unrelated distribution of B12 auxotrophy across the algal lineages suggests that the METE gene has been lost multiple times in evolution. Given that METE expression is repressed by the presence of B12, prolonged repression by a reliable source of the vitamin could lead to the accumulation of mutations and eventually gene loss. Here, we probe METE gene regulation by B12 and methionine/folate cycle metabolites in both marine and freshwater microalgal species. In addition, we identify a B12-responsive element of Chlamydomonas reinhardtii METE using a reporter gene approach. We show that complete repression of the reporter occurs via a region spanning -574 to -90 bp upstream of the METE start codon. A proteomics study reveals that two other genes (S-Adenosylhomocysteine hydrolase and Serine hydroxymethyltransferase2) involved in the methionine-folate cycle are also repressed by B12 in C. reinhardtii. The strong repressible nature and high sensitivity of the B12-responsive element has promising biotechnological applications as a cost-effective regulatory gene expression tool.
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Antiparasitic activity of natural and semi-synthetic tirucallane triterpenoids from Schinus terebinthifolius (Anacardiaceae): structure/activity relationships.
Molecules
PUBLISHED: 03-12-2014
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Leishmaniasis and Chagas are diseases caused by parasitic protozoans that affect the poorest population in the World, causing a high mortality and morbidity. As a result of highly toxic and long-term treatments, the discovery of novel, safe and more efficacious drugs is essential. In this work, the in vitro antiparasitic activity and mammalian cytotoxicity of three natural tirucallane triterpenoids, isolated from leaves of Schinus terebinthifolius (Anacardiaceae), and nine semi-synthetic derivatives were investigated against Leishmania (L.) infantum and Trypanosoma cruzi. Trypomastigotes of T. cruzi were the most susceptible parasites and seven compounds demonstrated a trypanocidal activity with IC50 values in the range between 15 and 58 µg/mL. Four compounds demonstrated selectivity towards the intracellular amastigotes of Leishmania, with IC50 values in the range between 28 and 97 µg/mL. The complete characterization of triterpenoids was afforded after thorough analysis of nuclear magnetic resonance (NMR) data as well as electrospray ionization mass spectrometry (ESI-MS). Additionally, structure-activity relationships were performed using Decision Trees.
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Enamel erosion and prevention efficacy characterized by confocal laser scanning microscope.
Microsc. Res. Tech.
PUBLISHED: 03-12-2014
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The aim of this study was to evaluate the erosion-inhibiting effect of two toothpastes on the development of erosion-like lesions, by a confocal laser scanning microscope (CLSM). Forty human enamel blocks were divided into five groups (n?=?8), in accordance to evaluate the GC MI Paste Plus and Oral B with stannous fluoride, applied as slurries and associated with toothbrush. Specimens were submitted to an erosion challenge from citric acid (0.5%, pH?=?2.8), for 5 min, six times a day, alternating in artificial saliva immersions. Reference group was not exposed to treatment. Part of specimens (Groups 02 and 03) was exposed twice daily just to slurries, for 2 min, therefore specimens from Groups 04 and 05 were also abraded, for 30 s. The enamel surfaces were morphological characterized using CLSM images, with mineral loss being measured using the resulting 3D images referenced to an un-challenged portion of the sample. Step values were compared using the one-way ANOVA test. CLSM was shown to be a viable, noncontact, and simple technique to characterize eroded surfaces. The statistical difference in the step size was significant between the groups (P?=?0.001) and using multiple comparisons a statistically significant protective effect of toothpastes was shown when these were applied as slurries. Although groups submitted to tooth brush showed mineral loss similar to reference control group, due to the damages of abrasion associated.
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First record of intestinal parasites in a wild population of jaguar in the Brazilian Atlantic Forest.
Rev Bras Parasitol Vet
PUBLISHED: 03-11-2014
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Small and isolated wildlife populations may be more susceptible to disease, which makes illness an important issue to investigate regarding the conservation of large carnivores. Here, we present the results of the first investigation of intestinal parasites in one of the last remaining populations of jaguars in the Brazilian Atlantic Forest. We studied parasites from fecal samples using three different techniques for parasitological examination: floatation in saturated sodium chloride solution, sedimentation and formalin-ether centrifugation. Intestinal parasites were detected in 70% of the analyzed samples, and seven taxa (mean = 3.7 taxa/sample) were identified. All the groups of parasites that were identified have been recorded in previous jaguar studies. However, the records of Class Trematoda and nematodes Trichuridae are the first evidence of these groups of worms in free-ranging jaguars in Brazil. Although our results do not provide conclusive evidence on the health of this jaguar population, given its very small size (approximately 20 animals) we stress the need to properly understand the dynamics of disease in this wild population and to evaluate the risk of contracting new diseases from domestic species inhabiting the neighboring areas. These represent imperative actions for the successful conservation of this threatened population of jaguar.
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Topical anesthesia in flexible nasofibrolaryngoscopy.
Acta Otorrinolaringol Esp
PUBLISHED: 03-06-2014
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The emergence of flexible nasofibrolaryngoscopy in recent decades has facilitated the development of Otolaryngology and the nasofibrolaryngoscope has become an essential diagnostic tool for the otolaryngologist. However, its use is not without discomfort for the patient, which is why various options for topical anaesthesia have been proposed during the development of the technique.
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THI1, a protein involved in the biosynthesis of thiamin in Arabidopsis thaliana: structural analysis of THI1(A140V) mutant.
Biochim. Biophys. Acta
PUBLISHED: 02-28-2014
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In eukaryotes, there are still steps of the vitamin B1 biosynthetic pathway not completely understood. In Arabidopsis thaliana, THI1 protein has been associated with the synthesis of the thiazole ring, a finding supported by the identification of a thiamine pyrophosphate (TPP)-like compound in its structure. Here, we investigated THI1 and its mutant THI1(A140V), responsible for the thiamin auxotrophy in a A. thaliana mutant line, aiming to clarify the impact of this mutation in the stability and activity of THI1. Recently, the THI1 orthologue (THI4) was revealed to be responsible for the donation of the sulfur atom from a cysteine residue to the thiazole ring in the thiamine intermediate. In this context, we carried out a cysteine quantification in THI1 and THI1(A140V) using electron spin resonance (ESR). These data showed that THI1(A140V) contains more sulfur-containing cysteines than THI1, indicating that the function as a sulfur donor is conserved, but the rate of donation reaction is somehow affected. Also, the bound compounds were isolated from both proteins and are present in different amounts in each protein. Unfolding studies presented differences in melting temperatures and also in the concentration of guanidine at which half of the protein unfolds, thus showing that THI1(A140V) has its conformational stability affected by the mutation. Hence, despite keeping its function in the early steps during the synthesis of TPP precursor, our studies have shown a decrease in the THI1(A140V) stability, which might be slowing down the biological activity of the mutant, and thus contributing to thiamin auxotrophy.
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In vitro studies of the activity of dithiocarbamate organoruthenium complexes against clinically relevant fungal pathogens.
Molecules
PUBLISHED: 02-26-2014
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The in vitro antifungal activity of nine dirutheniumpentadithiocarbamate complexes C1-C9 was investigated and assessed for its activity against four different fungal species with clinical interest and related to invasive fungal infections (IFIs), such as Candida spp. [C. albicans (two clinical isolates), C. glabrata, C. krusei, C. parapsolisis, C. tropicalis, C.dubliniensis (six clinical isolates)], Paracoccidioides brasiliensis (seven clinical isolates), Cryptococcus neoformans and Sporothrix schenckii. All synthesized complexes C1-C9 and also the free ligands L1-L9 were submitted to in vitro tests against those fungi and the results are very promising, since some of the obtained MIC (minimal inhibitory concentration) values were very low (from 10-6 mol mL-1 to 10-8 mol mL-1) against all investigated clinically relevant fungal pathogens, except for C. glabrata, that the MIC values are close to the ones obtained for fluconazole, the standard antifungal agent tested. Preliminary structure-activity relations (SAR) might be suggested and a strong influence from steric and lipophilic parameters in the antifungal activity can be noticed. Cytotoxicity assays (IC50) showed that the complexes are not as toxic (IC50 values are much higher-30 to 200 fold-than MIC values). These ruthenium complexes are very promising lead compounds for novel antifungal drug development, especially in IFIs, one of most harmful emerging infection diseases (EIDs).
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Astrocyte transforming growth factor beta 1 promotes inhibitory synapse formation via CaM kinase II signaling.
Glia
PUBLISHED: 02-21-2014
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The balance between excitatory and inhibitory synaptic inputs is critical for the control of brain function. Astrocytes play important role in the development and maintenance of neuronal circuitry. Whereas astrocytes-derived molecules involved in excitatory synapses are recognized, molecules and molecular mechanisms underlying astrocyte-induced inhibitory synapses remain unknown. Here, we identified transforming growth factor beta 1 (TGF-?1), derived from human and murine astrocytes, as regulator of inhibitory synapse in vitro and in vivo. Conditioned media derived from human and murine astrocytes induce inhibitory synapse formation in cerebral cortex neurons, an event inhibited by pharmacologic and genetic manipulation of the TGF-? pathway. TGF-?1-induction of inhibitory synapse depends on glutamatergic activity and activation of CaM kinase II, which thus induces localization and cluster formation of the synaptic adhesion protein, Neuroligin 2, in inhibitory postsynaptic terminals. Additionally, intraventricular injection of TGF-?1 enhanced inhibitory synapse number in the cerebral cortex. Our results identify TGF-?1/CaMKII pathway as a novel molecular mechanism underlying astrocyte control of inhibitory synapse formation. We propose here that the balance between excitatory and inhibitory inputs might be provided by astrocyte signals, at least partly achieved via TGF-?1 downstream pathways. Our work contributes to the understanding of the GABAergic synapse formation and may be of relevance to further the current knowledge on the mechanisms underlying the development of various neurological disorders, which commonly involve impairment of inhibitory synapse transmission. GLIA 2014;62:1917-1931.
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Women with cardiovascular risk after preeclampsia: is there follow-up within the Unified Health System in Brazil?
Rev Lat Am Enfermagem
PUBLISHED: 02-21-2014
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to identify women with cardiovascular risk, five years after a pre-eclampsic episode (PE), and identify the follow-up of these women within the Unified Health System (Sistema Único de Saúde--SUS), in the city of Natal/RN.
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[Impact of sexual and reproductive health theme insertion in the undergraduate medical].
Rev Bras Ginecol Obstet
PUBLISHED: 02-12-2014
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To evaluate the impact of sexual and reproductive health theme insertion in the undergraduate medical curriculum at a Brazilian public university.
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Brazilian adolescents' knowledge and beliefs about abortion methods: a school-based internet inquiry.
BMC Womens Health
PUBLISHED: 01-29-2014
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Internet surveys that draw from traditionally generated samples provide the unique conditions to engage adolescents in exploration of sensitive health topics.
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Mesenchymal stem cells as active prohealing and immunosuppressive agents in periapical environment: evidence from human and experimental periapical lesions.
J Endod
PUBLISHED: 01-28-2014
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Previous studies describe contrasting molecular profiles of active and inactive periapical granulomas characterized by distinct expression of cytokines, osteoclastogenic factors, and wound healing markers. Although the molecular mechanisms underlying such a dichotomy remain unknown, in this study we investigated the potential involvement of mesenchymal stem cells (MSCs) in determining human and murine periapical lesion activity and outcomes.
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Crystal structure of a Schistosoma mansoni septin reveals the phenomenon of strand slippage in septins dependent on the nature of the bound nucleotide.
J. Biol. Chem.
PUBLISHED: 01-24-2014
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Septins are filament-forming GTP-binding proteins involved in important cellular events, such as cytokinesis, barrier formation, and membrane remodeling. Here, we present two crystal structures of the GTPase domain of a Schistosoma mansoni septin (SmSEPT10), one bound to GDP and the other to GTP. The structures have been solved at an unprecedented resolution for septins (1.93 and 2.1 ?, respectively), which has allowed for unambiguous structural assignment of regions previously poorly defined. Consequently, we provide a reliable model for functional interpretation and a solid foundation for future structural studies. Upon comparing the two complexes, we observe for the first time the phenomenon of a strand slippage in septins. Such slippage generates a front-back communication mechanism between the G and NC interfaces. These data provide a novel mechanistic framework for the influence of nucleotide binding to the GTPase domain, opening new possibilities for the study of the dynamics of septin filaments.
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Clinical efficacy analysis of the mouth rinsing with pomegranate and chamomile plant extracts in the gingival bleeding reduction.
Complement Ther Clin Pract
PUBLISHED: 01-21-2014
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Medicinal plants represent important therapeutic resources to health restoration, including the use of herbal products in the mouth conditions treatment. A randomized controlled clinical trial was performed in order to evaluate the effectiveness of mouth rinse with pomegranate and chamomile plant extracts, against chlorhexidine 0.12% in the gingiva bleeding condition. The mouth rinses with the herbal products were effective for this case, showing thus, antimicrobial and anti-inflammatory properties similar to that of chlorhexidine 0.12%.
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Reversible paralysis of Schistosoma mansoni by forchlorfenuron, a phenylurea cytokinin that affects septins.
Int. J. Parasitol.
PUBLISHED: 01-20-2014
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Septins are guanosine-5'-triphosphate-binding proteins involved in wide-ranging cellular processes including cytokinesis, vesicle trafficking, membrane remodelling and scaffolds, and with diverse binding partners. Precise roles for these structural proteins in most processes often remain elusive. Identification of small molecules that inhibit septins could aid in elucidating the functions of septins and has become increasingly important, including the description of roles for septins in pathogenic phenomena such as tumorigenesis. The plant growth regulator forchlorfenuron, a synthetic cytokinin known to inhibit septin dynamics, likely represents an informative probe for septin function. This report deals with septins of the human blood fluke Schistosoma mansoni and their interactions with forchlorfenuron. Recombinant forms of three schistosome septins, SmSEPT5, SmSEPT7.2 and SmSEPT10, interacted with forchlorfenuron, leading to rapid polymerization of filaments. Culturing developmental stages (miracidia, cercariae, adult males) of schistosomes in FCF at 50-500 ?M rapidly led to paralysis, which was reversible upon removal of the cytokinin. The reversible paralysis was concentration-, time- and developmental stage-dependent. Effects of forchlorfenuron on the cultured schistosomes were monitored by video and/or by an xCELLigence-based assay of motility, which quantified the effect of forchlorfenuron on fluke motility. The findings implicated a mechanism targeting a molecular system controlling movement in these developmental stages: a direct effect on muscle contraction due to septin stabilization might be responsible for the reversible paralysis, since enrichment of septins has been described within the muscles of schistosomes. This study revealed the reversible effect of forchlorfenuron on both schistosome motility and its striking impact in hastening polymerization of septins. These novel findings suggested routes to elucidate roles for septins in this pathogen, and exploitation of derivatives of forchlorfenuron for anti-schistosomal drugs.
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Visual detection of DNA on paper chips.
Anal. Chem.
PUBLISHED: 01-17-2014
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On-site DNA analysis for diagnostic or forensic purposes is much anticipated in the future of molecular testing. Yet the challenges to achieve this goal remain large with rapid and inexpensive detection and visualization being key factors for any portable analysis system. We have developed a filter paper-based nucleic acid assay, which is able to identify and distinguish dog and human genomic and mitochondrial samples in a forensic setting. The filter paper material allows for transport by capillary force of the sample DNA through the detection surface, allowing the targets to hybridize specifically to their complementary capture sequences. Coupling micrometer-sized beads to DNA allows the results to be visualized by the naked eye, enabling instant, cost-efficient, and on-site detection, while eliminating the need for advanced expensive instrumentation.
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Acceptability, efficacy and safety of two treatment protocols for dental fluorosis: a randomized clinical trial.
J Dent
PUBLISHED: 01-10-2014
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This parallel randomized clinical trial evaluated the efficacy of two treatments for removing fluorosis stains.
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Human umbilical cord tissue-derived mesenchymal stromal cells attenuate remodeling after myocardial infarction by proangiogenic, antiapoptotic, and endogenous cell-activation mechanisms.
Stem Cell Res Ther
PUBLISHED: 01-10-2014
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Among the plethora of cells under investigation to restore a functional myocardium, mesenchymal stromal cells (MSCs) have been granted considerable interest. However, whereas the beneficial effects of bone marrow MSCs (BM-MSCs) in the context of the diseased heart are widely reported, data are still scarce on MSCs from the umbilical cord matrix (UCM-MSCs). Herein we report on the effect of UCM-MSC transplantation to the infarcted murine heart, seconded by the dissection of the molecular mechanisms at play.
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Antinociceptive and anti-inflammatory activities of the sesame oil and sesamin.
Nutrients
PUBLISHED: 01-06-2014
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Sesame oil is widely consumed as nutritious food, cooking oil, and in pharmaceuticals and food. In this study, the antinociceptive and anti-inflammatory properties of the sesame oil and sesamin were investigated. The sesame oil and sesamin reduced the number of abdominal contortions at the doses 100, 200, or 400 mg/kg. The first and second phases of the time paw licking were inhibited by sesame oil and sesamin (100, 200, or 400 mg/kg). After 90 min of treatment, sesame oil and sesamin increased the reaction time on a hot plate (200 or 400 mg/kg). Considering the tail-immersion assay, the sesame oil and sesamin produced significant effect after 60 min at the doses of 100, 200, or 400 mg/kg. After 4 h of application of the carrageenan, the sesame oil and sesamin were effective against the paw edema. The exudate volume and leucocyte migration were also reduced by sesame oil and sesamin. These results suggest that sesamin is one of the active compounds found in sesame oil and justify the antinociceptive and anti-inflammatory properties of this product.
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Antifungal ether diglycosides from Matayba guianensis Aublet.
Bioorg. Med. Chem. Lett.
PUBLISHED: 01-06-2014
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Since the 1960s, fungal infections have become a major worldwide public health problem. Antifungal treatments have many limitations, such as toxicity and resistance. Matayba guianensis Aublet (Sapindaceae) was chemically investigated as part of our ongoing search for lead molecules against fungi in the Brazilian Cerrado biome. The ethanolic extract of M. guianensis root bark revealed the presence of two previously unreported ether diglycosides: matayoside E (1) and F (2) with anti Candida activity, along with two known compounds: cupanioside (3) and stigmasterol (4).
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Clavanin bacterial sepsis control using a novel methacrylate nanocarrier.
Int J Nanomedicine
PUBLISHED: 01-01-2014
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Controlling human pathogenic bacteria is a worldwide problem due to increasing bacterial resistance. This has prompted a number of studies investigating peptides isolated from marine animals as a possible alternative for control of human pathogen infections. Clavanins are antimicrobial peptides isolated from the marine tunicate Styela clava, showing 23 amino acid residues in length, cationic properties, and also high bactericidal activity. In spite of clear benefits from the use of peptides, currently 95% of peptide properties have limited pharmaceutical applicability, such as low solubility and short half-life in the circulatory system. Here, nanobiotechnology was used to encapsulate clavanin A in order to develop nanoantibiotics against bacterial sepsis. Clavanin was nanostructured using EUDRAGIT(®) L 100-55 and RS 30 D solution (3:1 w:w). Atomic force, scanning electron microscopy and dynamic light scattering showed nanoparticles ranging from 120 to 372 nm in diameter, with a zeta potential of -7.16 mV and a polydispersity index of 0.123. Encapsulation rate of 98% was assessed by reversed-phase chromatography. In vitro bioassays showed that the nanostructured clavanin was partially able to control development of Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Furthermore, nanostructures did not show hemolytic activity. In vivo sepsis bioassays were performed using C57BL6 mice strain inoculated with a polymicrobial suspension. Assays led to 100% survival rate under sub-lethal sepsis assays and 40% under lethal sepsis assays in the presence of nanoformulated clavanin A until the seventh day of the experiment. Data here reported indicated that nanostructured clavanin A form shows improved antimicrobial activity and has the potential to be used to treat polymicrobial infections.
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Targeted inactivation of Cerberus like-2 leads to left ventricular cardiac hyperplasia and systolic dysfunction in the mouse.
PLoS ONE
PUBLISHED: 01-01-2014
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Previous analysis of the Cerberus like 2 knockout (Cerl2-/-) mouse revealed a significant mortality during the first day after birth, mostly due to cardiac defects apparently associated with randomization of the left-right axis. We have however, identified Cerl2-associated cardiac defects, particularly a large increase in the left ventricular myocardial wall in neonates that cannot be explained by laterality abnormalities. Therefore, in order to access the endogenous role of Cerl2 in cardiogenesis, we analyzed the embryonic and neonatal hearts of Cerl2 null mutants that did not display a laterality phenotype. Neonatal mutants obtained from the compound mouse line Cer2-/-::Mlc1v-nLacZ24+, in which the pulmonary ventricle is genetically marked, revealed a massive enlargement of the ventricular myocardium in animals without laterality defects. Echocardiography analysis in Cerl2-/- neonates showed a left ventricular systolic dysfunction that is incompatible with a long lifespan. We uncovered that the increased ventricular muscle observed in Cerl2-/- mice is caused by a high cardiomyocyte mitotic index in the compact myocardium which is mainly associated with increased Ccnd1 expression levels in the left ventricle at embryonic day (E) 13. Interestingly, at this stage we found augmented left ventricular expression of Cerl2 levels when compared with the right ventricle, which may elucidate the regionalized contribution of Cerl2 to the left ventricular muscle formation. Importantly, we observed an increase of phosphorylated Smad2 (pSmad2) levels in embryonic (E13) and neonatal hearts indicating a prolonged TGF?s/Nodal-signaling activation. Concomitantly, we detected an increase of Baf60c levels, but only in Cerl2-/- embryonic hearts. These results indicate that independently of its well-known role in left-right axis establishment Cerl2 plays an important role during heart development in the mouse, mediating Baf60c levels by exerting an important control of the TGF?s/Nodal-signaling pathway.
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Isolation, cultivation and genomic analysis of magnetosome biomineralization genes of a new genus of South-seeking magnetotactic cocci within the Alphaproteobacteria.
Front Microbiol
PUBLISHED: 01-01-2014
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Although magnetotactic bacteria (MTB) are ubiquitous in aquatic habitats, they are still considered fastidious microorganisms with regard to growth and cultivation with only a relatively low number of axenic cultures available to date. Here, we report the first axenic culture of an MTB isolated in the Southern Hemisphere (Itaipu Lagoon in Rio de Janeiro, Brazil). Cells of this new isolate are coccoid to ovoid in morphology and grow microaerophilically in semi-solid medium containing an oxygen concentration ([O2]) gradient either under chemoorganoheterotrophic or chemolithoautotrophic conditions. Each cell contains a single chain of approximately 10 elongated cuboctahedral magnetite (Fe3O4) magnetosomes. Phylogenetic analysis based on the 16S rRNA gene sequence shows that the coccoid MTB isolated in this study represents a new genus in the Alphaproteobacteria; the name Magnetofaba australis strain IT-1 is proposed. Preliminary genomic data obtained by pyrosequencing shows that M. australis strain IT-1 contains a genomic region with genes involved in biomineralization similar to those found in the most closely related magnetotactic cocci Magnetococcus marinus strain MC-1. However, organization of the magnetosome genes differs from M. marinus.
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Effects of exposure to a cafeteria diet during gestation and after weaning on the metabolism and body weight of adult male offspring in rats.
Br. J. Nutr.
PUBLISHED: 12-13-2013
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In the present study, we investigated whether maternal exposure to a cafeteria diet affects the metabolism and body composition of offspring and whether such an exposure has a cumulative effect during the lifetime of the offspring. Female rats were fed a control (CON) or a cafeteria (CAF) diet from their own weaning to the weaning of their offspring. At 21 d of age, male offspring were divided into four groups by diet during gestation and after weaning (CON-CON, CON-CAF, CAF-CON and CAF-CAF). Blood was collected from dams (after weaning) and pups (at 30 and 120 d of age) by decapitation. CAF dams had significantly greater body weight and adipose tissue weight and higher concentrations of total cholesterol, insulin and leptin than CON dams (Students t test). The energy intake of CAF rats was higher than that of CON rats regardless of the maternal diet (two-way ANOVA). Litters had similar body weights at weaning and at 30 d of age, but at 120 d, CON-CAF rats were heavier. At both ages, CAF rats had greater adipose tissue weight than CON rats regardless of the maternal diet, and the concentrations of TAG and cholesterol were similar between the two groups, as were blood glucose concentrations at 30 d of age. However, at 120 d of age, CAF rats were hyperglycaemic, hyperinsulinaemic and hyperleptinaemic regardless of the maternal diet. These findings suggest that maternal obesity does not modulate the metabolism of male offspring independently, modifying body weight only when associated with the intake of a cafeteria diet by the offspring.
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Septins of Platyhelminths: Identification, Phylogeny, Expression and Localization among Developmental Stages of Schistosoma mansoni.
PLoS Negl Trop Dis
PUBLISHED: 12-01-2013
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Septins are a family of eukaryotic GTP binding proteins conserved from yeasts to humans. Originally identified in mutants of budding yeast, septins participate in diverse cellular functions including cytokinesis, organization of actin networks, cell polarity, vesicle trafficking and many others. Septins assemble into heteroligomers to form filaments and rings. Here, four septins of Schistosoma mansoni are described, which appear to be conserved within the phylum Platyhelminthes. These orthologues were related to the SEPT5, SEPT10 and SEPT7 septins of humans, and hence we have termed the schistosome septins SmSEPT5, SmSEPT10, SmSEPT7.1 and SmSEPT7.2. Septin transcripts were detected throughout the developmental cycle of the schistosome and a similar expression profile was observed for septins in the stages examined, consistent with concerted production of these proteins to form heterocomplexes. Immunolocalization analyses undertaken with antibodies specific for SmSEPT5 and SmSEPT10 revealed a broad tissue distribution of septins in the schistosomulum and colocalization of septin and actin in the longitudinal and circular muscles of the sporocyst. Ciliated epidermal plates of the miracidium were rich in septins. Expression levels for these septins were elevated in germ cells in the miracidium and sporocyst. Intriguingly, septins colocalize with the protonephridial system of the cercaria, which extends laterally along the length of this larval stage. Together, the findings revealed that schistosomes expressed several septins which likely form filaments within the cells, as in other eukaryotes. Identification and localization demonstrating a broad distribution of septins across organs and tissues of schistosome contributes towards the understanding of septins in schistosomes and other flatworms.
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Saponin profile of green asparagus genotypes.
J. Agric. Food Chem.
PUBLISHED: 11-11-2013
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The main goal of this study was to determine the saponin profiles of different "triguero" asparagus genotypes and to compare them to green asparagus commercial hybrids. The samples consisted of 31 commercial hybrids and 58 genotypes from the Huétor-Tájar (HT) population variety ("triguero"). The saponin analysis by high-performance liquid chromatography-mass spectrometry allowed for the determination of 12 saponins derived from a furostan-type steroidal genin, 4 of which had never been described in the edible part of asparagus. The saponin profile of "triguero" asparagus was a combination of these new saponins and protodioscin. Although protodioscin was the major saponin found in commercial hybrids, some of these 12 saponins were detected as major components in some of the commercial hybrids. The total contents of saponins described in some of these HT genotypes reach values as high as 10-100 times higher than those found in commercial hybrids.
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Preeclampsia is associated with increased maternal body weight in a northeastern Brazilian population.
BMC Pregnancy Childbirth
PUBLISHED: 07-31-2013
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Preeclampsia is a disease with great variability in incidence across the world. The mortality is higher in lower income countries, where it is the leading cause of maternal mortality. This study aimed to determine the frequency of and risk factors for preeclampsia in a low income population from an urban area of Brazil.
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[Meanings and methods of territorialization in primary health care].
Cien Saude Colet
PUBLISHED: 07-31-2013
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Territorially-based participative analytical methodologies taking the environmental question and work into consideration are essential for effective primary health care. The study analyzed work and environment-related processes in the primary health care area and their repercussions on the health of workers and the community in a rural city in Ceará, whose economy is based on agriculture for export,. It sought to redeem the area and the proposal of actions focused on health needs by the social subjects through the making of social, environmental and work-related maps in workshops within the framework of action research. Examining the situation from a critical perspective, based on social participation and social determination of the health-disease process with regard to the relations between production, environment and health, was the most important step in the participative map-making process, with the qualitative material interpreted in light of discourse analysis. The process helped identify the health needs, the redemption of the area, strengthened the cooperation between sectors and the tie between the health of the worker and that of the environment, and represented an advance towards the eradication of the causes of poor primary health care services.
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Combination therapy with carboplatin and thalidomide suppresses tumor growth and metastasis in 4T1 murine breast cancer model.
Biomed. Pharmacother.
PUBLISHED: 07-18-2013
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Carboplatin, efficient cytostatics for cancer therapy, could induce apoptosis and inhibit the growth of vascular endothelium in several tumor cell lines and xenograft models. It has been suggested that the antitumor effect of chemotherapy could be increased by combining it with an antiangiogenesis agent in anticancer strategy. The present study explored the potential to increase the antitumor effect of carboplatin by combining it with thalidomide in mouse 4T1 breast cancer models, and the underlining mechanism was investigated. The systemic administration of carboplatin and thalidomide significantly decreased tumor growth through increased tumor cell apoptosis compared with either control group. Collectively, these findings suggest that combined treatment has shown synergistic suppression in tumor progression according to the analysis. Furthermore, also was observed reduction in number of lung metastases as compared to isolated treatments and increased survival of the animals. The present study may be important in future exploration of the potential application of the combined approach in the treatment of breast cancer.
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Molecular study of congenital erythrocytosis in 70 unrelated patients revealed a potential causal mutation in less than half of the cases (Where is/are the missing gene(s)?).
Eur. J. Haematol.
PUBLISHED: 07-14-2013
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Congenital erythrocytosis can be classified as primary, when the defect is intrinsic to the RBC progenitors and independent of the serum erythropoietin (Epo) concentration, or secondary, when the erythrocytosis is the result of an upregulation of Epo production. Primary erythrocytosis is associated with mutations in the EPOR gene, secondary CE can de due to mutations that stabilize the hemoglobin in the oxygenated form or to mutations in the genes that control the transcriptional activation of the EPO gene - VHL, EGLN1, EPAS1. Chuvash polycythemia, caused by mutations in VHL gene, shares features of both primary and secondary erythrocytosis, with increased Epo production but also hypersensitivity of progenitors to Epo.
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The repetitive component of the A genome of peanut (Arachis hypogaea) and its role in remodelling intergenic sequence space since its evolutionary divergence from the B genome.
Ann. Bot.
PUBLISHED: 07-04-2013
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Peanut (Arachis hypogaea) is an allotetraploid (AABB-type genome) of recent origin, with a genome of about 2·8 Gb and a high repetitive content. This study reports an analysis of the repetitive component of the peanut A genome using bacterial artificial chromosome (BAC) clones from A. duranensis, the most probable A genome donor, and the probable consequences of the activity of these elements since the divergence of the peanut A and B genomes.
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Zygomaticofacial foramen location accuracy and reliability in cone-beam computed tomography.
Acta Odontol. Scand.
PUBLISHED: 07-01-2013
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Abstract Objective. This study aimed to evaluate the possibility of detecting the zygomaticofacial foramen (ZFF) in cone-beam computed tomography (CBCT). Materials and methods. This study evaluated ZFFs in 151 macerated skulls (302 zygomatic bones, ZBs) by physical inspection, in which the presence and diameters of the ZFFs were determined. These data were compared with the CBCT images of the skulls to determine the accuracy of CBCT in detecting ZFFs. The diameters were measured by insertion of steel wires with known thicknesses into the ZFFs. The CBCT images were acquired by an i-CAT Classic® (International Imaging Sciences, Hatfield, PA) connected to a workstation (Model ITOX Midtower Workstation; Imaging Sciences International®) with a 20-inch Eizo monitor. The images were generated in coronal, sagittal and axial slices to evaluate the best tomographic plane for ZFF visualization. Results. The incidence of ZFF found by physical inspection was one foramen in 44% of ZBs (n = 133), two foramina in 28% (n = 86), three foramina in 8% (n = 24) and four foramina in 1% (n = 2). ZFF was absent in 19% (n = 57) of ZBs. The average diameter was 0.57 mm (± 0.27 mm). All foramina were observed in all tomography images. Conclusion. This preliminary study supports the conclusion that a CBCT scan has excellent accuracy in evaluating ZFFs.
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Identification of the acid/base catalyst of a glycoside hydrolase family 3 (GH3) beta-glucosidase from Aspergillus niger ASKU28.
Biochim. Biophys. Acta
PUBLISHED: 06-21-2013
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The commercially important glycoside hydrolase family 3 (GH3) beta-glucosidases from Aspergillus niger are anomeric-configuration-retaining enzymes that operate through the canonical double-displacement glycosidase mechanism. Whereas the catalytic nucleophile is readily identified across all GH3 members by sequence alignments, the acid/base catalyst in this family is phylogenetically variable and less readily divined.
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Dual role of IL-12 in the therapeutic efficacy or failure during combined PEG-Interferon-?2A and ribavirin therapy in patients with chronic hepatitis C.
Immunol. Lett.
PUBLISHED: 06-19-2013
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Several efforts have been made to establish novel biomarkers with relevant predictive values to monitor HCV-infected patients under pegilated Interferon-?2A-(PEG-IFN-?2A)/ribavirin therapy. The aim of this study was to monitor the kinetics of HCV viral load, serum levels of pro-inflammatory/regulatory cytokines and leukocyte activation status before and after PEG-IFN-?2A/ribavirin therapy in 52 volunteers, including 12 chronic HCV patients and 40 controls. The HCV viral load, serum levels of cytokines (IL-8/IL-6/TNF-?/IL-12/IFN-?/IL-4/IL-10) and the phenotype of peripheral blood leukocytes were evaluated before and after 4, 12 and 24 weeks following the PEG-IFN-?2A/ribavirin therapy. Our results demonstrated that sustained virological response-(SVR) is associated with early decrease in the viral load after 4 weeks of treatment. The presence of a modulated pro-inflammatory profile at baseline favors SVR, whereas a strong inflammatory response at baseline predisposes to therapeutic failure. Furthermore, a time-dependent increase on serum IL-12 levels in patients under treatment is critical to support the SVR, while the early predominance of IL-10 correlates to late virological relapse. On the other hand, a broad but unguided "cytokine storm" is observed in the non-responder HCV patients after 12 weeks of treatment. Corroborating these findings, monocyte/lymphocyte activation at baseline is associated with the non-responders to therapy whereas high CD8(+) T-cell numbers associate with SVR. All in all, these data suggest that the baseline pattern of serum pro-inflammatory/regulatory cytokines and the immunological activation status of chronic HCV patients undergoing PEG-IFN-?2A/ribavirin therapy are closely related with the therapeutic response.
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Asparagus byproducts as a new source of peroxidases.
J. Agric. Food Chem.
PUBLISHED: 06-18-2013
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Soluble peroxidase (POD) from asparagus byproducts was purified by ion exchange chromatographies, and its kinetic and catalytic properties were studied. The isoelectric point of the purified isoperoxidases was 9.1, and the optimum pH and temperature values were 4.0 and 25 °C, respectively. The cationic asparagus POD (CAP) midpoint inactivation temperature was 57 °C, which favors its use in industrial processes. The Km values of cationic asparagus POD for H?O? and ABTS were 0.318 and 0.634 mM, respectively. The purified CAP is economically obtained from raw materials using a simple protocol and possesses features that make it advantageous for the potential use of this enzyme in a large number of processes with demonstrated requirements of thermostable POD. The results indicate that CAP can be used as a potential candidate for removing phenolic contaminants.
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Optimization of a method for the profiling and quantification of saponins in different green asparagus genotypes.
J. Agric. Food Chem.
PUBLISHED: 06-18-2013
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The main goal of this study was the optimization of a HPLC-MS method for the qualitative and quantitative analysis of asparagus saponins. The method includes extraction with aqueous ethanol, cleanup by solid phase extraction, separation by reverse phase chromatography, electrospray ionization, and detection in a single quadrupole mass analyzer. The method was used for the comparison of selected genotypes of Huétor-Tájar asparagus landrace and selected varieties of commercial diploid hybrids of green asparagus. The results showed that while protodioscin was almost the only saponin detected in the commercial hybrids, eight different saponins were detected in the Huétor-Tájar asparagus genotypes. The mass spectra indicated that HT saponins are derived from a furostan type steroidal genin having a single bond between carbons 5 and 6 of the B ring. The total concentration of saponins was found to be higher in triguero asparagus than in commercial hybrids.
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Stress echocardiography and major cardiac events in patients with normal exercise test.
Arq. Bras. Cardiol.
PUBLISHED: 06-14-2013
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Exercise test (ET) is the preferred initial noninvasive test for the diagnosis and risk stratification of coronary artery disease (CAD), however, its lower sensitivity may fail to identify patients at greater risk of adverse events.
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Complete Genome of a Methanosarcina mazei Strain Isolated from Sediment Samples from an Amazonian Flooded Area.
Genome Announc
PUBLISHED: 05-25-2013
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Methanosarcina mazei is a strictly anaerobic methanogen from the Methanosarcinales order, which is known for its broad catabolic range among methanogens and is widespread throughout diverse environments. The draft genome of the strain presented here was cultivated from sediment samples collected from the Tucuruí hydroelectric power station reservoir.
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[Acute Panmyelosis with myelofibrosis: a rare cause of pancytopenia].
Acta Med Port
PUBLISHED: 05-22-2013
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Acute Panmyelosis with myelofibrosis is a rare and aggressive acute myeloid leukemia accounting for 2% of all acute leukemias. Clinically, it is characterized by an acute onset of cytopenias associated with extensive bone marrow fibrosis in the absence of splenomegaly and related morphological changes in the red blood cells such as dacryocytes. The presence of fibrosis further complicates the correct diagnosis of this rare disease, being important to differentiate this entity from other hematologic malignancies such as other acute myeloid leukemias and myelodysplastic syndromes associated with myelofibrosis. We report a case of a young adult of 16 years old, admitted with severe pancytopenia. The exposure of this case is justified by the rarity of this pathology associated with few cases reported in medical literature, allowing for a clinical sharing and a better understanding of the different clinical aspects of this entity of difficult diagnosis.
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[Perception of family members and health professionals on end of life care within the scope of primary health care].
Cien Saude Colet
PUBLISHED: 04-30-2013
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The objective of this study is to reflect on care for people with terminal illnesses in primary health care. This study is relevant in the current context of demographic and epidemiological transition, in which there is an increase in the aging population and a rise in the prevalence of chronic degenerative diseases. By means of qualitative research with a critical hermeneutic focus, individual interviews were conducted with seven family members, three professionals from the Family Health Strategy and two professionals from the Home Care Program. The analysis established two empirical categories: end of life care from the perception of family members and that of health professionals. The results showed that, despite the humanization discourse, reflected in the prerogative of living the final moments of life with the family, a serious problem in the discontinuity of care is concealed. It was concluded that, despite the efforts, little is achieved in improving the quality of life for those who die from terminal diseases at home in the Unified Health System in municipalities distant from large centers.
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Delivery cost of human papillomavirus vaccination of young adolescent girls in Peru, Uganda and Viet Nam.
Bull. World Health Organ.
PUBLISHED: 04-30-2013
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To estimate the incremental delivery cost of human papillomavirus (HPV) vaccination of young adolescent girls in Peru, Uganda and Viet Nam.
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Antioxidant, anti-inflammatory, and hypoglycemic effects of the leaf extract from Passiflora nitida Kunth.
Appl. Biochem. Biotechnol.
PUBLISHED: 04-29-2013
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Diabetes mellitus is a metabolic disease characterized by abnormally high plasma glucose levels, leading to major complications, such as insulin resistance, obesity, hyperlipidemia, and hypertension, also with alterations in the immune and neuronal systems. Brazilian plants have been studied as important sources for new molecules with medicinal properties. The genus Passiflora known as "Maracujá" has been used as a traditional folk medicine for a long time, so an investigation was performed regarding an endemic kind of passion fruit (Passiflora nitida Kunth) from Amazonas, Brazil. Here, we aimed to determine its potential biological activity against metabolic syndrome, oxidative stress, pain, and inflammation. The hydroethanol leaf extract revealed an in vitro ?-glucosidase inhibitory activity of 50 % inhibitory concentration (IC??) = 6.78 ± 0.31 ?g/mL and an ?-amylase inhibition of IC??= 93.36 ± 4.37. In vivo, experiments of different saccharide tolerance resulted in significant glycemia control and, with alloxan-diabetic mice, resulted in a decrease of total cholesterol, a hypoglycemic effect, and an antioxidant activity by thiobarbituric acid-reactive substances measurement. Also, it decreased the carrageenan-induced edema volume and the rate of writhing as a nociceptive response. These results indicate positive effects of P. nitida extract and its potential to inhibit metabolic syndrome.
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ON and OFF electroretinography and contrast sensitivity in Duchenne muscular dystrophy.
Invest. Ophthalmol. Vis. Sci.
PUBLISHED: 04-11-2013
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The study investigated possible asymmetric dysfunction of the ON and OFF visual mechanisms in DMD (Duchenne muscular dystrophy) patients associated with specific genetic alterations. Methods: nineteen DMD patients and 7 heterozygous dmd carriers were tested, as well as 19 age-matched controls.Full-field ergs were recorded using mesopic (1 cd/m(2)) and photopic (250 cd/m(2)) sawtooth luminance modulations as stimuli: rapid increase and ramping decrease (to isolate ON responses) or rapid decrease and ramping increase (for OFF responses). In addition, a psychophysical study comprised contrast sensitivity tests using two checkerboard stimuli at either higher (ON) or lower (OFF) luminance relative to the background: 0.3 cycles per degree (cpd) presented for 33 ms (low spatial frequency, short duration) and 2 cpd presented for 1500 ms (high spatial frequency, long duration).
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Insecticidal and repellence activity of the essential oil of Pogostemon cablin against urban ants species.
Acta Trop.
PUBLISHED: 04-09-2013
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Ants are highly abundant in neotropical regions, with certain species adapted to the urban environment, where they can cause damage to human health. The main method for controlling ants consists of using organosynthetic insecticides, which are potentially toxic to the environment. Essential plant oils are considered a viable alternative to the use of conventional insecticides. In this study, we analyze the bioinsecticidal activity and repellence of patchouli essential oil (Pogostemon cablin) against three species of urban ants: Camponotus melanoticus, Camponotus novograndensis, and Dorymyrmex thoracicus. The chemical composition of the essential oil was analyzed by GC-MS and GC-FID. The major compounds were patchoulol (36.6%) followed by ?-bulnesene (13.95%), and ?-guaiene (11.96%). Toxicity and repellency bioassays were performed using the essential oil over the ants, and mortality evaluations were performed at 4, 24, and 48 h after performing the bioassays. Mortality percentage of the ants on 7 ?g/mg was on average 84%. The essential oil of P. cablin displayed toxicity against all three species of urban ants, with the lowest LD?? being observed for D. thoracicus (2.02 ?g oil/mg insect) after 48 h of exposure compared to C. melanoticus (2.34 ?g oil/mg insect) and C. novogranadensis (2.95 ?g oil/mg insect). The essential oil of P. cablin was strongly repellent to the three species of ants in all concentrations tested (0.01% and 1% v/v). Considering the potential toxicity and repellency of the P. cablin essential oil to the urban ants, future studies could investigate the practical application of this oil to control of this insects.
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The structure and properties of septin 3: a possible missing link in septin filament formation.
Biochem. J.
PUBLISHED: 03-30-2013
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The human genome codes for 13 members of a family of filament-forming GTP-binding proteins known as septins. These have been divided into four different subgroups on the basis of sequence similarity. The differences between the subgroups are believed to control their correct assembly into heterofilaments which have specific roles in membrane remodelling events. Many different combinations of the 13 proteins are theoretically possible and it is therefore important to understand the structural basis of specific filament assembly. However, three-dimensional structures are currently available for only three of the four subgroups. In the present study we describe the crystal structure of a construct of human SEPT3 which belongs to the outstanding subgroup. This construct (SEPT3-GC), which includes the GTP-binding and C-terminal domains, purifies as a nucleotide-free monomer, allowing for its characterization in terms of GTP-binding and hydrolysis. In the crystal structure, SEPT3-GC forms foreshortened filaments which employ the same NC and G interfaces observed in the heterotrimeric complex of human septins 2, 6 and 7, reinforcing the notion of promiscuous interactions described previously. In the present study we describe these two interfaces and relate the structure to its tendency to form monomers and its efficiency in the hydrolysis of GTP. The relevance of these results is emphasized by the fact that septins from the SEPT3 subgroup may be important determinants of polymerization by occupying the terminal position in octameric units which themselves form the building blocks of at least some heterofilaments.
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Social representations of mothers about gestational hypertension and premature birth.
Rev Lat Am Enfermagem
PUBLISHED: 03-25-2013
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To identify the meanings attributed by mothers to hypertensive disorders of pregnancy (HDPs) and their consequences, such as premature birth and hospitalization of the infant in the neonatal intensive care unit (NICU).
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[Congenital syphilis in Ceará: epidemiological analysis of one decade].
Rev Esc Enferm USP
PUBLISHED: 03-22-2013
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This study evaluates the incidence of congenital syphilis in the Brazilian state of Ceará from 2000 to 2009, describes the epidemiologic profile of pregnant women whose newborns had congenital syphilis and verifies the receipt of prenatal care and treatment of their partners. It is a retrospective study that was conducted in July 2010 using a Center for Information and Analysis in Health database that contains information on National Notifiable Diseases. There were 2,930 reported cases of congenital syphilis, demonstrating an annual ascending historical series. The majority of the pregnant women were 20 to 34 years of age (n=1,836, 62.7%), illiterate or with little education (n=1623, 55.4%) and had received prenatal care (n=2077, 70.9%). The inadequate treatment of women and the lack of treatment for their partners is a reality in Ceará. The incidence of congenital syphilis is a quality indicator of prenatal care. Therefore, its increase in the last ten years highlights the necessity of syphilis control.
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Lifestyle and family history influence cancer prognosis in Brazilian individuals.
Pathol. Res. Pract.
PUBLISHED: 03-19-2013
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The aim of this research was to study prognostic parameters of CRC by analyzing clinical and pathological variables associated with cancer patients at a northeastern Brazilian Hospital. This was a retrospective study evaluating CRC-diagnosed patients across a 10-year period (1995-2005) at Dr. Luiz Antônio Hospital in Natal, RN, Brazil. Data were collected from patients medical files. A total of 358 patients were included over the 10-year period. The average age at diagnosis was 58.8 years (S.D.=15.26), 48.3% of the patients were males and 51.7% were females. Alcohol consumption significantly increased the chance of dying (p<0.023) from colorectal cancer; this increased risk of death was approximately 71%, compared to 52.2% of the non-alcoholics. In addition, tobacco increased the chance of developing high TNM stage tumors (level III, IV; p<0.001). Another risk factor for increased mortality was a family history for colorectal cancer (p<0.002). Our analysis found that patients with an unhealthy lifestyle and/or family history of colorectal cancer were more likely to develop advanced stage colorectal cancer and to have a poor disease prognosis compared to patients with healthy lifestyle and/or sporadic colorectal cancer. These data suggest that a mass screening program should be implemented in northeastern Brazil in order to better prevent and treat colorectal cancer.
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HABEBEE: habitability of eyeball-exo-earths.
Astrobiology
PUBLISHED: 03-19-2013
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Extrasolar Earth and super-Earth planets orbiting within the habitable zone of M dwarf host stars may play a significant role in the discovery of habitable environments beyond Earth. Spectroscopic characterization of these exoplanets with respect to habitability requires the determination of habitability parameters with respect to remote sensing. The habitable zone of dwarf stars is located in close proximity to the host star, such that exoplanets orbiting within this zone will likely be tidally locked. On terrestrial planets with an icy shell, this may produce a liquid water ocean at the substellar point, one particular "Eyeball Earth" state. In this research proposal, HABEBEE: exploring the HABitability of Eyeball-Exo-Earths, we define the parameters necessary to achieve a stable icy Eyeball Earth capable of supporting life. Astronomical and geochemical research will define parameters needed to simulate potentially habitable environments on an icy Eyeball Earth planet. Biological requirements will be based on detailed studies of microbial communities within Earth analog environments. Using the interdisciplinary results of both the physical and biological teams, we will set up a simulation chamber to expose a cold- and UV-tolerant microbial community to the theoretically derived Eyeball Earth climate states, simulating the composition, atmosphere, physical parameters, and stellar irradiation. Combining the results of both studies will enable us to derive observable parameters as well as target decision guidance and feasibility analysis for upcoming astronomical platforms.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.