13 research outputs found
Removal of the basal lamina in vivo reveals growth cone-basal lamina adhesive interactions and axonal tension in grasshopper embryos
No full textThe Ti1 afferent neurons are the first cells to undergo axonogenesis in embryonic grasshopper limbs. The Ti1 growth cones migrate between the limb epithelium and its basal lamina. We have investigated the nature of growth conebasal lamina interactions in vivo by removing the basal lamina with mild enzymatic digestion. Treatment with elastase, ficin, or papain removes the basal lamina when viewed in scanning electron microscopy. Trypsin and chymotrypsin leave the basal lamina intact. If the basal lamina is removed after the Ti1 growth cones have extended over intrasegmental epithelium but are not yet in contact with either differentiated segment boundaries or neurons, the growth cones retract to the cell somata. If the basal lamina is removed by elastase, and the Ti1 neurons are allowed to extend axons after treatment, a second elastase digestion does not cause the axons to retract. It is therefore unlikely that axon retraction is due to general proteolysis. These results suggest that if Ti1 growth cones have initially extended in the presence of an intact basal lamina, they are dependent on the lamina to remain extended over this region of the limb. The retraction of the Ti1 axons after removal of the basal lamina is inhibited by cytochalasin D, suggesting that microfilament-based cytoskeletal components underlie this event. This result indicates that the axons are under tension in vivo. The ability of the Ti1 growth cones to resist axonal tension suggests that adhesive interactions between the growth cones and the basal lamina underlie normal axon outgrowth in vivo.</jats:p
Pioneer growth cone adhesion in vivo to boundary cells and neurons after enzymatic removal of basal lamina in grasshopper embryos
No full textThe Ti1 pioneer neurons of embryonic grasshopper limbs extend axons between the limb epithelium and its basal lamina. Their growth cones exhibit high affinity for both limb segment boundaries and immature neurons. We have investigated the role of the basal lamina in growth cone adhesion to neurons and segment boundaries by removing the basal lamina with mild enzymatic digestion when the Ti1 growth cones are in contact with different cellular substrates. If the basal lamina is removed when the Ti1 growth cones are in contact with other neurons, the growth cones remain in contact with the neuronal somata, and the Ti1 cell bodies in contact with the neuronal somata, and the Ti1 cell bodies reposition proximally. This suggests that the basal lamina is involved in the adhesion of the Ti1 somata to the substrate but not in growth cone-neuronal adhesion. This is the first direct evidence that growth cones establish adhesive cell-cell interactions with other neurons in vivo. Enzymatic treatments that remove the basal lamina also cause embryonic limbs to elongate. If the Ti1 axons are strongly apposed to 2 segment boundaries prior to protease treatment, their somata reposition to the nearest segment boundary, yet their axons do not retract off of the segment boundaries, despite severe stretching by the enzyme-induced limb expansion. These results indicate that the affinity of the Ti1 cells for segment boundaries is due at least in part to adhesive cell-cell interactions that are resistant to proteolytic digestion and independent of the basal lamina.</jats:p
Patterns of migration and risks associated with leprosy among migrants in Maranhão, Brazil
No full textLeprosy remains a public health problem in Brazil with new case incidence exceeding World Health Organization (WHO) goals in endemic clusters throughout the country. Migration can facilitate movement of disease between endemic and non-endemic areas, and has been considered a possible factor in continued leprosy incidence in Brazil. A study was conducted to investigate migration as a risk factor for leprosy. The study had three aims: (1) examine past five year migration as a risk factor for leprosy, (2) describe and compare geographic and temporal patterns of migration among past 5-year migrants with leprosy and a control group, and (3) examine social determinants of health associated with leprosy among past 5-year migrants. The study implemented a matched case-control design and analysis comparing individuals newly diagnosed with leprosy (n = 340) and a clinically unapparent control group (n = 340) without clinical signs of leprosy, matched for age, sex and location in four endemic municipalities in the state of Maranhão, northeastern Brazil. Fishers exact test was used to conduct bivariate analyses. A multivariate logistic regression analysis was employed to control for possible confounding variables. Eighty cases (23.5%) migrated 5-years prior to diagnosis, and 55 controls (16.2%) migrated 5-years prior to the corresponding case diagnosis. Past 5 year migration was found to be associated with leprosy (OR: 1.59; 95% CI 1.07–2.38; p = 0.02), and remained significantly associated with leprosy after controlling for leprosy contact in the family, household, and family/household contact. Poverty, as well as leprosy contact in the family, household and other leprosy contact, was associated with leprosy among past 5-year migrants in the bivariate analysis. Alcohol consumption was also associated with leprosy, a relevant risk factor in susceptibility to infection that should be explored in future research. Our findings provide insight into patterns of migration to localize focused control efforts in endemic areas with high population mobility
Localisation of RNAs into the germ plasm of vitellogenic xenopus oocytes
No full textWe have studied the localisation of mRNAs in full-grown Xenopus laevis oocytes by injecting fluorescent RNAs, followed by confocal microscopy of the oocyte cortex. Concentrating on RNA encoding the Xenopus Nanos homologue, nanos1 (formerly Xcat2), we find that it consistently localised into aggregated germ plasm ribonucleoprotein (RNP) particles, independently of cytoskeletal integrity. This implies that a diffusion/entrapment-mediated mechanism is active, as previously reported for previtellogenic oocytes. Sometimes this was accompanied by localisation into scattered particles of the “late”, Vg1/VegT pathway; occasionally only late pathway localisation was seen. The Xpat RNA behaved in an identical fashion and for neither RNA was the localisation changed by any culture conditions tested. The identity of the labelled RNP aggregates as definitive germ plasm was confirmed by their inclusion of abundant mitochondria and co-localisation with the germ plasm protein Hermes. Further, the nanos1/Hermes RNP particles are interspersed with those containing the germ plasm protein Xpat. These aggregates may be followed into the germ plasm of unfertilized eggs, but with a notable reduction in its quantity, both in terms of injected molecules and endogenous structures. Our results conflict with previous reports that there is no RNA localisation in large oocytes, and that during mid-oogenesis even germ plasm RNAs localise exclusively by the late pathway. We find that in mid oogenesis nanos1 RNA also localises to germ plasm but also by the late pathway. Late pathway RNAs, Vg1 and VegT, also may localise into germ plasm. Our results support the view that mechanistically the two modes of localisation are extremely similar, and that in an injection experiment RNAs might utilise either pathway, the distinction in fates being very subtle and subject to variation. We discuss these results in relation to their biological significance and the results of others
A long way towards a scientific study about life satisfaction and subjective well-being
No full textPartiendo de la evidencia de que la bu?squeda de la satisfaccio?n con la vida y la discusio?n sobre los criterios de ordenar la propia hacia ese logro constituyen una constante literaria en nuestra historia, este texto inicia con un contraste entre lo que ya avanzaron los filo?sofos al respecto en los siglos pasados y lo que la reciente psicologi?a positiva o del bienestar subjetivo va encontrando. El nu?mero de pensadores elegidos entre los que trataron este tema en el pasado ha debido ser limitado a los que el autor entiende muy representativos de este reflexionar sobre las condiciones que influyen en la satisfaccio?n de la existencia por razones de espacio narrativo. Posteriormente, se describen resultados de investigaciones propias en el a?mbito de la psicologi?a positiva, situa?ndolos en el contexto de la discusio?n entre la perspectiva hedo?nica y la eudaemo?nica que hoy ocupan el reflexio- nar en el a?mbito de la psicologi?a de la satisfaccio?n con la vida y del bienestar subjetivo.
Abstract
This paper begins making a contrast between what philosophers stated in the last centuries about the search for satisfaction with life, as well as the discussion over the criteria to organize our own lives aimed to that purpose, and what the recent positive psychology or the subjective well-being are finding. The number of thinkers chosen among those who dealt with the subject in the past had to be limited to those the author considered the most representatives of this reflect on the conditions that influence the satisfaction with the existence having in mind limitations of narrative space. Subsequently, the author describes his research findings in the field of positive psychology placed in the context of a discussion between the hedonic and eudemonic perspective that occupy nowadays the thinking in the scope of the psychology of satisfaction with life and subjective well-being.
Resumo
Com base na evide?ncia de que a busca de satisfac?a?o com a vida e a discussa?o sobre os crite?rios para organiza?-la para essa conquista sa?o uma constante na nossa histo?ria litera?ria, este texto comec?a com um contraste entre o que os filo?sofos ja? avanc?aram sobre o tema nos se?culos passados e as descobertas da recente psicologia positiva ou do bem-estar subjetivo. O nu?mero de pensadores escolhidos entre aqueles que abordaram esta questa?o no passado teve de ser limitado, segundo a opinia?o do autor ao que ele entende ser mais representativo deste pensar sobre as condic?o?es que influenciam a satisfac?a?o de existe?ncia, por razo?es de espac?o narrativo. Posteriormente, sa?o descritos os resultados de pesquisas no campo da psicologia positiva, co- locando-os no contexto da discussa?o entre a perspectiva hedo?nica e a eudaimo?nica que ocupam hoje o pensamento no campo da psicologia de satisfac?a?o com a vida e do bem-estar subjetivo. Partiendo de la evidencia de que la bu?squeda de la satisfaccio?n con la vida y la discusio?n sobre los criterios de ordenar la propia hacia ese logro constituyen una constante literaria en nuestra historia, este texto inicia con un contraste entre lo que ya avanzaron los filo?sofos al respecto en los siglos pasados y lo que la reciente psicologi?a positiva o del bienestar subjetivo va encontrando. El nu?mero de pensadores elegidos entre los que trataron este tema en el pasado ha debido ser limitado a los que el autor entiende muy representativos de este reflexionar sobre las condiciones que influyen en la satisfaccio?n de la existencia por razones de espacio narrativo. Posteriormente, se describen resultados de investigaciones propias en el a?mbito de la psicologi?a positiva, situa?ndolos en el contexto de la discusio?n entre la perspectiva hedo?nica y la eudaemo?nica que hoy ocupan el reflexio- nar en el a?mbito de la psicologi?a de la satisfaccio?n con la vida y del bienestar subjetivo.
Abstract
This paper begins making a contrast between what philosophers stated in the last centuries about the search for satisfaction with life, as well as the discussion over the criteria to organize our own lives aimed to that purpose, and what the recent positive psychology or the subjective well-being are finding. The number of thinkers chosen among those who dealt with the subject in the past had to be limited to those the author considered the most representatives of this reflect on the conditions that influence the satisfaction with the existence having in mind limitations of narrative space. Subsequently, the author describes his research findings in the field of positive psychology placed in the context of a discussion between the hedonic and eudemonic perspective that occupy nowadays the thinking in the scope of the psychology of satisfaction with life and subjective well-being.
Resumo
Com base na evide?ncia de que a busca de satisfac?a?o com a vida e a discussa?o sobre os crite?rios para organiza?-la para essa conquista sa?o uma constante na nossa histo?ria litera?ria, este texto comec?a com um contraste entre o que os filo?sofos ja? avanc?aram sobre o tema nos se?culos passados e as descobertas da recente psicologia positiva ou do bem-estar subjetivo. O nu?mero de pensadores escolhidos entre aqueles que abordaram esta questa?o no passado teve de ser limitado, segundo a opinia?o do autor ao que ele entende ser mais representativo deste pensar sobre as condic?o?es que influenciam a satisfac?a?o de existe?ncia, por razo?es de espac?o narrativo. Posteriormente, sa?o descritos os resultados de pesquisas no campo da psicologia positiva, co- locando-os no contexto da discussa?o entre a perspectiva hedo?nica e a eudaimo?nica que ocupam hoje o pensamento no campo da psicologia de satisfac?a?o com a vida e do bem-estar subjetivo. 
Isolation, characterization, and substrate properties of the external limiting membrane from the avian embryonic optic tectum
No full textThe external limiting membrane of the avian embryonic optic tectum is isolated by mechanically separating the neuronal mesencephalon from the overlying mesenchymal tissue. The preparation consists of a basal lamina which is covered on its neural side by endfeet of neuroepithelial cells and has attached to it on its meningeal side a collageneous stroma, containing blood vessels. The external limiting membrane can be flat-mounted on a piece of nitrocellulose filter as mechanical support. It covers an area between 0.3 and 1 the cm2, depending on the age of me donor embryo. The endfeet can be removed together with all cellular components of the meninges by treatment with 2% Triton-X-100 or with distilled water. The basal lamina itself is approximately 80 nm thick and consists of two laminae rarae and a central lamina densa. Immunohistochemical staining reveals that the basal lamina in the embryo, after isolation and after detergent extraction of the isolated preparation, contains type IV collagen, nidogen, laminin, and low density heparan sulfate proteoglycan as do other basement membranes. Antibodies against the neural cell adhesion molecule (N-CAM), chondroitin sulfate proteoglycan, and fibronectin fail to stain the external limiting membrane, but these proteins were clearly identified in the blood vessel-containing meninges or in the optic tectum.
The flat-mounted external limiting membrane preparation was used as substrate to culture several different neural tissues of central and peripheral origin. Explants of neural crest cells, dorsal root ganglia, and sympathetic ganglia can be cultured on the external limiting membrane. All explants grow well on the basal lamina preparations whether the endfeet are attached or detergent-extracted prior to explantation; however, neurite outgrowth from sympathetic ganglia is reduced in the presence of the endfeet. Although the endfoot-lined external limiting membrane represents at least part of the immediate environment encountered by retinal axons as they invade the optic tectum and despite its excellent properties as a substrate for retinal axons in vitro, cues guiding the orientation of axons were not detected in the flat-mounted preparation
FIGURE 5 from Immune Profiling of Vulvar Squamous Cell Cancer Discovers a Macrophage-rich Subtype Associated with Poor Prognosis
No full textCharacterization of in vitro polarized TAMs. A, Cytokines and chemokines detected in the supernatant of high-density VSCC cultures by ELISA. B, Healthy donor monocytes were differentiated into macrophages using GMCSF and exposed to different culture conditions: medium alone (monocyte-derived macrophage), VSCC supernatant (in vitro polarized TAM), or 20 ng/mL IFNγ (M1). Flow cytometry on day 5 determined surface antigen expression, presented as d-gMFIs. C, Macrophages were generated as in B; TNFα and IL10 levels were quantified in the supernatant by ELISA on day 5; background levels were subtracted. Data represent results from two individuals and/or experiments and are presented as mean ± SEM; significance was determined by two-sided Student t test.</p
Neurogenesis and apoptosis in the developmentally regulated loss of spinal cord regeneration.
No full textUnlike the adult mammal, the chick can successfully regenerate its spinal cord until embryonic day (E) 13. Multiple factors may contribute to the subsequent loss of regenerative capacity, although most research has concentrated on axonal re-growth inhibition as a key issue. The number of viable cells remaining in the spinal cord could also be important and may be affected by cell survival and cell replacement. In this thesis the early response of the chick spinal cord to injury has been investigated, focusing on cell death and the potential to replace lost cells by neurogenesis. Pharmacological reduction of haemorrhage after injury at El5 resulted in reduced apoptosis and cavitation, suggesting that blood-borne factors, such as the serine protease thrombin, may cause apoptosis. Endogenous thrombin expression and activity after injury was investigated. Thrombin was not up-regulated after injury at El5 however, evidence suggests that the activity of other serine proteases was increased. In parallel, in organotypic slice cultures, exogenous thrombin treatment did not increase apoptosis. These results provide new information about the contribution of serine proteases to apoptosis in the chick, suggesting that, although thrombin is not of primary importance, other serine proteases could play a greater role. Next, the contribution of neurogenesis to regeneration at Ell was examined. Changes in the expression and phosphorylation of the early neuronal marker, doublecortin, in response to injury were observed. Although increased proliferation in the grey matter was observed, no increase in neurogenesis after injury was detected. Surprisingly, ongoing neurogenesis was discovered in the normal spinal cord at Ell. These results challenge established views about the timing of neurogenesis in the chick spinal cord and suggest that ongoing proliferation and neurogenesis may contribute to the regenerative capacity at this stage. This thesis presents insights into factors involved in the early response of the chick spinal cord to injury, providing new information about the contribution of neurogenesis and cell survival to regenerative capacity
Self-reported Diabetes In Older People: Comparison Of Prevalences And Control Measures
No full textObjective: The objective of this study was to analyze the prevalence of diabetes in older people and the adopted control measures. Methods: Data regarding older diabetic individuals who participated in the Health Surveys conducted in the Municipality of Sao Paulo, SP, ISA-Capital, in 2003 and 2008, which were cross-sectional studies, were analyzed. Prevalences and confidence intervals were compared between 2003 and 2008, according to sociodemographic variables. The combination of the databases was performed when the confidence intervals overlapped. The Chi-square (level of significance of 5%) and the Pearson's Chi-square (Rao-Scott) tests were performed. The variables without overlap between the confidence intervals were not tested. Results: The age of the older adults was 60-69 years. The majority were women, Caucasian, with an income of between < 0.5 and 2.5 times the minimum salary and low levels of schooling. The prevalence of diabetes was 17.6% (95%CI 14.9;20.6) in 2003 and 20.1% (95%CI 17.3;23.1) in 2008, which indicates a growth over this period (p at the limit of significance). The most prevalent measure adopted by the older adults to control diabetes was hypoglycemic agents, followed by diet. Physical activity was not frequent, despite the significant differences observed between 2003 and 2008 results. The use of public health services to control diabetes was significantly higher in older individuals with lower income and lower levels of education. Conclusions: Diabetes is a complex and challenging disease for patients and the health systems. Measures that encourage health promotion practices are necessary because they presented a smaller proportion than the use of hypoglycemic agents. Public health policies should be implemented, and aimed mainly at older individuals with low income and schooling levels. 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