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1.
Ecol Evol ; 14(4): e11304, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38628919

RESUMO

Mammals in arid zones have to trade off thermal stress, predation pressure, and time spent foraging in a complex thermal landscape. We quantified the relationship between the environmental heat load and activity of a mammal community in the hot, arid Kalahari Desert. We deployed miniature black globe thermometers within the existing Snapshot Safari camera trap grid on Tswalu Kalahari Reserve, South Africa. Using the camera traps to record species' activity throughout the 24-h cycle, we quantified changes in the activity patterns of mammal species in relation to heat loads in their local environment. We compared the heat load during which species were active between two sites with differing predator guilds, one where lion (Panthera leo) biomass dominated the carnivore guild and the other where lions were absent. In the presence of lion, prey species were generally active under significantly higher heat loads, especially during the hot and dry spring. We suggest that increased foraging under high heat loads highlights the need to meet nutritional requirements while avoiding nocturnal activity when predatory pressures are high. Such a trade-off may become increasingly costly under the hotter and drier conditions predicted to become more prevalent as a result of climate change within the arid and semi-arid regions of southern Africa.

2.
Pediatr Obes ; 13 Suppl 1: 27-35, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30209890

RESUMO

BACKGROUND: The Healthy Communities Study (HCS) was a national study of community programs and policies that aimed to address childhood obesity; it necessitated recruitment of a large sample of children from communities throughout the United States. OBJECTIVE: The HCS aimed to complete visits with an average of 45 children and 12 key informants from at least 120 communities, diverse with respect to region of the country, urbanicity, socioeconomic status, race, ethnicity and intensity of community programs and policies that aim to address childhood obesity. METHODS: Purchased address lists were utilized to select households for recruitment during Wave 1 of the study, and recruitment of families through schools was employed for Wave 2. RESULTS: The HCS successfully obtained approval from 149 school districts and 478 schools in 130 communities, recruited 5138 families, and interviewed 1421 key informants to allow for characterization of overall intensity of obesity prevention/treatment efforts in each community. CONCLUSIONS: Lessons learned are presented. Future studies should plan for inclusion of the following in development of recruitment strategies: literature review, formative research, pilot testing, and ongoing monitoring and adjustment.


Assuntos
Seleção de Pacientes , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/métodos , Saúde Pública/métodos , Adolescente , Criança , Feminino , Humanos , Masculino , Instituições Acadêmicas , Estados Unidos
3.
Pediatr Obes ; 13 Suppl 1: 46-55, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29992765

RESUMO

BACKGROUND: The influence of community characteristics on the effectiveness of childhood obesity prevention efforts is not well understood. OBJECTIVE: Examine the interaction of community characteristics with the relationship between community programmes and policies (CPPs) and dietary intake. METHODS: An observational study of 5138 children in grades K-8 in 130 US communities was conducted in 2013-2015. Key informant interviews identified and characterized CPPs. CPP scores were generated for the number of target behaviours (CPP-Behav) and the number of behaviour change strategies (CPP-Strat) addressed by all CPPs and CPPs with nutrition goals over the prior 6 years in each community. Dietary intake was assessed by dietary screener and included intake of sugar from sugar-sweetened beverages; energy-dense foods; fruits and vegetables; whole grains; and fibre. Multivariate statistical models assessed the interactions between US region, urbanicity, community-level income, and community-level race/ethnicity and CPP scores in relation to dietary intake. RESULTS: CPP-Strat was positively associated with healthier dietary intakes in the Northeast and West, and in high Hispanic communities; the reverse was true in the South, and in high African-American and low-income communities. The CPP-Behav was positively associated with healthier dietary intakes in the South and rural areas, and the reverse was true in the West. CONCLUSION: The relationships between CPP index scores and dietary intake were most strongly influenced by region and urbanicity and to a lesser extent by community-level race/ethnicity and income. Findings suggest that different considerations may be needed for childhood obesity prevention efforts in communities with different characteristics.


Assuntos
Dieta/estatística & dados numéricos , Comportamento Alimentar , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/estatística & dados numéricos , Saúde Pública/estatística & dados numéricos , Características de Residência/estatística & dados numéricos , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Classe Social , Estados Unidos
4.
Pediatr Obes ; 13 Suppl 1: 14-26, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29992795

RESUMO

BACKGROUND: The impact of community-based obesity prevention efforts on child nutrition has not been adequately studied. OBJECTIVE: Examine relationships between number, type and intensity of community programs and policies (CPPs) and child nutrition. METHODS: An observational study of 5138 children (grades K-8) in 130 U.S. communities was conducted in 2013-2015. CPPs were identified by 10-14 key informant interviews per community. CPPs were characterized based on: count, intensity, number of different strategies used and number of different behaviours targeted. Scores for the prior 6 years were calculated separately for CPPs that addressed primarily nutrition, primarily physical activity (PA) or total combined. Child intakes were calculated from a dietary screener and dietary behaviours were based on survey responses. Multi-level statistical models assessed associations between CPP indices and nutrition measures, adjusting for child and community-level covariates. RESULTS: Implementing more types of strategies across all CPPs was related to lower intakes of total added sugar (when CPPs addressed primarily PA), sugar-sweetened beverages (for nutrition and PA CPPs) and energy-dense foods of minimal nutritional value (for total CPPs). Addressing more behaviours was related to higher intakes of fruit and vegetables (for nutrition and total CPPs) and fibre (total CPPs). Higher count and intensity (PA and total CPPs) were related to more consumption of lower fat compared with higher fat milk. A higher count (PA CPPs) was related to fewer energy-dense foods and whole grains. No other relationships were significant at P < 0.05. CONCLUSION: Multiple characteristics of CPPs to prevent obesity appear important to improve children's diets.


Assuntos
Comportamento Alimentar , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/métodos , Saúde Pública/estatística & dados numéricos , Criança , Fenômenos Fisiológicos da Nutrição Infantil/fisiologia , Pré-Escolar , Dieta/estatística & dados numéricos , Exercício Físico , Feminino , Humanos , Masculino , Estados Unidos
5.
Pediatr Obes ; 13 Suppl 1: 103-112, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29923334

RESUMO

BACKGROUND: Rational planning of community policies and programs (CPPs) to prevent obesity requires an understanding of CPP objectives associated with dietary behaviours. OBJECTIVE: The objective of the study is to identify objectives of CPPs associated with healthful dietary behaviours. METHODS: An observational study identified 4026 nutrition CPPs occurring in 130 communities in the prior 6 years. Dietary intakes of fruits and vegetables, added sugar and sugar-sweetened beverages, among others, were reported among 5138 children 4-15 years of age from the communities, using a Dietary Screener Questionnaire with children age 9 years and older (parent assisted) or parent proxies for younger children. CPPs were documented through key informant interviews and characterized by their intensity, count, and objectives including target dietary behaviour and food environment change strategy. Associations between dietary intakes and CPP objectives were assessed using hierarchical statistical models. RESULTS: CPPs with the highest intensity scores that targeted fast food or fat intake or provided smaller portions were associated with greater fruit and vegetable intake (0.21, 0.19, 0.23 cup equivalents/day respectively with p values <0.01, 0.04, 0.03). CPPs with the highest intensity scores that restricted the availability of less healthful foods were associated with lower child intakes of total added sugar (-1.08 tsp/day, p < 0.01) and sugar from sugar-sweetened beverages (-1.63 tsp/day, p = 0.04). Similar associations were observed between CPP count and dietary outcomes. No other significant associations were found between CPP target behaviours or environmental strategies and dietary intakes/behaviours. CONCLUSION: CPPs that targeted decreases in intakes of less healthful foods and/or aimed to modify the availability of less healthful foods and portions were associated with healthier child dietary behaviours.


Assuntos
Dieta Saudável/estatística & dados numéricos , Comportamento Alimentar , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/estatística & dados numéricos , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Política Nutricional , Saúde Pública/estatística & dados numéricos , Inquéritos e Questionários , Estados Unidos
6.
Pediatr Obes ; 13 Suppl 1: 93-102, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29921032

RESUMO

BACKGROUND: Little is known about whether characteristics of communities are associated with differential implementation of community programmes and policies to promote physical activity and healthy eating. This study examines associations between community characteristics (e.g. region and race/ethnicity) and the intensity of community programmes and policies implemented to prevent childhood obesity. It explores whether community characteristics moderate the intensity of community efforts to prevent childhood obesity. OBJECTIVE: The objective of this study is to investigate associations between community characteristics and the intensity of community policies and programmes to prevent childhood obesity documented in the Healthy Communities Study that engaged a diverse sample of US communities. METHOD: Programmes and policies were documented in 130 communities across the USA, reporting over 9000 different community programmes and policies to prevent obesity among children ages 4-15. We examined associations between community characteristics and the intensity of community programmes and policies implemented (i.e. their amount and reach, duration and strength of change strategy). CONCLUSION: Community characteristics explain 25% of the variability in the intensity of community programmes and policies implemented in communities. Particular characteristics - urbanicity, region, being a large county and the per cent of African-Americans in a community - contributed to more (over 18% of the 25%) of the observed variability.


Assuntos
Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/estatística & dados numéricos , Saúde Pública/estatística & dados numéricos , Características de Residência/estatística & dados numéricos , Adolescente , Criança , Pré-Escolar , Estudos Transversais , Exercício Físico , Disparidades em Assistência à Saúde/estatística & dados numéricos , Humanos , Estados Unidos
7.
Pediatr Obes ; 13 Suppl 1: 72-81, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29900697

RESUMO

BACKGROUND: Community initiatives to promote physical activity in children are common, but evidence supporting their effectiveness is limited. OBJECTIVES: The objective of this study is to examine the relationships between community programmes and policies and children's physical activity in a large and diverse sample of US communities. METHODS: Programmes and policies to promote children's physical activity were assessed in 130 communities by key informant interviews, and physical activity behaviours were measured by self-report and parental report in samples of children in each community (total n = 5138). Associations between composite indices of community programmes and policies and indicators of total and moderate-to-vigorous physical activity were examined without and with adjustment for demographic factors. RESULTS: An index reflecting the 6-year history of the number of behaviour change strategies used in community programmes and policies was positively associated with children's moderate-to-vigorous physical activity. This association was attenuated with adjustment for demographic factors. Effect modification analyses found that the association was positive among non-Hispanic children but was negative for Hispanic children. CONCLUSIONS: Community initiatives to promote physical activity in children were positively associated with children's physical activity in non-Hispanic children. Such initiatives were negatively associated with physical activity in Hispanic children, suggesting that future research should consider unique cultural factors when designing community initiatives to promote activity in this population sub-group.


Assuntos
Exercício Físico , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/estatística & dados numéricos , Saúde Pública/estatística & dados numéricos , Criança , Estudos Transversais , Feminino , Humanos , Estudos Longitudinais , Masculino , Estudos Retrospectivos , Autorrelato , Estados Unidos
8.
Pediatr Obes ; 13 Suppl 1: 82-92, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29493122

RESUMO

BACKGROUND: Although a national epidemic of childhood obesity is apparent, how community-based programmes and policies (CPPs) affect this outcome is not well understood. OBJECTIVES: This study examined the longitudinal relationship between the intensity of CPPs in 130 communities over 10 years and body mass index (BMI) of resident children. We also examined whether these relationships differ by key family or community characteristics. METHODS: Five thousand one hundred thirty-eight children in grades K-8 were recruited through 436 schools located within 130 diverse US communities. Measures of height, weight, nutrition, physical activity and behavioural and demographic family characteristics were obtained during in-home visits. A subsample of families consented to medical record review; these weight and height measures were used to calculate BMI over time for 3227 children. A total of 9681 CPPs were reported during structured interviews of 1421 community key informants, and used to calculate a time series of CPP intensity scores within each community over the previous decade. Linear mixed effect models were used to assess longitudinal relationships between childhood BMI and CPP intensity. RESULTS: An average BMI difference of 1.4 kg/m2 (p-value < 0.01) was observed between communities with the highest and lowest observed CPP intensity scores, after adjusting for community and child level covariates. BMI/CPP relationships differed significantly by child grade, race/ethnicity, family income and parental education; as well as community-level race/ethnicity. CONCLUSIONS: These results indicate that, over time, more intense CPP interventions are related to lower childhood BMI, and that there are disparities in this association by sociodemographic characteristics of families and communities.


Assuntos
Índice de Massa Corporal , Obesidade Infantil/prevenção & controle , Serviços Preventivos de Saúde/estatística & dados numéricos , Saúde Pública/estatística & dados numéricos , Peso Corporal , Criança , Pré-Escolar , Exercício Físico , Características da Família , Feminino , Disparidades em Assistência à Saúde/estatística & dados numéricos , Humanos , Estudos Longitudinais , Masculino , Obesidade Infantil/epidemiologia , Instituições Acadêmicas/estatística & dados numéricos , Estados Unidos/epidemiologia
9.
Conserv Physiol ; 5(1): cow078, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29383253

RESUMO

Some mammals have the ability to lower their hypothalamic temperature below that of carotid arterial blood temperature, a process termed selective brain cooling. Although the requisite anatomical structure that facilitates this physiological process, the carotid rete, is present in members of the Cetartiodactyla, Felidae and Canidae, the carotid rete is particularly well developed in the artiodactyls, e.g. antelopes, cattle, sheep and goats. First described in the domestic cat, the seemingly obvious function initially attributed to selective brain cooling was that of protecting the brain from thermal damage. However, hyperthermia is not a prerequisite for selective brain cooling, and selective brain cooling can be exhibited at all times of the day, even when carotid arterial blood temperature is relatively low. More recently, it has been shown that selective brain cooling functions primarily as a water-conservation mechanism, allowing artiodactyls to save more than half of their daily water requirements. Here, we argue that the evolutionary success of the artiodactyls may, in part, be attributed to the evolution of the carotid rete and the resulting ability to conserve body water during past environmental conditions, and we suggest that this group of mammals may therefore have a selective advantage in the hotter and drier conditions associated with current anthropogenic climate change. A better understanding of how selective brain cooling provides physiological plasticity to mammals in changing environments will improve our ability to predict their responses and to implement appropriate conservation measures.

10.
J Comp Physiol B ; 186(4): 527-40, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26920796

RESUMO

The use of selective brain cooling, where warm arterial blood destined for the brain is cooled in the carotid rete via counter-current heat exchange when in close proximity to cooler venous blood, contributes to the conservation of body water. We simultaneously measured carotid blood and hypothalamic temperature in four gemsbok, five red hartebeest and six blue wildebeest to assess the extent to which these free-living animals, with varying water dependency, routinely rely on selective brain cooling. We investigated the hypothesis that innate differences in selective brain cooling exist in large, sympatric artiodactyls with varying water dependency. All three species used selective brain cooling, without any discernible differences in three selective brain cooling indices. GLMMs revealed no species differences in the threshold temperature for selective brain cooling (z = 0.79, P = 0.43), the magnitude (z = -0.51, P = 0.61), or the frequency of selective brain cooling use (z = -0.47, P = 0.64), after controlling for carotid blood temperature and black globe temperature. Comparison of anatomical attributes of the carotid retes of the three species revealed that the volume (F 2,9 = 5.54, P = 0.03) and height (F 2,9 = 5.43, P = 0.03) of the carotid rete, per kilogram body mass, were greater in the red hartebeest than in the blue wildebeest. Nevertheless, intraspecific variability in the magnitude, the frequency of use, and the threshold temperature for selective brain cooling exceeded any interspecific variability in the three indices of selective brain cooling. We conclude that the three species have similar underlying ability to make use of selective brain cooling in an environment with freely available water. It remains to be seen to what extent these three species would rely on selective brain cooling, as a water conservation mechanism, when challenged by aridity, a condition likely to become prevalent throughout much of southern Africa under future climate change scenarios.


Assuntos
Antílopes/fisiologia , Regulação da Temperatura Corporal/fisiologia , Encéfalo/fisiologia , Animais , Temperatura Corporal/fisiologia , Água Corporal/fisiologia , Artérias Carótidas/anatomia & histologia , Artérias Carótidas/fisiologia , Feminino , África do Sul , Especificidade da Espécie
11.
PLoS One ; 10(2): e0115514, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25675092

RESUMO

In artiodactyls, arterial blood destined for the brain can be cooled through counter-current heat exchange within the cavernous sinus via a process called selective brain cooling. We test the hypothesis that selective brain cooling, which results in lowered hypothalamic temperature, contributes to water conservation in sheep. Nine Dorper sheep, instrumented to provide measurements of carotid blood and brain temperature, were dosed with deuterium oxide (D2O), exposed to heat for 8 days (40 ◦C for 6-h per day) and deprived of water for the last five days (days 3 to 8). Plasma osmolality increased and the body water fraction decreased over the five days of water deprivation, with the sheep losing 16.7% of their body mass. Following water deprivation, both the mean 24h carotid blood temperature and the mean 24h brain temperature increased, but carotid blood temperature increased more than did brain temperature resulting in increased selective brain cooling. There was considerable inter-individual variation in the degree to which individual sheep used selective brain cooling. In general, sheep spent more time using selective brain cooling, and it was of greater magnitude, when dehydrated compared to when they were euhydrated. We found a significant positive correlation between selective brain cooling magnitude and osmolality (an index of hydration state). Both the magnitude of selective brain cooling and the proportion of time that sheep spent selective brain cooling were negatively correlated with water turnover. Sheep that used selective brain cooling more frequently, and with greater magnitude, lost less water than did conspecifics using selective brain cooling less efficiently. Our results show that a 50 kg sheep can save 2.6L of water per day (~60% of daily water intake) when it employs selective brain cooling for 50% of the day during heat exposure. We conclude that selective brain cooling has a water conservation function in artiodactyls.


Assuntos
Regulação da Temperatura Corporal , Água Corporal/metabolismo , Encéfalo , Desidratação , Ovinos , Animais , Temperatura Corporal , Artérias Carótidas , Seio Cavernoso , Concentração Osmolar , Temperatura
12.
Zoology (Jena) ; 115(6): 411-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23036437

RESUMO

Predicting whether behaviour could buffer the effects of climate change on long-lived mammals requires a better understanding of the long-term behavioural responses of mammals to environmental stress. Using biologging, we measured locomotor activity and microclimate selection, over eight months, in five Arabian oryx (Oryx leucoryx) living free in a Saudi Arabian desert. The oryx displayed seasonal flexibility in activity patterns, shifting from a continuous 24-h activity pattern with crepuscular peaks in cooler months to a predominantly nocturnal activity pattern during the hottest months, without reducing the total 24-h activity level. The proportion of total 24-h activity that occurred during daylight hours was just 29±8% during the hottest months, versus 53±8% (mean±SD, n=5 oryx) in the other months. The attenuation in diurnal activity levels during the hot months was accompanied by the selection of cooler microclimates, presumably via shade seeking, during the heat of the day. Analysis of miniature black globe (miniglobe) temperature from a remote sensor on the collar of two female animals revealed that oryx selected microclimates cooler than the microclimates in direct sun at higher environmental heat loads across all periods, but with enhanced efficiency during the dry periods. We have quantified activity re-assignment and microclimate selection as responses to hot arid conditions in a free-living artiodactyl. Such flexible behavioural processes may act to buffer the adverse effects of the progressively hotter and drier conditions predicted to occur with climate change.


Assuntos
Comportamento Animal , Temperatura Alta , Atividade Motora , Ruminantes/psicologia , Animais , Mudança Climática , Feminino , Homeostase , Masculino , Microclima
13.
J Exp Biol ; 215(Pt 22): 3917-24, 2012 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-22899527

RESUMO

Selective brain cooling is a thermoregulatory effector proposed to conserve body water and, as such, may help artiodactyls cope with aridity. We measured brain and carotid blood temperature, using implanted data loggers, in five Arabian oryx (Oryx leucoryx) in the desert of Saudi Arabia. On average, brain temperature was 0.24±0.05°C lower than carotid blood temperature for four oryx in April. Selective brain cooling was enhanced in our Arabian oryx compared with another species from the same genus (gemsbok Oryx gazella gazella) exposed to similar ambient temperatures but less aridity. Arabian oryx displayed a lower threshold (37.8±0.1°C vs 39.8±0.4°C), a higher frequency (87±6% vs 15±15%) and a higher maximum magnitude (1.2±0.2°C vs 0.5±0.3°C) of selective brain cooling than did gemsbok. The dominant male oryx displayed less selective brain cooling than did any of the other oryx, but selective brain cooling was enhanced in this oryx as conditions became hotter and drier. Enhanced selective brain cooling in Arabian oryx supports the hypothesis that selective brain cooling would bestow survival advantages for artiodactyl species inhabiting hot hyper-arid environments.


Assuntos
Antílopes/fisiologia , Regulação da Temperatura Corporal/fisiologia , Encéfalo/fisiologia , Clima Desértico , Ar , Animais , Artérias Carótidas/fisiologia , Ritmo Circadiano/fisiologia , Conservação dos Recursos Naturais , Feminino , Modelos Lineares , Masculino , Especificidade de Órgãos , Arábia Saudita
14.
Eur J Surg Oncol ; 38(4): 352-60, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22284346

RESUMO

AIMS: We aimed to demonstrate that Hypericin, a component of St. Johns Wort, selectively visualizes malignant gliomas. Hypericin is known as one of the most powerful photosensitizers in nature with excellent fluorescent properties. METHODS: In five patients with a recurrence of a malignant glioma a newly developed water soluble formulation of hypericin was given intravenously (0.1 mg/kg body weight) 6 h before the surgical procedure. Tumor resection was performed under white light and fluorescence mode. The intensity grade of the tissue fluorescence was categorisized by the surgeon in three grades, highly fluorescent, weakly fluorescent and not fluorescent. In these areas tissue samples were taken and investigated by two blinded independent neuropathologists. Tissue samples were histologically classified differentiating between tumor tissue, tumor necrosis, tissue with scattered tumor cells and normal brain tissue. RESULTS: In all patients tumor tissue was clearly distinguishable by its typically red fluorescence color from normal brain tissue which was colored blue under a special fluorescent filter. Histological evaluation of the 110 tissue samples showed a specificity of 100% and sensitivity of 91% for one of the two neuropathologists, whereas specificity for second pathologist was 90% and sensitivity 94%. The i.v. application of Hypericin proofed to be safe in all cases and there were no side effects observed. CONCLUSION: Hypericin in its water soluble form is a well tolerated drug. In addition to its high photosensitizing properties hypericin will open up interesting new therapeutic possibilities especially when used in combination with fluorescence detection and simultaneously photodynamic therapy.


Assuntos
Neoplasias Encefálicas/diagnóstico , Glioma/diagnóstico , Perileno/análogos & derivados , Fármacos Fotossensibilizantes , Idoso , Antracenos , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/cirurgia , Feminino , Fluorescência , Glioma/patologia , Glioma/cirurgia , Humanos , Injeções Intravenosas , Masculino , Pessoa de Meia-Idade , Perileno/administração & dosagem , Fármacos Fotossensibilizantes/administração & dosagem , Valor Preditivo dos Testes , Sensibilidade e Especificidade
15.
J Microsc ; 245(3): 311-8, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22126439

RESUMO

Test systems for measuring cell viability in optical microscopy (based on colony formation ability or lysosomal integrity) were established and applied to native cells as well as to cells incubated with fluorescence markers or transfected with genes encoding for fluorescent proteins. Human glioblastoma and Chinese hamster ovary cells were irradiated by various light doses, and maximum doses where at least 90% of the cells survived were determined. These tolerable light doses were in the range between 25 J cm⁻² and about 300 J cm⁻² for native cells (corresponding to about 250-3000 s of solar irradiance and depending on the wavelength as well as on the mode of illumination, e.g. epi- or total internal reflection illumination) and decreased to values between 50 J cm⁻² and less than 1 J cm⁻² upon application of fluorescent markers, fluorescent proteins or photosensitizers. In high-resolution wide field or laser scanning microscopy of single cells, typically 10-20 individual cell layers needed for reconstruction of a 3D image could be recorded with tolerable dose values. Tolerable light doses were also maintained in fluorescence microscopy of larger 3D samples, e.g. cell spheroids exposed to structured illumination, but may be exceeded in super-resolution microscopy based on single molecule detection.


Assuntos
Corantes Fluorescentes/efeitos adversos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Animais , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Cricetinae , Cricetulus , Humanos , Imageamento Tridimensional/métodos , Luz , Microscopia de Fluorescência/efeitos adversos
16.
Pharmazie ; 66(11): 894-8, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22204137

RESUMO

A set of human tumor cell lines was characterized in terms of the GPx isoenzymes GPx1, -2, -3 and -4. Semiquantitative PCR was used to investigate the GPx mRNA transcripts and the GPx activity was determined photometrically. As a result of culturing under standard conditions, diverse distribution of GPx mRNA and basic GPx activity was found in the investigated cell lines. PCR results showed nearly ubiquitous existence of the isoenzymes GPx1 and GPx4. GPx2 mRNA transcript was only detected in the colonic cell line CaCo-2. After detection of the GPx3 mRNA transcripts in most of the tested cell lines, an ELISA was performed to investigate if the GPx3 protein is present as well. However, the GPx3 protein could not be detected. Glutathione peroxidases contain the amino acid selenocysteine in their active centre. Selenocysteine contains selenium instead of sulfur in cysteine. Therefore, the influence of selenium on GPx activity and GPx isoenzyme distribution was investigated. Cell culturing with additional selenium showed a clear elevation of GPx activity in Mono Mac 6 cells but no gain of mRNA transcripts or any change in the isoenzyme's distribution.


Assuntos
Linhagem Celular Tumoral/enzimologia , Glutationa Peroxidase/metabolismo , Antioxidantes/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Humanos , Isoenzimas/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Espécies Reativas de Oxigênio/metabolismo , Padrões de Referência , Compostos de Selênio/farmacologia , Óxidos de Selênio
18.
J Microsc ; 211(Pt 1): 19-29, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12839547

RESUMO

A novel compact illumination device in variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) is described. This device replaces the standard condensor of an upright microscope. Light from different laser sources is delivered via a monomode fibre and focused onto identical parts of a sample under variable angles of total internal reflection. Thus, fluorophores in close proximity to a cell-substrate interface are excited by an evanescent wave with variable penetration depth, and localized with high (nanometre) axial resolution. In addition to quantitative measurements in solution, fluorescence markers of the cytoplasm and the plasma membrane, i.e. calcein and laurdan, were examined using cultivated endothelial cells. Distances between the glass substrate and the plasma membrane were determined using the mathematical algorithm of a four-layer model, as well as a Gaussian-shaped intensity profile of the illumination spot on the samples. Distances between 0 and 30 nm in focal contacts and between 100 and 300 nm in other parts of the cell were thus determined. In addition to measurements of cell-substrate topology, the illumination device appears appropriate for numerous applications in which high axial resolution is required, e.g. experiments on endocytosis or exocytosis, as well as measurements of ion concentrations proximal to the plasma membrane. The compact illumination device is also suitable for combining TIRFM with further innovative techniques, e.g. time-resolved fluorescence spectroscopy, fluorescence lifetime imaging (FLIM) or fluorescence resonance energy transfer (FRET).


Assuntos
2-Naftilamina/análogos & derivados , Luz , Microscopia de Fluorescência/instrumentação , Microscopia de Fluorescência/métodos , 2-Naftilamina/química , Animais , Simulação por Computador , Endotélio/ultraestrutura , Fluoresceínas/química , Lauratos/química , Proteínas Luminescentes/análise , Matemática , Modelos Estruturais , Refratometria/instrumentação
19.
J Microsc ; 211(Pt 1): 30-6, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12839548

RESUMO

A novel setup for total internal reflection fluorescence microscopy with spectral and temporal (nanosecond) resolution was used to measure the emission spectra of the membrane marker laurdan either selectively within the plasma membrane or in whole living cells, depending on the incident angle of the excitation light. With increasing temperature, the intensity of the fluorescence band around 490 nm increased in comparison with the band around 440 nm, which has previously been assigned to a phase transition of membrane lipids from gel to liquid crystalline phase. For a better separation of the overlapping spectral bands, time-gated detection with a delay of 10-15 ns with respect to the exciting laser pulse was used. As a parameter of membrane dynamics the so-called generalized polarization GP = (I440 - I490)/(I440 + I490) was evaluated at temperatures between 24 and 41 degrees C and variable angles of the incident light permitting to excite laurdan molecules either within the plasma membrane or in the whole cell. A decrease of the GP values by approximately 0.2 units between 28 and 41 degrees C indicated an increase in membrane fluidity or a decrease in membrane stiffness with increasing temperature. In addition, higher GP values were observed for the plasma membrane as compared with intracellular membranes, probably due to a higher amount of cholesterol. Because properties of the plasma membrane have a large influence on the uptake or release of certain pharmaceutical agents or metabolites, the direct assessment of the dynamics of the plasma membrane by total internal reflection fluorescence spectroscopy appears to be important for pharmacology.


Assuntos
2-Naftilamina/análogos & derivados , 2-Naftilamina/química , Membrana Celular/química , Corantes Fluorescentes/química , Lauratos/química , Espectrometria de Fluorescência/métodos , Animais , Membrana Celular/ultraestrutura , Endotélio/citologia , Endotélio/ultraestrutura , Microscopia de Fluorescência , Temperatura
20.
Nucl Med Commun ; 23(12): 1171-5, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12464781

RESUMO

Secondary lymphedema is a localized, acquired lymphatic microcirculatory disturbance that affects large numbers of patients after breast cancer therapy. There is a paucity of objective methods to quantitate lymphatic function and to anticipate the response to therapeutic interventions. We applied radionuclide lymphoscintigraphy to evaluate lymphatic transport and axillary lymph node visualization in women following breast cancer therapy to determine the utility of these data in these patients. Lymphoscintigraphy was performed after subcutaneous injection of 0.25 mCi of Tc-filtered sulfur colloid. Subcutaneous accumulation of radiotracer ('dermal backflow') and the visualization of axillary lymph nodes were graded using our own scoring system. The ratio of radioactivity within the affected to normal axillae (ARR) was also quantified. Nineteen patients with lymphedema after breast cancer therapy were evaluated. The disease severity was documented by serial measurements of the limb volume using the truncated cone formula. Responses to therapy were quantified after completion of the therapy. There was a correlation between the ARR and the percentage reduction in edema volume. The lymphoscintigraphic score correlated with the initial arm volume excess and with the durationof lymphedema. It can be concluded that quantitative and semi-quantitative assessment by radionuclide lymphoscintigraphy represents a potentially useful tool for the clinical assessment of upper extremity lymphedema.


Assuntos
Neoplasias da Mama/complicações , Linfedema/diagnóstico por imagem , Linfedema/terapia , Linfocintigrafia , Modalidades de Fisioterapia , Extremidade Superior , Idoso , Feminino , Humanos , Linfonodos/diagnóstico por imagem , Linfedema/etiologia , Mastectomia , Pessoa de Meia-Idade , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/terapia , Valor Preditivo dos Testes , Compostos Radiofarmacêuticos , Coloide de Enxofre Marcado com Tecnécio Tc 99m , Extremidade Superior/patologia
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