31 resultados para Fiducial marker
em Deakin Research Online - Australia
Resumo:
Cell culture analyses of growth, morphology and apoptosis commonly require counting of different cell types stained with antibodies to discriminate between them. Previously, we reported the use of l-Leucine methyl ester (l-LME) to prepare purified cultures of type 1 astrocytes with minimal microglia, and staining by GFAP and CD antibodies, respectively. Here, we demonstrate a novel use of acridine orange (AO) for rapid discrimination between these cell types using fluorescence microscopy. AO accumulates in the lysosomes and also binds strongly to nuclear DNA and cytoplasmic/nucleolar RNA. Microglia may contain abundant lysosomes due to known roles in homeostasis and immune response. AO staining of lysosomes was tested at a range of concentrations, and 2.5 μg/mL was most suitable. In agreement with previous reports, microglia treated with AO showed very intense yellow, orange or red granular cytoplasmic staining of lysosomes. Microglia contain a substantially higher number of lysosomes than astrocytes, which have a variable amount. We measured the microglia population at 5.14 ± 0.50% in mixed cultures. Thus, these results show AO is a novel discriminatory marker, as microglia were easily observed and counted in clumps on top of the monolayer of astrocytes, providing a rapid alternative to time-consuming and costly antibody-based assays.
Resumo:
The aim of this study was to explore the effects of diets containing saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and ω-3 and ω-6 polyunsaturated fatty acids (ω-3 and ω-6 PUFA, respectively) on the passive and active transport properties of rat jejunum using marker compounds. Rats were fed diets supplemented with 18.4% (w/w) lipid (4 groups) or standard rat chow (1 group) for a period of 30 days. At the end of the dietary period, mucosal scrapings were taken for the determination of membrane phospholipids, and the apparent jejunal permeability of radiolabelled marker compounds was determined using modified Ussing chambers. Changes in the phospholipid content of the brush border membrane reflected the different lipid content of the diets. The passive paracellular permeability of mannitol was not significantly affected by the fatty acid composition of the diet, although there was a trend toward decreased mannitol permeability in the rats fed both the ω-3 and ω-6 PUFA diets. In comparison, the transcellular diffusion of diazepam was reduced by 20% (P < 0.05) in rats fed diets supplemented with ω-3 and ω-6 PUFA. In the lipid-fed rats, the serosal to mucosal flux of digoxin, an intestinal P-glycoprotein substrate, was reduced by 20% (P < 0.05) relative to the chow-fed group, however there were no significant differences between the different lipid groups. The active absorption of D-glucose via the Na+-dependent transport pathway was highest in the SFA, MUFA and PUFA ω-3 dietary groups, intermediate in the low-fat chow group and lowest in the PUFA ω-6 group, and was positively correlated with short-circuit current. These studies indicate that dietary fatty acid changes can result in moderate changes to the active and passive transport properties of excised rat jejunum.
Resumo:
The Jak-Stat-Socs pathway is an important component of cytokine receptor signaling. Not surprisingly, perturbation of this pathway is implicated in diseases of hematopoietic and immune origin, including leukemia, lymphoma and immune deficiencies. This review examines the role of a key component of this pathway, Stat5. This has been shown to be activated in a variety of leukemias and myeloproliferative disorders, including downstream of a range of key oncogenes where it has been shown to play an important role in mediating their effects. Therefore, Stat5 represents a useful pan-leukemia/myeloproliferative disorder diagnostic marker and key therapeutic end point, as well as representing an attractive therapeutic target for these disorders.
Resumo:
Background: Television (TV) viewing time is associated with abnormal glucose metabolism, the metabolic syndrome, and risk of type 2 diabetes; associations are stronger and more consistent in women. One explanation of this difference may be that TV viewing is a marker of an overall pattern of sedentary behavior in women.
Purpose: We sought to examine associations of TV viewing time with other sedentary behaviors and with leisure-time physical activity in a large sample of Australian adults.
Methods: Adults aged between 20 and 65 years (n=2,046) completed a self-administered questionnaire on TV viewing, five other leisure-time sedentary behaviors, and leisure-time physical activity. Mean adjusted time spent in other sedentary behaviors and in physical activity was compared across TV-time categories previously shown to be associated with abnormal glucose metabolism.
Results: After adjustment for body mass index and sociodemographic variables, women’s time spent watching TV was associated positively with time in other sedentary behaviors and negatively with leisure-time physical activity, but no such associations were observed in men.
Conclusions: TV viewing time may be a robust marker of a sedentary lifestyle in women but not in men. Gender differences in the pattern of sedentary behaviors may explain at least in part the gender differences in the previously reported associations of TV viewing time with biological attributes related to type 2 diabetes.
Resumo:
Objective
Foot temperature has long been advocated as a reliable noninvasive measure of cardiac output despite equivocal evidence. The aim of this pilot study was to investigate the relationship between noninvasively measured skin temperature and the more invasive core-peripheral temperature gradients (CPTGs), against cardiac output, systemic vascular resistance, serum lactate, and base deficit.
Research methodology
The study was of a prospective, observational and correlational design. Seventy-six measurements were recorded on 10 adults postcardiac surgery. Haemodynamic assessments were made via bolus thermodilution. Skin temperature was measured objectively via adhesive probes, and subjectively using a three-point scale.
Setting
The study was conducted within a tertiary level intensive care unit.
Results
Cardiac output was a significant predictor for objectively measured skin temperature and CPTG (p = .001 and p = .004, respectively). Subjective assessment of skin temperature was significantly related to cardiac output, systemic vascular resistance, and serum lactate (p < .001, respectively).
Conclusions
These results support the utilisation of skin temperature as a noninvasive marker of cardiac output and perfusion. The use of CPTG was shown to be unnecessary, given the parallels in results with the less invasive skin temperature parameters. A larger study is however required to validate these findings.
Resumo:
Background – Epidemiological studies have shown low folate status is associated with colorectal cancer. Colonic tissue folate levels at different stages of cancer development should give important information, but different methodologies to extract the colonic tissue folates have been used. This has hampered progress in defining the relationship between systemic and tissue folate levels.
Objective – To evaluate two methods of colonic tissue preparation for estimation of total folate content.
Design – Whole tissue punch biopsy samples were obtained from the descending colon of 31individuals following a normal colonoscopy. Blood samples were obtained for the determination of plasma homocysteine (Hcy), red cell folate (RCF), methylenetetrahydrofolate reductase 677C>T genotype, and serum vitamin B12 and folate. Colonic tissue folate was measured both in washed whole tissue biopsies and in epithelial cells isolated from tissue biopsies.
Outcomes - Whole biopsy and epithelial cell folate concentrations were significantly correlated (R=.375; P=.038). Hcy was inversely correlated with both measures (R=-.365; P=.043 and R=-.364; P=.044 respectively). RCF was significantly correlated with isolated epithelial cell folate (R=.477; P=.007) but not with whole tissue biopsy folate (R=.264; P=.151). There were no significant associations between serum and colonic folate in this study.
Conclusion - Both methods are useful for comparing systemic and localised tissue folate status but epithelial cells may provide more reliable data.
Resumo:
Insulin-regulated aminopeptidase (IRAP), a marker of glucose transporter 4 (GLUT4) storage vesicles (GSVs), is the only protein known to traffic with GLUT4. In the basal state, GSVs are sequestered from the constitutively recycling endosomal system to an insulin-responsive, intracellular pool. Insulin induces a rapid translocation of GSVs to the cell surface from this pool, resulting in the incorporation of IRAP and GLUT4 into the plasma membrane. We sought to identify proteins that interact with IRAP to further understand this GSV trafficking process. This study describes our identification of a novel interaction between the amino terminus of IRAP and the Akt substrate, AS160 (Akt substrate of 160 kDa). The validity of this interaction was confirmed by coimmunoprecipitation of both overexpressed and endogenous proteins. Moreover, confocal microscopy demonstrated colocalization of these proteins. In addition, we demonstrate that the IRAP-binding domain of AS160 falls within its second phosphotyrosine-binding domain and the interaction is not regulated by AS160 phosphorylation. We hypothesize that AS160 is localized to GLUT4-containing vesicles via its interaction with IRAP where it inhibits the activity of Rab substrates in its vicinity, effectively tethering the vesicles intracellularly.
Resumo:
Background: There is evidence to suggest that β-blockers used in the management of cardiovascular disease may also modulate bone metabolism and reduce bone fragility.
Aim: The study aimed to determine the association between β-blocker use, serum markers of bone turnover and bone loss in early postmenopausal women.
Subjects and methods: In this observational study, we evaluated β-blocker exposure in association with serum levels of C-telopeptide and bone-specific alkaline phosphatase, and rates of bone loss. β-blocker use, concomitant therapy and lifestyle were documented for 197 women (50–59 years), 175 of whom had changes in whole body bone mineral density monitored over a 2–year period.
Results: Twenty-four β-blocker users were identified at baseline. After controlling for concomitant use of hormone therapy, C-telopeptide levels were 6.7% lower among β-blocker users (p = 0.02). No association was detected between bone-specific alkaline phosphatase and β-blocker use. Analysis of 15 β-blocker users and 152 non-users identified 2 years post-baseline showed that levels of C-telopeptide but not bone-specific alkaline phosphatase were predictors of adjusted rates of bone loss (p = 0.008 and p>0.05, respectively). Adjusted rates of bone loss were −0.001 ± 0.026 g cm−2 over 2 years for the users and −0.004 ± 0.025 g cm−2 over 2 years for non-users, but this difference was not significant.
Conclusion: β-blockers might suppress bone resorption with relative preservation of bone formation. A study with greater power is required to determine whether β-blocker use is associated with lower rates of bone loss.
Resumo:
The filling of printed forms has always been an issue for the visually impaired. Though optical character recognition technology has helped many blind people to ‘read’ the world, there is not a single device that allows them to fill out a paper-based form without a human assistant. The task of filling forms is however an essential part of their daily lives, for example, for access to social security or benefits. This paper describes a solution that allows a blind person to complete paper-based forms, pervasively and independently, using only off-the-shelf equipment including a Smartphone, a clipboard with sliding ruler, and a ballpoint pen. A dynamic color fiduciary (point of reference) marker is designed so that it can be moved by the user to any part of the form such that all regions can be “visited”. This dynamic color fiduciary marker is robust to camera focus and partial occlusion, allowing flexibility in handling the Smartphone with embedded camera. Feedback is given to the blind user via both voice and tone to facilitate efficient guidance in filling out the form. Experimental results have shown that this prototype can help visually impaired people to fill out a form independently.
Resumo:
Background: There is evidence that myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is characterized by activation of immune, inflammatory, oxidative and nitrosative stress (IO&NS) pathways. The present study was carried out in order to examine whether ME/CFS is accompanied by increased levels of plasma peroxides and serum oxidized LDL (oxLDL) antibodies, two biomarkers of oxidative stress.
Material/Methods: Blood was collected from 56 patients with ME/CFS and 37 normal volunteers. Severity of ME/CFS was measured using the Fibromyalgia and Chronic Fatigue Syndrome (FF) Rating Scale.
Results: Plasma peroxide concentrations were significantly higher in patients with ME/CFS than in normal controls. There was a trend towards significantly higher serum oxLDL antibodies in ME/CFS than in controls. Both biomarkers contributed significantly in discriminating between patients with ME/CFS and normal controls. Plasma peroxide and serum oxLDL antibody levels were both significantly related to one of the FF symptoms.
Conclusions: The results show that ME/CFS is characterized by increased oxidative stress.