A rapid cell counting method utilising acridine orange as a novel discriminating marker for both cultured astrocytes and microglia

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.

Conceptualization, measurement, and application of quality of life for persons with intellectual disabilities: report of an international panel of experts

In this article a number of issues involving the concept of quality of life as applied to persons with intellectual disabilities are summarized, and a number of agreed-upon principles regarding its conceptualization, measurement, and application are presented. We realize that the concepts and models presented in this article will vary potentially from country to country, and even from area to area within countries. The cross-cultural understanding of the concept of quality of life is in its infancy, and we hope that the discourses resulting from the material presented in this article will facilitate both cross-cultural understanding and collaborative work. The article reflects current thought about the conceptualization, measurement, and application of this increasingly important and widely used concept in the field of intellectual disabilities and sets the stage for its continuing development.

Effect of dietary fatty acids on the intestinal permeability of marker drug compounds in excised rat jejunum

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.

'Class' and political theatre : the case of Melbourne workers theatre

Traditionally, class has been an important category of identity in discussions of political theatre. However, in recent years the concept has fallen out of favour, partly because of changes in the forces and relations of capitalist production. The conventional Marxist use of the term, which defined an individual's class position in relation to the position they occupied in the capitalist production process, seemed anachronistic in an era of globalization. Moreover, the rise of identity politics, queer theory, feminism, and post-colonialism have proffered alternative categories of identity that have displaced class as the primary marker of self. Glenn D'Cruz reconsiders the role of class in the cultural life of Australia by examining the recent work of Melbourne Workers Theatre, a theatre company devoted to promoting class-consciousness, in relation to John Frow's more recent re-conceptualization of class. He looks specifically at two of the company's plays, the award-winning Who's Afraid of the Working Class? and The Waiting Room, with reference to Frow's work on class, arguing that these productions articulate a more complex and sophisticated understanding of class and its relation to politics of race and gender today.

Stat5 as a diagnostic marker for leukemia

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.

Is television viewing time a marker of a broader pattern of sedentary behavior?

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.

Skin temperature as a noninvasive marker of haemodynamic and perfusion status in adult cardiac surgical patients : an observational study

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.

Assessment and case conceptualization in sex offender treatment

The assessment of sexual offenders consists of the systematic collection of clinically relevant information in order to detect clinical phenomena or problems and to provide clear treatment targets. The result of this process is a conceptual model, or case formulation, representing the client’s various problems, the hypothesized underlying mechanisms, and their interrelationships. The focus of this article is on the importance of psychological assessment and case formulation in the rehabilitation and management of individuals convicted of sexual offences. First, we make a number of general points about the importance of evidence based assessment and clinical reasoning in case formulation. Second, we review key elements of contemporary sexual offender theory that highlights the heterogeneity evident among sex offenders and the implications for case formulation and treatment planning. Third, we discuss the role of case formulation for risk assessment and management. Finally, we illustrate our major points with a brief case study and conclude with a brief consideration of the value of case formulations.

Epithelial cell folate is an accurate marker when compared with whole tissue biopsy folate for examining the role of folate status in colorectal cancer

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.

Do we need to flick the switch? The need for a broader conceptualization of iatrogenic course aggravation in clinical trials of bipolar disorder?

The term ‘switching’ is often used in bipolar disorder when describing polarity changes in bipolar disorder, but this term is ambiguous and imprecise, and is sometimes used interchangeably with the term ‘cycling’. Furthermore, polarity changes in bipolar disorder can be understood in different ways, because their clinical manifestations range from the emergence of subthreshold symptoms to a full episode of the opposite pole. Besides the need to tighten the meaning of the term ‘switching’, this paper also argues that switching does not adequately describe the complex phenomena that occur with course aggravation of bipolar disorder, such as alteration in episode frequency or amplitude. A more-fine grained approach to course aggravation in bipolar disorder is proposed, which incorporates trans-polar switching, index polarity aggravation, as well as alterations in episodic amplitude, episodic duration, and interepisode length. This approach has the potential to capture a broader, more fine-grained and clinically relevant picture of the process of aggravation of the bipolar cycle.

Interaction of the Akt substrate, AS160, with the glucose transporter 4 vesicle marker protein, insulin-regulated aminopeptidase

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.

β-blockers reduce bone resorption marker in early postmenopausal women

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.