Aerosolization of lipoplexes using AERx Pulmonary Delivery System

The lung represents an attractive target for delivering gene therapy to achieve local and potentially systemic delivery of gene products. The objective of this study was to evaluate the feasibility of the AERx Pulmonary Delivery System for delivering nonviral gene therapy formulations to the lung. We found that "naked" DNA undergoes degradation following aerosolization through the AERx nozzle system. However, DNA formulated with a molar excess of cationic lipids (lipoplexes) showed no loss of integrity. In addition, the lipoplexes showed no significant change in particle size, zeta (zeta) potential, or degree of complexation following extrusion. The data suggest that complexation with cationic lipids had a protective effect on the formulation following extrusion. In addition, there was no significant change in the potency of the formulation as determined by a transfection study in A-549 cells in culture. We also found that DNA formulations prepared in lactose were aerosolized poorly. Significant improvements in aerosolization efficiency were seen when electrolytes such as NaCl were added to the formulation. In conclusion, the data suggest that delivery of lipoplexes using the AERx Pulmonary Delivery System may be a viable approach for pulmonary gene therapy.     

Environment,phenotype and genetics: risk factors associated with BCC of the skin

Basal cell carcinoma is the most common cancer in Caucasians and its incidence is rising. Whilst not life threatening, the tumor can cause substantial morbidity and because of long follow-up, it places a major burden on healthcare agencies worldwide. Patients with the disease demonstrate wide phenotypic diversity with respect to tumor numbers, rate of tumor appearance and site. The factors involved in patient susceptibility to basal cell carcinoma are not well understood although exposure to ultraviolet radiation in sunlight appears critical. In this review we discuss the role of environmental and genetic factors on predisposition to basal cell carcinoma and illustrate how stratification of the basal cell carcinoma cohort into high-risk subgroups is helpful in identifying factors significant in disease susceptibility.     

The rate of increase in the numbers of primary sporadic basal cell carcinomas during follow up is associated with age at first presentation

Basal cell carcinoma (BCC) patients demonstrate marked variation in tumour numbers and site. Previous studies also show an association between age at first BCC presentation and development of BCC on the trunk. In this study we have investigated the association between age at first presentation and the rate of development of truncal and non-truncal tumours in 747 patients with BCC. We used negative binomial regression analysis to show that increasing age at first presentation was associated with an increased rate of BCC development (rate ratio 1.01/year, 95% CI 1.01-1.02, P < 0.001). In particular, development of tumours was greater in cases aged 60.0-69.9, 70.0-79.9 and 80.0-89.9 years than in those 40.0-49.9 years (P = 0.05, 0.01 and 0.039, respectively). While few cases aged over 70 years of age first present with a truncal BCC, the numbers of BCC/year were greater than in those with a head/neck BCC. The data suggest different genetic factors mediate the appearance of BCC in patients of different ages particularly those aged above and below 60 years.     

Visualizing reaction pathways in photoactive yellow protein from nanoseconds to seconds

Determining 3D intermediate structures during the biological action of proteins in real time under ambient conditions is essential for understanding how proteins function. Here we use time-resolved Laue crystallography to extract short-lived intermediate structures and thereby unveil signal transduction in the blue light photoreceptor photoactive yellow protein (PYP) from Halorhodospira halophila. By analyzing a comprehensive set of Laue data during the PYP photocycle (forty-seven time points from one nanosecond to one second), we track all atoms in PYP during its photocycle and directly observe how absorption of a blue light photon by its p-coumaric acid chromophore triggers a reversible photocycle. We identify a complex chemical mechanism characterized by five distinct structural intermediates. Structural changes at the chromophore in the early, red-shifted intermediates are transduced to the exterior of the protein in the late, blue-shifted intermediates through an initial "volume-conserving" isomerization of the chromophore and the progressive disruption of hydrogen bonds between the chromophore and its surrounding binding pocket. These results yield a comprehensive view of the PYP photocycle when seen in the light of previous biophysical studies on the system.     

Prediction of mortality in an Indian intensive care unit

Objective To compare hospital outcome prediction using an artificial neural network model, built on an Indian data set, with the APACHE II (Acute Physiology and Chronic Health Evaluation II) logistic regression model.Design Analysis of a database containing prospectively collected data.Setting Medical-neurological ICU of a university hospital in Mumbai, India.Subjects Two thousand sixty-two consecutive admissions between 1996 and1998.Interventions None.Measurements and results The 22 variables used to obtain day-1 APACHE II score and risk of death were recorded. Data from 1,962 patients were used to train the neural network using a back-propagation algorithm. Data from the remaining 1,000 patients were used for testing this model and comparing it with APACHE II. There were 337 deaths in these 1,000 patients; APACHE II predicted 246 deaths while the neural network predicted 336 deaths. Calibration, assessed by the Hosmer-Lemeshow statistic, was better with the neural network (=22.4) than with APACHE II (=123.5) and so was discrimination (area under receiver operating characteristic curve =0.87 versus 0.77, p=0.002). Analysis of information gain due to each of the 22 variables revealed that the neural network could predict outcome using only 15 variables. A new model using these 15 variables predicted 335 deaths, had calibration (=27.7) and discrimination (area under receiver operating characteristic curve =0.88) which was comparable to the 22-variable model (p=0.87) and superior to the APACHE II equation (p<0.001).Conclusion Artificial neural networks, trained on Indian patient data, used fewer variables and yet outperformed the APACHE II system in predicting hospital outcome.Electronic Supplementary Material Supplementary material is available in the online version of this article at Part of this work was presented at the Sixth Annual Critical Care Congress of the Indian Society for Critical Care Medicine, Bangalore, India     

Control of autoimmune diseases by the B7-CD28 family molecules

The roles of B7-CD28 family molecules in the regulation of immune responses have been intensively studied over the past decade. The findings resulting from these studies not only broaden our understanding in the control of immune responses at the molecular level, but also lead to identification of molecular targets for future manipulation and potential treatment of human diseases. There is convincing evidence that the B7-CD28 family molecules play critical roles in the control of initiation, progression and pathogenesis of autoimmune diseases, which is the focus of this review. Currently, five molecular pathways within this family have been identified and each of them appears to overlap but have distinct functions in the control of priming, activation, maturation and amplification of cellular and humoral immune responses. Rationale-based design of intervention, targeting on multiple pathways should lead to new methods and approaches for management of autoimmune diseases.     

Development and evaluation of cultural food frequency questionnaires for South Asians,Chinese, and Europeans in North America

We developed three ethnic food frequency questionnaires (FFQs) to characterize the diets of South Asian, Chinese, and European immigrants. FFQs were developed from foods reported in the diet records and recalls of 29 South Asians, 25 Chinese, and 20 Europeans participating in a pilot study from 1995-1996 in Hamilton, Ontario, Canada. The FFQ and a seven-day diet record were then administered to 342 South Asians, 317 Chinese, and 346 Europeans participating in the Study of Health Assessment and Risk in Ethnic groups (SHARE) in three Canadian centers from 1996-1998. For FFQ validation, a subset of these participants completed a second seven-day diet record and second FFQ 8 to 10 months later. The FFQ generally underestimated macronutrient and overestimated micronutrient intake compared with the records. Consumption of most macronutrients was lower among South Asians. Energy-adjusted deattenuated correlation coefficients between the records and second FFQ ranged from 0.32 to 0.73 (South Asians), 0.17 to 0.84 (Chinese), and 0.30 to 0.83 (Europeans). The FFQs generally performed well and will be used to investigate diet-disease relations in SHARE. Lower correlations for dietary fats among Chinese persons (0.17 to 0.31) may be improved with more direct questions on the FFQ regarding brand, type, and amount of oil consumed in stirfry servings.     

Yoga therapy for breast cancer patients: A prospective cohort study

We sought to study the impact of yoga therapy on anxiety, depression and physical health in breast cancer patients.Stage I–III post-operative breast cancer patients were recruited with twelve 1-h weekly yoga sessions completed with an experienced yoga instructor. Before and after each module completion, assessments were obtained with the Hospital Anxiety and Depression scale (HADS), the Dallas pain scale and shoulder flexibility measurements.Fourteen patients completed the entire yoga session with 42.8% having a total mastectomy and 15.4% having breast reconstruction. Both right and left shoulder abduction flexibility significantly improved (p = 0.004; p = 0.015 respectively) as well as left shoulder flexion (p = 0.046). An improvement trend in scores for the HADS and Dallas questionnaires pre- and post-intervention was found, although it was not statistically significant.Our data indicates an improvement in physical function in addition to a consistent amelioration in anxiety, depression and pain symptoms after a yoga intervention.