Human Induced Pluripotent Stem Cell-Derived Models to Investigate Human Cytomegalovirus Infection in Neural Cells

Human cytomegalovirus (HCMV) infection is one of the leading prenatal causes of congenital mental retardation and deformities world-wide. Access to cultured human neuronal lineages, necessary to understand the species specific pathogenic effects of HCMV, has been limited by difficulties in sustaining primary human neuronal cultures. Human induced pluripotent stem (iPS) cells now provide an opportunity for such research. We derived iPS cells from human adult fibroblasts and induced neural lineages to investigate their susceptibility to infection with HCMV strain Ad169. Analysis of iPS cells, iPS-derived neural stem cells (NSCs), neural progenitor cells (NPCs) and neurons suggests that (i) iPS cells are not permissive to HCMV infection, i.e., they do not permit a full viral replication cycle; (ii) Neural stem cells have impaired differentiation when infected by HCMV; (iii) NPCs are fully permissive for HCMV infection; altered expression of genes related to neural metabolism or neuronal differentiation is also observed; (iv) most iPS-derived neurons are not permissive to HCMV infection; and (v) infected neurons have impaired calcium influx in response to glutamate.     

Regulatory factors that determine growth and phenotype of normal human melanocytes

Normal human melanocytes, unlike malignant melanoma cells, required at least three growth-promoting agents, i.e., phorbol ester for protein kinase C activation and the growth factors basic fibroblast growth factor (bFGF) and insulin, for growth in chemically defined W489 medium. Cell growth was further stimulated by addition of agents that increase intracellular levels of cyclic adenosine 3',5'-monophosphate (cAMP) to the medium. Among these agents, the pituitary hormones alpha-melanocyte-stimulating hormone (alpha-MSH) and follicle-stimulating hormone were the most potent, whereas bacterial toxins, including cholera, tetanus, and pertussis toxin and their subunits either were less mitogenic or gave variable results depending on the culture tested. Medium containing phorbol ester PMA, growth factors bFGF and insulin (or insulin-like growth factor-I), and synthetic alpha-MSH supported melanocyte growth for more than 5 months with doubling times between 5 and 8 days. Two copper-binding proteins, ceruloplasmin and tyrosinase, were mitogenic when added to medium and ceruloplasmic induced a long bi- to tripolar-shape of cells. Addition of 1 mM dibutyryl cAMP to the medium led to the formation of dendrites in all cells, with an average of 28 extensions per cell. Although cell growth was inhibited by dibutyryl cAMP, cells were not terminally differentiated and continued to proliferate. Dendritic melanocytes showed a 2.2-fold increase in activity of the tyrosine kinase pp60c-src. The induction of dendritic processes in melanocytes by dibutyryl cAMP or sodium butyrate was reversible and appears to reflect the expression of the mature melanocytic phenotype in situ.     

Potential involvement of MYC- and p53-related pathways in tumorigenesis in human oral squamous cell carcinoma revealed by proteomic analysis

Oral squamous cellular carcinoma is a malignant tumor with poor prognosis. Discovery of early markers to discriminate between malignant and normal cells is of high importance in clinical diagnosis. Subcellular fractions from 10 oral squamous cell carcinoma and corresponding control samples, enriched in mitochondrial and cytosolic proteins, as well as blood from the tumor were analyzed by proteomics, two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Three-hundred and fifty different gene products were identified. Twenty proteins showed deranged levels in oral squamous cell carcinoma in comparison with the control samples and are potentially involved in tumor growth and metastasis. Of these, 16 proteins were upregulated. By applying pathway analysis, we found 8 of the upregulated gene products to be linked to three main locus genes, p53, MYC, and MYCN, and could be candidate biomarkers for OSCC. The findings of this pilot study show that OSCC gene ontology combined with proteomic analysis is a powerful tool in systems biology for the elucidation of the complexity of expression profiles in cellular processes. Application of such pathway analysis has the potential to generate new insights into complex molecular mechanisms underlying disease related processes and could therefore significantly contribute to the efficient performance of the entire discovery process.     

The effect of analgesics and physical therapy on respiratory function after open and laparoscopic cholecystectomy

In this study we present prospective clinical trial included 100 patients. One half of the patients underwent open cholecystectomy, whereas laparoscopic cholecystectomy was performed in the other half Spirometric parameters, arterial blood gases, acid-base balance, were determined preoperatively, and then at 6 h, 24 h, 72 h and 144 h postoperatively. The impact of physical therapy on the respiratory parameter patterns, VAS-pain score and use of tramadol were studied after cholecystectomy. Significantly lower VAS-pain score and less tramadol use, higher values and faster recovery of ventilation parameters and PaO2 were recorded after laparoscopic cholecystectomy than after open cholecystectomy (p = 0.001 for both). Physical therapy resulted in a significant improvement in the values of respiratory parameters in the open cholecystectomy group within a short time (30 min) after therapy was performed. Physical therapy failed to produce any improvement of respiratory parameters in laparoscopic cholecystectomy, whereas in open cholecystectomy group who had a favorable although transient effect, strictly limited to the short time from its application. (p = 0.005). The patients operated on by open cholecystectomy had statistically significantly more pronounced disturbances including hypoxia, hypocapnia and hyperventilation when compared to the group submitted to laparoscopic cholecystectomy. It is recommended that physical therapy be more frequently performed during the postoperative period in patients submitted to open cholecystectomy.     

The effect of the oxidation state of a terthiophene-conducting polymer and of the presence of a redox probe on its gene-sensing properties

The gene-sensing properties of sensor films made of a terthiophene-conducting polymer, poly(3-((2':2', 5':2'-terthiophene)-3'-yl)acrylic acid) (PTAA), were evaluated using electrochemical impedance spectroscopy for films in their reduced and oxidised states with and without the Fe(CN)(6)(3-/4-) redox probe (RP) in dilute tris-EDTA buffer. Porous films of PTAA were prepared and attached to an oligonucleotide sequence specific to the Salmonella virulence gene InvA. These films could be described with a dual transmission line model in which the polymer conductivity was increased as a consequence of surface binding of complementary DNA. The effect is analogous to that reported for silicon nanowires and field-effect transistors in dilute electrolyte modified by charge exchange across the polymer-electrolyte interface. As a result, gene sensing could be conveniently observed as a change in the impedance phase angle at a fixed frequency.     

Influence of passive smoking on basic anthropometric characteristics and respiratory function in young athletes

The primary objective of this study is to investigate the maintenance difference in basic anthropometric characteristics and to outline the dynamics of respiratory function change in youngsters athletes exposed to passive smoking (PS) and athletes not exposed to passive smoking in their families (NPS). High and weight were determined as basis anthropometric characteristics. Measured parameters for respiratory function were vital capacity (VC), forced expiratory volume in the first second (FEV1), maximum expiratory flow (PEF), forced expiratory flow at 50% forced vital capacity (MEF 50) and forced expiratory flow at 25% forced vital capacity (MEF 25). Significant statistical differences in separate spirometric variable were found in three variables (FEV1, MEF50, and MEF25) for group older youngsters. Analysis of variance showed statistical differences between athletes unexposed to passive smoking (NPS) and athletes exposed to passive smoking (PS) in even four spirometric variables (VC, FEV1, MEF50 and MEF25).     

The impact of novel antipsychotic drugs on quality of life among people suffering from schizophrenia

More attention is increasingly being paid to quality of life of people suffering from schizophrenia. The results of numerous clinical trials indicate that novel antipsychotic drugs are as efficient (if not more so) than the conventional drugs. Novel drugs also cause fewer side effects and allow for better quality of life. In order to confirm these thesis we have studied the quality of life of 80 female outpatients in good social remission that have been under psychiatric evaluation for at least six months and were on antipsychotic drugs. Of those 80 outpatients, half were on the conventional medication, while the other half were on the novel antipsychotic drugs. Their life quality was evaluated with the questionnaire "Heinrichs-Hanlon-Carpenter--Quality of life questionnaire", which is one of the most frequently used QL scales. The results demonstrate that the only difference between the two groups lies in the field of social activity. While leaving the question of different influence of novel and conventional drugs open, the authors are trying to find the possible reasons for such results.     

A 3D visualization and simulation of the individual human jaw

A new biomechanical three-dimensional (3D) model for the human mandible based on computer-generated virtual model is proposed. Using maps obtained from the special kinds of photos of the face of the real subject, it is possible to attribute personality to the virtual character, while computer animation offers movements and characteristics within the confines of space and time of the virtual world. A simple two-dimensional model of the jaw cannot explain the biomechanics, where the muscular forces through occlusion and condylar surfaces are in the state of 3D equilibrium. In the model all forces are resolved into components according to a selected coordinate system. The muscular forces act on the jaw, along with the necessary force level for chewing as some kind of mandible balance, preventing dislocation and loading of nonarticular tissues. In the work is used new approach to computer-generated animation of virtual 3D characters (called "Body SABA"), using in one object package of minimal costs and easy for operation.     

Does treatment of subsyndromal depression improve depression-related and diabetes-related outcomes? A randomised controlled comparison of psychoeducation,physical exercise and enhanced treatment as usual

Background

Elevated depressive symptoms that do not reach criteria for a clinical diagnosis of depression are highly prevalent in persons with diabetes. This study was aimed at determining the efficacy of psychoeducation and physical exercise compared with enhanced treatment as usual on 1-year changes in depressive symptoms, diabetes distress and self-management, and quality of life and metabolic control in type 2 diabetes patients with subsyndromal depression.

Methods

Adult type 2 diabetes patients who screened positively for depression and expressed a need for professional help with mood-related issues were eligible. Exclusion criteria were clinical depression, current psychiatric treatment and advanced diabetes complications. Out of 365 eligible patients 209 consented to either 6 weekly sessions of psychoeducation (A) and physical exercise (B), or to enhanced treatment as usual (C). Computer-generated sequences for block randomisation stratified by gender were used. Depressive symptoms (primary outcome) and diabetes distress, diabetes self-care, metabolic control and health-related quality of life (secondary outcomes) were analysed at 6-month and 12-month follow-up using repeated-measures ANOVAs.

Results

Out of the 74 patients randomised into group A, 66 into B and 69 into group C, 203 completed the interventions, and 179 patients with all 3 assessments were analysed. Depressive symptoms in participants from the psychoeducational, physical exercise and the enhanced treatment as usual groups improved equally from baseline to 12-month follow-up (time versus time x group effect; F = 12.51, p < 0.001, η² = 0.07 and F = 0.609, p = 0.656, η² = 0.007 respectively), as did diabetes distress and quality of life (all p < 0.001), diabetes self-care (p < 0.001 to < 0.05), triglycerides, and total cholesterol and LDL-cholesterol (p < 0.001).

Conclusions

The employed interventions had comparable positive effects on 12-month psychological and diabetes-related outcomes suggesting that even minimal intervention addressing patients’ diabetes-related problems and concerns had favourable clinical implications and might be sufficient to treat subsyndromal depression. Further investigation is warranted to clarify possible mechanisms of improvement.

Trial registration

Current Controlled Trials ISRCTN05673017The message on assigning the above mentioned ISRCTN was received on 11 August 2010

     

Six-Minute Walk Test in Renal Failure Patients: Representative Results,Performance Analysis and Perceived Dyspnea Predictors

Objectives

Six-minute walk test in dialysis population hasn’t been consistently evaluated for the isolated impact of renal failure and other predictive factors. We measured six-minute walk distance in patients representative for low level of comorbidity and searched for potentially modifiable predictive factors of performance and dyspnea.

Methods

This was a cross-sectional study with hemodialysis patients (N = 90) and control subjects (N = 140). Main outcome measures: six-minute walk test distance and dyspnea severity using the 10-item Borg scale.

Results

Median distance decreased from 600m below the 6^th decade to 420m in the 8^th decade of age. Dialysis dependence predicted 101.5m shorter distance in the adjusted model that explained 70% of variability in results. Adjusted for significant covariates of age, height and spontaneous gait speed, fat mass (but not lean body mass) and serum total iron binding capacity were significantly associated with distance (95% CI for B coefficients -4.6 to –1.4 m/kg and 0.1 to 5 m/μmol/l, respectively). Serum total iron binding capacity as an explanatory variable was superior to C-reactive protein and albumin. Dialysis dependence, odds ratio (OR) 2.97 (1.11–7.94), spontaneous gait speed, OR 0.08 (0.02–0.41), rate-pressure product, OR 1.15 (1.08–1.23) and hemoglobin, OR 0.95 (0.92–0.98) predicted dyspnea in the adjusted model.

Conclusions

Renal failure without the confounding effect of comorbidity is a significant negative predictor of performance at six-minute walk test and perceived level of dyspnea. Body fat mass and serum total iron binding capacity are the main potentially modifiable predictors of performance, total iron binding capacity being superior to C-reactive protein and albumin. Although hemoglobin is not associated with test performance, it negatively predicts perceived shortness of breath.